pheophytin-a and chlorophyll-b

Nanomaterials are being used increasingly in various areas such as electronic devices manufacture, medicine, mechanical devices production, and even food industry. Therefore, the evaluation of their toxicity is mandatory. Graphene oxide (GO) has been shown to have both positive as well as negative impact on different crop plants, depending on species, dose, and duration of exposure. The current study evaluated the impact of GO sheets at different concentrations (500, 1000 and 2000 mg/L) on physiological, biochemical and genetic levels to determine the possible toxic action. Wheat caryopses were treated with GO for 48 h and 7 days. The germination rate and roots elongation decreased in a dose-response manner, except the sample treated with GO at a concentration of 1000 mg/L. Mitotic index has ascendant trend; its increase may be due to the accumulation of prophases GO induced significant accumulation of the cells with aberrations, their presence suggests a clastogenic/aneugenic effect of these carbon nanomaterials. Regarding enzymatic and non-enzymatic antioxidant system defence, the activity varied depending on the dose of GO. Thus, chlorophyll a pigments content decreased significantly at high dose (2000 mg/L), while the carotenoid pigments had lower content at 500 mg/L of GO, and no statistical difference encountered in case of chlorophyll b amount. The antioxidant enzyme activity (CAT, POD, and SOD) was higher at low dose of GO, indicating the presence of oxidative stress generated as a response to the GO treatment. Also, the free radical scavenging activity of the polyphenolic compounds was enhanced upon GO exposure. The GO accumulation has been identified by transmission electron microscopy only at plumules level, near the intercellular space.

Topics: Antioxidants; Chlorophyll; Chlorophyll A; Germination; Graphite; Nanostructures; Oxidative Stress; Oxides; Plant Roots; Seedlings; Triticum

Passive optical hyperspectral remote sensing of plant pigments offers potential for understanding plant ecophysiological processes across a range of spatial scales. Following a number of decades of research in this field, this paper undertakes a systematic meta-analysis of 85 articles to determine whether passive optical hyperspectral remote sensing techniques are sufficiently well developed to quantify individual plant pigments, which operational solutions are available for wider plant science and the areas which now require greater focus. The findings indicate that predictive relationships are strong for all pigments at the leaf scale but these decrease and become more variable across pigment types at the canopy and landscape scales. At leaf scale it is clear that specific sets of optimal wavelengths can be recommended for operational methodologies: total chlorophyll and chlorophyll a quantification is based on reflectance in the green (550-560nm) and red edge (680-750nm) regions; chlorophyll b on the red, (630-660nm), red edge (670-710nm) and the near-infrared (800-810nm); carotenoids on the 500-580nm region; and anthocyanins on the green (550-560nm), red edge (700-710nm) and near-infrared (780-790nm). For total chlorophyll the optimal wavelengths are valid across canopy and landscape scales and there is some evidence that the same applies for chlorophyll a.

Topics: Anthocyanins; Carotenoids; Chlorophyll; Chlorophyll A; Light; Pigments, Biological; Plant Leaves; Plants; Publications; Remote Sensing Technology; Spectrum Analysis

Chlorophylls are magnesium-tetrapyrrole molecules that play essential roles in photosynthesis. All chlorophylls have similar five-membered ring structures, with variations in the side chains and/or reduction states. Formyl group substitutions on the side chains of chlorophyll a result in the different absorption properties of chlorophyll b, chlorophyll d, and chlorophyll f. These formyl substitution derivatives exhibit different spectral shifts according to the formyl substitution position. Not only does the presence of various types of chlorophylls allow the photosynthetic organism to harvest sunlight at different wavelengths to enhance light energy input, but the pigment composition of oxygenic photosynthetic organisms also reflects the spectral properties on the surface of the Earth. Two major environmental influencing factors are light and oxygen levels, which may play central roles in the regulatory pathways leading to the different chlorophylls. I review the biochemical processes of chlorophyll biosynthesis and their regulatory mechanisms.

Topics: Carbon-Oxygen Ligases; Chlorophyll; Chlorophyll A; Light; Lyases; Magnesium; Oxygen; Photosynthesis; Plant Physiological Phenomena; Protoporphyrins

Chlorophyll a and chlorophyll b are the major constituents of the photosynthetic apparatus in land plants and green algae. Chlorophyll a is essential in photochemistry, while chlorophyll b is apparently dispensable for their photosynthesis. Instead, chlorophyll b is necessary for stabilizing the major light-harvesting chlorophyll-binding proteins. Chlorophyll b is synthesized from chlorophyll a and is catabolized after it is reconverted to chlorophyll a. This interconversion system between chlorophyll a and chlorophyll b refers to the chlorophyll cycle. The chlorophyll b levels are determined by the activity of the three enzymes participating in the chlorophyll cycle, namely, chlorophyllide a oxygenase, chlorophyll b reductase, and 7-hydroxymethyl-chlorophyll reductase. This article reviews the recent progress on the analysis of the chlorophyll cycle and its enzymes. In particular, we emphasize the impact of genetic modification of chlorophyll cycle enzymes on the construction and destruction of the photosynthetic machinery. These studies reveal that plants regulate the construction and destruction of a specific subset of light-harvesting complexes through the chlorophyll cycle. This article is part of a Special Issue entitled: Regulation of Electron Transport in Chloroplasts.

Topics: Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Plant Proteins; Plants

A limiting factor for photosynthetic organisms is their light-harvesting efficiency, that is the efficiency of their conversion of light energy to chemical energy. Small modifications or variations of chlorophylls allow photosynthetic organisms to harvest sunlight at different wavelengths. Oxygenic photosynthetic organisms usually utilize only the visible portion of the solar spectrum. The cyanobacterium Acaryochloris marina carries out oxygenic photosynthesis but contains mostly chlorophyll d and only traces of chlorophyll a. Chlorophyll d provides a potential selective advantage because it enables Acaryochloris to use infrared light (700-750 nm) that is not absorbed by chlorophyll a. Recently, an even more red-shifted chlorophyll termed chlorophyll f has been reported. Here, we discuss using modified chlorophylls to extend the spectral region of light that drives photosynthetic organisms.

Topics: Chlorophyll; Chlorophyll A; Cyanobacteria; Oxygen; Photosynthesis; Plants; Sunlight

The prochlorophytes are a diverse group of photosynthetic prokaryotes that fall within the cyanobacterial lineage, yet lack phycobilisomes as light harvesting structures. Instead, the prochlorophytes have a light-harvesting apparatus composed of the higher plant pigments chlorophylls a and b. This review discusses the evolutionary relationships among these bacteria, with focus on the structure and function of the photosynthetic apparatus. This analysis yields a consensus from studies both on Prochloron sp. and Prochlorothrix hollandica as to how the thylakoid membrane is organized. Overall, we propose that the structure of the light-harvesting complexes (LHC) from prochlorophytes is very different from those of chloroplast systems, and is evolutionarily very ancient. The functional association of the light-harvesting apparatus with photosystem I (PSI) in both Prochlorothrix and Prochloron, as well as a demonstrated capacity for PSI-dependent anoxygenic photosynthesis in Prochlorothrix, may indicate that there is an increased dependence on cyclic photophosphorylation in these organisms. Finally, the structure of the prochlorophyte thylakoid membrane is discussed with respect to the forces that drive thylakoid membrane stacking in prochlorophytes and chloroplasts. We suggest that the light-harvesting structures in prochlorophytes play little, if any, role in this process.

Topics: Chlorophyll; Chlorophyll A; Gram-Negative Bacteria; Photosynthesis; Phycobilisomes; Phylogeny; Pigments, Biological

A large proportion of the chlorophyll in a plant is engaged in harvesting light energy and transferring it to the photochemical reaction centres. These 'antenna' chlorophylls are non-covalently bound to specific proteins to form chlorophyll-protein complexes. The chlorophyll a/b-binding (CAB) polypeptides are encoded by an extended family of nuclear genes. It has recently been discovered that other proteins not known to bind chlorophyll, the early light-inducible proteins (ELIPs), are also related and could be considered part of this family. We suggest that the latter proteins may be involved in pigment biosynthesis or in assembly of the thylakoid membrane.

Topics: Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Plant Proteins; Protein Binding

Three different LED spectra (W: White light; WFR: W + far-red light; WB: W + blue light) with similar photosynthetic photon flux density (PPFD) were designed to explore the effects of supplementary far-red and blue lights on leaf color, biomass and phytochemicals of two cultivars of red-leaf lettuce ("Yanzhi" and "Red Butter") in an artificial lighting plant factory. Lettuce plants under WB had redder leaf color and significantly higher contents of pigments, such as chlorophyll a, chlorophyll b, chlorophyll (a + b) and anthocyanins. The accumulation of health-promoting compounds, such as vitamin C, vitamin A, total phenolic compounds, total flavonoids and anthocyanins in the two lettuce cultivars were obviously enhanced by WB. Lettuce under WFR showed remarkable increase in fresh weight and dry weight; meanwhile, significant decreases of pigments, total phenolic compounds, total flavonoids and vitamin C were found. Thus, in the plant factory system, the application of WB can improve the coloration and quality of red leaf lettuce while WFR was encouraged for the purpose of elevating the yield of lettuce.

Topics: Anthocyanins; Ascorbic Acid; Biomass; Chlorophyll; Chlorophyll A; Flavonoids; Lactuca; Lighting; Phenols; Photosynthesis; Phytochemicals; Pigments, Biological; Vitamin A

Imbalanced light absorption by photosystem I (PSI) and photosystem II (PSII) in oxygenic phototrophs leads to changes in interaction of photosystems altering the linear electron flow. In plants and green algae, this imbalance is mitigated by a partial migration of the chlorophyll a/b containing light-harvesting antenna between the two photosystem core complexes. This migration is registered as fluorescence changes of the pigment apparatus and is termed the reverse transitions between States 1 and 2. By contrast, the molecular mechanism of State 1/2 transitions in phycobilisome (PBS)-containing photosynthetics, cyanobacteria and red algae, is still insufficiently understood. The suggested hypotheses - PBS movement along the surface of thylakoid membrane between PSI and PSII complexes, reversible PBS detachment from the dimeric PSII complex, and spillover - have some limitations as they do not fully explain the accumulated data. Here, we have recorded changes in the stationary fluorescence emission spectra of red algae and cyanobacteria in States 1/2 at room temperature, which allowed us to offer an explanation of the existing contradictions. The change of room temperature fluorescence of chlorophyll belonged to PSII was revealed, while the fluorescence of PBS associated with the PSII complexes remained during States 1/2 transitions at the stable level. Only the reversible dissociation of PBS from the monomeric PSI was revealed earlier which implied different degree of surface contact of PBS with the two photosystems. The detachment of PBS from the PSI corresponds to ferredoxin oxidation as electron carrier and the increase of cyclic electron transport in the pigment apparatus in State I.

Topics: Chlorophyll; Chlorophyll A; Cyanobacteria; Electron Transport; Microalgae; Oxidation-Reduction; Photosynthesis; Photosystem I Protein Complex; Photosystem II Protein Complex; Phycobilisomes; Spectrometry, Fluorescence; Thylakoids

Chlorophyll is the most abundant pigment on Earth, essential for the capture of light energy during photosynthesis. During senescence, chlorophyll degradation is highly regulated in order to diminish toxicity of the free chlorophyll molecule due to its photoactivity. The first step in the chlorophyll degradation pathway is the conversion of chlorophyll b to chlorophyll a by means of two consecutive reactions catalyzed by enzymes coded by NYC1 (NON-YELLOW COLORING 1), NOL (NYC1-LIKE) and HCAR.. In this work, we studied the expression of NOL and HCAR genes during postharvest senescence of broccoli. We found that the expression of BoNOL increase during the first days of storage and then decrease. In the case of BoHCAR, its expression is maintained during the first days and then it also diminishes. Additionally, the effect of different postharvest treatments on the expression of these genes was also analyzed. It was observed that the expression of BoNOL is lower in the treatments performed with 1-methylcyclopropene (1-MCP), 6-benzylaminopurine (6-BAP) and modified atmospheres, while BoHCAR expression showed an increase in these same treatments, and a decrease in the treatment with ethylene. There were no variations in the expression of both genes in heat treatment, UV-C treatment and visible light treatment.. These results suggest that both BoHCAR and BoNOL show a lower regulation of their expression than other genes involved in chlorophyll degradation during senescence. © 2020 Society of Chemical Industry.

Topics: Brassica; Chlorophyll; Chlorophyll A; Gene Expression Regulation, Plant; Plant Proteins

The fate of antibiotics and their effects on plant growth may be changed by the application of fertilizers. The present study was carried out to investigate the effect of sulfadiazine (SDZ), rice husk compost (RHC), rice husk biochar (RHB), and mycorrhiza (MR) on the growth attributes of Iranian Echium amoenum Fisch & C.A. Mey. A greenhouse experiment as a completely randomized design with six treatments of bio/organic-fertilizers (no bio-fertilizer (NF), RHB, RHC, MR, RHB+MR, and RHC+MR) and three levels of SDZ application (0, 100, and 200 mg kg

Topics: Charcoal; Chlorophyll; Chlorophyll A; Echium; Environmental Pollution; Fertilizers; Iran; Oryza; Soil; Soil Pollutants; Sulfadiazine

Soils contaminated with heavy metals such as Chromium (Cr) and Cadmium (Cd) severely impede plant growth. Several rhizospheric microorganisms support plant growth under heavy metal stress. In this study, Cr and Cd stress was applied to in vitro germinating seedlings of a Legume plant species, Sesbania sesban, and investigated the plant growth potential in presence and absence of Bacillus anthracis PM21 bacterial strain under heavy metal stress. The seedlings were exposed to different concentrations of Cr (25-75 mg/L) and Cd (100-200 mg/L) in Petri plates. Growth curve analysis of B. anthracis PM21 revealed its potential to adapt Cr and Cd stress. The bacteria supported plant growth by exhibiting ACC-deaminase activity (1.57-1.75 μM of α-ketobutyrate/h/mg protein), producing Indole-3-acetic acid (99-119 μM/mL) and exopolysaccharides (2.74-2.98 mg/mL), under heavy metal stress condition. Analysis of variance revealed significant differences in growth parameters between the seedlings with and without bacterial inoculation in metal stress condition. The combined Cr+Cd stress (75 + 200 mg/L) significantly reduced root length (70%), shoot length (24%), dry weight (54%) and fresh weight (57%) as compared to control. Conversely, B. anthracis PM21 inoculation to seedlings significantly increased (p ≤ 0.05) seed germination percentage (5%), root length (31%), shoot length (23%) and photosynthetic pigments (Chlorophyll a: 20%; Chlorophyll b: 16% and total chlorophyll: 18%), as compared to control seedlings without B. anthracis PM21 inoculation. The B. anthracis PM21 inoculation also enhanced activities of antioxidant enzymes, including superoxide dismutase (52%), peroxidase (66%), and catalase (21%), and decreased proline content (56%), electrolyte leakage (50%), and malondialdehyde concentration (46%) in seedlings. The B. anthracis PM21 inoculated seedlings of S. sesban exhibited significantly high (p ≤ 0.05) tissue deposition of Cr (17%) and Cd (16%) as compared to their control counterparts. Findings of the study suggested that B. anthracis PM21 endured metal stress through homeostasis of antioxidant activities, and positively impacted S. sesban growth and biomass. Further experiments in controlled conditions are necessary for investigating phytoremediation potential of S. sesban in metal-contaminated soils in presence of B. anthracis PM21 bacterial strain.

Topics: Bacillus anthracis; Biodegradation, Environmental; Cadmium; Catalase; Chlorophyll; Chlorophyll A; Chromium; Germination; Indoleacetic Acids; Metals, Heavy; Seedlings; Sesbania; Soil; Soil Pollutants

Chlorophyllase catalyzes the hydrolysis of chlorophyll and produces chlorophyllide and phytol. Cyanobacterial chlorophyllases are likely to be more highly heterologously expressed than plant chlorophyllases. A novel recombinant chlorophyllase from the cyanobacterium Oscillatoria acuminata PCC 6304 was successfully expressed in Escherichia coli BL21(DE3).. The putative N-terminal 28-amino-acid signal peptide sequence of O. acuminata chlorophyllase (OaCLH) is essential for its activity, but may confer poor solubility on OaCLH. The C-terminal fusion of a 6 × His tag caused a partial loss of activity in recombinant OaCLH, but an N-terminal 6 × His tag did not destroy its activity. The optimal pH and temperature for recombinant OaCLH activity are 7.0 and 40 °C, respectively. Recombinant OaCLH has hydrolysis activities against chlorophyll a, chlorophyll b, bacteriochlorophyll a, and pheophytin a, but prefers chlorophyll b and chlorophyll a as substrates. The results of site-directed mutagenesis experiments indicated that the catalytic triad of OaCLH consists of Ser159, Asp226, and His258.. The high-level expression and broad substrate specificity of recombinant OaCLH make it suitable for genetically engineering and a promising biocatalyst for industrial production, with applications in vegetable oil refining and laundry detergents.

Topics: Amino Acid Sequence; Bacterial Proteins; Biocatalysis; Carboxylic Ester Hydrolases; Chlorophyll; Chlorophyll A; Cloning, Molecular; Escherichia coli; Hydrogen-Ion Concentration; Hydrolysis; Kinetics; Models, Molecular; Oscillatoria; Protein Structure, Tertiary; Recombinant Proteins; Sequence Homology, Amino Acid; Substrate Specificity; Temperature

The CRISPR/Cas9-based targeted genome editing has emerged as a versatile technique, widely employed in plant genome engineering, both to decipher gene function and as an alternative to classical breeding technique for traits improvement in plants. However, to date, no such platform has been developed for hop (Humulus lupulus L.), which is an economically important crop producing valuable secondary metabolites utilized in the brewing and pharmaceutical industries. Here, we present the first report on the successful establishment of efficient CRISPR/Cas9-based genome editing using the visible endogenous marker gene phytoene desaturase (PDS) involved in carotenoid biosynthesis to demonstrate successful genome editing in hop. Agrobacterium tumefaciens-mediated transformation of in vitro generated internodal explants was used for the stable integration of constructs expressing plant codon-optimized Cas9 and a pair of co-expressed guide RNAs to target the distinct genomic sites of the PDS gene of hop. Analysis of RNA-guided genome-editing events, including mutant lines screening and homozygosity assessment using the T7 endonuclease assay showed that 33.3% of transformed plants were successfully edited at the target site, displaying albino and mosaic regenerants. Intriguingly, the detected mutations were ranges of deletions (16 bp to 39 bp) which led to disruption of the exon-intron boundary, few base substitutions, and a 1 bp insertion at 3 bp upstream of the PAM region of the target site. The decrease in chlorophyll a/b, and carotenoid content in the mutant lines further confirmed the functional disruption of the HlPDS gene. Taken together, our results demonstrate that the CRISPR/Cas9 system can precisely edit the targeted genome sequences, which may revolutionize our way to overcome some of the obstacles that have plagued the traits improvement in hop.

Topics: Agrobacterium tumefaciens; Chlorophyll; Chlorophyll A; CRISPR-Cas Systems; Gene Editing; Genome, Plant; Humulus; Mutagenesis; Oxidoreductases; Plants, Genetically Modified; RNA, Guide, Kinetoplastida

In the methyl-D-erythritol-4-phosphate (MEP) pathway, 1-deoxy-D-xylose-5-phosphate synthase (DXS) is considered the key enzyme for the biosynthesis of terpenoids. In this study,

Topics: Acetates; Chlorophyll; Chlorophyll A; Computational Biology; Cyclopentanes; Escherichia coli; Gene Expression Profiling; Nicotiana; Oxylipins; Pentosephosphates; Pigmentation; Pinus; Plant Leaves; Plant Stems; Promoter Regions, Genetic; Recombinant Proteins; Salicylic Acid; Terpenes; Transferases; Xylose

The siphonaxanthin-siphonein-chlorophyll-a/b-binding protein (SCP), a trimeric light-harvesting complex isolated from photosystem II of the siphonous green alga Codium fragile, binds the carotenoid siphonaxanthin (Sx) and/or its ester siphonein in place of lutein, in addition to chlorophylls a/b and neoxanthin. SCP exhibits a higher content of chlorophyll b (Chl-b) than its counterpart in green plants, light-harvesting complex II (LHCII), increasing the relative absorption of blue-green light for photosynthesis. Using low temperature absorption and resonance Raman spectroscopies, we reveal the presence of two non-equivalent Sx molecules in SCP, and assign their absorption peaks at 501 and 535 nm. The red-absorbing Sx population exhibits a significant distortion that is reminiscent of lutein 2 in trimeric LHCII. Unexpected enhancement of the Raman modes of Chls-b in SCP allows an unequivocal description of seven to nine non-equivalent Chls-b, and six distinct Chl-a populations in this protein.

Topics: Chlorophyll; Chlorophyll A; Chlorophyta; Light-Harvesting Protein Complexes; Photosynthesis; Photosystem II Protein Complex; Pigments, Biological; Xanthophylls

Improving the growth and pigment accumulation of microalgae by electrochemical approaches was considered a novel and promising method. In this research, we investigated the effect of conductive polymer poly (3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) dispersible in water on growth and pigment accumulation of Haematococcus lacustris and Euglena gracilis. The results revealed that effect of PEDOT:PSS was strongly cell-dependent and each cell type has its own peculiar response. For H. lacustris, the cell density in the 50 mg·l

Topics: Bridged Bicyclo Compounds, Heterocyclic; Cell Proliferation; Chlorophyceae; Chlorophyll; Chlorophyll A; Electric Conductivity; Electrochemical Techniques; Euglena gracilis; Polymers; Polystyrenes; Thiophenes; Xanthophylls

Chlorophylls function in photosynthesis, and are critical to plant developmental processes and responses to environmental stimuli. Chlorophyll b is synthesized from chlorophyll a by chlorophyll a oxygenase (CAO). Here, we characterize a yellow-green leaf (ygl) mutant and identify the causal gene which encodes a chlorophyll a oxygenase in maize (ZmCAO1). A 51 bp Popin transposon insertion in ZmCAO1 strongly disrupts its transcription. Low enzyme activity of ZmCAO1 leads to reduced concentrations of chlorophyll a and chlorophyll b, resulting in the yellow-green leaf phenotype of the ygl mutant. The net photosynthetic rate, stomatal conductance, and transpiration rate are decreased in the ygl mutant, while concentrations of δ-aminolevulinic acid (ALA), porphobilinogen (PBG) and protochlorophyllide (Pchlide) are increased. In addition, a ZmCAO1 mutation results in down-regulation of key photosynthetic genes, limits photosynthetic assimilation, and reduces plant height, ear size, kernel weight, and grain yield. Furthermore, the zmcao1 mutant shows enhanced reactive oxygen species production leading to sensitivity to waterlogging. These results demonstrate the pleiotropy of ZmCAO1 function in photosynthesis, grain yield, and waterlogging tolerance in maize.

Topics: Chlorophyll; Chlorophyll A; Floods; Oxygenases; Photosynthesis; Plant Leaves; Plant Proteins; Seeds; Zea mays

Photosynthetic organisms adjust to fluctuating natural light under physiological ambient conditions through flexible light-harvesting ability of light-harvesting complex II (LHCII). A process called state transition is an efficient regulation mechanism to balance the excitations between photosystem II (PSII) and photosystem I (PSI) by shuttling mobile LHCII between them. However, in situ observation of the migration of LHCII in vivo remains limited. In this study, we investigated the in vivo reversible changes in the intracellular distribution of the chlorophyll (Chl) fluorescence during the light-induced state transitions in Chlamydomonas reinhardtii. The newly developed noninvasive excitation-spectral microscope provided powerful spectral information about excitation-energy transfer between Chl-a and Chl-b. The excitation spectra were detected through the fluorescence emission in the 700-750-nm spectral range, where PSII makes the main contribution, though PSI still makes a non-negligible contribution at room temperature. The technique is sensitive to the Chl-b spectral component specifically bound to LHCII. Using a PSI-specific 685-nm component also provided visualization of the local relative concentration of PSI within a chloroplast at room temperature. The decrease in the relative intensity of the Chl-b band in state 2 was more conspicuous in the PSII-rich region than in the PSI-rich region, reflecting the dissociation of LHCII from PSII. We observed intracellular redistributions of the Chl-b-related light-harvesting abilities within a chloroplast during the state transitions. This observation implies the association of the state transitions with the morphological changes in the thylakoid membrane.

Topics: Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Chloroplasts; Lasers; Light; Light-Harvesting Protein Complexes; Microscopy; Photosystem I Protein Complex; Photosystem II Protein Complex; Spectrometry, Fluorescence

Fast growing woody plants are proposed for potential application for phytoremediation of contaminated soil. In this study, the plant growth, physiological responses, mineral element uptake, and phytoremediation potential of the woody plant Morus alba L. were studied in different levels of Cd-contaminated soil through dynamic sampling (30, 60, 120, and 180 d). The results indicated that M. alba L. had strong physiological coordination, tolerance and detoxification capacity in response to Cd in contaminated soil. Compared with the control, the photosynthetic pigment content in M. alba L. leaves was significantly suppressed during initial cultivation (30-60 d) and the malonaldehyde (MDA) content and electrolyte leakage (EL) were increased from 30 to 120 d of cultivation. Furthermore, the uptake of Cu, Mn, and Zn in plant tissues was imbalanced throughout cultivation (30-180 d) under 55 mg·kg

Topics: Antioxidants; Biodegradation, Environmental; Biomass; Cadmium; Chlorophyll; Chlorophyll A; Malondialdehyde; Minerals; Morus; Photosynthesis; Plant Leaves; Plant Roots; Soil Pollutants; Stress, Physiological

Topics: Agrobacterium; Bacterial Proteins; beta-Glucosidase; Chlorophyll; Chlorophyll A; Gene Expression; Gene Expression Regulation, Plant; Heat-Shock Proteins, Small; Light; Photosynthesis; Photosystem II Protein Complex; Plant Leaves; Plant Proteins; Plants, Genetically Modified; Solanum lycopersicum

Water stress is one of the main abiotic factors that reduces plant growth, mainly due to high evaporative demand and low water availability. In order to evaluate the effects of drought stress on certain morphological and physiological characteristics of two canola cultivars, we conducted a factorial experiment based on a completely randomized design. The findings show that drought stress exacerbations result in the plant's response to stress due to increased canola resistance caused by changes in plant pigments, proline, catalase, ascorbate peroxidase, peroxidase, superoxide dismutase and malondialdehyde, glucose, galactose, rhamnose and xylose. These in turn ultimately influence the morphological characteristics of canola. Drought stress reduces the concentration of carotenoids, chlorophyll a, chlorophyll b, total chlorophylls; however, glucose, galactose, rhamnose, xylose, proline, catalase, ascorbate peroxidase, peroxidase, superoxide dismutase, malondialdehyde (in leaves and roots) and the chlorophyll a and b ratios were increased. Reduction of plant height, stem height, root length, fresh and dry weight of canola treated with 300 g/l PEG compared to non-treatment were 0.264, 0.236, 0.394, 0.183 and 0.395, respectively. From the two canola cultivars, the morphological characteristics of the NIMA increased compared to the Ks7 cultivar. Interaction effects of cultivar and drought stress showed that NIMA cultivar without treatment had the highest number of morphological characteristics such as carotenoid concentration, chlorophyll a, chlorophyll b, total chlorophylls a and b, whereas the cultivar with 300 g/l PEG (drought stress) had the highest amount of proline, malondialdehyde, soluble sugars and enzymes in leaves and roots. Increasing activity of oxidative enzymes and soluble sugars in canola under drought stress could be a sign of their relative tolerance to drought stress.

Topics: Brassica napus; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Droughts; Glucose; Plant Leaves; Plant Roots; Rhamnose; Stress, Physiological; Superoxide Dismutase

The interplay between active molecules and the protein environment in light-harvesting complexes tunes the photophysics and the dynamical properties of pigment-protein complexes in a subtle way, which is not fully understood. Here we characterized the photophysics and the ultrafast dynamics of four variants of the water-soluble chlorophyll protein (WSCP) as an ideal model system to study the behavior of strongly interacting chlorophylls. We found that when coordinated by the WSCP protein, the presence of the formyl group in chlorophyll

Topics: Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Models, Molecular; Temperature; Thermodynamics; Water

Cell immobilization on the magnetic nanoparticles (MNPs) and magnetic harvesting is a novel approach for microalgal cells separation. To date, the effect of these nanoparticles on microalgal cells was only studied over a short period of time. More studies are hence needed for a better understanding of the magnetic harvesting proposes or environmental concerns relating to long-term exposure to nanoparticles. In this study, the impact of various concentrations of MNPs on the microalgal cells growth and their metabolic status was investigated over 12 days. More than 60% reduction in mitochondrial activity and pigments (chlorophyll a, chlorophyll b, and carotenoids) content occurred during the first 6 days of exposure to ≥50 µg/mL nanoparticles. However, more than 50% growth inhibitory effect was seen at concentrations higher than 400 µg/mL. Exposure to MNPs gradually induced cellular adaptation and after about 6 days of exposure to stress generating concentrations (˂400 µg/mL) of IONs, microalgae could overcome the imposed damages. This work provides a better understanding regarding the environmental impact of MNPs and appropriate concentrations of these particles for future algal cells magnetic immobilization and harvesting.

Topics: Cells, Immobilized; Chlorella vulgaris; Chlorophyll; Chlorophyll A; Magnetic Phenomena; Microalgae; Nanoparticles

Overexpression of phytoglobins (formerly plant hemoglobins) increases the survival rate of plant tissues under hypoxia stress by the following two known mechanisms: (1) scavenging of nitric oxide (NO) in the phytoglobin/NO cycle and (2) mimicking ethylene priming to hypoxia when NO scavenging activates transcription factors that are regulated by levels of NO and O

Topics: Anaerobiosis; Chlorophyll; Chlorophyll A; Gene Expression Regulation, Plant; Hemoglobins; Hordeum; Metabolome; Metabolomics; Nitric Oxide; Oxygen; Plant Proteins; Proteome; Proteomics; Reactive Oxygen Species; Seedlings

A thorough characterization of the early time sub-100 fs relaxation dynamics of biologically relevant chromophores is of crucial importance for a complete understanding of the mechanisms regulating the ultrafast dynamics of the relaxation processes in more complex multichromophoric light-harvesting systems. While chlorophyll

Topics: Chlorophyll; Chlorophyll A; Diffusion; Energy Transfer; Kinetics; Spectrophotometry, Infrared; Spinacia oleracea; Vibration

The coexistence of polycyclic aromatic hydrocarbons (PAHs) and heavy metals has deleterious effects on environmental quality. Few reports have studied the mechanisms of plant inoculation with Piriformospora indica to remediate PAH-metal co-contaminated soil by analyzing the chemical speciation of the contaminants. This study investigated the influence of the inoculation of Medicago sativa with P. indica to remediate soil co-contaminated with phenanthrene (a kind of PAH) and cadmium (a heavy metal) by analyzing plant growth, physiological parameters and chemical speciation in rhizosphere and nonrhizosphere soils.. The presence of P. indica significantly increased plant tolerance, chlorophyll a, chlorophyll b, maximum quantum efficiency of PSII photochemistry and electron transport rate values in phenanthrene- and/or cadmium-contaminated soil. P. indica inoculation in M. sativa roots increased fluorescein diacetate activities in soils contaminated with phenanthrene, cadmium or both, especially in the nonrhizosphere. The presence of phenanthrene prevented the inoculated plant from accumulating cadmium to some extent, whereas the presence of cadmium did not prevent the degradation of phenanthrene in either the rhizosphere or the nonrhizosphere after P. indica colonization. Although the low bioavailability of cadmium in the rhizosphere restricted its transportation into the stem, P. indica colonization in plants effectively increased cadmium accumulation in roots in soil co-contaminated with cadmium and phenanthrene.. In conclusion, this work provides a theoretical basis for the use of P. indica combined with M. sativa for the remediation of PAH-metal co-contaminated soil.

Topics: Basidiomycota; Biodegradation, Environmental; Biomass; Cadmium; Catechol Oxidase; Chlorophyll; Chlorophyll A; Medicago sativa; Phenanthrenes; Plant Development; Plant Roots; Soil Microbiology; Soil Pollutants

Topics: Antineoplastic Agents; Antioxidants; Carboplatin; Chlorella vulgaris; Chlorophyll; Chlorophyll A; Cisplatin; Fresh Water; Lipid Peroxidation; Malondialdehyde; Microalgae; Oxaliplatin; Photosynthesis; Reactive Oxygen Species; Water Pollutants, Chemical

Sulfonamides (SAs) are antibiotics widely used in clinical practice, livestock and poultry production, and the aquaculture industry. The compounds enter the soil environment largely through livestock and poultry manure application to farmland. SAs not only affect plant growth, but also pose a potential threat to human health through SA residues in plant tissues. In particular, sulfamethoxazole (SMZ) has been classified as a Category 3 carcinogen by the World Health Organization, and thus its soil ecological toxicity and possible health risks are of concern. Using A. thaliana as a model plant, stress responses and biological residues of sulfadiazine (SD), sulfametoxydiazine (SMD), and SMZ were investigated in the present study. Root length and aboveground plant biomass were significantly inhibited by the three types of SA, whereas lateral roots exposed to SMD grew vigorously. The contents of chlorophyll a and chlorophyll b and photosystem II maximum photochemical quantum yield declined with increase in drug concentration, which indicated that exposure to SAs affected photosynthesis and inhibited chlorophyll synthesis in A. thaliana. With increase in drug concentration, reactive oxygen species (ROS) accumulation in the leaves increased significantly. Activities of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) were activated at low SA concentrations, but increased lipid peroxidation occurred with increase in SA concentration. Of the three compounds, SMZ was the most toxic to A. thaliana, followed by SD, and SMD was the least toxic. The results indicated that the risk of SMD entering an organism through the food chain is greater than that for SMZ and SD.

Topics: Anti-Bacterial Agents; Antioxidants; Arabidopsis; Chlorophyll; Chlorophyll A; Lipid Peroxidation; Oxidative Stress; Photosynthesis; Photosystem II Protein Complex; Plant Leaves; Plant Roots; Reactive Oxygen Species; Soil Pollutants; Sulfanilamides

Microalgae application in agriculture is an alternative measure that could be highly beneficial to plants. The application of microalgae

Topics: Agriculture; Beta vulgaris; Chlorella vulgaris; Chlorophyll; Chlorophyll A; Photosynthesis; Plant Leaves; Symbiosis

Selenium (Se) is a beneficial element to higher plants. Application of Se at low concentrations enhances the antioxidant metabolism reducing the reactive oxygen species (ROS) generated by plant membrane cells. This study aimed to evaluate how the application of Se in the forms sodium selenate and sodium selenite regulates ROS scavenging in field-grown cowpea plants. Seven Se application rates (0; 2.5; 5; 10; 20; 40 and 60 g ha

Topics: Antioxidants; Ascorbate Peroxidases; Catalase; Chlorophyll; Chlorophyll A; Glutathione Reductase; Photosynthesis; Plant Leaves; Reactive Oxygen Species; Selenic Acid; Selenious Acid; Selenium; Sodium Selenite; Vigna

The aim of this study is to optimize and evaluate the effectiveness of vitrification, droplet-vitrification, and encapsulation-vitrification techniques in the cryopreservation of

Topics: Chlorophyll; Chlorophyll A; Cryopreservation; Cryoprotective Agents; Desiccation; Genetic Markers; Genetic Variation; Glycerol; Papaveraceae; Plant Shoots; Plants, Medicinal; Polymerase Chain Reaction; Random Amplified Polymorphic DNA Technique; Sucrose; Vitrification

Hydrilla verticillata (L.f.) Royle has a great ability to accumulate large amounts of arsenic (As). We studied the influence of phosphorus (P), nitrogen (N), pH, and arsenite (As(III)) on As transformation and fate in the H. verticillata - medium system via orthogonal experimental design. The results showed highest plant growth was under intermediate As(III) in the medium, with Chlorophyll a and Chlorophyll b contents in plant diminishing after 96 h treatment. Exposure to high N, high As(III), intermediate P, and low pH in the medium, the highest total arsenic uptake by plants were 169.1 ± 5.5 μg g

Topics: Arsenates; Arsenic; Arsenites; Biodegradation, Environmental; Chlorophyll; Chlorophyll A; Hydrocharitaceae; Oxidation-Reduction

Microplastics pollution in farmlands has become a major concern. However, few studies have assessed the effects of microplastics on higher plants. In this study, we investigated the influence of polystyrene nanoplastics (PSNPs, 50 mg L

Topics: Antioxidants; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Cucumis sativus; Hydrogen Peroxide; Malondialdehyde; Peroxidases; Photosynthesis; Plant Leaves; Plastics; Polystyrenes; Soil Pollutants; Superoxide Dismutase

Cross-talk between exogenous salicylic acid (SA) and endogenous phytohormone pathways affects the antioxidant defense system and its response to salt stress. The study presented here investigated the effects of SA treatment before and during salt stress on the levels of endogenous plant growth regulators in three barley cultivars with different salinity tolerances: Hordeum vulgare L. cvs. Akhisar (sensitive), Erginel (moderate), and Kalaycı (tolerant). The cultivars' relative leaf water contents, growth parameters, proline contents, chlorophyll a/b ratios, and lipid peroxidation levels were measured, along with the activities of enzymes involved in detoxifying reactive oxygen species (ROS) including superoxide-dismutase, peroxidase, catalase, ascorbate-peroxidase, and glutathione-reductase. In addition, levels of several endogenous phytohormones (indole-3-acetic-acid, cytokinins, abscisic acid, jasmonic acid, and ethylene) were measured. Barley is known to be more salt tolerant than related plant species. Accordingly, none of the studied cultivars exhibited changes in membrane lipid peroxidation under salt stress. However, they responded differently to salt-stress with respect to their accumulation of phytohormones and antioxidant enzyme activity. The strongest and weakest increases in ABA and proline accumulation were observed in Kalaycı and Akhisar, respectively, suggesting that salt-stress was more effectively managed in Kalaycı. The effects of exogenous SA treatment depended on both the timing of the treatment and the cultivar to which it was applied. In general, however, where SA helped mitigate salt stress, it appeared to do so by increasing ROS scavenging capacity and antioxidant enzyme activity. SA treatment also induced changes in phytohormone levels, presumably as a consequence of SA-phytohormone salt-stress cross-talk.

Topics: Antioxidants; Biomass; Chlorophyll; Chlorophyll A; Hordeum; Plant Growth Regulators; Plant Leaves; Plant Shoots; Proline; Reactive Oxygen Species; Salicylic Acid; Salt Stress; Thiobarbituric Acid Reactive Substances; Time Factors; Water

Pecan is one of the most famous nut species in the world. The phenotype of mutants with albino leaves was found in the process of seeding pecan, providing ideal material for the study of the molecular mechanisms leading to the chlorina phenotype in plants. Both chlorophyll a and chlorophyll b contents in albino leaves (ALs) were significantly lower than those in green leaves (GLs). A total of 5171 differentially expression genes (DEGs) were identified in the comparison of ALs vs. GLs using high-throughput transcriptome sequencing; 2216 DEGs (42.85%) were upregulated and 2955 DEGs (57.15%) were downregulated. The expressions of genes related to chlorophyll biosynthesis (

Topics: Carya; Chlorophyll; Chlorophyll A; Gene Expression Profiling; Gene Expression Regulation, Plant; High-Throughput Nucleotide Sequencing; Mutation; Phenotype; Photosynthesis; Plant Leaves; Plant Proteins; Quantitative Trait Loci; Sequence Analysis, RNA

Chlorophyll (Chl) pigments are responsible for vital mechanisms in photosynthetic proteins: light harvesting, energy transfer and charge separation. A complex interplay between the Chl molecule and its microenvironment determines its transition energy. Interactions such as excitonic coupling with one or more pigments (Chls or carotenoids), axial ligation to the magnesium center, or electrostatic interactions between Chl and nearby amino-acid residues all influence the photophysical properties. Here we use time-resolved photodissociation action spectroscopy to determine transition energies of Chla/b complexes in vacuo to directly compare the impact of a negatively charged axial ligand (formate) to that of exciton coupling between two Chls. Experiments carried out at the electrostatic ion storage ring ELISA allow dissociation to be sampled on hundreds of milliseconds time scale. Absorption-band maxima of Chla-formate complexes are found at 433 ± 4 nm/2.86 ± 0.03 eV (Soret band) and in the region 654-675 nm/1.84-1.90 eV (Q band) and those of Chla dimers tagged by a quaternary ammonium ion at 419 ± 5 nm/2.96 ± 0.04 eV (Soret band) and 647 nm/1.92 eV (Q band). The axial ligand strongly affects the Chla transition energies causing redshifts of 0.21 eV of the Soret band and 0.04-0.1 eV of the Q band compared to Chla tagged by a quaternary ammonium. Slightly smaller shifts were found in case of Chlb. The redshifts are approximately twice that induced by excitonic coupling between two Chlas, also tagged by a quaternary ammonium ion. Axial ligation brings the absorption by isolated Chls very close to that of photosynthetic proteins.

Topics: Chlorophyll; Chlorophyll A; Color; Formates; Quaternary Ammonium Compounds; Spectrum Analysis

Canavalia ensiformis is a legume native to Central and South America that has historically been a source of protein. Its main proteins, urease, and lectin have been extensively studied and are examples of bioactive compounds. In this work, the effect of pH and light effects on the growth of C. ensiformis were analyzed. Also, the bioactive compounds such as phenols, carotenoids, chlorophyll a/b, and the growth of callus biomass of C. ensiformis from the effect of different types of light treatments (red, blue and mixture) were evaluated. Likewise, the antioxidative activity of C. ensiformis extracts were studied and related to the production of bioactive compounds. For this, a culture of calluses obtained from seeds were carried out. For the light experiments, polypropylene boxes with red, blue, combination (1/3, 3/1 and 1/1 R-B, respectively) lights and white LED were used as control. In each treatment, three glass containers with 25 ml of MS salts containing 0.25 g of fresh callus were seeded.. The results have shown that the pH of the culture medium notably affects the increase in callogenic biomass. It shows that the pH of 5.5 shows better results in the callogenic growth of C. ensiformis with an average increase of 1.3051 g (198.04%), regarding the initial weight. It was found that the pH 5.5 and the 1/3 R-B LED combination had higher production of bioactive compounds and better antioxidant activity. At the same time, the red-light treatment was least effective.. It was possible to find the ideal conditions of important growth under conditions of pH and light of C. ensiformis. Likewise, it is evaluated whether the production of compounds of interest, such as phenolic compounds and carotenoids, occurs under these conditions. The highest production of calluses occurs in the 1/3 R-B LED combined light treatment, which showed a significant increase in biomass, followed by B. From this study, it could be demonstrated that C. ensiformis produces compounds such as phenols and carotenoids in vitro culture that are essential for the antioxidant activity of the plant.

Topics: Antioxidants; Biomass; Canavalia; Chlorophyll; Chlorophyll A; Culture; Hydrogen-Ion Concentration; Light; Phenols; Phytochemicals

The use of higher plants for the production of plant growth biostimulants is receiving increased attention among scientists, farmers, investors, consumers and regulators. The aim of the present study was to examine the possibility of converting plants commonly occurring in Europe (St. John's wort, giant goldenrod, common dandelion, red clover, nettle, valerian) into valuable and easy to use bio-products. The biostimulating activity of botanical extracts and their effect on the chemical composition of celeriac were identified. Plant-based extracts, obtained by ultrasound-assisted extraction and mechanical homogenisation, were tested in field trials. It was found that the obtained formulations increased the total yield of leaves rosettes and roots, the dry weight of leaves rosettes and roots, the content of chlorophyll

Topics: Antioxidants; Apium; Ascorbic Acid; Benzothiazoles; Biphenyl Compounds; Carotenoids; Chemistry, Pharmaceutical; Chlorophyll; Chlorophyll A; Europe; Fatty Acids; Horticulture; Nitrates; Photosynthesis; Picrates; Pigmentation; Plant Extracts; Plant Leaves; Plant Roots; Polyphenols; Sulfonic Acids; Ultrasonics

The importance of green light for driving natural photosynthesis has long been underappreciated, however, under the presence of strong illumination, green light actually drives photosynthesis more efficiently than red light. This green light is absorbed by mixed vibronic Q

Topics: Chlorophyll; Chlorophyll A; Color; Energy Transfer; Light-Harvesting Protein Complexes; Photons; Photosynthesis; Plant Leaves; Spectrum Analysis; Thylakoids

Heavy losses by dark leaf spot disease in oilseed Brassica have incited research towards identifying sources of genetic tolerance against causal pathogen, Alternaria brassicicola. Several morpho-molecular parameters were evaluated to test the performance of field mustard and rapeseed genotypes under artificial inoculation with this pathogen. During Brassica-Alternaria interaction, physio-biochemical defense response was witnessed in tolerant genotypes. Two tolerant genotypes (one for field mustard and one for rapeseed), i.e., EC250407 and EC1494 were identified. However, necrotic lesions were more prominent in susceptible genotypes with minimum chlorophyll (chlorophyll a, chlorophyll b and total chlorophyll) and carotenoids contents. Contrary to photosynthetic pigments, increase in total soluble protein (TSP) contents was observed with disease progression in susceptible genotypes. Tolerant genotypes of field mustard and rapeseed displayed remarkable increase in the activities of redox enzyme in infected leaves with least yield loss (6.47% and 5.74%) and disease severity index (DSI) of 2.9 and 2.1, respectively. However, yield/plant showed close association with other morpho-yield parameters, photosynthetic pigments and redox enzymes (superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD)) activities except silique length and TSP. Based on the results of morpho-biochemical analyses, redox enzymes and morphological parameters; their interplay is proposed to determine the tolerance outcome of the Brassica-A. brassicicola interaction.

Topics: Alternaria; Antioxidants; Brassica napus; Brassica rapa; Brassicaceae; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Disease Resistance; Genotype; Oxidation-Reduction; Photosynthesis; Plant Diseases; Superoxide Dismutase

The drought-tolerant crop plants, including rice, are required for fulfilling food requirements when drought occurs in Indonesia. The response to drought stress in rice cultivars could be studied based on the morphological, anatomical and physiological characteristics. This study evaluated the drought tolerance based on the physiological characteristics at the vegetative phase in six rice cultivars cultivated in North Sulawesi, Indonesia.. The Completely Randomized Design experiment was conducted in the greenhouse by using six rice cultivars (cv. Superwin, Sultan, Ciherang, Serayu, Cigeulis and IR 64) grown in the soil mixture at the vegetative phase. The treatments in this experiment were water deficit (without water for up to 22 days) and well-watered (watering until field capacity). The evaluated physiological characteristics consisted of leaf water content, leaf relative water content, concentrations of chlorophylls (total, a and b) at 0, 7, 14, 17 and 22 days after treatment.. Withholding water for 22 days at the vegetative phase resulted in a decrease of water content and the increase of concentration of chlorophylls (total, a and b) in leaf. There were two categories of drought tolerance in rice observed in this study, i.e. semi tolerant for Cigeulis, Superwin, Serayu, IR 64, Sultan and non-tolerant for Ciherang.. Rice cv. Cigeulis, Superwin, Serayu, IR 64 and Sultan were semi tolerant, whereas cv. Ciherang was non-tolerant rice cultivar. Leaf water content was a potential physiological indicator for drought tolerance in rice.

Topics: Adaptation, Physiological; Chlorophyll; Chlorophyll A; Crops, Agricultural; Droughts; Indonesia; Oryza; Plant Leaves; Stress, Physiological; Time Factors; Water

Sucrose metabolism contributes to the growth and development of plants and helps plants cope with abiotic stresses, including stress from Cd. Many of these processes are not well-defined, including the mechanism underlying the response of sucrose metabolism to Cd stress. In this study, we investigated how sucrose metabolism in maize varieties with low (FY9) and high (SY33) sensitivities to Cd changed in response to different levels of Cd (0 (control), 5, 10, and 20 mg L-1 Cd). The results showed that photosynthesis was impaired, and the biomass decreased, in both varieties of maize at different Cd concentrations. Cd inhibited the activities of sucrose phosphate synthase (SPS) and sucrose synthase (SS) (sucrose synthesis), and stimulated the activities of acid invertase (AI) and SS (sucrose hydrolysis). The total soluble sugar contents were higher in the Cd-treated seedlings than in the control. Also, Cd concentrations in the shoots were higher in SY33 than in FY9, and in the roots were lower in SY33 than in FY9. The decreases in the photosynthetic rate, synthesis of photosynthetic products, enzyme activity in sucrose synthesis direction, and increases in activity in hydrolysis direction were more obvious in SY33 (the sensitive variety) than in FY9 (the tolerant variety), and more photosynthetic products were converted into soluble sugar in SY33 than in FY9 as the Cd stress increased. The transcript levels of the sugar transporter genes also differed between the two varieties at different concentrations of Cd. These results suggest that sucrose metabolism may be a secondary response to Cd additions, and that the Cd-sensitive variety used more carbohydrates to defend against Cd stress rather than to support growth than the Cd-tolerant variety.

Topics: Biomass; Cadmium; Chlorophyll; Chlorophyll A; Gene Expression Profiling; Gene Expression Regulation, Plant; Glucosyltransferases; Photosynthesis; Plant Proteins; Plant Roots; Plant Shoots; RNA, Messenger; Solubility; Stress, Physiological; Sucrose; Sugars; Zea mays

Five-day exposure of clary sage (

Topics: Cadmium; Chlorophyll; Chlorophyll A; Chloroplasts; Hormesis; Hydroponics; Microscopy, Electron, Transmission; Photochemistry; Photosynthesis; Photosystem II Protein Complex; Plant Leaves; Plant Roots; Reactive Oxygen Species; Salvia

The origin of chlorophyll

Topics: Arabidopsis; Arabidopsis Proteins; Chlorophyll; Chlorophyll A; Chlorophyllides; Fluorescence; Gene Expression Regulation, Plant; Mutation; Oxygenases; Phenotype; Photosynthesis; Photosystem I Protein Complex; Photosystem II Protein Complex; RNA, Messenger; Thylakoids

This study comparatively examined spectroscopic features, photosynthetic parameters and energy partitioning in plants of Fittonia albivenis cv. Agyroneura and Fittonia albivenis cv. Verschaffeltii with different pigmentation. Fittonia albivenis cv. Verschaffeltii, rich in anthocyanins, presented lower values than the green variety (cv. Agyroneura) for several parameters: the ratio chlorophyll a/b, the carotenoid content, the heat dissipation by nonphotochemical quenching (NPQ) and the energy-dependent component of the quantum yield of NPQ. Additionally, the red plant displayed higher resistance to water shortage. The spectral distribution of the chlorophyll a fluorescence, free from distortions due to light reabsorption processes, was obtained for both varieties by application of a physical model previously developed in our group. From this modeling, a higher ratio photosystem II/photosystem I was inferred for the red variety, in agreement with the screening effect of anthocyanins. From a thorough analysis of the fluorescence, the different operating strategies adopted by these plants with dissimilar pigmentation could be elucidated. These strategies were related to the photosystem stoichiometry, the distribution of the absorbed energy and the dissipation of heat under increasing light intensities.

Topics: Acanthaceae; Anthocyanins; Carotenoids; Chlorophyll; Chlorophyll A; Pigmentation; Plant Leaves

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Chloroplasts; Color; Gene Expression Profiling; Gene Expression Regulation, Developmental; Gene Expression Regulation, Plant; Microscopy, Electron, Transmission; Mutation; Photosynthesis; Pigments, Biological; Plant Leaves; Thylakoids; Ulmus

An investigation of the photoexcited triplet state of chlorophyll (Chl)

Topics: Chlorophyll; Chlorophyll A; Density Functional Theory; Electron Spin Resonance Spectroscopy; Models, Molecular; Molecular Conformation

Stereocaulon sorediiferum is expected to be a Cu-hyperaccumulator lichen and has fluorescent substances. To clarify the relationship between the fluorescence (FL) of the lichen and its Cu concentration, we collected S. sorediiferum samples at Cu-contaminated and uncontaminated sites in Japan, determined the concentration of Cu, K, Mg, Al, Ca, Mn, Fe, Zn, chlorophyll a,b, and total carotenoids in them, analyzed lichen secondary metabolites and fluorescent substances extracted from them, and measured the FL of them and their extracts. We found that the FL intensity of S. sorediiferum samples is significantly negatively correlated with their Cu concentration. The application of its FL for Cu monitoring may allow a new nondestructive quantitative method for assessing Cu contamination. The spectroscopic and chromatographic analysis shows that the fluorescent substances negatively correlated with Cu concentration are not major lichen secondary metabolites (lobaric acid and atranorin) and remain after immersion in acetone. The correlation analysis and the comparison with the causal relationship between Cu concentration and the chlorophyll a/b ratio suggest that the reason for the decrease in FL intensity with increasing Cu concentration is a structural change of the fluorescent substances by accumulated Cu. These findings lead to a better understanding of the relationship between the FL of S. sorediiferum and its Cu concentration and provide new insights into fluorescent lichen substances.

Topics: Air Pollutants; Ascomycota; Chlorophyll; Chlorophyll A; Copper; Depsides; Fluorescence; Hydroxybenzoates; Japan; Lactones; Lichens; Salicylates

The Water-Soluble Chlorophyll Protein (WSCP) of Brassicaceae is a remarkably stable tetrapyrrole-binding protein that, by virtue of its simple design, is an exceptional model to investigate the interactions taking place between pigments and their protein scaffold and how they affect the photophysical properties and the functionality of the complexes. We investigated variants of WSCP from Lepidium virginicum (Lv) and Brassica oleracea (Bo), reconstituted with Chlorophyll (Chl) b, to determine the mechanisms by which the different Chl binding sites control their Chl a/b specificities. A combined Raman and crystallographic investigation has been employed, aimed to characterize in detail the hydrogen-bond network involving the formyl group of Chl b. The study revealed a variable degree of conformational freedom of the hydrogen bond networks among the WSCP variants, and an unexpected mixed presence of hydrogen-bonded and not hydrogen-bonded Chls b in the case of the L91P mutant of Lv WSCP. These findings helped to refine the description of the mechanisms underlying the different Chl a/b specificities of WSCP versions, highlighting the importance of the structural rigidity of the Chl binding site in the vicinity of the Chl b formyl group in granting a strong selectivity to binding sites.

Topics: Brassicaceae; Chlorophyll; Chlorophyll A; Crystallography, X-Ray; Hydrogen Bonding; Photosynthesis; Plant Proteins; Protein Conformation; Solubility; Spectrum Analysis, Raman; Water

Alkalinity is a known threat to crop plant growth and production, yet the role of exogenous silicon (Si) and salicylic acid (SA) application has been largely unexplored. Here, we sought to understand the beneficial impacts of Si and SA on tomato seedlings during high-pH (9.0) stress. Results showed that Si- and SA-treated plants displayed higher biomass, chlorophyll contents, relative leaf water and better root system than none-treated plants under alkaline conditions. Both Si and SA counteracted the alkaline stress-induced oxidative damage by lowering the accumulation of reactive oxygen species and lipid peroxidation. The major antioxidant defence enzyme activities were largely stimulated by Si and SA, and these treatments caused significantly increased K

Topics: Abscisic Acid; Antioxidants; Biomass; Chlorophyll; Chlorophyll A; Hydrogen-Ion Concentration; Lipid Peroxidation; Oxidative Stress; Plant Leaves; Plant Roots; Plant Shoots; Rhizosphere; Salicylic Acid; Seedlings; Silicon; Soil; Solanum lycopersicum; Stress, Physiological

Photosynthetic pigments perform critical physiological functions in tea plants. Their content is an essential indicator of photosynthetic efficiency and nutritional status. The present study aimed to predict chlorophyll a (Chl a), chlorophyll b (Chl b), total chlorophyll (total Chl), and carotenoid (Car) content in tea leaves under different levels of nitrogen treatment using hyperspectral imaging (HSI) in combination with variable selection algorithms.. A total of 150 samples were collected and scanned using the HSI system. The mean spectrum in the region of interest (ROI) was extracted, and the pigment content was measured by traditional chemical methods. Five and seven optimal wavelengths (OWs) were selected using the regression coefficients (RCs) of partial least squares regression (PLSR) and the second-derivative (2-Der), respectively. The optimal 2-Der-PLSR models for Chl a, Chl b, total Chl, and Car performed remarkably well based on seven OWs with correlation coefficients of prediction (R. The results of this study revealed that HSI combined with variable selection method can be employed as a rapid and accurate method for predicting the content of pigments in tea plants. © 2018 Society of Chemical Industry.

Topics: Algorithms; Camellia sinensis; Carotenoids; Chlorophyll; Chlorophyll A; Color; Fertilizers; Least-Squares Analysis; Nitrogen; Pigments, Biological; Plant Leaves; Spectrum Analysis

It is reported that the accumulated polycyclic aromatic hydrocarbons (PAHs) can cause wheat leaf chlorosis, and we identified that carotenoid (Car) and superoxide dismutase (SOD) are the two most active factors in antioxidant system in the previous study. Herein, we applied Car as an exogenous chemical added to alleviate the toxicity triggered by phenanthrene (a model PAH) in wheat seedlings. In the exogenous Car addition groups, we found that the leaf number would grow three, and the relative biomass and the relative root length of 20 mg L

Topics: Antioxidants; Carotenoids; Chlorophyll; Chlorophyll A; Malondialdehyde; Oxidative Stress; Phenanthrenes; Plant Leaves; Seedlings; Soil Pollutants; Superoxide Dismutase; Triticum

The role of the environment in excitation energy transport in the pigment-protein complexes (PPCs) of photosynthetic organisms is a widely investigated topic. The spectral density is a key component in understanding this protein-pigment interaction; however, the typical approach for calculating spectral density, combining molecular dynamics with quantum chemistry (QC) calculations, suffers from the geometry mismatch problem, arising from the structural inconsistency between the force field (FF) and the QC calculation. Existing parameterization methods demand much time-consuming manual inputs, limiting the number of systems that can be studied. We present a method, utilizing force matching for the autoparameterization of new pigment FFs for the use in spectral density calculations of PPCs, and apply the method to three pigments. The use of these optimized FFs in spectral density computation results in a notable difference in comparison to the original FF.

Topics: Bacteria; Bacterial Proteins; Bacteriochlorophyll A; Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Light-Harvesting Protein Complexes; Molecular Dynamics Simulation; Quantum Theory

We tested the potential of Raman microspectroscopy to determine carotenoid pigments - both primary (lutein, beta-carotene) and secondary (astaxanthin) carotenoids - in the different species and life-cycle stages of snow algae from the order Chlamydomonadales (Chlorophyta). We compared the performance of Raman spectrometry to a reference method of biological pigment analysis, high-performance liquid chromatography (HPLC). The three main carotenoid Raman bands of the astaxanthin-rich red cysts were located at 1520, 1156 and 1006 cm

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Europe; Geography; Microalgae; Microspectrophotometry; Snow; Spectrum Analysis, Raman

Thorium (Th) is a natural radioactive element present in the environment and has the potential to be used as a clean nuclear fuel. Relatively little is known about the aquatic toxicity of Th, especially in nanoparticulate form, which may be the main chemical species of Th in the natural waters. In this study, impacts of ThO

Topics: Cell Membrane Permeability; Chlorella; Chlorophyll; Chlorophyll A; Nanoparticles; Oxidation-Reduction; Oxidative Stress; Particle Size; Reactive Oxygen Species; Thorium Dioxide; Toxicity Tests; Water Pollutants, Chemical

The purpose of this experiment was to study the effects of different shading conditions on the growth,physiological characteristics and biomass allocation of Polygonatum cyrtonema,which offered a theoretical basis for its cultivation.Different light environments(100%,80%,60% and 35% light transmittance) were simulated with shading treatments.Growth and photosynthetic indexes of P.cyrtonema were measured and the variances were analyzed.The results show that shading decreased superoxide anion radical(O-·2)production rate and hydrogen peroxide(H_2O_2) accumulation,kept the activity of SOD,POD and CAT enzyme at a high level.Furthermore,The content of chlorophyll a and chlorophyll b,net photosynthetic rate(Pn),stomatal conductance(Gs),transpiration rate(Tr),maximal photochemical efficiency of photosystem Ⅱ(Fv/Fm),photochemical quenching index(q P) and effective quantum yield of photosystem II(ΦPSⅡ) of P.cyrtonema were increased while the intercellular CO2 concentration(Ci),Foand NPQ were decreased by shading.Shading is beneficial to P.cyrtonema growth,can increase the total biomass P.cyrtonema.The allocation proportion of biomass on the aerial portion of P.cyrtonema increased but underground parts decreased with increasing shading conditions.In this study,P.cyrtonema can grow well in shading conditions,shading is beneficial to the formation of the yield and quality of the rhizomes of P.cyrtonema,especially in 65% light transmittance.

Topics: Biomass; Chlorophyll; Chlorophyll A; Photosynthesis; Plant Leaves; Plant Stomata; Plant Transpiration; Polygonatum; Sunlight

Arsenic (As) is a significant contaminant in the environment and its detection through macrophytes can provide a powerful tool. Myriophyllum alterniflorum constitutes a good candidate by virtue of its ability to accumulate contaminants, and moreover its biomarkers can respond to the presence of trace metals and metalloids. The objective of this study therefore is to evaluate the watermilfoil response to As exposure under several hydrodynamic conditions since it is well known that hydrodynamics affect plant functioning. For this purpose, fresh watermilfoil plants are subjected to three hydrodynamic conditions, namely laminar, turbulent and calm, in a synthetic medium either enriched or not by 100 μg.L

Topics: Arsenic; Biodegradation, Environmental; Biomarkers; Carotenoids; Chlorophyll; Chlorophyll A; Environmental Monitoring; Hydrodynamics; Hydrogen Peroxide; Oxidative Stress; Photosynthesis; Plant Roots; Plant Shoots; Saxifragales; Trace Elements; Water Quality

Aluminum (Al) is one of the most important stress factors that reduce plant productivity in acidic soils. Present work thereby analyzed Al-induced genomic alterations in Bryophyllum daigremontianum clones using RAPD and ISSR markers, and investigated responding changes in photosynthetic pigment (chlorophyll a, b, a/b, total chlorophyll and carotenoid) contents and total soluble protein amounts in plant leaves. The main reason for the use of bulbiferous spurs originated clone plants was to increase reliability and acceptability of RAPD and ISSR techniques in DNA fingerprinting. Raised 40 clone plants were divided into five separate groups each with eight individuals and each experimental group was watered with 0 (control), 0 (acid control), 50, 100 and 200 µM AlCl

Topics: Aluminum; Carotenoids; Chlorophyll; Chlorophyll A; DNA Fingerprinting; DNA, Plant; Genetic Markers; Genetic Variation; Kalanchoe; Microsatellite Repeats; Plant Leaves; Polymorphism, Genetic; Random Amplified Polymorphic DNA Technique; Reproducibility of Results

In some cultivars, yellowing resulting from chlorophyll breakdown has a direct and negative effect on food supply and health. The 'Zaosu' pear (Pyrus bretschneideri Rehd.), a commercial Asian pear cultivar in China, rapidly turns yellow when stored at room temperature after harvest. To develop techniques that delay or suppress chlorophyll degradation, the effects of methyl salicylate (MeSA) on yellowing in 'Zaosu' pear fruit during storage were evaluated.. Compared with the untreated fruit, the application of 0.05 mmol L. Methyl salicylate could delay chlorophyll breakdown in the fruit. The results also suggested that the conversion from chlorophyll a to pheophorbide a could proceed via two pathways, and that alternative pathways for the breakdown of chlorophyll a exist in 'Zaosu' pears. © 2019 Society of Chemical Industry.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Food Preservation; Food Preservatives; Food Storage; Fruit; Pyrus; Salicylates

Chlorophyll is a vital photosynthetic pigment that plays a key role in plant development, participating in light energy capture and energy conversion. In this study, a novel wucai ( Brassica campestris L.) germplasm with green outer leaves and yellow inner leaves at the adult stage (W7-2) was used to examine chlorophyll metabolism response to cold acclimation. A green leaf wucai genotype without leaf color changes named W7-1 was selected as the control to evaluate the chlorophyll metabolism changes of W7-2. Compared to W7-1, the contents of chlorophyll a (Chl a) and chlorophyll b (Chl b) in W7-2 were significantly reduced at five developmental stages (13, 21, 29, 37, and 45 days after planting (DAP)). An iTRAQ-based quantitative proteomic analysis was carried out at 21 and 29 DAP according to the leaf color changes in both of genotypes. 1409 proteins were identified, while 218 of them displayed differential accumulations between W7-2 and W7-1 during the two developmental stages. The differentially expressed proteins (DEPs) mainly assigned to chlorophyll biosynthesis, photosynthesis, carbohydrate metabolism, ribosome metabolism and posttranslational modification. Among these DEPs, NADPH-protochlorophyllide oxidoreductase (PORB) and Mg-protoporphyrin IX chelatase 1 (CHLI1) were the key enzymes participating in chlorophyll (Chl) biosynthesis, which was down-regulated at 21 DAP and up-regulated at 29 DAP in W7-2 compared with W7-1, respectively. The expression analysis of genes of three subunits of Mg-chelatase ( CHLI1, CHLD, and CHLH), Genomes Uncoupled 4 ( GUN4), and Thioredoxin ( TRX3) associated with chlorophyll metabolism also displayed significant down-regulation in W7-2. In particular, PORB showed significant up-regulation in W7-2, significantly affecting chlorophyll biosynthesis. Additionally, differences in chlorophyll metabolism between W7-2 and W7-1 were in terms of altered photosynthesis, carbohydrate, and energy metabolism. We found that the transcription levels of most photosynthesis proteins showed significantly lower levels, and the genes expression level, associated with carbohydrate and energy metabolism, were lower in W7-2 than in W7-1. Therefore, the present study results help understand the physiological and molecular mechanisms underlying leaf coloring responding to cold acclimation.

Topics: Acclimatization; Brassica; Chlorophyll; Chlorophyll A; Cold Temperature; Pigments, Biological; Plant Leaves; Proteomics

Topics: Araceae; Carotenoids; Chlorophyll; Chlorophyll A; Gamma Rays; Lipid Peroxidation; Oxidative Stress; Photosynthesis; Photosystem II Protein Complex; Radiation, Ionizing; Reactive Oxygen Species

Chlorophylls a and b (Chla/b) are responsible for light-harvesting by photosynthetic proteins in plants. They display broad absorption in the visible region with multiple bands, due to the asymmetry of the macrocycle and strong vibronic coupling. Their photophysics relies on the microenvironment, with regard to transition energies as well as quenching of triplet states. Here, we firmly establish the splitting of the Q and Soret bands into x- and y- polarized bands for the isolated molecules in vacuo, and resolve vibronic features. Storage-ring experiments reveal that dissociation of photoexcited charge-tagged complexes occurs over several milliseconds, but with two different time constants. A fast decay is ascribed to dissociation after internal conversion and a slow decay to the population of a triplet state that acts as a bottleneck. Support for the latter is provided by pump-probe experiments, where a second laser pulse probes the long-lived triplet state.

Topics: Chlorophyll; Chlorophyll A; Energy Transfer; Light-Harvesting Protein Complexes; Photosynthesis; Plants; Quantum Theory; Spectrophotometry

Although iron oxide occurs naturally in the environment, iron oxide nanoparticles have distinct mobility, reactivity, and toxicity, which can harm the human health and nature. This scenario has motivated the investigation of the toxic effects of iron oxide nanoparticles (akaganeite predominance + hematite) on the aquatic plant Lemna minor. First, nanoparticles were synthesized and characterized; then, different iron oxide NP concentrations were added to Lemna minor culture. After 7 days, all the Lemna minor leaves died, irrespective of the added NP concentration. The iron oxide NP impact on the plant was evaluated based on malondialdehyde (MDA) production from thiobarbituric acid reactive substances (TBARS), which was dose-dependent; i.e., lipid peroxidation in the plant increased with rising iron oxide NP concentration. The chlorophyll content decreased at high iron oxide NP concentrations, which disrupted the light absorption mechanism. Fe accumulation in Lemna minor roots also occurred, which can harm nutrient uptake. Therefore, the iron oxide NP toxic impact on plants and related ecosystems requires further studies in order to prevent environmental damage.

Topics: Araceae; Chlorophyll; Chlorophyll A; Ferric Compounds; Iron; Lipid Peroxidation; Malondialdehyde; Nanoparticles; Plant Leaves; Reactive Oxygen Species; Thiobarbituric Acid Reactive Substances; Water Pollutants, Chemical

Chlamydomonas reinhardtii (WT 2137) P. A. Dang. (Volvocales, Chlorophyceae) is a green microalgae serving as a suitable model in scientific research and a promising industrial biotechnology platform for production of biofuel, hydrogen and recombinant proteins. Fullerenes (C60) are allotropic carbon nanoparticles discovered in 1985 and used in biomedical studies since the early 1990s, when water solubilization methodologies were developed. Recently, surface-modified hydroxylated derivatives of fullerenes were proven to enhance algal growth and drought tolerance in plants. Here, a novel type of water-soluble [60]fullerene derivative with 12 glycine residues (GF) has been synthesized and tested for acute toxicity (up to 50 µg/ml) and as a potential biostimulant of algal growth. The effects of GF on pigment composition and growth rate of Chlamydomonas reinhardtii were systematically investigated. Our results suggest that GF was not toxic, and no negative change in the pigment content and no stress symptoms were observed. No changes in the photosynthetic parameters based on the fluorescence of chlorophyll a in Photosystem II (NPQ, Fv/Fm, Fv/F0, PI and RC/ABS) were observed. The GF had no effect on cell size and growth rate. At a concentration of 20 µg/ml, GF stimulated chlorophyll accumulation in 3-day-old cultures.

Topics: Biofuels; Biotechnology; Carotenoids; Cell Proliferation; Cell Size; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Fullerenes; Hydrogen; Nanoparticles; Photosynthesis; Recombinant Proteins; Solubility; Water

Topics: Air Pollutants; Air Pollution; Chlorophyll; Chlorophyll A; Coal; Dust; Environmental Exposure; Environmental Monitoring; India; Plant Leaves; Plants; Power Plants; Species Specificity; Water

The antioxidant balance, photochemical activity of photosystem II (PSII), and photosynthetic pigment content, as well as the expression of genes involved in the light signalling of callus lines of Eutrema salsugineum plants (earlier Thellungiella salsuginea) under different spectral light compositions were studied. Growth of callus in red light (RL, maximum 660 nm), in contrast to blue light (BL, maximum 450 nm), resulted in a lower H

Topics: Antioxidants; Brassicaceae; Carotenoids; Cell Survival; Chlorophyll; Chlorophyll A; Enzymes; Gene Expression Regulation, Plant; Hydrogen Peroxide; Light; Photoreceptors, Plant; Photosystem II Protein Complex; Plant Proteins; Signal Transduction; Thiobarbituric Acid Reactive Substances; Tissue Culture Techniques

Glyphosate is a broad-spectrum herbicide that is frequently detected in water bodies and is harmful to aquatic systems. We conducted an experiment to explore the ecological sensitivity of Hydrilla verticillata (L.f.) Royle and Vallisneria natans (Lour.) Hara to glyphosate. Our research focused on the physiological responses of H. verticillata and V. natans after exposure to various concentrations of glyphosate (0, 1, 10, 20, 30, 40, 50 and 80 mg/L) in hydroponic culture after one day (1D) and seven days (7D). The results show that after 1D, the soluble protein content of H. verticillata was significantly stimulated under low herbicide concentrations. Other indices for H. verticillata and V. natans had no remarkable changes at 1D. After 7D of treatment, the soluble protein content of H. verticillata showed no significant differences, while the malondialdehyde (MDA), pigment contents and catalase (CAT) activity significantly increased at low glyphosate concentrations. Guaiacol peroxidase (POD) activity in H. verticillata significantly increased with increasing herbicide concentrations. The chlorophyll a/b ratio of H. verticillata sharply decreased above 10 mg/L. For V. natans, soluble protein, chlorophyll a, and carotenoid content; and CAT activity declined significantly after glyphosate application, while other indicators showed no significant changes. Our results indicate that glyphosate concentrations from 0 to 80 mg/L can induce oxidative stress in H. verticillate and may impede metabolism processes for protein and pigments without causing oxidative stress in V. natans. Taken together, our results suggest that the sensitivity of H. verticillata to glyphosate exposure is higher than that of V. natans.

Topics: Animals; Catalase; Chlorophyll; Chlorophyll A; Glycine; Glyphosate; Herbicides; Hydrocharitaceae; Malondialdehyde; Oxidative Stress

The abundance of calcareous soils makes bicarbonate-induced iron (Fe) deficiency a major problem for plant growth and crop yield. Therefore, Fe-efficient plants may constitute a solution for use on calcareous soils. We investigated the ability of the forage legume Sulla carnosa (Desf.) to maintain integrity of its photosynthetic apparatus under Fe deficiency conditions. Three treatments were applied: control, direct Fe deficiency and bicarbonate-induced Fe deficiency. At harvest, all organs of deficient plants showed severe growth inhibition, the effect being less pronounced under indirect Fe deficiency. Pigment analysis of fully expanded leaves revealed a reduction in concentrations of chlorophyll a, chlorophyll b and carotenoids under Fe deficiency. Electron transport rate, maximum and effective quantum yield of photosystem II (PSII), photochemical quenching (qP), non-photochemical quenching (qN) as well as P700 activity also decreased significantly in plants exposed to direct Fe deficiency, while qN was not affected. The effects of indirect Fe deficiency on the same parameters were less pronounced in bicarbonate-treated plants. The relative abundances of thylakoid proteins related to PSI (PsaA, Lhca1, Lhca2) and PSII (PsbA, Lhcb1) were also more affected under direct than indirect Fe deficiency. We conclude that S. carnosa can maintain the integrity of its photosynthetic apparatus under bicarbonate-induced Fe deficiency, preventing harmful effects to both photosystems under direct Fe deficiency. This suggests a high capacity of this species not only to take up Fe in the presence of bicarbonate (HCO

Topics: Bicarbonates; Carotenoids; Chlorophyll; Chlorophyll A; Electron Transport; Fabaceae; Iron Deficiencies; Photosynthesis; Photosystem I Protein Complex; Photosystem II Protein Complex; Plant Leaves

Present study was conducted to evaluate the effect of lead tolerant plant growth promoting rhizobacteria (LTPGPR) on growth, physiology, yield, antioxidant activities and lead uptake in sunflower in soil contaminated with lead under pot conditions. Three pre-characterized LTPGP strains (S2 (Pseudomonas gessardii strain BLP141), S5 (Pseudomonas fluorescens A506) and S10 (Pseudomonas fluorescens strain LMG 2189)) were used to inoculate sunflower growing in soil contaminated with different levels (300, 600 and 900 mg kg

Topics: Antioxidants; Ascorbate Peroxidases; Catalase; Chlorophyll; Chlorophyll A; Environmental Pollution; Glutathione Reductase; Helianthus; Lead; Nitrates; Plant Development; Plant Roots; Proline; Pseudomonas; Soil; Soil Microbiology; Soil Pollutants; Superoxide Dismutase

The first step in chlorophyll a degradation is the extraction of the central Mg. This reaction is catalyzed by Mg-dechelatase encoded by Stay-Green (SGR) in land plants. SGR extracts Mg from chlorophyll a but not from chlorophyll b, and chlorophyll b must be converted to chlorophyll a before degradation. The first reaction of the chlorophyll b to chlorophyll a conversion is catalyzed by chlorophyll b reductase. Non-Yellow Coloring 1 (NYC1) and NYC1 like (NOL) are isozymes of chlorophyll b reductase. When SGR was transiently overexpressed in Arabidopsis, both chlorophyll a and b were degraded, suggesting that the chlorophyll b to chlorophyll a conversion is activated by SGR overexpression. To examine the involvement of chlorophyll b reductases in SGR-induced chlorophyll b degradation, SGR was transiently overexpressed in nyc1, nol, and nyc1 nol double mutants by dexamethasone treatment. It was found that in the wild type and nol mutant, chlorophyll a and b were degraded and all the chlorophyll-binding proteins decreased. Meanwhile, in nyc1 and nyc1 nol mutants, chlorophyll b degradation was suppressed and the light-harvesting complex of photosystem II remained. The mRNA and protein levels of NYC1 increased after SGR overexpression in wild type plants. These results suggest that Mg-dechelation of chlorophyll a by SGR activates chlorophyll b degradation by inducing the expression of NYC1. This is an effective regulation of a metabolic pathway.

Topics: Arabidopsis; Arabidopsis Proteins; Chlorophyll; Chlorophyll A; Chloroplast Proteins; Magnesium; Membrane Proteins; Oxidoreductases

Information on metallophytes during reclamation of land contaminated with heavy metals is sparse. We investigated the response of D. carthusianorum calamine ecotype to Pb and Cd stress. We focused on in vitro selection of tolerant plant material for direct use in chemically degraded areas. Shoot cultures were treated with various concentrations of Pb or Cd ions. Plantlet status was estimated as micropropagation efficiency, growth tolerance index (GTI) and through physiological analysis. Moreover, determination of plant Pb, Cd and other elements was performed. The application of Pb(NO

Topics: Amides; Cadmium; Carotenoids; Chlorophyll; Chlorophyll A; Dianthus; Ecotype; In Vitro Techniques; Ropivacaine

The aim of this study is to determine the combined effect of copper and hydrodynamic conditions on the response of certain biomarkers of an aquatic macrophyte, namely Myriophyllum alterniflorum. Watermilfoil biomarkers are monitored in a synthetic medium enriched or not with copper (100 μg.L

Topics: Biomarkers; Cell Respiration; Chlorophyll; Chlorophyll A; Copper; Environmental Monitoring; Hydrodynamics; Hydrogen Peroxide; Photosynthesis; Plant Roots; Plant Shoots; Water Quality

Standard chemical toxicity testing guidelines using aquatic plant Lemna minor have been developed by several international standardisation organisations. Although being highly useful for regulatory purposes by focusing on traditional adverse endpoints, these tests provide limited information about the toxic mechanisms and modes of action (MoA). The present study aimed to use selected functional assays in L. minor after exposure to 3,5-dichlorophenol (3,5-DCP) as a model to characterise the toxic mechanisms causing growth inhibition and lethality in primary producers. The results demonstrated that 3,5-DCP caused concentration-dependent effects in chloroplasts and mitochondria. Uncoupling of oxidative phosphorylation (OXPHOS), reduction in chlorophyll (Chlorophyll a and b) content, reproduction rate and frond size were the most sensitive endpoints, followed by formation of reactive oxygen species (ROS), lipid peroxidation (LPO), reduction of carotenoid content and impairment of photosynthesis efficiency. Suppression of photosystem II (PSII) efficiency, electron transport rate (ETR), chlorophyll (a and b) contents and oxidative phosphorylation (OXPHOS) were closely correlated while ROS production and LPO were negative correlated with ETR, carotenoid content and growth parameters. A network of conceptual Adverse Outcome Pathways (AOPs) was developed to decipher the causal relationships between molecular, cellular, and apical adverse effects occurring in L. minor to form a basis for future studies with similar compounds.

Topics: Aerobiosis; Araceae; Carotenoids; Chlorophenols; Chlorophyll; Chlorophyll A; Electron Transport; Fluorescence; Lipid Peroxidation; Membrane Potential, Mitochondrial; Oxidative Phosphorylation; Oxidative Stress; Photosynthesis; Photosystem II Protein Complex; Principal Component Analysis; Reactive Oxygen Species; Water Pollutants, Chemical

Salicylic acid (SA), Epi-brassinolide (EBL) and calcium (Ca) play crucial roles in plant development and mediate plant response to biotic and abiotic stress. This study was aimed to investigate the possible mediatory role of SA, EBL, Ca or their combination in protecting tomato plants from cadmium (Cd) toxicity. According to the results, Cd stress resulted in a significant reduction of plant dry mass, photosynthetic pigment content as well as photosynthetic rate. Exogenous application of SA decreased the malondialdehyde (MDA) level by 39.27% and increased catalase (CAT) activity by 81.17%. SA and EBL treatment significantly increased chlorophyll a (Chl a), chlorophyll b (Chl b) content, photosynthetic rate (Pn) as well as water use efficiency (WUE). SA+EBL (1:1)/Ca+SA+EBL (1:1:1) treatment obviously alleviated Cd-induced growth inhibition, the dry mass of different tomato organs were significantly increased (p < 0.05). Especially in Ca+SA+EBL treated plants, the dry mass of roots, stems and leaves increased by 141.18%, 128.57% and 118.52%, respectively. Besides, SA+EBL and Ca+SA+EBL treatments reduced the MDA level, but increased photosynthetic pigment concentration and photosynthetic efficiency. CAT activity was increased by 62.92% in Ca+SA+EBL treated plants, the WUE was increased by 557.76% in SA+EBL pretreated plants. Moreover, exogenous application of SA, SA+EBL and Ca+SA+EBL significantly decreased Cd accumulation in tomato organs (p < 0.05) compared with Cd-stressed plants. Taken together, our results indicated that exogenous application of SA, EBL and Ca individually or in combination could alleviate Cd toxicity in tomato plants, although the extent varies.

Topics: Brassinosteroids; Cadmium; Calcium; Catalase; Chlorophyll; Chlorophyll A; Malondialdehyde; Photosynthesis; Plant Leaves; Plant Roots; Salicylic Acid; Seedlings; Solanum lycopersicum; Steroids, Heterocyclic; Stress, Physiological

Naphthenic acids (NAs) account for 1-2% of crude oil and represent its main acidic component. However, the aquatoxic effects of NAs on marine phytoplankton and their ecological risks have remained largely unknown. Using the marine microalgae Phaeodactylum tricornutum and Platymonas helgolandica var. tsingtaoensis as the target, we studied the effects of NAs on their growth, cell morphology and physiological characteristics. The cell density decreased as the concentrations of NAs increased, indicating that they had an adverse effect on growth of the investigated algae in a concentration-dependent manner. Moreover, scanning electron microscopy revealed NAs exposure caused damage such as deformed cells, shrunken surface and ruptured cell structures. Exposure to NAs at higher concentrations for 48 h significantly increased the content of chlorophyll (Chl) a and b in P. tricornutum, but decreased their levels in P. helgolandica var. tsingtaoensis. NAs with concentrations no higher than 4 mg/L gradually enhanced the Chl fluorescence (ChlF) parameters and decreased the ChlF parameters at higher concentrations for the two marine microalgae. Additionally, NAs induced hormesis on photosynthetic efficiency of the two microalgae and also have the species difference in their aquatic toxicity. Overall, the results of this study provide a better understanding of the physiological responses of phytoplankton and will enable better risk assessments of NAs.

Topics: Carboxylic Acids; Chlorophyll; Chlorophyll A; Chlorophyta; Diatoms; Microalgae; Petroleum; Photosynthesis; Water Pollutants, Chemical

Dandelion (Taraxacum officinale) var. Garnet Stem was harvested from Texas and New Jersey for identification, quantification of phytochemicals, measurement of free radical scavenging activity, and bile acid binding capacity. The red midrib and petioles were extracted with methanol or ethanol and with or without water in combination with four different acids such as formic, hydrochloric, acetic, and citric acid. LC-ESI-HR-QTOF-MS was used to identify four anthocyanins including cyanidin-3-glucoside, cyanidin-3-(6-malonyl)-glucoside (A-1), cyanidin-3-(6-malonyl)-glucoside (A-2), and peonidin-3-(malonyl)-glucoside for the 1st time. In New Jersey samples, vitamin C and β-carotene were highest in the leaf blades versus whole leaf and petioles. Samples from Texas had highest amount of lutein, violaxanthin, and chlorophyll a and b in leaf blades versus whole leaf and petioles. Maximum DPPH free scavenging activity was found in MeOH: water: acid (80:19:1) and the combination of FA with EtOH: water: acid (80:19:1) demonstrated the higher level of total phenolic. Among six bile acids, sodium chenodeoxycholate was bound maximum in both Texas and New Jersey samples. This is the first report of anthocyanin identification from the midvein and petiole of Garnet Stem dandelion and results suggested that the phytochemicals and nutrients are highest in the leaf but may vary the amount depending on harvest location.. Four anthocyanins in the red midrib and petioles of Garnet Stem could be a potential source for antioxidants and can be used as a source of natural food color.

Topics: Anthocyanins; Antioxidants; Ascorbic Acid; beta Carotene; Bile Acids and Salts; Chenodeoxycholic Acid; Chlorophyll; Chlorophyll A; Glucosides; Lutein; New Jersey; Phytochemicals; Plant Leaves; Plant Stems; Taraxacum; Texas; Xanthophylls

Severity of plant diseases is often influenced by the availability of nutrients, particularly N; however, its effect on the phyllosphere microbiome in foliar pathogen challenged plants is less investigated in rice. The tripartite interaction among the fungal pathogen (Magnaporthe oryzae), rice cultivars (basmati and non-basmati, blast resistant or susceptible) and nitrogen (N) fertilization (0, 120 and 180 N) was investigated. Plant growth, elicitation of defense responses and abundance of microbial members in the rice phyllosphere were monitored using biochemical and molecular methods. In general, photosynthetic pigments were distinct for each cultivar, and optimal N doses led to higher values. The susceptible var. CO-39 and resistant CO-39I exhibited higher contents of photosynthetic pigments and micronutrients such as zinc in leaves in response to N doses. Elicitation of defense and hydrolytic enzymes was significantly influenced by pathogen inoculation and modulated by N doses, but varietal effects were distinct. Scoring indices emphasized the pathogen susceptibility of var. CO-39 and PB-1, which showed almost 40-60% higher values than the resistant cultivars; the interactions of cultivars and N doses was also significant. Characteristic changes were recorded in the abundances of the gene copies, particularly, with an overall increase in the number of cyanobacterial 16S rRNA, and bacterial amoA in pathogen-challenged treatments, while nifH gene copies exhibited a reducing trend with increasing N doses, in the presence or absence of pathogen. The varietal differences in the cause and effect relationships can be valuable in crop protection for more effective foliar application of pesticides or biocontrol agents.

Topics: Biological Control Agents; Carotenoids; Chlorophyll; Chlorophyll A; Cyanobacteria; Fertilizers; Magnaporthe; Microbiota; Nitrogen; Oryza; Oxidoreductases; Plant Diseases; Plant Leaves; RNA, Ribosomal, 16S

The orchids GL and YL are two cultivars of Cymbidium longibracteatum. YL displays an obviously yellowing rhizome and yellow leaves, while GL ('Longchangsu') shows dark green leaves and greenish rhizome. But the molecular mechanism for the differences between the two cultivars is poorly understood. In the present study, we showed that the structure of chloroplasts was significantly damaged in YL. Biochemical analysis uncovered the contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid were notably decreased in YL. Using RNA-Seq technology, more than 38 million clean reads were generated in each pool, and 116,422 unigenes were assembled de novo. 6,660 unigenes with differential expression patterns (FDR≤0.01 and |log2 ratio|≥1) were totally identified between the two cultivars. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of differentially expressed unigenes (DEGs) suggested 33 KEGG pathways were notably enriched, including biological processes such as "phenylpropanoid biosynthesis", "phagosome", "starch and sucrose metabolism", "drug metabolism-cytochrome P450", "fatty acid elongation", and "flavone and flavonol biosynthesis". Further analysis revealed that chlorophyll degeneration related unigene (c48794_g1) and flavonoid biosynthesis related unigenes (c16388_g1, c48963_g1, c63571_g1, c4492_g1, c52282_g1, c78740_g1, c4645_g1) were up-regulated while carotenoid biosynthesis related unigene (c7212_g1) were down-regulated in YL. Additionally, six of NAC, R2R3-MYB, bHLH transcription factors (c42861_g1, c105949_g1, c61265_g1, c42659_g1, c82171_g1, c19158_g1) might be involved in regulation of pigment biosynthesis. The chlorophyll degeneration and the flavonoid biosynthesis related unigenes up-regulation together with the carotenoid biosynthesis related unigenes down-regulation may contribute to the yellowing phenotype of YL.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Chloroplasts; Gene Expression Profiling; Gene Expression Regulation, Plant; Molecular Sequence Annotation; Orchidaceae; Plant Leaves; Plant Proteins; Sequence Analysis, RNA; Transcriptome

In the photic zone, phytoplankton experience diurnal variation in light intensity. However, prolonged exposure to aphotic condition influences their physiological state. Pigment composition is a useful biomarker to decipher cells physiological state and adaptive response to changing environmental conditions. Chlorophyll, a natural pigment, is biosynthesised even in darkness and studies have shown this ability is determined by genetic characteristics of an organism. The purpose of this study was to examine the influence of darkness on pigments and chlorophyll autofluorescence of Tetraselmis indica. Dark exposure (up to 6 months) had no significant impact on chlorophyll a and b concentration, whereas carotenoids were enhanced. Upon re-illumination pigments gradually recovered to pre-dark phase condition. These adaptive survival strategies of T. indica by altering pigment concentration in response to prolonged darkness are interesting. The absence of loroxanthin and loroxanthin esters in T. indica is reported in a first Tetraselmis species so far. In addition, the evaluation of autofluorescence and cellular chlorophyll concentration pointed out that they are not interdependent in this species. Hence, careful consideration of these two factors is needed when either of them is used as a proxy for other. The results obtained encourage a thorough study of pigment analysis, especially when subjected to darkness, to elucidate potential role in the evolution, chemotaxonomy, and survivability of species.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Chlorophyta; Chromatography, High Pressure Liquid; Darkness; Pigments, Biological; Spectrometry, Fluorescence; Time Factors; Xanthophylls

Spatial distribution of potentially toxic metals (PTMs) and their accumulation in mangrove Avicennia officinalis L. were studied along 8 locations in and around Sundarban mangrove wetland, India. Among 8 locations, S3 (Chemaguri) and S5 (Ghushighata) showed higher concentration of PTMs (Cd, Cr, Cu, Ni, Pb, Zn) characterized by higher enrichment factors (3.45-10.03), geo-accumulation indices (0.04-1.22), contamination factors (1.14-3.51) and pollution load indices (1.3-1.45) indicating progressive deterioration of estuarine quality and considerable ecotoxicological risk. Metal concentration in A. officinalis leaves showed significant correlation with sediment metals implying elevated level of bioaccumulation. Significant statistical correlation between photosynthetic pigments (Chlorophyll a, Chlorophyll b), antioxidant response (free radical scavenging and reducing ability) and stress enzymatic activity (Peroxidase, Catalase, Super-oxide dismutase) of A. officinalis with increasing metal concentration in the contaminated locations reflects active detoxification mechanism of the plant. The study indicates the potentiality of biomonitoring metal pollution using studied biochemical markers in mangrove habitats.

Topics: Avicennia; Biomarkers; Catalase; Chlorophyll; Chlorophyll A; Environmental Monitoring; Geologic Sediments; India; Metals, Heavy; Peroxidases; Plant Leaves; Superoxide Dismutase; Water Pollutants, Chemical; Water Pollution, Chemical; Wetlands

The decolourization and detoxification of azo dyes (Orange 2, Acid Orange 6) by fungal laccase from Trametes versicolor were evaluated. For laccase catalysed reaction, the azonaphthol Orange 2, with 72.8% decolourization, was degraded more rapidly than the azobenzene Acid Orange 6, with 45.3%. The presence of hydroxyl group at o-position to azo bond in the structure of Orange 2 was more preferable than the presence of two hydroxyl groups at o- and p-positions to azo bond in Acid Orange 6. Although the laccase treatment was more effective for the Orange 2 decolourization, the toxicity of both monoazo dye solutions became less toxic for the prokaryote growth. The phytotoxicity of Orange 2 and Acid Orange 6 solutions after laccase treatment was decreased in the range of 41.2-64.3 %. Also, the photoxicity, as measured by the production of chlorophylls a and b by Chlorella vulgaris and Microcystis aeruginosa, was decreased by laccase treatment of selected monoazo dyes. Our results show that different dyes can be decolorized and detoxified by laccase from T. versicolor in a single step.

Topics: Avena; Azo Compounds; Bacteria; Benzenesulfonates; Chlorella vulgaris; Chlorophyll; Chlorophyll A; Color; Coloring Agents; Fungi; Hordeum; Laccase; Microcystis; Trametes; Triticum; Water Pollutants, Chemical; Water Purification

Chlorophyll (Chl) triplet states generated in photosynthetic light-harvesting complexes (LHCs) can be quenched by carotenoids to prevent the formation of reactive singlet oxygen. Although this quenching occurs with an efficiency close to 100% at physiological temperatures, the Chl triplets are often observed at low temperatures. This might be due to the intrinsic temperature dependence of the Dexter mechanism of excitation energy transfer, which governs triplet quenching, or by temperature-induced conformational changes. Here, we report about the temperature dependence of Chl triplet quenching in two LHCs. We show that both the effects contribute significantly. In LHC II of higher plants, the core Chls are quenched with a high efficiency independent of temperature. A different subpopulation of Chls, which increases with lowering temperature, is not quenched at all. This is probably caused by the conformational changes which detach these Chls from the energy-transfer chain. In a membrane-intrinsic LHC of dinoflagellates, similarly two subpopulations of Chls were observed. In addition, another part of Chl triplets is quenched by carotenoids with a rate which decreases with temperature. This allowed us to study the temperature dependence of Dexter energy transfer. Finally, a part of Chls was quenched by triplet-triplet annihilation, a phenomenon which was not observed for LHCs before.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Cold Temperature; Dinoflagellida; Energy Transfer; Light; Light-Harvesting Protein Complexes; Spinacia oleracea

One of the most powerful line-narrowing techniques used to unravel the homogeneous lineshapes of inhomogeneously broadened systems is difference fluorescence line-narrowing spectroscopy. When this spectroscopy was applied to multichromophoric systems so far, the spectra were analyzed by an effective two-level system approach, composed of the electronic ground state and the lowest exciton state. An effective Huang-Rhys factor was assigned for the coupling of this state to the vibrations. Here, we extend this approach by including a multilevel line shape theory, which takes into account the excitonic coupling between pigments and thereby the effect of the delocalization of the excited states explicitly. In this way, it becomes possible to extract the spectral density of the local exciton-vibrational coupling. The theory is applied to the recombinant water-soluble chlorophyll binding protein reconstituted with chlorophyll a or b and reveals a significant decrease of the Huang-Rhys factor of the local exciton-vibrational coupling with decreasing transition energy of the chlorophylls. This decrease could be due to the increase in steric interactions reducing the flexibility of the environment and red-shifting the site energy of the pigments.

Topics: Brassica; Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Cold Temperature; Fluorescence; Light; Models, Chemical; Recombinant Proteins; Spectrometry, Fluorescence

We altered the chlorophyll (Chl) binding sites in various versions of water-soluble chlorophyll protein (WSCP) by amino acid exchanges to alter their preferences for either Chl a or Chl b. WSCP is ideally suited for this mutational analysis since it forms a tetrameric complex with only four identical Chl binding sites. A loop of 4-6 amino acids is responsible for Chl a versus Chl b selectivity. We show that a single amino acid exchange within this loop changes the relative Chl a/b affinities by a factor of 40. We obtained crystal structures of this WSCP variant binding either Chl a or Chl b. The Chl binding sites in these structures were compared with those in the major light-harvesting complex (LHCII) of the photosynthetic apparatus in plants to search for similar structural features involved in Chl a/b binding specificity.

Topics: Amino Acid Sequence; Binding Sites; Brassica; Chlorophyll; Chlorophyll A; Lepidium; Protein Binding; Protein Structure, Tertiary; Sequence Alignment; Water

Carotenoids are widely distributed in plants and algae, and their biosynthesis has attracted widespread interest. Carotenoid-related research has mostly focused on model species, and there is a lack of data on the carotenoid biosynthetic pathway in U. prolifera that is the main species leading to green tide, a harmful plague of floating green algae.. The carotenoid content of U. prolifera samples, that is the main species leading to green tide, a harmful plague of floating green algae at different temperatures revealed that its terpenoid was highest in the samples subjected to high temperature at 28 °C (H), followed by the samples subjected to low temperature at 12 °C (L). Its terpenoid was lowest in the samples subjected to medium temperature at 20 °C (M). We conducted transcriptome sequencing (148.5 million raw reads and 49,676 unigenes in total) of samples that were subjected to different temperatures to study the carotenoid biosynthesis of U. prolifera. There were 1125-3164 significant differentially expressed genes between L, M and H incubation temperatures, of which 11-672 genes were upregulated and 453-3102 genes were downregulated. A total of 3164 genes were significantly differentially expressed between H and M, of which 62 genes were upregulated and 3102 genes were downregulated. A total of 2669 significant differentially expressed genes were observed between L and H, of which 11 genes were upregulated and 2658 genes were downregulated. A total of 13 genes were identified to be involved in carotenoid biosynthesis in U. prolifera, and the expression levels of the majority were highest at H and lowest at M of incubation temperature. Both the carotenoid concentrations and the expression of the analysed genes were lowest in the normal temperature group, while low temperature and high temperature seemed to activate the biosynthesis of carotenoids in U. prolifera.. In this study, transcriptome sequencing provided critical information for understanding the accumulation of carotenoids and will serve as an important reference for the study of other metabolic pathways in U. prolifera.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Gene Expression Profiling; Gene Expression Regulation, Plant; Genes, Plant; Phylogeny; RNA, Plant; Sequence Analysis, RNA; Temperature; Ulva

Oxidative stress is considered to be involved in growth retardation of plants when they are exposed to a variety of biotic and abiotic stresses. Despite its potential importance in improving crop production, comparative studies on oxidative stress tolerance between rice (Oryza sativa L.) cultivars are limited. This work describes the difference in term of oxidative stress tolerance between 72 rice cultivars.. 72 rice cultivars grown under naturally lit greenhouse were used in this study. Excised leaf discs were subjected to a low concentration of methyl viologen (paraquat), a chemical reagent known to generate reactive oxygen species in chloroplast. Chlorophyll fluorescence analysis using a two-dimensional fluorescence meter, ion leakage analysis as well as the measurement of chlorophyll contents were used to evaluate the oxidative stress tolerance of leaf discs. Furthermore, fluorescence intensities were finely analyzed based on new fluorescence theories that we have optimized.. Treatment of leaf discs with methyl viologen caused differential decrease of maximum quantum yield of photosystem II (Fv/Fm) between cultivars. Decrease of Fv/Fm was also closely correlated with increase of ion leakage and decrease of chlorophyll a/b ratio. Fv/Fm was factorized into photochemical and non-photochemical parameters to classify rice cultivars into sensitive and tolerant ones. Among the 72 compared rice cultivars, the traditional cultivar Co13 was identified as the most tolerant to oxidative stress. Koshihikari, a dominant modern Japonica cultivar in Japan as well as IR58, one of the modern Indica breeding lines exhibited a strong tolerance to oxidative stress.. Close correlation between Fv/Fm and chlorophyll a/b ratio provides a simple method to estimate oxidative stress tolerance, without measurement of chlorophyll fluorescence with special equipment. The fact that modern cultivars, especially major cultivars possessed tolerance to oxidative stress suggests that oxidative stress tolerance is one of the agricultural traits prerequisite for improvement of modern rice cultivars. Data presented in this study would enable breeding of rice cultivars having strong tolerance to oxidative stress.

Topics: Adaptation, Physiological; Chlorophyll; Chlorophyll A; Crops, Agricultural; Hydrogen Peroxide; Ion Transport; Oryza; Oxidants; Oxidative Stress; Paraquat; Photosynthesis; Photosystem II Protein Complex; Plant Breeding; Plant Leaves; Quantitative Trait, Heritable; Spectrometry, Fluorescence

Identification of tree species that can biologically monitor air pollution and can endure air pollution is very much important for a sustainable green belt development around any polluted place. To ascertain the species, ten tree species were selected on the basis of some previous study from the campus of the University of Burdwan and were studied in the pre-monsoon and post-monsoon seasons. The study has been designed to investigate biochemical and physiological activities of selected tree species as the campus is presently exposed to primary air pollutants and their impacts on plant community were observed through the changes in several physical and biochemical constituents of plant leaves. As the plant species continuously exchange different gaseous pollutants in and out of the foliar system and are very sensitive to gaseous pollutants, they serve as bioindicators. Due to air pollution, foliar surface undergoes different structural and functional changes. In the selected plant species, it was observed that the concentration of primary air pollutants, proline content, pH, relative water holding capacity, photosynthetic rate, and respiration rate were higher in the pre-monsoon than the post-monsoon season, whereas the total chlorophyll, ascorbic acid, sugar, and conductivity were higher in the post-monsoon season. From the entire study, it was observed that the concentration of sulfur oxide (SO

Topics: Adaptation, Physiological; Air Pollutants; Air Pollution; Chlorophyll; Chlorophyll A; Environmental Monitoring; Particulate Matter; Photosynthesis; Plant Leaves; Plants; Seasons; Trees; Water

Photosynthetic organisms utilize carotenoids for photoprotection as well as light harvesting. Our previous study revealed that high-intensity light increases the expression of the gene for phytoene synthase (EgcrtB) in Euglena gracilis (a unicellular phytoflagellate), the encoded enzyme catalyzes the first committed step of the carotenoid biosynthesis pathway. To examine carotenoid synthesis of E. gracilis in response to light stress, we analyzed carotenoid species and content in cells grown under various light intensities. In addition, we investigated the effect of suppressing EgcrtB with RNA interference (RNAi) on growth and carotenoid content.. Our results indicate that suppression of EgcrtB causes a significant decrease in carotenoid and chlorophyll content in E. gracilis accompanied by changes in intracellular structures, suggesting that Dtx (de-epoxidized form of diadinoxanthin cycle pigments) contributes to photoprotection of this alga during the long-term acclimation to light-induced stress.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Euglena gracilis; Gene Silencing; Genes, Protozoan; Geranylgeranyl-Diphosphate Geranylgeranyltransferase; Light

The PROSPECT leaf optical model has, to date, well-separated the effects of total chlorophyll and carotenoids on leaf reflectance and transmittance in the 400-800 nm. Considering variations in chlorophyll a:b ratio with leaf age and physiological stress, a further separation of total plant-based chlorophylls into chlorophyll a and chlorophyll b is necessary for advanced monitoring of plant growth. In this study, we present an extended version of PROSPECT model (hereafter referred to as PROSPECT-MP) that can combine the effects of chlorophyll a, chlorophyll b and carotenoids on leaf directional hemispherical reflectance and transmittance (DHR and DHT) in the 400-800 nm. The LOPEX93 dataset was used to evaluate the capabilities of PROSPECT-MP for spectra modelling and pigment retrieval. The results show that PROSPECT-MP can both simultaneously retrieve leaf chlorophyll a and b, and also performs better than PROSPECT-5 in retrieving carotenoids concentrations. As for the simulation of DHR and DHT, the performances of PROSPECT-MP are similar to that of PROSPECT-5. This study demonstrates the potential of PROSPECT-MP for improving capabilities of remote sensing of leaf photosynthetic pigments (chlorophyll a, chlorophyll b and carotenoids) and for providing a framework for future refinements in the modelling of leaf optical properties.

Topics: Algorithms; Calibration; Carotenoids; Chlorophyll; Chlorophyll A; Models, Biological; Plant Leaves; Remote Sensing Technology

Cadmium (Cd) pollution in the soil threatens the quality of environmental health, and deleteriously affects physiological activities of crops. Symbiosis of endophytic fungi with various plants is a promising manner to improving numerous plant characteristics and remediating heavy metal-polluted soils. In this pot experiment, the influence of root endophyte fungus Piriformospora indica on growth, physiological parameters and organs Cd accumulation in sunflower cv. Zaria plants under the toxic levels of Cd (0, 40, 80 and 120mg/kg soil) were studied. Increasing Cd concentration in the soil reduced growth parameters, chlorophyll (Chl) a and Chl b contents, and Fv/Fm and ETR (electron transport rate) values, but increased root, stem and leaf Cd accumulation, and proline content. The presence of P. indica significantly enhanced growth, Chl a, Chl b and proline contents, and Fv/Fm and ETR values. Compared to non-inoculated ones, P. indica-inoculated plants had higher Cd accumulation in root, whereas lower Cd accumulation in stem and leaf. The present study strongly supports the established ability of P. indica to alleviate Cd toxicity by improving the physiological status in sunflower. Furthermore, this endophyte fungus can be useful for Cd phyto-stabilization in sunflower roots in contaminated soils.

Topics: Basidiomycota; Biodegradation, Environmental; Cadmium; Chlorophyll; Chlorophyll A; Fluorescence; Fluorometry; Helianthus; Nigeria; Plant Roots; Soil Pollutants

The effects of field dodder on physiological and anatomical processes in untreated sugar beet plants and the effects of propyzamide on field dodder were examined under controlled conditions. The experiment included the following variants: N-noninfested sugar beet plants (control); I - infested sugar beet plants (untreated), and infested plants treated with propyzamide (1500 g a.i. ha

Topics: Animals; Benzamides; Beta vulgaris; Chlorophyll; Chlorophyll A; Cuscuta; Herbicides; Photosynthesis; Plant Leaves

The light-harvesting mechanisms in the three strains of Prochlorococcus marinus, CCMP1986, CCMP1375, and CCMP2773, grown under blue and red light-emitting diodes (LEDs) at two intensity levels were investigated. The blue LED was divinyl chlorophyll b (DV-Chl b) selective and the red LED was DV-Chl a selective. Under the red LED, the relative amount of DV-Chl b in CCMP1375 and CCMP2773 decreased and the relative amount of zeaxanthin increased in CCMP1375. Furthermore, the pigment composition of cells of CCMP1375 grown under red LED was remodified when they were transplanted under the blue LED. Picosecond-time-resolved fluorescence of the LED-grown Prochlorococcus was measured, and the excitation-energy-transfer efficiency between DV-Chl a did not significantly change for the different LED colors or intensities; however, a change in the pigment composition of the DV-Chl b-rich strains (CCMP1375 and CCMP2773) was observed. It appears that peripheral antenna responds to light conditions, with small modifications in the photosystems.

Topics: Absorption, Physiological; Chlorophyll; Chlorophyll A; Light; Pigments, Biological; Prochlorococcus

Effects of benzophenone-3 (BP-3) on the green alga, Chlamydomonas reinhardtii, and the cyanobacterium, Microcystis aeruginosa, were investigated. The tested organisms were exposed to environmental levels of BP-3 for 10 days, at nominal concentrations from 0.01 to 5000μgL

Topics: Benzophenones; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Microcystis; Photosynthesis; Pigments, Biological; Sunscreening Agents; Water Pollutants, Chemical

Kinetics of acid-induced chlorophyll demetallation was recorded in microdroplets by fusing a stream of microdroplets containing 40 µM chlorophyll a or b dissolved in methanol with a stream of aqueous microdroplets containing 35 mM hydrochloric acid (pH = 1·46). The kinetics of the demetallation of chlorophyll in the fused microdroplets (14 ± 6 µm diameter; 84 ± 18 m s-1 velocity) was recorded by controlling the traveling distance of the fused microdroplets between the fusion region and the inlet of a mass spectrometer. The rate of acid-induced chlorophyll demetallation was about 960 ± 120 times faster in the charged microdroplets compared with that reported in bulk solution. If no voltage was applied to the sprayed microdroplets, then the acceleration factor was about 580 ± 90, suggesting that the applied voltage is not a major factor determining the acceleration. Chlorophyll a was more rapidly demetallated than chlorophyll b by a factor of ~26 in bulk solution and ~5 in charged microdroplets. The demetallation kinetics was second order in the H+ concentration, but the acceleration factor of microdroplets compared with bulk solution appeared to be unchanged in going from pH = 1·3 to 7·0. The water:methanol ratio of the fused microdroplets was varied from 7:3 to 3:7 causing an increase in the reaction rate of chlorophyll a demetallation by 20%. This observation demonstrates that the solvent composition, which has different evaporation rates, does not significantly affect the acceleration. We believe that a major portion of the acceleration can be attributed to confinement effects involving surface reactions rather than either to evaporation of solvents or to the introduction of charges to the microdroplets.

Topics: Chlorophyll; Chlorophyll A; Hydrogen-Ion Concentration; Kinetics; Lab-On-A-Chip Devices; Mass Spectrometry; Metals

Plant canopy pigment concentration is a critical variable for agricultural remote sensing due to its close relationship to leaf nitrogen content. The aims of this study were to: (1) compare the prediction performances on chlorophyll, chlorophyll-a and b, and carotenoid concentration in jujube leaf at canopy scale between partial least squares regression (PLSR) and support vector machine (SVM), (2) develop quantitative models to estimate pigment concentration in jujube canopy using hyperspectral data and provide theoretical and technical support for rapidly, non-destructive, less expensive and eco-friendly measuring the concentration. Results from correlation analysis showed that jujube canopy pigment concentration correlated strongly with hyperspectral data. What’s more, the hyperspectral data was better correlated by chlorophyll and chlorophyll-a than chlorophyll-b and carotenoid. Results of independent samples tested in predicting performance indicated that both of the PLSR and SVM models could effectively estimate pigment concentration, however, with different prediction precisions. Additionally, the precision of SVM outperformed PLSR for predicting chlorophyll and carotenoid. Whereas chlorophyll-a and chlorophyll-b were better predicted using PLSR than SVM. Compared among all the pigments’ prediction precisions with corresponding optimal inversion models showed that prediction precisions on chlorophyll, chlorophyll-a and carotenoid were superior to chlorophyll-b. The determination coefficients and residual prediction deviation from predicting chlorophyll, chlorophyll-a and carotenoid were higher than 0.8 and 2.0, respectively, while the mean relative error values were lower than 13%. And the corresponding values from predicting chlorophyll-b were 0.60%, 20.79% and 1.79% respectively.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Fruit; Least-Squares Analysis; Models, Theoretical; Pigmentation; Plant Leaves; Spectrum Analysis; Support Vector Machine; Ziziphus

Excess rainfalls may be the cause of waterlogging in soil, which affects the growth and development of wheat. Therefore, the objectives of this study were to examine the effects of waterlogging on shoot and root growth and physiological characteristics of wheat. Three experiments were conducted: experiment 1 (E1): evaluation of seedling growth on ten Iranian winter wheat varieties with waterlogging periods (1-4, 4-8, 8-12, and 12-16 days starting from seed germination). Seminal roots and plumule were investigated at seedling. The others are E2: pretreatment of waterlogging (15 days) at tillering and stem elongation stages and its effects on shoot and root growth at anthesis stage and experiment 3 (E3): pretreatment of waterlogging (15 days) at tillering and jointing stages and its effects on yield and yield components and also evaluation of stress tolerance indexes. The results of the seedling growth test (E1) showed that 1-4- and 4-8-day waterlogging severity reduced seminal root length (94.5 to 93.7 %) and plumule length (86.2 to 50.0 %) compared to control. Results of E2 indicated that waterlogging stress decreased shoot dry weight, root dry weight, total secondary root length, and chlorophyll a + b content of flag leaf by 28-31, 44-35, 20-31, and 28-35 %, respectively. Also, result of E3 showed that the grain yields of wheat varieties at two conditions of stress were different in base tolerance indexes. In general, the responses of wheat varieties to waterlogging were different at the three experiments. The varieties that had the most of dry weight and length of the root were tolerant. Thus, it is possible to use these characteristics as an index for selecting the varieties with tolerance to waterlogging.

Topics: Chlorophyll; Chlorophyll A; Plant Roots; Plant Shoots; Rain; Seedlings; Triticum; Water

Topics: Carotenoids; Chemical Fractionation; Chlorophyll; Chlorophyll A; Hibiscus; Lasers; Luminescent Measurements; Pigments, Biological; Plant Extracts; Plant Leaves; Refractometry; Spectrophotometry, Ultraviolet; Temperature

The Chlamydomonas reinhardtii truncated light-harvesting antenna 4 (tla4) DNA transposon mutant has a pale green phenotype, a lower chlorophyll (Chl) per cell and a higher Chl a/b ratio in comparison with the wild type. It required a higher light intensity for the saturation of photosynthesis and displayed a greater per chlorophyll light-saturated rate of oxygen evolution than the wild type. The Chl antenna size of the photosystems in the tla4 mutant was only about 65% of that measured in the wild type. Molecular genetic analysis revealed that a single plasmid DNA insertion disrupted two genes on chromosome 11 of the mutant. A complementation study identified the "chloroplast signal recognition particle 54" gene (CpSRP54), as the lesion causing the tla4 phenotype. Disruption of this gene resulted in partial failure to assemble and, therefore, lower levels of light-harvesting Chl-binding proteins in the C. reinhardtii thylakoids. A comparative in silico 3-D structure-modeling analysis revealed that the M-domain of the CpSRP54 of C. reinhardtii possesses a more extended finger loop structure, due to different amino acid composition, as compared to that of the Arabidopsis CpSRP54. The work demonstrated that CpSRP54 deletion in microalgae can serve to generate tla mutants with a markedly smaller photosystem Chl antenna size, improved solar energy conversion efficiency, and photosynthetic productivity in high-density cultures under bright sunlight conditions.

Topics: Amino Acid Sequence; Amino Acids; Arabidopsis Proteins; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Chloroplasts; DNA Transposable Elements; Genes, Chloroplast; Light; Light-Harvesting Protein Complexes; Microalgae; Phenotype; Photosynthesis; Sequence Alignment; Signal Recognition Particle; Thylakoids

There is increasing evidence that the circadian clock is a significant driver of photosynthesis that becomes apparent when environmental cues are experimentally held constant. We studied whether the composition of photosynthetic pigments is under circadian regulation, and whether pigment oscillations lead to rhythmic changes in photochemical efficiency. To address these questions, we maintained canopies of bean and cotton, after an entrainment phase, under constant (light or darkness) conditions for 30-48 h. Photosynthesis and quantum yield peaked at subjective noon, and non-photochemical quenching peaked at night. These oscillations were not associated with parallel changes in carbohydrate content or xanthophyll cycle activity. We observed robust oscillations of Chl a/b during constant light in both species, and also under constant darkness in bean, peaking when it would have been night during the entrainment (subjective nights). These oscillations could be attributed to the synthesis and/or degradation of trimeric light-harvesting complex II (reflected by the rhythmic changes in Chl a/b), with the antenna size minimal at night and maximal around subjective noon. Considering together the oscillations of pigments and photochemistry, the observed pattern of changes is counterintuitive if we assume that the plant strategy is to avoid photodamage, but consistent with a strategy where non-stressed plants maximize photosynthesis.

Topics: Carbohydrate Metabolism; Chlorophyll; Chlorophyll A; Circadian Rhythm; Gossypium; Phaseolus; Photosynthesis; Pigments, Biological; Plant Leaves

Dryland vegetation developed morphological and physiological strategies to cope with drought. However, as aridity increases, vascular plant coverage gets sparse and microbially-dominated surface communities (MSC), comprising cyanobacteria, algae, lichens and bryophytes together with heterotropic bacteria, archaea and fungi, gain relevance. Nevertheless, the relevance of MSC net primary productivity has only rarely been considered in ecosystem scale studies, and detailed information on their contribution to the total photosynthetic biomass reservoir is largely missing. In this study, we mapped the spatial distribution of two different MSC (biological soil crusts and quartz fields hosting hypolithic crusts) at two different sites within the South African Succulent Karoo (Soebatsfontein and Knersvlakte). Then we characterized both types of MSC in terms of chlorophyll content, and combining these data with the biocrust and quartz field maps, we estimated total biomass values of MSCs and their spatial patterns within the two different ecosystems. Our results revealed that MSC are important vegetation components of the South African Karoo biome, revealing clear differences between the two sites. At Soebatsfontein, MSC occurred as biological soil crusts (biocrusts), which covered about one third of the landscape reaching an overall biomass value of ~480gha

Topics: Biomass; Chlorophyll; Chlorophyll A; Ecosystem; Remote Sensing Technology; Soil; Soil Microbiology; South Africa

Water-soluble chlorophyll proteins (WSCPs) of class IIa from Brassicaceae form tetrameric complexes containing one chlorophyll (Chl) per apoprotein but no carotenoids. The complexes are remarkably stable toward dissociation and protein denaturation even at 100 °C and extreme pH values, and the Chls are partially protected against photooxidation. There are several hypotheses that explain the biological role of WSCPs, one of them proposing that they function as a scavenger of Chls set free upon plant senescence or pathogen attack. The biochemical properties of WSCP described in this paper are consistent with the protein acting as an efficient and flexible Chl scavenger. At limiting Chl concentrations, the recombinant WSCP apoprotein binds substoichiometric amounts of Chl (two Chls per tetramer) to form complexes that are as stable toward thermal dissociation, denaturation, and photodamage as the fully pigmented ones. If more Chl is added, these two-Chl complexes can bind another two Chls to reach the fully pigmented state. The protection of WSCP Chls against photodamage has been attributed to the apoprotein serving as a diffusion barrier for oxygen, preventing its access to triplet excited Chls and, thus, the formation of singlet oxygen. By contrast, the sequential binding of Chls by WSCP suggests a partially open or at least flexible structure, raising the question of how WSCP photoprotects its Chls without the help of carotenoids.

Topics: Apoproteins; Brassica; Chlorophyll; Chlorophyll A; Gene Expression; Hot Temperature; Hydrogen-Ion Concentration; Light; Light-Harvesting Protein Complexes; Models, Molecular; Oxidation-Reduction; Oxygen; Pisum sativum; Plant Proteins; Protein Binding; Protein Denaturation; Protein Domains; Protein Multimerization; Protein Stability; Protein Structure, Secondary; Recombinant Fusion Proteins; Singlet Oxygen; Solubility; Thylakoids; Water

In this experiment, the effects of different lead (Pb) concentrations (0, 200, 600, 1000, 1400 mg kg

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Fluorescence; Photosynthesis; Plant Leaves; Robinia; Seedlings

The plastidial methylerythritol phosphate(MEP) pathway provides 5-carbon precursors to the biosynthesis of isoprenoid (including artemisinin). 2-C-Methyl-D-erythritol-4-phosphate cytidylyltransferase (MCT) is the third enzyme of the MEP pathway, which catalyzes 2-C-methyl-D-erythritol-4-phosphate to form 4-(cytidine 5’-diphospho)-2-C-methyl-D-erythritol. The full-length MCT cDNA sequence (AaMCT) was cloned and characterized for the first time from Artemisia annua L. Analysis of tissue expression pattern revealed that AaMCT was highly expressed in glandular secretory trichome and poorly expressed in leaf, flower, root and stem. AaMCT was found to be a methyl jasmonate (Me JA)-induced genes, the expression of AaMCT was significantly increased after MeJA treatment. Subcellular localization indicated that the GFP protein fused with AaMCT was targeted specifically in chloroplasts. The transgenic plants of Arabidopsis thaliana with AaMCT overexpression exhibited a significantly increase in the content of chlorophyll a, chlorophyll b and carotenoids, demonstrating that AaMCT kinase plays an influential role in isoprenoid biosynthesis.

Topics: Acetates; Arabidopsis; Artemisia annua; Artemisinins; Carotenoids; Chlorophyll; Chlorophyll A; Cloning, Molecular; Cyclopentanes; DNA, Complementary; Gene Expression Regulation, Plant; Nucleotidyltransferases; Oxylipins; Plant Proteins; Plants, Genetically Modified

Artemisinin is the first choice for malaria treatment. The plastidial MEP pathway provides 5-carbon precursors (IPP and its isomer DMAPP) for the biosynthesis of isoprenoid (including artemisinin). Hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (HDR) is the last enzyme involved in the MEP pathway, which catalyzes HMBPP to form IPP and DMAPP. In this study, we isolated the full-length cDNA of HDR from Artemisia annua\ L. (AaHDR2) and performed functional analysis. According to gene expression analysis of AaHDR2 (GenBank: KX058541) and AaHDR1 reported ever (GenBank: ADC84348.1) by qPCR, we found that AaHDR1 and AaHDR2 had much higher expression level in trichomes than that in roots, stems, leaves and flowers. AaHDR2 had much higher expression level in flowers than that in leaves. Further, the plant hormones such as Me JA and ABA respectively up-regulated the expression level of AaHDR1 and AaHDR2 significantly, but GA3 up-regulated the expression level of AaHDR2 only. The gene expression analysis of AaHDR1 and AaHDR2 showed that AaHDR2 had a greater contribution than AaHDR1 to isoprenoid biosynthesis(including artemisinin). We used AaHDR2 for the following experiments. Bioinformatic analysis indicated that AaHDR2 belonged to the HDR family and the functional complementation assay showed that AaHDR2 did have the enzymatic function of HDR, using E. coli mutant MG1655(ara)<>HDR as host cell. The subcellular localization assay showed that AaHDR2 fused with GFP at its N-terminal specifically targeted in chloroplasts. Finally, AaHDR2 was overexpressed in Arabidopsis thaliana. The AaHDR2-overexpressing plants produced the isoprenoids including chlorophyll a, chlorophyll b and carotenoids at significantly higher levels than the wild-type Arabidopsis plants. In summary, AaHDR2 might be a candidate gene for genetic improvement of the isoprenoid biosynthesis.

Topics: Amino Acid Sequence; Arabidopsis; Artemisia annua; Carotenoids; Chlorophyll; Chlorophyll A; Chloroplasts; Cloning, Molecular; DNA, Complementary; Escherichia coli; Oxidoreductases; Plant Growth Regulators; Plant Proteins; Terpenes

Alzheimer's disease (AD) and amyotrophic lateral sclerosis (ALS) are progressive neurodegenerative diseases that affect the neurons in the brain and the spinal cord. Neuroinflamation and apoptosis are key players in the progressive damage of the neurons in AD and ALS. Currently, there is no drug to offer complete cure for both these diseases. Riluzole is the only available drug that can prolong the life time of the ALS patients for nearly 3 months. Molecules that offer good HIT to the molecular targets of ALS will help to treat AD and ALS patients. P53 kinase receptor (4AT3), EphA4 (3CKH) and histone deacetylase (3SFF) are the promising disease targets of AD and ALS. This paper discusses on a new approach to combat neurodegenerative diseases using photosynthetic pigments. The docking studies were performed with the Autodock Vina algorithm to predict the binding of the natural pigments such as β carotene, chlorophyll a, chlorophyll b, phycoerythrin and phycocyanin on these targets. The β carotene, phycoerythrin and phycocyanin had higher binding energies indicating the antagonistic activity to the disease targets. These pigments serve as a potential therapeutic molecule to treat neuroinflammation and apoptosis in the AD and ALS patients.

Topics: Alzheimer Disease; Amyotrophic Lateral Sclerosis; beta Carotene; Chlorophyll; Chlorophyll A; Histone Deacetylases; Humans; Models, Molecular; Phycocyanin; Phycoerythrin; Protein Conformation; Receptor, EphA4

To elucidate the physiological and morphological responses generated by vanadium (V) in plants, hydroponic culture experiments were performed with swamp morning glory (Ipomoea aquatica Forsk) exposed to 0 mg L(-1) to 2.50 mg L(-1) pentavalent V [V(V)] in Hoagland nutrient solutions. The concentration of chlorophyll a, chlorophyll b, and carotene peaked at a V(V) concentration of 0.05 mg L(-1) and gradually decreased at higher V(V) concentrations. Similarly, the plant biomass was stimulated at low levels of V(V) and was inhibited when V(V) concentrations exceeded 0.1 mg L(-1). Pentavalent V had negative effects on the uptake of phosphorus (P) by roots, shoots, and leaves. The biological absorption coefficients of V of the roots were higher than those of the aerial parts. Under low concentrations of V(V) exposure, the predominant species of V in the aerial parts was tetravalent V [V(IV)], whereas V(V) became more prevalent when concentrations of V(V) in the solution was higher than 0.50 mg L(-1). In the roots, however, the concentrations of V(V) were always higher than those of the V(IV), except in the control group. Organelles in the V(V)-treated leaves were distorted, and the periplasmic space became wider. These results indicate V(V) has concentration-dependent effects on the physiological properties of swamp morning glory, whereas the plant has the ability to develop self-protective function to adapt to the toxicity of V(V).

Topics: Biomass; Carotenoids; Chlorophyll; Chlorophyll A; Hydroponics; Ipomoea; Phosphorus; Photosynthesis; Plant Leaves; Plant Roots; Plant Shoots; Vanadium

Babyleaf salads such as spinach (Spinacia oleracea L.) and spinach beet (Beta vulgaris L. subsp. cicla var. cicla) are an important dietary source of antioxidants such as ascorbic acid (vitamin C). Such compounds may be important in disease prevention in consumers but the level of these compounds in leaves frequently declines after harvest. As such, methods to maintain antioxidant levels in fresh produce are being sought.. Irrigation deficits were used to apply water stress to S. oleracea and B. vulgaris plants. This treatment prevented postharvest decline of leaf ascorbic acid content in S. oleracea but not in B. vulgaris. Ascorbic acid levels in leaves at harvest were unaffected by the treatment in both species compared to well-watered controls.. We have shown that restricted irrigation provides a viable means to maintain leaf vitamin content after harvest in S. oleracea, an important finding for producers, retailers and consumers alike. © 2015 Society of Chemical Industry.

Topics: Agricultural Irrigation; Antioxidants; Ascorbic Acid; Beta vulgaris; Carotenoids; Chlorophyll; Chlorophyll A; Crop Production; Food Quality; Food Storage; Humans; Nutritive Value; Pigments, Biological; Plant Leaves; Refrigeration; Species Specificity; Spinacia oleracea

This study investigated the effects of selected four pharmaceutically active compounds (PhACs) (carbamazepine, sulfamethoxazole, ofloxacin, and roxithromycin) on the photosynthesis and antioxidant enzymes of Cyperus alternifolius in constructed wetlands (CWs). Moreover, the removal and kinetics of PhACs in CWs were evaluated to explore the related removal mechanisms. Results showed that C. alternifolius can uptake and withstand certain PhACs. The PhAC tolerance of C. alternifolius might be attributed to their capacity to maintain relatively normal photosynthetic activity and elevated antioxidative defense. CWs offered comparable or even higher removal efficiencies for the selected PhACs compared with conventional WWTPs. The removal of the target PhACs was enhanced in the planted CWs versus the unplanted CWs mostly because of plant uptake and rhizosphere effects. In particular, carbamazepine, which is considered the most recalcitrant of the PhACs, was significantly reduced (p<0.05). The removal of target PhACs fitted into two distinct periods. The initial fast step (within the first 2 h) was essentially attributed to the adsorption onto the CW medium surface. The subsequent slow process (2-12 h) closely followed first-order kinetics probably because of the interaction between microorganisms and plants. The obtained results indicate that C. alternifolius can phytoremediate PhAC-contaminated waters in CWs.

Topics: Biodegradation, Environmental; Carbamazepine; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Cyperus; Ofloxacin; Peroxidase; Roxithromycin; Sulfamethoxazole; Superoxide Dismutase; Waste Disposal, Fluid; Water Pollutants, Chemical; Wetlands

In recent years, the response of fern gametophytes to environment has raised much attention. However, studies on the influence of plant invasion to fern gametophytes are scarce. Allelopathy plays an important role in biological invasion. Hence, it is necessary to study the allelopathic effects of invasive plants on fern gametophytes and elucidate the mechanisms by which invasive plants cause phytotoxicity. As one of the main invasive plants in China, Bidens pilosa exhibits allelopathic effects on spermatophyte growth. Field investigation shows that many ferns are threatened by the invasion of B. pilosa. The distribution of Pteris multifida overlaps with that of B. pilosa in China. To examine the potential involvement of allelopathic mechanisms of B. pilosa leaves, changes in the physiology in P. multifida gametophytes are analyzed. We found that cell membrane and antioxidant enzyme activities as well as photosynthesis pigment contents of the gametophytes were affected by B. pilosa leachates. Gametophytes of P. multifida exposed to B. pilosa had increased damages to cell membranes, expressed in thiobarbituric acid reacting substance (TBARS) concentrations, malondialdehyde (MDA), electrolyte leakage (membrane permeability), and degree of injury. Enzyme activities, assessed by superoxide dismutase (SOD) and catalase (CAT) as well as guaiacol peroxidase (GPX) enhanced with the increase in leachate concentration after 2-day exposure. Meanwhile, lower chlorophyll a (Chl a), chlorophyll b (Chl b), carotenoid (Car), and the total chlorophyll were measured as leachate concentrations increased. At day 10, leaf leachates of B. pilosa exhibited the greatest inhibition. These results suggest that the observed inhibitory or stimulatory effects on the physiology studied can have an adverse effect on P. multifida and that allelopathic interference seems to have involved in this process.

Topics: Allelopathy; Bidens; Catalase; China; Chlorophyll; Chlorophyll A; Germ Cells, Plant; Introduced Species; Malondialdehyde; Pheromones; Photosynthesis; Plant Leaves; Pteris; Superoxide Dismutase

Little effort has been made to understand the influence of Mg on cellular processes of plant cell during Cu and Cd toxicities. The present work demonstrates the influence of magnesium (Mg) on copper (Cu) and cadmium (Cd) toxicity on Triticum aestivum (Wheat). We measured a range of parameters related to oxidative stress in wheat exposed to Cu or Cd toxicity in media with different concentrations of Mg. Decreasing Mg concentration significantly exacerbated Cu and Cd toxicity and optimum supply of Mg improved the growth and decreased the toxicity-induced oxidative stress (a substantial decline in the amount of hydrogen peroxide (H2O2) and malondialdehyde (MDA) in root and shoot tissues). Activity of antioxidant enzymes-superoxide dismutase (SOD), ascorbae peroxidase (APX), catalase (CAT) was restored upon optimum Mg concentration in the presence of Cu and Cd toxicity. An increase in proline concentration in roots and shoots that was triggered by Cu and Cd exposure was partly reversed. This was due to decline in pyrroline-5-carboxylate synthetase (P5CS) and pyrroline-5-carboxylate reductase (P5CR) activity and enhanced proline dehydrogenase (PDH) activity. In conclusion, decreasing supply of Mg effectively exacerbated the toxicities of Cu and Cd in wheat.

Topics: Ascorbate Peroxidases; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Hydrogen Peroxide; Magnesium; Malondialdehyde; Metals, Heavy; Oxidation-Reduction; Plant Roots; Plant Shoots; Proline; Pyrroles; Superoxide Dismutase; Triticum

This paper discusses the impact of four quaternary ammonium salts (QAS) such as tetraethylammonium tetrafluoroborate [TEA][BF4], tetrabutylammonium tetrafluoroborate [TBA][BF4], tetrahexylammonium tetrafluoroborate [THA][BF4], and tetraoctylammonium tetrafluoroborate [TOA][BF4] on the growth and development of spring barley and common radish. Analogous tests were performed with the inorganic salt ammonium tetrafluoroborate [A][BF4] for comparison purposes. Results indicated that the phytotoxicity of the QAS applied is dependent on the concentration of the substance and their number of carbon atoms. The most toxic compound was [TBA][BF4], causing the greatest drop in fresh weight of both study plants, similar to the phytotoxic effects of [A][BF4]. All the tested compounds caused oxidative stress in spring barley and common radish seedlings due to a drop in the chlorophyll content. Stress was also observed in plants, which was indicated by the increased level of ROS (reactive oxygen species) such as H2O2 and lipid peroxidation of MDA (malondialdehyde). Due to the stress, both plants displayed changes in the activity of antioxidative enzymes such as superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD). Based on the results of the study, it was concluded that changes in chlorophyll levels and peroxidase activity are the best biomarkers to determine oxidative stress in plants.

Topics: Ammonium Compounds; Borates; Boric Acids; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Hordeum; Hydrogen Peroxide; Lipid Peroxidation; Malondialdehyde; Oxidative Stress; Peroxidase; Raphanus; Superoxide Dismutase

Jatropha curcas, a multipurpose plant attracting a great deal of attention due to its high oil content and quality for biofuel, is recognized as a drought-tolerant species. However, this drought tolerance is still poorly characterized. This study aims to contribute to uncover the molecular background of this tolerance, using a combined approach of transcriptional profiling and morphophysiological characterization during a period of water-withholding (49 d) followed by rewatering (7 d). Morphophysiological measurements showed that J. curcas plants present different adaptation strategies to withstand moderate and severe drought. Therefore, RNA sequencing was performed for samples collected under moderate and severe stress followed by rewatering, for both roots and leaves. Jatropha curcas transcriptomic analysis revealed shoot- and root-specific adaptations across all investigated conditions, except under severe stress, when the dramatic transcriptomic reorganization at the root and shoot level surpassed organ specificity. These changes in gene expression were clearly shown by the down-regulation of genes involved in growth and water uptake, and up-regulation of genes related to osmotic adjustments and cellular homeostasis. However, organ-specific gene variations were also detected, such as strong up-regulation of abscisic acid synthesis in roots under moderate stress and of chlorophyll metabolism in leaves under severe stress. Functional validation further corroborated the differential expression of genes coding for enzymes involved in chlorophyll metabolism, which correlates with the metabolite content of this pathway.

Topics: Adaptation, Physiological; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Cluster Analysis; Desiccation; Droughts; Galactose; Gases; Gene Expression Profiling; Gene Expression Regulation, Plant; Genes, Plant; Jatropha; Metabolic Networks and Pathways; Models, Biological; Photosystem II Protein Complex; Plant Leaves; Plant Roots; Plant Stomata; Reproducibility of Results; Sequence Analysis, RNA; Starch; Stress, Physiological; Water

The comparative study of biochemical and ultrastructure features in senescing sugar beet (Beta vulgaris L.) leaves was carried out. One group of plants was grown under normal conditions in washed river sand and poured in turn with nitrate-containing mineral solution or water (N plants). Another group of plants, after 1 month of normal growth, was further grown with nitrate omitted in the nutritive solution (defN plants). The starting point of normal leaf senescence in N plants was identified by the maximal content of soluble protein. Soluble carbohydrate pools were statistically constant in senescing N plants, whereas glucose pools varied noticeably. A decrease in the contents of soluble protein and chlorophyll (a + b) in the course of senescing was typical for N plant leaves. The cell membrane in N plant leaves remained mostly intact; the central vacuoles in the leaf cells were large, and their membranes remained intact. The chloroplasts and mitochondria in senescing N plant leaves became swollen. The vesicles that were present in the cytoplasm of N plant leaves were especially large in the oldest leaves. It was concluded that senescing of sugar beet leaves at sufficient nitrate nutrition occurs according to a "vacuolar" scenario. In the case of nitrate deficiency, the content of soluble carbohydrates in defN leaves first reached maximum and then decreased in older leaves; the protein and chlorophyll (a + b) contents were totally lower than those in normal leaves and continuously decreased during the experiments. Chloroplasts in mesophyll cells of defN plant leaves became more rounded; starch grains in chloroplasts degraded and the number and size of lipid globules increased. The multitude of membrane impairments and lots of large vesicles-"crystals" appeared during the experiment. The results showed the controlling action of nitrogen nutrition in the senescing of sugar beet leaves.

Topics: Beta vulgaris; Chlorophyll; Chlorophyll A; Chloroplasts; Nitrates; Nitrogen; Plant Leaves; Stress, Physiological

Studies assessing chromium phytoextration from natural waters rarely consider potential implications of chromium speciation in the presence of ubiquitous humic substances. Therefore, the present study investigated the influence of environmentally relevant concentration of humic acid (TOC = 10 mg L(-1)) on chromium speciation (Cr = 0.15 mg L(-1)) and consequently on phytoextraction by aquatic macrophyte duckweed Lemna minor. In absence of humic acid, only hexavalent chromium was present in water samples and easily taken up by L. minor. Chromium uptake resulted in a significant reduction of growth rate by 22% and decrease of chlorophyll a and chlorophyll b contents by 48% and 43%, respectively. On the other hand, presence of humic acid significantly reduced chromium bioavailability (57% Cr uptake decrease) and consequently it did not cause any measurable effect to duckweed. Such effect was related to abiotic reduction of hexavalent chromium species to trivalent. Hence, findings of our study suggest that presence of humic acid and chromium speciation cannot be neglected during phytoextraction studies.

Topics: Araceae; Chlorophyll; Chlorophyll A; Chromium; Humic Substances; Water Pollutants, Chemical

The phytohormone cytokinin (CK) is known to delay senescence in plants. We studied the effect of a CK analog, 6-benzyl adenine (BA), on rice leaves to understand the possible mechanism by which CK delays senescence in a drought- and heat-tolerant rice cultivar Nagina22 (N22) using dark-induced senescence (DIS) as a surrogate for natural senescence of leaves. Leaves of N22-H-dgl162, a stay-green mutant of N22, and BA-treated N22 showed retention of chlorophyll (Chl) pigments, maintenance of the Chl a/b ratio, and delay in reduction of both photochemical efficiency and rate of oxygen evolution during DIS. HPLC analysis showed accumulation of 7-hydroxymethyl chlorophyll (HmChl) during DIS, and the kinetics of its accumulation correlated with progression of senescence. Transcriptome analysis revealed that several plastid-localized genes, specifically those associated with photosystem II (PSII), showed higher transcript levels in BA-treated N22 and the stay-green mutant leaves compared with naturally senescing N22 leaves. Real-time PCR analyses showed that genes coding for enzymes associated with Chl a/b interconversion and proteins associated with light-harvesting complexes maintained higher transcript levels up to 72h of DIS following BA treatment. The pigment-protein complexes analyzed by green gel remained intact in both N22-H-dgl162 and BA-treated N22 leaves even after 96h of DIS. Thus, CK delays senescence by accumulation of HmChl and up-regulating genes in the Chl cycle, thereby maintaining the Chl a/b ratio. Also, CK treatment retains higher transcript levels of PSII-related genes, resulting in the stability of photosynthetic pigment complexes and functional stay-greenness in rice.

Topics: Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Cytokinins; Darkness; Gene Expression Profiling; Gene Expression Regulation, Plant; Genes, Plant; Mass Spectrometry; Metabolic Networks and Pathways; Oligonucleotide Array Sequence Analysis; Oryza; Photosynthesis; Plant Leaves; RNA, Messenger

We present two-dimensional electronic-vibrational (2DEV) spectra of isolated chlorophyll a and b in deuterated ethanol. We excite the Q-band electronic transitions and measure the effects on the carbonyl and C ═ C double-bond stretch region of the infrared spectrum. With the aid of density functional theory calculations, we provide assignments for the major features of the spectrum. We show how the 2DEV spectra can be used to readily distinguish different solvation states of the chlorophyll, with features corresponding to the minority pentacoordinate magnesium (Mg) species being resolved along each dimension of the 2DEV spectra from the dominant hexacoordinate Mg species. These assignments represent a crucial first step toward the application of 2DEV spectroscopy to chlorophyll-containing pigment-protein complexes.

Topics: Chlorophyll; Chlorophyll A; Magnesium; Spectrophotometry, Infrared

Changes in leaf soluble proteome were explored in 3-month-old plants of metallicolous (M) and nonmetallicolous (NM) Agrostis capillaris L. populations exposed to increasing Cu concentrations (1-50 μM) to investigate molecular mechanisms underlying plant responses to Cu excess and tolerance of M plants. Plants were cultivated on perlite (CuSO4 spiked-nutrient solution). Soluble proteins, extracted by the trichloroacetic acid/acetone procedure, were separated with 2-DE (linear 4-7 pH gradient). Analysis of CCB-stained gels (PDQuest) reproducibly detected 214 spots, and 64 proteins differentially expressed were identified using LC-MS/MS. In both populations, Cu excess impacted both light-dependent (OEE, cytochrome b6-f complex, and chlorophyll a-b binding protein), and -independent (RuBisCO) photosynthesis reactions, more intensively in NM leaves (ferredoxin-NADP reductase and metalloprotease FTSH2). In both populations, upregulation of isocitrate dehydrogenase and cysteine/methionine synthases respectively suggested increased isocitrate oxidation and enhanced need for S-containing amino-acids, likely for chelation and detoxification. In NM leaves, an increasing need for energetic compounds was indicated by the stimulation of ATPases, glycolysis, pentose phosphate pathway, and Calvin cycle enzymes; impacts on protein metabolism and oxidative stress increase were respectively suggested by the rise of chaperones and redox enzymes. Overexpression of a HSP70 may be pivotal for M Cu tolerance by protecting protein metabolism. All MS data have been deposited in the ProteomeXchange with the dataset identifier PXD001930 (http//proteomecentral.proteomexchange.org/dataset/PXD001930).

Topics: Adaptation, Physiological; Agrostis; Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Copper Sulfate; Cytochrome b6f Complex; Energy Metabolism; Gene Expression Profiling; Gene Expression Regulation, Plant; Gene Ontology; Molecular Sequence Annotation; Photosynthesis; Plant Leaves; Proteome; Ribulose-Bisphosphate Carboxylase; Solubility; Stress, Physiological

The present study was developed to respond to the need for an increase in crop yield in the mountain region of Caparaó (southern Espírito Santo State, Brazil), an area of traditional coffee production. This study aimed to analyze the diversity and characterize the crop yield of genotypes of Coffea arabica L. with potential for cultivation in high plant density systems. In addition, it also aimed to quantify the expression of agronomic traits in this cultivation system and provide information on the genotypes with the highest cultivation potential in the studied region. The experiment followed a randomized block design with 16 genotypes, four repetitions, and six plants per experimental plot. Plant spacing was 2.00 x 0.60 m, with a total of 8333 plants per hectare, representing a high-density cultivation system. Coffee plants were cultivated until the start of their reproductive phenological cycles and were evaluated along four complete reproductive cycles. Genotypes with high crop yield and beverage quality, short canopy, and rust resistance were selected. C. arabica genotypes showed variability in almost all characteristics. It was possible to identify different responses among genotypes grown in a high plant density cultivation system. Although the chlorophyll a content was similar among genotypes, the genotypes Acauã, Araponga MG1, Sacramento MG1, Tupi, and Catuaí IAC 44 showed a higher chlorophyll b content than the other genotypes. Among these, Sacramento MG1 also showed high leafiness and growth of vegetative structures, whereas Araponga MG1, Pau-Brasil MG1, and Tupi showed high fruit production. In addition, Araponga MG1 had also a higher and more stable crop yield over the years.

Topics: Agriculture; Biometry; Chlorophyll; Chlorophyll A; Coffea; Genetic Variation; Genotype; Phenotype; Plant Breeding; Plant Leaves; Quantitative Trait, Heritable; Seeds

Photosensitization, also known as photodermatitis, occurs when phototoxic or photoactive substances accumulate in the skin and interact with sunlight to result in an often severe, crusting, itching or painful dermatitis in unpigmented and/or lightly haired areas of the skin. Primary photosensitization, caused by direct ingestion of photosensitizing agents, has been reported anecdotally in horses after ingestion of alfalfa hay. Between 2004 and 2014, several large outbreaks of primary photosensitization in horses fed primarily alfalfa hay were investigated in California. Alfalfa hay samples were collected and carefully examined for the presence of known photosensitizing plants and pesticide residues but none were identified. Select hay samples were evaluated for unusual fungal infestation and for phototoxicity assay using a specific Candida albicans assay; results were negative. In the 2004 outbreak, a feeding study was conducted with three horses exclusively fed alfalfa hay that was suspected to have caused the outbreak. Two weeks after ingestion of alfalfa hay, two horses developed several lesions in non-pigmented skin characterized as chronic ulcerative and necrotizing dermatitis with superficial vasculitis, which was consistent with photosensitization. In the 2014 outbreak, seven different implicated alfalfa hay samples were analyzed for chlorophyll a and b, and pheophorbide a. These compounds had been suspected to play a role in alfalfa-induced primary photosensitization. The chlorophyll contents ranged from 0.90 to 2.30 mg/g in the alfalfa hay samples, compared to 1.37 and 2.94 mg/g in locally grown alfalfa and orchard grass hay. The pheophorbide a levels ranged from 3.36 to 89.87 µg/g in alfalfa samples compared to 81.39 and 42.33 µg/g in control alfalfa and orchard grass hay samples. These findings eliminate chlorophyll a, chlorophyll b, and pheophorbide a as possible causes for alfalfa-hay induced primary photosensitization.

Topics: Animal Feed; Animals; California; Chlorophyll; Chlorophyll A; Diet; Disease Outbreaks; Female; Horse Diseases; Horses; Male; Medicago sativa; Photosensitivity Disorders; Silage

The foliar and biochemical traits of Azadirachta indica A. Juss from fly ash (FA) dumping site in Badarpur thermal power plant (BTPP) New Delhi, India was studied. Three different experimental sites were selected at different distances from the thermal power plant. Ambient suspended particulate matter (SPM) and plant responses such as leaf pigments (chlorophyll a, chlorophyll b, and carotenoids), total chlorophyll, net photosynthetic rate, stomatal index (SI), stomatal conductance (SC), intercellular carbon dioxide concentration [CO2]i, net photosynthetic rate (NPR), nitrogen, nitrate, nitrate reductase activity, proline, protein, reducing sugar and sulphur content were measured. Considerable reduction in pigments (chlorophyll a, chlorophyll b and carotenoids), and total chlorophyll was observed at fly ash dumping site. Fly ash stress revealed the inhibitory effect on Nitrate reductase activity (NRA), Nitrate, soluble protein, and reducing sugar content, whereas stimulatory effect was found for the stomatal index, nitrogen, proline, antioxidants and sulphur content in the leaves. Under fly ash stress, stomatal conductance was low, leading to declining in photosynthetic rate and increase in the internal CO2 concentration of leaf. Single leaf area (SLA), leaf length and leaf width also showed a declining trend from control to the polluted site. Antioxidant enzymes increased in leaves reflecting stress and extenuation of reactive oxygen species (ROS).

Topics: Air Pollutants; Antioxidants; Azadirachta; Carbon Dioxide; Carotenoids; Chlorophyll; Chlorophyll A; Coal; Coal Ash; India; Nitrogen; Oxidative Stress; Photosynthesis; Plant Leaves; Power Plants; Proline; Sulfur

Biological soil crusts, comprising assemblages of cyanobacteria, fungi, lichens and mosses, are common in dryland areas and are important elements in these ecosystems. Increasing N deposition has led to great changes in community structure and function in desert ecosystems worldwide. However, it is unclear how moss crusts respond to increased atmospheric N deposition, especially in terms of growth and physiological parameters. The aim of this study was to understand how Syntrichia caninervis, a dominant species in moss crusts in many northern hemisphere desert ecosystems, responds to added N.. The population and shoot growth, and physiological responses of S. caninervis to six different doses of simulated N deposition (0, 0·3, 0·5, 1·0, 1·5 and 3·0 g N m(-2) year(-1)) were studied over a 3 year period.. Low amounts of added N increased shoot length and leaf size, whereas high doses reduced almost all growth parameters. Moss shoot density increased, but population biomass decreased with high N. Low N augmented chlorophyll b, total chlorophyll content and soluble protein concentrations, but not chlorophyll a or chlorophyll fluorescence. High N was detrimental to all these indices. Soluble sugar concentration declined with increased N, but proline concentration was not affected significantly. Antioxidant enzyme activities generally decreased with low N additions and increased with high doses of simulated N deposition.. Low amounts of added N (0-0·5 g N m(-2) year(-1)) may enhance moss growth and vitality, while higher amounts have detrimental effects.

Topics: Antioxidants; Bryopsida; China; Chlorophyll; Chlorophyll A; Desert Climate; Ecosystem; Environmental Pollutants; Enzymes; Nitrogen; Plant Leaves; Plant Shoots; Proline; Soil

The analysis of compounds from single cells is a major challenge in analytical life science. Labeling strategies, for instance fluorescence detection, are well established for measuring proteins with single cell sensitivity, but they mostly fail to detect small molecules. More recently mass spectrometry has entered the realm of single cell sensitivity and enables the label-free and highly parallelized detection of small biomolecules from single cells. The assignment of signals detected in single cells, however, generally has to rely on measurements in whole cell culture extracts. Isobaric structures, contaminations, higher noise levels and the high variability in the abundance of peaks between single cells complicate the assignment of peaks in single-cell spectra. Tandem mass spectrometry would be very useful for compound identification via mass spectrometry directly in single-cell analyses. Here we present the first single cell tandem mass spectra collected using matrix-assisted laser-desorption/ionization. The spectra obtained allow the assignment of most compounds detected in the spectra. We also show that the fragmentation is not restricted to the most abundant peaks in the spectra, but over a dynamic range of more than one order of magnitude.

Topics: Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Hydroxybenzoates; Lipids; Single-Cell Analysis; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tandem Mass Spectrometry

LHCSR3 is a member of the Light-Harvesting Complexes (LHC) family, which is mainly composed of pigment-protein complexes responsible for collecting photons during the first steps of photosynthesis. Unlike related LHCs, LHCSR3 is expressed in stress conditions and has been shown to be essential for the fast component of photoprotection, non-photochemical quenching (NPQ), in the green alga Chlamydomonas reinhardtii. In plants, which do not possess LHCSR homologs, NPQ is triggered by the PSBS protein. Both PSBS and LHCSR3 possess the ability to sense pH changes but, unlike PSBS, LHCSR3 binds multiple pigments. In this work we have analyzed the properties of the pigments bound to LHCSR3 and their excited state dynamics. The data show efficient excitation energy transfer between pigments with rates similar to those observed for the other LHCs. Application of an exciton model based on a template of LHCII, the most abundant LHC, satisfactorily explains the collected steady state and time-resolved spectroscopic data, indicating that LHCSR3 has a LHC-like molecular architecture, although it probably binds less pigments. The model suggests that most of the chlorophylls have similar energy and interactions as in LHCII. The most striking difference is the localization of the lowest energy state, which is not on the Chlorophyll a (Chl a) 610-611-612 triplet as in all the LHCB antennas, but on Chl a613, which is located close to the lumen and to the pH-sensing region of the protein.

Topics: Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Photosystem II Protein Complex

Unicellular cyanobacteria are ubiquitous photoautotrophic microbes that contribute substantially to global primary production. Picocyanobacteria such as Synechococcus and Prochlorococcus depend on chlorophyll a-binding protein complexes to capture light energy. In addition, Synechococcus has accessory pigments organized into phycobilisomes, and Prochlorococcus contains chlorophyll b. Across a surface water transect spanning the sparsely studied tropical Indian Ocean, we examined Synechococcus and Prochlorococcus occurrence, taxonomy and habitat preference in an evolutionary context. Shotgun sequencing of size fractionated microbial communities from 0.1 μm to 20 μm and subsequent phylogenetic analysis indicated that cyanobacteria account for up to 15% of annotated reads, with the genera Prochlorococcus and Synechococcus comprising 90% of the cyanobacterial reads, even in the largest size fraction (3.0-20 mm). Phylogenetic analyses of cyanobacterial light-harvesting genes (chl-binding pcb/isiA, allophycocyanin (apcAB), phycocyanin (cpcAB) and phycoerythin (cpeAB)) mostly identified picocyanobacteria clades comprised of overlapping sequences obtained from Indian Ocean, Atlantic and/or Pacific Oceans samples. Habitat reconstructions coupled with phylogenetic analysis of the Indian Ocean samples suggested that large Synechococcus-like ancestors in coastal waters expanded their ecological niche towards open oligotrophic waters in the Indian Ocean through lineage diversification and associated streamlining of genomes (e.g. loss of phycobilisomes and acquisition of Chl b); resulting in contemporary small celled Prochlorococcus. Comparative metagenomic analysis with picocyanobacteria populations in other oceans suggests that this evolutionary scenario may be globally important.

Topics: Biological Evolution; Chlorophyll; Chlorophyll A; Cyanobacteria; Ecosystem; Genome, Bacterial; Indian Ocean; Likelihood Functions; Metagenome; Metagenomics; Phycobilisomes; Phylogeny; Prochlorococcus; Seawater; Synechococcus; Temperature

By using 77 families of 3 year-old Reaumuria soongorica seedlings that grew well without injection of any pests and diseases as experimental material, the contents of soluble protein (SP), soluble sugar (SS), proline (Pro) and chlorophyll (Chl) were measured, and their drought resistance was evaluated with principal component analysis and subordinate function method. The results showed that there were great differences in leaf osmolytes and chlorophyll among the 77 families. The soluble protein content varied from 2.14 to 8.60 mg · g⁻¹ FM, the soluble sugar content was from 6.82 to 21.86 mg · g⁻¹ FM, the proline content was from 118.73 to 1494.30 µg · g⁻¹ FM, the chlorophyll a content was from 321.88 to 897.37 µg · g⁻¹ FM, the chlorophyll b content was from 53.65 to 249.04 µg · g⁻¹ FM, chlorophyll (a+b) was from 387.39 to 1146.40 µg · g⁻¹ FM, and the chlorophyll a/b was from 3.46 to 6.42. All drought-resistant indices had significant difference among R. soongorica families, among which the proline content varied most, followed by the soluble sugar content. Evaluated by using the synthesized multi-index, it was found that 12 families showed good drought resistance, with Zhazigou 1-2 and Zhazigou 1-1 performing the best.

Topics: Carbohydrates; Chlorophyll; Chlorophyll A; Droughts; Osmoregulation; Plant Leaves; Proline; Seedlings; Tamaricaceae

Resonance Raman spectroscopy was used to evaluate the structure of light-harvesting chlorophyll (Chl) a/b complexes of photosystem II (LHCII), reconstituted from wild-type (WT) and mutant apoproteins over-expressed in Escherichia coli. The point mutations involved residue S123, exchanged for either P (S123P) or G (S123G). In all reconstituted proteins, lutein 2 displayed a distorted conformation, as it does in purified LHCII trimers. Reconstituted WT and S123G also exhibited a conformation of bound neoxanthin (Nx) molecules identical to the native protein, while the S123P mutation was found to induce a change in Nx conformation. This structural change of neoxanthin is accompanied by a blue shift of the absorption of this carotenoid molecule. The interactions assumed by (and thus the structure of the binding sites of) the bound Chls b were found identical in all the reconstituted proteins, and only marginally perturbed as compared to purified LHCII. The interactions assumed by bound Chls a were also identical in purified LHCII and the reconstituted WT. However, the keto carbonyl group of one Chl a, originally free-from-interactions in WT LHCII, becomes involved in a strong H-bond with its environment in LHCII reconstituted from the S123P apoprotein. As the absorption in the Qy region of this protein is identical to that of the LHCII reconstituted from the WT apoprotein, we conclude that the interaction state of the keto carbonyl of Chl a does not play a significant role in tuning the binding site energy of these molecules.

Topics: Binding Sites; Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Lutein; Mutation; Photosystem II Protein Complex; Spectrum Analysis, Raman; Xanthophylls

Podophyllotoxin (PPT) and its derivatives, isolated from the rhizome of Podophyllum hexandrum Royle (P. hexandrum), are typically used in clinical settings for anti-cancer and anti-virus treatments. Empirical studies have verified that P. hexandrum had stronger tolerance to chilling, due to involving PPT accumulation in rhizome induced by cold stress. However, the cold-adaptive mechanism and its association with PPT accumulation at a molecular level in P. hexandrum are still limited. In this study, the morpho-physiological traits related to plant growth, PPT accumulation and key gene expressions controlling PPT biosynthesis were assessed by exposing P. hexandrum seedlings to different temperatures (4 °C and 10 °C as chilling stress and 22 °C as the control). The results showed that chilling significantly increased chlorophyll content, net photosynthetic rate, stomatal conductance, and plant biomass, whereas it greatly decreased transpiration rates and intercellular CO2 concentration. Compared to the control, the chilling treatments under 4 °C and 10 °C conditions induced a 5.00- and 3.33-fold increase in PPT contents, respectively. The mRNA expressions of six key genes were also up-regulated by chilling stresses. The findings are useful in understanding the molecular basis of P. hexandrum response to chilling.

Topics: Biosynthetic Pathways; Chlorophyll; Chlorophyll A; Cold Temperature; Gene Expression Regulation, Plant; Genes, Plant; Photosynthesis; Podophyllotoxin; Podophyllum; Rhizome; Seedlings; Stress, Physiological

Although silicon (Si) has been widely reported to alleviate plant nutrient deficiency, the alleviating effect of Si on potassium (K) deficiency and its underlying mechanism are poorly understood. Here, we examined whether Si-regulated putrescine (Put) metabolisms are involved in Si-alleviated K deficiency.. Sorghum seedlings were grown in K deficiency solution with and without Si for 15 d. The influence of K deficiency and Si on leaf chlorosis symptoms, K(+) concentration, polyamine (PA) levels, amine oxidase activities, the transcription of Put synthesis genes, antioxidant enzyme activities and H2O2 accumulation were measured.. Under K-sufficient conditions, plant growth was not affected by Si application. Si application significantly alleviated the growth inhibition induced by K-deficient stress, however. K deficiency induced leaf chlorosis and reduction in several leaf chlorosis-related metrics, including photosynthesis, efficiency of photosystem II photochemistry, chlorophyll content and chlorophyll a/b ratio; all of these changes were moderated by Si application. Si application did not influence the K(+) concentration in leaves under K-sufficient or K-deficient conditions. It did, however, decrease the excessive accumulation of Put that was otherwise induced by K deficiency. Simultaneously, Put synthesis gene transcription and activation of amine oxidases were down-regulated by Si application under K-deficient conditions. In addition, Si reduced K-deficiency-enhanced antioxidant enzyme activities and decreased K-deficiency-induced H2O2 accumulation.. These results indicate that Si application could reduce K-deficiency-induced Put accumulation by inhibiting Put synthesis and could decrease H2O2 production via PA oxidation. Decreased H2O2 accumulation contributes to the alleviation of cell death, thereby also alleviating K-deficiency-induced leaf chlorosis and necrosis.

Topics: Antioxidants; Carboxy-Lyases; Chlorophyll; Chlorophyll A; Hydrogen Peroxide; Photosynthesis; Plant Leaves; Plant Proteins; Polyamines; Potassium Deficiency; Putrescine; Seedlings; Silicon; Sorghum

The photosystem I/II ratio increased when antenna size was enlarged by transient induction of CAO in chlorophyll b -less mutants, thus indicating simultaneous regulation of antenna size and photosystem I/II stoichiometry. Regulation of antenna size and photosystem I/II stoichiometry is an indispensable strategy for plants to acclimate to changes to light environments. When plants grown in high-light conditions are transferred to low-light conditions, the peripheral antennae of photosystems are enlarged. A change in the photosystem I/II ratio is also observed under the same light conditions. However, our knowledge of the correlation between antenna size modulation and variation in photosystem I/II stoichiometry remains limited. In this study, chlorophyll a oxygenase was transiently induced in Arabidopsis thaliana chlorophyll b-less mutants, ch1-1, to alter the antenna size without changing environmental conditions. In addition to the accumulation of chlorophyll b, the levels of the peripheral antenna complexes of both photosystems gradually increased, and these were assembled to the core antenna of both photosystems. However, the antenna size of photosystem II was greater than that of photosystem I. Immunoblot analysis of core antenna proteins showed that the number of photosystem I increased, but not that of photosystem II, resulting in an increase in the photosystem I/II ratio. These results clearly indicate that antenna size adjustment was coupled with changes in photosystem I/II stoichiometry. Based on these results, the physiological importance of simultaneous regulation of antenna size and photosystem I/II stoichiometry is discussed in relation to acclimation to light conditions.

Topics: Arabidopsis; Arabidopsis Proteins; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Electrophoresis, Polyacrylamide Gel; Fluorescence; Gene Expression Regulation, Plant; Immunoblotting; Light-Harvesting Protein Complexes; Models, Biological; Oxygenases; Photosynthesis; Photosystem I Protein Complex; Photosystem II Protein Complex; RNA, Messenger; Substrate Specificity; Temperature; Transformation, Genetic

Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas.

Topics: Agrobacterium; Carotenoids; Chenopodiaceae; Chlorophyll; Chlorophyll A; Cuminum; Gene Expression Regulation, Plant; Osmotic Pressure; Plants, Genetically Modified; Salinity; Salt Tolerance; Sodium Chloride; Sodium-Hydrogen Exchangers

Confocal fluorescence microscopy and electron microscopy (EM) are complementary methods for studying the intracellular localization of proteins. Confocal fluorescence microscopy provides a rapid and technically simple method to identify the organelle in which a protein localizes but only EM can identify the suborganellular compartment in which that protein is present. Confocal fluorescence microscopy, however, can provide information not obtainable by EM but required to understand the dynamics and interactions of specific proteins. In addition, confocal fluorescence microscopy of cells transfected with a construct encoding a protein of interest fused to a fluorescent protein tag allows live cell studies of the subcellular localization of that protein and the monitoring in real time of its trafficking. Immunostaining methods for confocal fluorescence microscopy are also faster and less involved than those for EM allowing rapid optimization of the antibody dilution needed and a determination of whether protein antigenicity is maintained under fixation conditions used for EM immunogold labeling. This chapter details a method to determine by confocal fluorescence microscopy the intracellular localization of a protein by transfecting the organism of interest, in this case Giardia lamblia, with the cDNA encoding the protein of interest and then processing these organisms for double label immunofluorescence staining after chemical fixation. Also presented is a method to identify the organelle targeting information in the presequence of a precursor protein, in this case the presequence of the precursor to the Euglena light harvesting chlorophyll a/b binding protein of photosystem II precursor (pLHCPII), using live cell imaging of mammalian COS7 cells transiently transfected with a plasmid encoding a pLHCPII presequence fluorescent protein fusion and stained with organelle-specific fluorescent dyes.

Topics: Animals; Chlorocebus aethiops; Chlorophyll; Chlorophyll A; COS Cells; DNA, Complementary; Euglena; Fluorescent Antibody Technique; Gene Expression; Giardia lamblia; Green Fluorescent Proteins; Microscopy, Confocal; Microscopy, Fluorescence; Photosystem II Protein Complex; Plasmids; Protein Transport; Protozoan Proteins; Recombinant Fusion Proteins

Road dust is one of the most common pollutants and causes a series of negative effects on plant physiology. Dust's impacts on plants can be regarded as a combination of load, composition and grain size impacts on plants; however, there is a lack of integrated dust effect studies involving these three aspects. In our study, Sophora japonica seedlings were artificially dusted with road dust collected from the road surface of Beijing so that we could study the impacts of this dust on nitrogen/carbon allocation, biomass allocation and photosynthetic pigments from the three aspects of composition, load and grain size. The results showed that the growth characteristics of S. japonica seedlings were mostly influenced by dust composition and load. Leaf N, root-shoot ratio and chlorophyll a/b were significantly affected by dust composition and load; leaf C/N, shoot biomass, total chlorophyll and carotenoid were significantly affected by dust load; stem N and stem C/N were significantly affected by dust composition; while the dust grain size alone did not affect any of the growth characteristics. Road dust did influence the growth characteristics more extensively than loam. Therefore, a higher dust load could increase the differences between road dust and loam treatments. The elements in dust are well correlated to the shoot N, shoot C/N, and root-shoot ratio of S. japonica seedlings. This knowledge could benefit the management of urban green spaces.

Topics: Beijing; Carbon; Chlorophyll; Chlorophyll A; Dust; Environmental Monitoring; Nitrogen; Particulate Matter; Photosynthesis; Plant Leaves; Plant Stems; Seedlings; Sophora

Microalgae are fast-growing photosynthetic organisms which have the potential to be exploited as an alternative source of liquid fuels to meet growing global energy demand. The cultivation of microalgae, however, still needs to be improved in order to reduce the cost of the biomass produced. Among the major costs encountered for algal cultivation are the costs for nutrients such as CO₂, nitrogen and phosphorous. In this work, therefore, different microalgal strains were cultivated using as nutrient sources three different anaerobic digestates deriving from municipal wastewater, sewage sludge or agro-waste treatment plants. In particular, anaerobic digestates deriving from agro-waste or sewage sludge treatment induced a more than 300% increase in lipid production per volume in

Topics: Anaerobiosis; Biofuels; Biomass; Carotenoids; Chlorella vulgaris; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Sewage; Wastewater

The microenvironments in photosynthetic proteins affect the absorption by chlorophyll (Chl) pigments. It is, however, a challenge to disentangle the impact on the transition energies of different perturbations, for example, the global electrostatics of the protein (nonbonded environmental effects), exciton coupling between Chl's, conformational variations, and binding of an axial ligand to the magnesium center. This is needed to distinguish between the two most commonly proposed mechanisms for energy transport in photosynthetic proteins, relying on either weakly or strongly coupled pigments. Here, on the basis of photodissociation action spectroscopy, we establish that the redshift of the Soret absorption band due to binding of a negatively charged carboxylate (as present in aspartic acid and glutamic acid residues) is 0.1-0.2 eV for Chl a and b. This effect is almost enough to reproduce the well-known green color of plants and can account for the strong spectral variation between Chl's. The experimental data serve to benchmark future high-level calculations of excited-state energies. Finally, we demonstrate that complexes between Chl a and histidine, tagged by a quaternary ammonium ion, can be made in the gas phase by electrospray ionization, but more work is needed to produce enough ions for gas-phase spectroscopy.

Topics: Acetylcholine; Aspartic Acid; Chlorophyll; Chlorophyll A; Energy Transfer; Formates; Glutamic Acid; Histidine; Kinetics; Ligands; Magnesium; Models, Molecular; Photosystem II Protein Complex; Pisum sativum; Protein Structure, Secondary; Spectrum Analysis; Static Electricity; Thermodynamics

Common duckweed (Lemna minor L.) is ideally suited to test the impact of metals on freshwater vascular plants. Literature on cadmium (Cd) and uranium (U) oxidative responses in L. minor are sparse or, for U, non-existent. It was hypothesised that both metals impose concentration-dependent oxidative stress and growth retardation on L. minor. Using a standardised 7-day growth inhibition test, the adverse impact of these metals on L. minor growth was confirmed, with EC50 values for Cd and U of 24.1 ± 2.8 and 29.5 ± 1.9 μm, respectively, and EC10 values of 1.5 ± 0.2 and 6.5 ± 0.9 μm, respectively. The metal-induced oxidative stress response was compared through assessing the activity of different antioxidative enzymes [catalase, glutathione reductase, superoxide dismutase (SOD), ascorbate peroxidase (APOD), guaiacol peroxidase (GPOD) and syringaldizyne peroxidase (SPOD)]. Significant changes in almost all antioxidative enzymes indicated their importance in counteracting the U- and Cd-imposed oxidative burden. However, some striking differences were also observed. For activity of APODs and SODs, a biphasic but opposite response at low Cd compared to U concentrations was found. In addition, Cd (0.5-20 μm) strongly enhanced plant GPOD activity, whereas U inhibited it. Finally, in contrast to Cd, U up to 10 μm increased the level of chlorophyll a and b and carotenoids. In conclusion, although U and Cd induce similar growth arrest in L. minor, the U-induced oxidative stress responses, studied here for the first time, differ greatly from those of Cd.

Topics: Antioxidants; Araceae; Cadmium; Carotenoids; Chlorophyll; Chlorophyll A; Glutathione; Hydrogen-Ion Concentration; Oxidative Stress; Photosynthesis; Uranium

Chromium is a highly toxic non-essential metal, which causes a variety of metabolic activities in plants. Pyxine cocoes a well known toxitolerant lichen species was considered to evaluate the possible physiological, biochemical, and genetic changes that occur due to chromium Cr (+VI) stress. The physiological (chlorophyll a, chlorophyll b, total chlorophyll, carotenoid, protein, and Fv/Fm) and genetic (ISSR-PCR and ITS) parameters were used to estimate the changes in P. cocoes. Different concentrations of Cr (+VI) (0, 10, 25, 50, 75, and 100 μM) for 10, 20, 30, and 45 days were employed on transplanted lichen species. The results revealed that the exposure of Cr (+VI) for 10, 20, 30, and 45 days under controlled conditions caused a significant decline in physiological processes with increasing metal stress. Amino acid profile at different concentrations on the 45th day too indicated prevailing stress condition as proline content significantly increased at 100 μM concentration. Inter-simple sequence repeat (ISSR) and internal transcribed spacer (ITS) techniques were used to evaluate the genotoxicity induced by chromium stress. ISSR profiles showed a consistent increase in appearance and disappearance of bands with increasing concentration of the chromium. ISSR technique, therefore, is more sensitive and reproducible to study polymorphism induced by environmental stress. The present study revealed that the physiological and genetic changes induced by the Cr (+VI) can be used as a tool to study environmental stress and polymorphisms due to genotoxicity. To the best of our knowledge, application of ISSR-PCR and ITS sequences in toxitolerant species (P. cocoes) appears to be the maiden attempt to evaluate the genotoxicity.

Topics: Amino Acids; Ascomycota; Carotenoids; Chlorophyll; Chlorophyll A; Chromium; DNA, Intergenic; Dose-Response Relationship, Drug; Lichens; Microsatellite Repeats; Mutagenicity Tests; Sensitivity and Specificity; Stress, Physiological

Although considered as "green" solvents, the toxic effects of ionic liquids (ILs) on organisms have been widely investigated in recent years. However, studies on the toxic effects of ILs on plants all focus on toxicity in nutrient solution. In the present paper, the toxic effects of 1-butyl-3-methylimidazolium chloride ([C4mim]Cl) on Vicia faba seedlings in soil at 0, 500, 1500, and 2500 mg kg(-1) on day 10 were studied. The present results showed that the growth of V. faba seedlings may be seriously inhibited when the IL concentrations were higher than 500 mg kg(-1). The EC50 values for shoot length, root length, and dry weight were 3886, 2483, and 3359 mg kg(-1), respectively. In addition, [C4mim]Cl caused lipid peroxidation and DNA damage at 500 mg kg(-1) and oxidative stress at 1500 mg kg(-1), which indicated that [C4mim]Cl may have genotoxicity and cytotoxicity on V. faba seedlings. Moreover, oxidative damage may be the main toxic mechanism of ILs.

Topics: Ascorbate Peroxidases; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; DNA Damage; Glutathione; Imidazoles; Ionic Liquids; Lipid Peroxidation; Peroxidase; Plant Leaves; Plant Roots; Plant Shoots; Proline; Reactive Oxygen Species; Seedlings; Soil Pollutants; Superoxide Dismutase; Vicia faba

Understanding sub-lethal effects of nanomaterial may be particularly important to determining ecosystem responses as current levels of nanomaterial release are low compared to levels projected for the future. In this work, the sub-lethal effects of water stable, nanocrystalline fullerenes as C60 (termed nC60) were studied on Scenedesmusobliquus, a globally distributed phytoplankton. Sub-lethal concentration for S. obliquus was firstly determined as 0.09mgL(-1) using the standard 72h exposure tests (OECD Guideline 201). Subsequent sub-lethal experiment of nC60 on the S. obliquus was carried out for 60d and focused on the photosynthesis processes. The results demonstrate that upon sub-lethal exposure, the photosynthetic products of polysaccharide, soluble protein and total lipid were decreased with exposure time. The photosynthetic pigments of chlorophyll a and chlorophyll b were negatively impacted. Further investigations indicate that the decrements in photosynthetic products and pigments were mainly due to the algal Mg(2+) decrement (by 40%) at the sub-lethal concentration (0.09mgL(-1)) of nC60. The decrement in Mg(2+) of S. obliquus was due to the inhibition of Mg(2+)-ATPase activity caused by nC60. Sum up, these results not only describe the sub-lethal effects but also provide the probably mechanism for sub-lethal effects of nC60 on exposed S. obliquus.

Topics: Ca(2+) Mg(2+)-ATPase; Chlorophyll; Chlorophyll A; Fullerenes; Nanoparticles; Photosynthesis; Plant Proteins; Scenedesmus; Water Pollutants, Chemical

Photosynthetic characteristics of two broadleaved evergreen trees, Quercus myrsinaefolia and Machilus thunbergii, were compared in autumn and winter. The irradiance was similar in both seasons, but the air temperature was lower in winter. Under the winter conditions, net photosynthesis under natural sunlight (Anet) in both species dropped to 4 μmol CO2 m(-2) s(-1), and the quantum yield of photosystem II (PSII) photochemistry in dark-adapted leaves (Fv/Fm) also dropped to 0.60. In both species the maximum carboxylation rates of Rubisco (V(cmax)) decreased, and the amount of Rubisco increased in winter. A decline in chlorophyll (Chl) concentration and an increase in the Chl a/b ratio in winter resulted in a reduction in the size of the light-harvesting antennae. From measurements of Chl a fluorescence parameters, both the relative fraction and the energy flux rates of thermal dissipation through other non-photochemical processes were markedly elevated in winter. The results indicate that the photosynthetic apparatus in broadleaved evergreen species in warm temperate regions responds to winter through regulatory mechanisms involving the downregulation of light-harvesting and photosynthesis coupled with increased photoprotective thermal energy dissipation to minimize photodamage in winter. These mechanisms aid a quick restart of photosynthesis without the development of new leaves in the following spring.

Topics: Algorithms; Carbon Dioxide; Chlorophyll; Chlorophyll A; Japan; Lauraceae; Lipid Peroxidation; Photochemical Processes; Photosynthesis; Photosystem II Protein Complex; Plant Leaves; Quercus; Ribulose-Bisphosphate Carboxylase; Seasons; Sunlight; Temperature; Thermodynamics; Time Factors

Algae are the dominant primary producers in aquatic ecosystems. Since algae are highly varied group organisms, which have important functions in ecosystem, and their biomass is an essential biological resource. Currently, algae have been applied increasingly to diverse range of biomass applications. Therefore, this study was aimed to investigate the ecological algae features of microalgal production by natural medium, ecological function by lab scale of the symbiotic reactor which is imitated nature ecosystem, and atmospheric CO2 absorption that was related the algal growth of biomass to understand algae in natural water body better. Consequently, this study took advantages of using the unsupplemented freshwater natural medium to produce microalgae. Algal biomass by direct measurement of total suspended solids (TSS) and volatile suspended solids (VSS) resulted as 0.14g/L and 0.08g/L respectively. The biomass measurements of TSS and VSS are the sensible biomass index for algae production. The laboratory results obtained in the present study proved the production of algae by the natural water medium is potentially feasible.

Topics: Biomass; Carbon Dioxide; Chlorophyll; Chlorophyll A; Microalgae; Water

The exact color of light absorbed by chlorophyll (Chl) pigments, the light-harvesters in photosynthesis, is tuned by the protein microenvironment, but without knowledge of the intrinsic color of Chl it remains unclear how large this effect is. Experimental first absorption energies of Chl a and b isolated in vacuo and tagged with quaternary ammonium cations are reported. The energies are largely insensitive to details of the tag structure, a finding supported by first-principles calculations using time-dependent density functional theory. Absorption is significantly blue-shifted compared to that of Chl-containing proteins (by 30-70 nm). A single red-shifting perturbation, such as axial ligation or the protein medium, is insufficient to account even for the smallest shift; the largest requires pigment-pigment interactions.

Topics: Ammonium Compounds; Chlorophyll; Chlorophyll A; Color; Coloring Agents; Models, Molecular; Spectrophotometry

A challenge for the theory of optical spectra of pigment-protein complexes is the equal strength of the pigment-pigment and the pigment-protein couplings. Treating both on an equal footing so far can only be managed by numerically costly approaches. Here, we exploit recent results on a normal mode analysis derived spectral density that revealed the dominance of the diagonal matrix elements of the exciton-vibrational coupling in the exciton state representation. We use a cumulant expansion technique that treats the diagonal parts exactly, includes an infinite summation of the off-diagonal parts in secular and Markov approximations, and provides a systematic perturbative way to include non-secular and non-Markov corrections. The theory is applied to a model dimer and to chlorophyll (Chl) a and Chl b homodimers of the reconstituted water-soluble chlorophyll-binding protein (WSCP) from cauliflower. The model calculations reveal that the non-secular/non-Markov effects redistribute oscillator strength from the strong to the weak exciton transition in absorbance and they diminish the rotational strength of the exciton transitions in circular dichroism. The magnitude of these corrections is in a few percent range of the overall signal, providing a quantitative explanation of the success of time-local convolution-less density matrix theory applied earlier. A close examination of the optical spectra of Chl a and Chl b homodimers in WSCP suggests that the opening angle between Qy transition dipole moments in Chl b homodimers is larger by about 9(∘) than for Chl a homodimers for which a crystal structure of a related WSCP complex exists. It remains to be investigated whether this change is due to a different mutual geometry of the pigments or due to the different electronic structures of Chl a and Chl b.

Topics: Brassica; Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Circular Dichroism; Dimerization; Models, Molecular; Rotation; Temperature; Vibration; Water

Abundance of CaCO3 rich soil dust is a typical feature of atmospheric environment in the Indian region. During prevailing dry weather conditions, dustfall is deposited onto the foliar surfaces of plant affecting their morphology, stomata and the levels of biochemical constituents. This study reports the chemical characteristics of dustfall, its effect on foliar morphology and biochemical constituents of a medicinal plant (Morus alba) at two sites which are differentiated on the basis of landuse pattern, viz., (i) residential, Jawaharlal Nehru University (JNU), and (ii) industrial, Sahibabad (SB), located in the National Capital Region (NCR) of Delhi. Dustfall was characterized for major anions (F(-), Cl(-), NO3 (-) and SO4 (--)) and cations (Na(+), NH4 (+), K(+), Mg(++) and Ca(++)). Biochemical parameters such as chlorophyll a, chlorophyll b, total chlorophyll, carotenoid, proline and ascorbic acid were determined in foliar samples. The results showed that the dustfall fluxes of all the major ions were found to be higher at the industrial site (SB) as compared to the residential site (JNU). Foliar analysis revealed that the levels of biochemical parameters were more affected at SB site due to higher levels of dust SO4 (--) contributed by various anthropogenic sources resulting in more stressful conditions affecting the biochemistry of the plant. The possible entry pathways for dust SO4 (--) into foliar cells are also discussed in the paper. It was noticed that the deposition of urban dust was responsible for the damage of trichome, epidermis, cuticle and stomatal guard cells significantly affecting foliar morphology. SB exhibited more damage to these morphological parts suggesting that industrial dust is harmful to the plants.

Topics: Air Pollutants; Chlorophyll; Chlorophyll A; Dust; Environmental Monitoring; Industry; Ions; Morus; Plants; Soil; Sulfates

A unique CO2-Responsive CONSTANS, CONSTANS-like, and Time of Chlorophyll a/b Binding Protein1 (CCT) Protein (CRCT) containing a CCT domain but not a zinc finger motif is described, which is up-regulated under elevated CO2 in rice (Oryza sativa). The expression of CRCT showed diurnal oscillation peaked at the end of the light period and was also increased by sugars such as glucose and sucrose. Promoter β-glucuronidase analysis showed that CRCT was highly expressed in the phloem of various tissues such as leaf blade and leaf sheath. Overexpression or RNA interference knockdown of CRCT had no appreciable effect on plant growth and photosynthesis except that tiller angle was significantly increased by the overexpression. More importantly, starch content in leaf sheath, which serves as a temporary storage organ for photoassimilates, was markedly increased in overexpression lines and decreased in knockdown lines. The expressions of several genes related to starch synthesis, such as ADP-glucose pyrophospholylase and α-glucan phospholylase, were significantly changed in transgenic lines and positively correlated with the expression levels of CRCT. Given these observations, we suggest that CRCT is a positive regulator of starch accumulation in vegetative tissues, regulating coordinated expression of starch synthesis genes in response to the levels of photoassimilates.

Topics: Adenosine Diphosphate Glucose; Carbohydrate Metabolism; Carbon Dioxide; Chlorophyll; Chlorophyll A; Gene Expression; Gene Expression Regulation, Plant; Gene Knockdown Techniques; Glucose-1-Phosphate Adenylyltransferase; Glucuronidase; Oligonucleotide Array Sequence Analysis; Organ Specificity; Oryza; Phloem; Phosphorylases; Photosynthesis; Plant Leaves; Plant Proteins; Starch

Critical influence of different short chain fatty acids as organic carbon source, during growth (GP) and nutrient stress lipogenic phase (NSLP) was investigated on biomass and lipid productivity, in mixotrophic fed-batch microalgae cultivation. Nutrient deprivation induced physiological stress stimulated highest lipid productivity with acetate (total/neutral lipids, 35/17) with saturation index of 80.53% by the end of NSLP followed by butyrate (12/7%; 78%). Biomass growth followed the order of acetate (2.23 g/l) >butyrate (0.99 g/l) >propionate (0.77 g/l). VFA removal (as COD) was maximum with acetate (87%) followed by butyrate (55.09%) and propionate (10.60%). Palmitic acid was the most dominant fatty acid found in the fatty acid composition of all variants and butyrate fed system yielded a maximum of 44% palmitic acid. Protein profiling illustrated prominence of acetyl CoA-synthetase activity in acetate system. Thus, fatty acids provide a promising alternative feedstock for biodiesel production with integrated microalgae-biorefinery.

Topics: Acetate-CoA Ligase; Acetates; Biofuels; Biological Oxygen Demand Analysis; Biomass; Biotechnology; Butyrates; Chlorophyll; Chlorophyll A; Fatty Acids, Volatile; Gasoline; Lipid Metabolism; Lipids; Microalgae; Nitrates; Palmitic Acid; Phosphates; Propionates; Wastewater; Water Purification

Cadmium (Cd) is well known as one of the most toxic metals affecting the environment and can severely restrict plant growth and development. In this study, Cd toxicities were studied in strawberry cv. Camarosa using pot experiment. Chlorophyll and malondialdehyde (MDA) contents, catalase (CAT), superoxide dismutase (SOD), ascorbate peroxidase (APX) activities and mineral nutrient concentrations were investigated in both roots and leaves of strawberry plant after exposure Cd.. Cd content in both roots and leaves was increased with the application of increasing concentrations of Cd. We found higher Cd concentration in roots rather than in leaves. Chlorophyll a and b was decreased in leaves but MDA significantly increased under increased Cd concentration treatments in both roots and leaves. SOD and CAT activities was also increased with the increase Cd concentrations. K, Mn and Mg concentrations were found higher in leaves than roots under Cd stress. In general, increased Cd treatments increased K, Mg, Fe, Ca, Cu and Zn concentration in both roots and leaves. Excessive Cd treatments reduced chlorophyll contents, increased antioxidant enzyme activities and changes in plant nutrition concentrations in both roots and leaves.. The results presented in this work suggested that Cd treatments have negative effect on chlorophyll content and nearly decreased 30% of plant growth in strawberry. Strawberry roots accumulated higher Cd than leaves. We found that MDA and antioxidant enzyme (CAT, SOD and APX) contents may have considered a good indicator in determining Cd tolerance in strawberry plant.

Topics: Antioxidants; Ascorbate Peroxidases; Cadmium; Catalase; Chlorophyll; Chlorophyll A; Fragaria; Lipid Peroxidation; Magnesium; Malondialdehyde; Manganese; Micronutrients; Plant Extracts; Plant Leaves; Plant Roots; Potassium; Superoxide Dismutase

Cultivated orchids are the most abundantly attacked by polyphagous mealybugs. This study documented how different density of mealybug Pseudococcus longispinus (Targioni Tozzetti) infestation is associated with a response of antioxidative systems of Phalaenopsis × hybridum 'Innocence'. The degree of cell damage, estimated by electrolyte leakage measurement and the level of thiobarbituric acid reactive substances (TBARS), the content of pigments as well as the activity of antioxidative enzymes and proline level, as measurements of stress and stress compensation in moth orchid were examined. The highest electrolyte leakage (E L) value among samples from colonized plants was found in the orchids from series III (50 individuals/plant), whereas the lowest in the plants from series II (20 individuals/plant). The TBARS content reached the highest level at the lowest number of feeding insects (series I). Peroxidase activity toward guaiacol was significantly increased in series I (5 individuals/plant). The highest catalase activity was recorded in plants colonized by the highest number of scale insects (series III). Whereas, the highest value of proline was in series II. The content of individual photosynthetic pigments (chlorophyll a, chlorophyll b and carotenoids) in plant tissues did not vary significantly between control and colonized orchids. The results have not confirmed hypothesis that the increasing number of mealybugs occurring on plant enhanced plant physiological response. The degree of longtailed mealybug infestation on plants was positively correlated only with electrolyte leakage and catalase activity in leaf tissues.

Topics: Analysis of Variance; Animals; Antioxidants; Carotenoids; Catalase; Cell Membrane; Chlorophyll; Chlorophyll A; Electrolytes; Hemiptera; Hybridization, Genetic; Orchidaceae; Peroxidase; Plant Leaves; Population Density; Proline; Thiobarbituric Acid Reactive Substances

We have performed gas-phase absorption spectroscopy in the Soret-band region of chlorophyll (Chl) a and b tagged by quaternary ammonium ions together with time-dependent density functional theory (TD-DFT) calculations. This band is the strongest in the visible region of metalloporphyrins and an important reporter on the microenvironment. The cationic charge tags were tetramethylammonium, tetrabutylammonium, and acetylcholine, and the dominant dissociation channel in all cases was breakage of the complex to give neutral Chl and the charge tag as determined by photoinduced dissociation mass spectroscopy. Two photons were required to induce fragmentation on the time scale of the experiment (microseconds). Action spectra were recorded where the yield of the tag as a function of excitation wavelength was sampled. These spectra are taken to represent the corresponding absorption spectra. In the case of Chl a we find that the tag hardly influences the band maximum which for all three tags is at 403 ± 5 nm. A smaller band with maximum at 365 ± 10 nm was also measured for all three complexes. The spectral quality is worse in the case of Chl b due to lower ion beam currents; however, there is clear evidence for the absorption being to the red of that of Chl a (most intense peak at 409 ± 5 nm) and also a more split band. Our results demonstrate that the change in the Soret-band spectrum when one peripheral substituent (CH3) is replaced by another (CHO) is an intrinsic effect. First principles TD-DFT calculations agree with our experiments, supporting the intrinsic nature of the difference between Chl a and b and also displaying minimal spectral changes when different charge tags are employed. The deviations between theory and experiment have allowed us to estimate that the Soret-band absorption maxima in vacuo for the neutral Chl a and Chl b should occur at 405 nm and 413 nm, respectively. Importantly, the Soret bands of the isolated species are significantly blueshifted compared to those of solvated Chl and Chl-proteins. The protein microenvironment is certainly not innocent of perturbing the electronic structure of Chls.

Topics: Chlorophyll; Chlorophyll A; Mass Spectrometry; Quaternary Ammonium Compounds; Spinacia oleracea

Microcystins (MCs) produced by cyanobacteria in natural environments are a potential risk to the integrity of ecosystems. In this study, the effects of cyanobacterial cell-free crude extracts from a Microcystis aeruginosa bloom containing three MC-congeners MC-LR, -RR, and -YR at environmental relevant concentrations of 49.3±2.9, 49.8±5.9, and 6.9±3.8μg/L, respectively, were evaluated on Ceratophyllum demersum (L.), Egeria densa (Planch.), and Hydrilla verticillata (L.f.). Effects on photosynthetic pigments (total chlorophyll (chl), chl a, chl b, and carotenoids), enzymatic defense led by catalase (CAT), peroxidase (POD) and glutathione reductase (GR), and biotransformation enzyme glutathione S-transferase (GST) were measured after 1, 4, and 8h and after 1, 3, 7, and 14 days of exposure. Results show that in all exposed macrophytes, photosynthetic pigments were negatively affected. While chl a and total chl decreased with increasing exposure time, a parallel increase in chl b was observed after 8h. Concomitant increase of ∼5, 16, and 34% of antioxidant carotenoid concentration in exposed C. demersum, E. densa, and H. verticillata, respectively, was also displayed. Enzymatic antioxidant defense systems in all exposed macrophytes were initiated within the first hour of exposure. In exposed E. densa, highest values of CAT and GR activities were observed after 4 and 8h, respectively, while in exposed H. verticillata highest value of POD activity was observed after 8h. An early induction with a significant increase of biotransformation enzyme GST was observed in E. densa after 4h and in C. demersum and H. verticillata after 8h. These results are the first to show rapid induction of stress and further possible MC biotransformation (based on the activation of GST enzymatic activity included in MC metabolization during the biotransformation mechanism) in macrophytes exposed to crude extract containing a mixture of MCs.

Topics: Antioxidants; Catalase; Chlorophyll; Chlorophyll A; Complex Mixtures; Cyanobacteria; Glutathione Reductase; Glutathione Transferase; Guatemala; Lakes; Magnoliopsida; Microcystins; Peroxidase; Photosynthesis; Time Factors

In this study, we isolated a full-length cDNA and named ZmBDF from zea mays. ZmBDF encoded a protein of 356 amino acids and phylogenetic analysis showed that it belongs to a closely related subgroup with B3 domain factors in plants. The transcript level of ZmBDF could be induced by ABA, MeJA, salt or drought treatments. To further investigated the function of ZmBDF, ZmBDF over-expression transgenic lines were got by transforming it into Arabidopsis thaliana. ZmBDF over-expression transgenic plants in Arabidopsis could increase drought and salt tolerant in germination assay. Under drought condition, net photosynthetic rates (PN), stomatal conductance (gs), and internal leaf CO2 concentration (Ci) were less affected in transgenic plants compared with wild type. Besides, the chlorophyll a and chlorophyll b (chl a/chl b) ratio decreased in WT plants than the transgenic plants and total carotenoid content show opposite trends. Moreover, transgenic plants could also reduce the stomatal density and changed the stomatal shape. Taken together, our data suggested that ZmBDF could improve stress tolerance to drought and salt in maize.

Topics: Amino Acid Sequence; Arabidopsis; Carbon Dioxide; Chlorophyll; Chlorophyll A; DNA, Complementary; DNA, Plant; Droughts; Molecular Sequence Data; Photosynthesis; Phylogeny; Plant Proteins; Plant Stomata; Plant Transpiration; Plants, Genetically Modified; Protein Structure, Secondary; Salt Tolerance; Sequence Analysis, DNA; Stress, Physiological; Zea mays

The amount of light plants can tolerate during different phases of ontogenesis remains largely unknown. This was addressed here employing a novel methodology that uses the coefficient of photochemical quenching (qP) to assess the intactness of photosystem II reaction centres. Fluorescence quenching coefficients, total chlorophyll content and concentration of anthocyanins were determined weekly during the juvenile, adult, reproductive and senescent phases of plant ontogenesis. This enabled quantification of the protective effectiveness of non-photochemical fluorescence quenching (NPQ) and determination of light tolerance. The light intensity that caused photoinhibition in 50% of leaf population increased from ∼70 μmol m(-2)  s(-1) , for 1-week-old seedlings, to a maximum of 1385 μmol m(-2)  s(-1) for 8-week-old plants. After 8 weeks, the tolerated light intensity started to gradually decline, becoming only 332 μmol m(-2)  s(-1) for 13-week-old plants. The dependency of light tolerance on plant age was well-related to the amplitude of protective NPQ (pNPQ) and the electron transport rates (ETRs). Light tolerance did not, however, show a similar trend to chlorophyll a/b ratios and content of anthocyanins. Our data suggest that pNPQ is crucial in defining the capability of high light tolerance by Arabidopsis plants during ontogenesis.

Topics: Arabidopsis; Arabidopsis Proteins; Chlorophyll; Chlorophyll A; Electron Transport; Fluorescence; Light; Photosystem II Protein Complex; Plant Leaves

Chloroplasts have means to manage excess reducing power but these mechanisms may become restricted by rates of ATP turnover. Alternative oxidase (AOX) is a mitochondrial terminal oxidase that uncouples the consumption of reducing power from ATP synthesis. Physiological and biochemical analyses were used to compare respiration and photosynthesis of Nicotiana tabacum wild-type (WT) plants with that of transgenic lines overexpressing AOX, under both well-watered and drought stress conditions. With increasing drought severity, AOX overexpression acted to increase respiration in the light (RL ) relative to WT. CO2 and light response curves indicated that overexpression also improved photosynthetic performance relative to WT, as drought severity increased. This was not due to an effect of AOX amount on leaf water status or the development of the diffusive limitations that occur due to drought. Rather, AOX overexpression dampened photosystem stoichiometry adjustments and losses of key photosynthetic components that occurred in WT. The results indicate that AOX amount influences RL , particularly during severe drought, when cytochrome pathway respiration may become increasingly restricted. This impacts the chloroplast redox state, influencing how the photosynthetic apparatus responds to increasing drought severity. In particular, the development of biochemical limitations to photosynthesis are dampened in plants with increased nonenergy conserving RL .

Topics: Carbon Dioxide; Cell Respiration; Chlorophyll; Chlorophyll A; Droughts; Electron Transport; Electrons; Light; Nicotiana; Photosynthesis; Photosystem I Protein Complex; Photosystem II Protein Complex; Plant Proteins; Plant Stomata; Plants, Genetically Modified; RNA, Messenger

The health of several East Antarctic moss-beds is declining as liquid water availability is reduced due to recent environmental changes. Consequently, a noninvasive and spatially explicit method is needed to assess the vigour of mosses spread throughout rocky Antarctic landscapes. Here, we explore the possibility of using near-distance imaging spectroscopy for spatial assessment of moss-bed health. Turf chlorophyll a and b, water content and leaf density were selected as quantitative stress indicators. Reflectance of three dominant Antarctic mosses Bryum pseudotriquetrum, Ceratodon purpureus and Schistidium antarctici was measured during a drought-stress and recovery laboratory experiment and also with an imaging spectrometer outdoors on water-deficient (stressed) and well-watered (unstressed) moss test sites. The stress-indicating moss traits were derived from visible and near infrared turf reflectance using a nonlinear support vector regression. Laboratory estimates of chlorophyll content and leaf density were achieved with the lowest systematic/unsystematic root mean square errors of 38.0/235.2 nmol g(-1) DW and 0.8/1.6 leaves mm(-1) , respectively. Subsequent combination of these indicators retrieved from field hyperspectral images produced small-scale maps indicating relative moss vigour. Once applied and validated on remotely sensed airborne spectral images, this methodology could provide quantitative maps suitable for long-term monitoring of Antarctic moss-bed health.

Topics: Antarctic Regions; Bryophyta; Chlorophyll; Chlorophyll A; Dehydration; Droughts; Geography; Imaging, Three-Dimensional; Plant Leaves; Spectrum Analysis; Stress, Physiological; Water

Sea grasses are foundation species for estuarine ecosystems. The available light for sea grasses diminishes rapidly during pollutant spills, effluent releases, disturbances such as intense riverine input, and tidal changes. We studied how sea grasses' remote-sensing signatures and light-capturing ability respond to short term light alterations. In vivo responses were measured over the entire visible-light spectra to diminishing white-light on whole-living-plants' spectral reflectance, including 6h of full oceanic-light fluences from 10% to 100%. We analyzed differences by various reflectance indices. We compared the sea grasses species responses of tropical vs. temperate and intertidals (Halodule wrightii, and Zostera marina) vs. subtidal (Thalassia testudinum). Reflectance diminished with decreasing light intensity that coincided with greater accessory pigment stimulation (anthocyanin, carotenoids, xanthins). Chlorophyll a and Chlorophyll b differed significantly among species (Thalassia vs. Halodule). Photosynthetic efficiency diminished at high light intensities. The NDVI index was inadequate to perceive these differences. Our results demonstrate the leaf-level utility of data to remote sensing for mapping sea grass and sea grass stress.

Topics: Alismatales; Atlantic Ocean; Chlorophyll; Chlorophyll A; Ecosystem; Environmental Monitoring; Hydrocharitaceae; Light; Photosynthesis; Pigments, Biological; Plant Leaves; Remote Sensing Technology; Zosteraceae

Thalli of the lichen Pseudevernia furfuracea were transplanted for 3 months (November 2010-January 2011) at 61 monitoring sites around a cement plant near Castrovillari (Calabria, southern Italy). NH3, NO x and SO2 concentrations were monitored monthly in a subarea of 10 sites (SA10) where the cement plant was located. At the end of the exposure period, the integrity of cell membranes; membrane lipid peroxidation (thiobarbituric acid reactive substances, TBARS level); vitality (cell respiration); chlorophyll a; chlorophyll b; carotenoids; phaeophytization quotient; photosynthetic efficiency and thalli concentrations of Al, Ca, Mg, V and Fe were measured. NO x concentrations correlated with the site distance from the cement plant while NH3 concentrations correlated with lichen vitality within SA10. For the monitoring area as a whole, only Fe and Mg concentrations correlated with membrane lipid peroxidation, while TBARS levels showed a significant increase and chlorophyll a, chlorophyll b and carotenoids a significant decrease with respect to the lichen origin area. Multivariate analysis (detrended correspondence analysis, cluster analysis and multi-response permutation procedure) of the eco-physiological parameters × monitoring sites data set resulted in four clusters termed C1, C2, C3 and C4. The eco-physiological parameters were compared among the four clusters and lichen origin area by one-way ANOVA. An index of environmental favourableness (IEF) to lichens was calculated to evaluate the spatial recovery of impaired values of TBARS, chlorophyll a, chlorophyll b, xanthophylls + carotenoids and phaeophytization quotient. The results indicate that there is no clear spatial trend in mycobiont impairment even though the IEF values suggest a higher number of sites with low levels of membrane lipid peroxidation in the 2--3-km distance band from the cement plant (the outermost) than in the two other distance bands (0-1 and 1-2 km). The photobiont seems to be damaged mainly in the inner distance band of the study area as suggested by the gradual but significant recovery trend of pigment levels and phaeophytization quotient from the inner distance band to the outer one (as shown by the IEF values). Conversion of chlorophyll to phaeophytin probably is not the only process affecting pigment levels.

Topics: Adsorption; Ammonia; Ascomycota; Carotenoids; Chlorophyll; Chlorophyll A; Construction Industry; Environmental Monitoring; Environmental Pollution; Italy; Lichens; Lipid Peroxidation; Nitrogen Oxides; Photosynthesis; Stress, Physiological; Sulfur Dioxide

Chlamydomonas sp. UWO 241 (UWO 241) is a psychrophilic green alga isolated from Antarctica. A unique characteristic of this algal strain is its inability to undergo state transitions coupled with the absence of photosystem II (PSII) light-harvesting complex protein phosphorylation. We show that UWO 241 preferentially phosphorylates specific polypeptides associated with an approximately 1,000-kD pigment-protein supercomplex that contains components of both photosystem I (PSI) and the cytochrome b₆/f (Cyt b₆/f) complex. Liquid chromatography nano-tandem mass spectrometry was used to identify three major phosphorylated proteins associated with this PSI-Cyt b₆/f supercomplex, two 17-kD PSII subunit P-like proteins and a 70-kD ATP-dependent zinc metalloprotease, FtsH. The PSII subunit P-like protein sequence exhibited 70.6% similarity to the authentic PSII subunit P protein associated with the oxygen-evolving complex of PSII in Chlamydomonas reinhardtii. Tyrosine-146 was identified as a unique phosphorylation site on the UWO 241 PSII subunit P-like polypeptide. Assessment of PSI cyclic electron transport by in vivo P700 photooxidation and the dark relaxation kinetics of P700(+) indicated that UWO 241 exhibited PSI cyclic electron transport rates that were 3 times faster and more sensitive to antimycin A than the mesophile control, Chlamydomonas raudensis SAG 49.72. The stability of the PSI-Cyt b₆/f supercomplex was dependent upon the phosphorylation status of the PsbP-like protein and the zinc metalloprotease FtsH as well as the presence of high salt. We suggest that adaptation of UWO 241 to its unique low-temperature and high-salt environment favors the phosphorylation of a PSI-Cyt b₆/f supercomplex to regulate PSI cyclic electron transport rather than the regulation of state transitions through the phosphorylation of PSII light-harvesting complex proteins.

Topics: Amino Acid Sequence; Antimycin A; Chemical Fractionation; Chlamydomonas; Chlorophyll; Chlorophyll A; Cytochrome b6f Complex; Diuron; Electron Transport; Electrophoresis, Gel, Two-Dimensional; Immunoblotting; Molecular Sequence Data; Multiprotein Complexes; Phosphorylation; Photosystem I Protein Complex; Protein Stability; Sequence Alignment; Sodium Chloride; Thylakoids

The light-harvesting-like (LIL) proteins are a family of membrane proteins that share a chlorophyll a/b-binding motif with the major light-harvesting antenna proteins of oxygenic photoautotrophs. LIL proteins have been associated with the regulation of tetrapyrrol biosynthesis, and plant responses to light-stress. Here, it was found in a native PAGE approach that chlorophyllide, and chlorophyllide plus geranylgeraniolpyrophosphate trigger assembly of Lil3 in three chlorine binding fluorescent protein bands, termed F1, F2, and F3. It is shown that light and chlorophyllide trigger accumulation of protochlorophyllide-oxidoreductase, and chlorophyll synthase in band F3. Chlorophyllide and chlorophyll esterified to geranylgeraniol were identified as basis of fluorescence recorded from band F3. A direct interaction between Lil3, CHS and POR was confirmed in a split ubiquitin assay. In the presence of light or chlorophyllide, geranylgeraniolpyrophosphate was shown to trigger a loss of the F3 band and accumulation of Lil3 and geranylgeranyl reductase in F1 and F2. No direct interaction between Lil3 and geranylgeraniolreductase was identified in a split ubiquitin assay; however, accumulation of chlorophyll esterified to phytol in F1 and F2 corroborated the enzymes assembly. Chlorophyll esterified to phytol and the reaction center protein psbD of photosystem II were identified to accumulate together with psb29, and APX in the fluorescent band F2. Data show that Lil3 assembles with proteins regulating chlorophyll synthesis in etioplasts from barley (Hordeum vulgare L.).

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Hordeum; Light; Light-Harvesting Protein Complexes; Oxidoreductases; Photosystem II Protein Complex; Phytol; Protein Binding

Spartina maritima is an ecosystem engineer that has shown to be useful for phytoremediation purposes. A glasshouse experiment using soil from a metal-contaminated estuary was designed to investigate the effect of a native bacterial consortium, isolated from S. maritima rizhosphere and selected owing to their plant growth promoting properties and multiresistance to heavy metals, on plant growth and metal accumulation. Plants of S. maritima were randomly assigned to three soil bioaugmentation treatments (without inoculation, one inoculation and repeated inoculations) for 30 days. Growth parameters and photosynthetic traits, together with total concentrations of several metals were determined in roots and/or leaves. Bacterial inoculation improved root growth, through a beneficial effect on photosynthetic rate (AN) due to its positive impact on functionality of PSII and chlorophyll concentration. Also, favoured intrinsic water use efficiency of S. maritima, through the increment in AN, stomatal conductance and in root-to-shoot ratio. Moreover, this consortium was able to stimulate plant metal uptake specifically in roots, with increases of up to 19% for As, 65% for Cu, 40% for Pb and 29% for Zn. Thus, bioaugmentation of S. maritima with the selected bacterial consortium can be claimed to enhance plant adaptation and metal rhizoaccumulation during marsh restoration programs.

Topics: Arsenic; Bacillus; Biodegradation, Environmental; Chlorophyll; Chlorophyll A; Estuaries; Metals, Heavy; Plant Leaves; Plant Roots; Poaceae; Rhizosphere; Soil Pollutants

Narrowleaf cottonwood (Populus angustifolia James) is an obligate riparian poplar that is a foundation species in river valleys along the Rocky Mountains, spanning 16° of latitude from southern Arizona, USA to southern Alberta, Canada. Its current distribution is fragmented, and genetic variation shows regional population structure consistent with the effects of geographic barriers and past climate. It is thus very well-suited for investigating ecophysiological adaptation associated with latitude. In other section Tacamahaca poplar species, genotypes from higher latitudes show evidence of short-season adaptation with foliar traits that contribute to higher photosynthetic capacity. We tested for similar adaptation in three populations of narrowleaf cottonwoods: from Arizona (south), Alberta (north) and Utah, near the centre of the latitudinal distribution. We propagated 20 genotypes from each population in a common garden in Alberta, and measured foliar and physiological traits after 3 years. Leaves of genotypes from the northern population had higher leaf mass per area (LMA), increased nitrogen (N) content and higher carotenoid and chlorophyll content, and these were associated with higher light-saturated net photosynthesis (Asat). In leaves of all populations the majority of stomata were abaxial, with the proportion of abaxial stomata highest in the southern population. Stomatal conductance (gs) and transpiration rates were higher in the northern population but water-use efficiency (Asat/gs) and leaf carbon isotope composition (δ(13)C) did not differ across the populations. These results (i) establish links between Asat and gs, N, chlorophyll and LMA among populations within this species, (ii) are consistent with the discrimination of populations from prior investigation of genetic variation and (iii) support the concept of latitudinal adaptation, whereby deciduous trees from higher latitudes display higher photosynthetic capacity, possibly compensating for a shorter and cooler growth season and reduced insolation.

Topics: Altitude; Carotenoids; Chlorophyll; Chlorophyll A; Climate; Gases; Geography; Light; Linear Models; Photosynthesis; Plant Leaves; Plant Stomata; Populus; United States; Water

The correct interplay of interactions between protein, pigment and lipid molecules is highly relevant for our understanding of the association behavior of the light harvesting complex (LHCII) of green plants. To cover the relevant time and length scales in this multicomponent system, a multi-scale simulation ansatz is employed that subsequently uses a classical all atomistic (AA) model to derive a suitable coarse grained (CG) model which can be backmapped into the AA resolution, aiming for a seamless conversion between two scales. Such an approach requires a faithful description of not only the protein and lipid components, but also the interaction functions for the indispensable pigment molecules, chlorophyll b and chlorophyll a (referred to as chl b/chl a). In this paper we develop a CG model for chl b and chl a in a dipalmitoylphosphatidyl choline (DPPC) bilayer system. The structural properties and the distribution behavior of chl within the lipid bilayer in the CG simulations are consistent with those of AA reference simulations. The non-bonded potentials are parameterized such that they fit to the thermodynamics based MARTINI force-field for the lipid bilayer and the protein. The CG simulation shows chl aggregation in the lipid bilayer which is supported by fluorescence quenching experiments. It is shown that the derived chl model is well suited for CG simulations of stable, structurally consistent, trimeric LHCII and can in the future be used to study their large scale aggregation behavior.

Topics: Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Lipid Bilayers; Models, Molecular; Protein Multimerization; Protein Structure, Quaternary

Oxygenic photosynthetic organisms evolved mechanisms for thermal dissipation of energy absorbed in excess to prevent formation of reactive oxygen species. The major and fastest component, called non-photochemical quenching, occurs within the photosystem II antenna system by the action of two essential light-harvesting complex (LHC)-like proteins, photosystem II subunit S (PSBS) in plants and light-harvesting complex stress-related (LHCSR) in green algae and diatoms. In the evolutionary intermediate Physcomitrella patens, a moss, both gene products are active. These proteins, which are present in low amounts, are difficult to purify, preventing structural and functional analysis. Here, we report on the overexpression of the LHCSR1 protein from P. patens in the heterologous systems Nicotiana benthamiana and Nicotiana tabacum using transient and stable nuclear transformation. We show that the protein accumulated in both heterologous systems is in its mature form, localizes in the chloroplast thylakoid membranes, and is correctly folded with chlorophyll a and xanthophylls but without chlorophyll b, an essential chromophore for plants and algal LHC proteins. Finally, we show that recombinant LHCSR1 is active in quenching in vivo, implying that the recombinant protein obtained is a good material for future structural and functional studies.

Topics: Bryopsida; Chlorophyll; Chlorophyll A; Light; Light-Harvesting Protein Complexes; Lutein; Nicotiana; Photochemistry; Photosynthesis; Photosystem II Protein Complex; Plant Leaves; Protein Binding; Recombinant Proteins; Subcellular Fractions; Thylakoids; Xanthophylls

Light-harvesting chlorophyll a/b-binding proteins (LHCB) have been implicated in the stress response. In this study, a gene encoding LHCB in the pigeon pea was cloned and characterized. Based on the sequence of a previously obtained 327 bp Est, a full-length 793 bp cDNA was cloned using the rapid amplification of cDNA ends (RACE) method. It was designated CcLHCB1 and encoded a 262 amino acid protein. The calculated molecular weight of the CcLHCB1 protein was 27.89 kDa, and the theoretical isoelectric point was 5.29. Homology search and sequence multi-alignment demonstrated that the CcLHCB1 protein sequence shared a high identity with LHCB from other plants. Bioinformatics analysis revealed that CcLHCB1 was a hydrophobic protein with three transmembrane domains. By fluorescent quantitative real-time polymerase chain reaction (PCR), CcLHCB1 mRNA transcripts were detectable in different tissues (leaf, stem, and root), with the highest level found in the leaf. The expression of CcLHCB1 mRNA in the leaves was up-regulated by drought stimulation and AM inoculation. Our results provide the basis for a better understanding of the molecular organization of LCHB and might be useful for understanding the interaction between plants and microbes in the future.

Topics: Amino Acid Sequence; Cajanus; Chlorophyll; Chlorophyll A; Cloning, Molecular; Computational Biology; Light-Harvesting Protein Complexes; Molecular Sequence Data

Nonradiative relaxation of high-energy excited states to the lowest excited state in chlorophylls marks the first step in the process of photosynthesis. We perform ultrafast transient absorption spectroscopy measurements, that reveal this internal conversion dynamics to be slightly slower in chlorophyll B than in chlorophyll A. Modeling this process with non-adiabatic excited state molecular dynamics simulations uncovers a critical role played by the different side groups in the two molecules in governing the intramolecular redistribution of excited state wavefunction, leading, in turn, to different time-scales. Even given smaller electron-vibrational couplings compared to common organic conjugated chromophores, these molecules are able to efficiently dissipate about 1 eV of electronic energy into heat on the timescale of around 200 fs. This is achieved via selective participation of specific atomic groups and complex global migration of the wavefunction from the outer to inner ring, which may have important implications for biological light-harvesting function.

Topics: Chlorophyll; Chlorophyll A; Light; Models, Theoretical; Photosynthesis; Spectrum Analysis

To study the impacts of lead (Pb) stress on the leaf photosynthetic pigment and the growth of Rabdosia rubescens,in order to provide a basis for planting area selection and growth regulation.. Taking chlorophyll a, chlorophyll b, carotenoids, growth rate, biomass and Pb content as the indexes, the Ramets hydroponic experiments at Pb concentration levels (135,270 and 540 mg/L) in the time(20, 35 and 50 d) were carried out. Photosynthetic pigment content was determined by spectrophotometer, and Pb mass fraction was detected with plasma emission spectrometer.. There was uncertain effect on chlorophyll and carotenoid synthesis in different Pb concentrations in-early period (20 and 35 d). At the time of 50 d, the chlorophyll content was higher in the low-mid Pb concentrations, significantly lower in the high Pb concentration compared with the control group, and there were no significant differences on carotenoid contents in different Pb concentrations.. Low-mid Pb concentrations can promote chlorophyll synthesis, and the bioaccumulation of high Pb concentration can inhibit the chlorophyll synthesis, and then restrict the growth of Rabdosia rubescens.

Topics: Biomass; Carotenoids; Chlorophyll; Chlorophyll A; Isodon; Lead; Photosynthesis; Plant Leaves

Over the past decade, both experimentalists and theorists have worked to develop methods to describe pigment-protein coupling in photosynthetic light-harvesting complexes in order to understand the molecular basis of quantum coherence effects observed in photosynthesis. Here we present an improved strategy based on the combination of quantum mechanics/molecular mechanics (QM/MM) molecular dynamics (MD) simulations and excited-state calculations to predict the spectral density of electronic-vibrational coupling. We study the water-soluble chlorophyll-binding protein (WSCP) reconstituted with Chl a or Chl b pigments as the system of interest and compare our work with data obtained by Pieper and co-workers from differential fluorescence line-narrowing spectra (Pieper et al. J. Phys. Chem. B 2011, 115 (14), 4042-4052). Our results demonstrate that the use of QM/MM MD simulations where the nuclear positions are still propagated at the classical level leads to a striking improvement of the predicted spectral densities in the middle- and high-frequency regions, where they nearly reach quantitative accuracy. This demonstrates that the so-called "geometry mismatch" problem related to the use of low-quality structures in QM calculations, not the quantum features of pigments high-frequency motions, causes the failure of previous studies relying on similar protocols. Thus, this work paves the way toward quantitative predictions of pigment-protein coupling and the comprehension of quantum coherence effects in photosynthesis.

Topics: Brassica; Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Molecular Dynamics Simulation; Plant Proteins; Protein Structure, Tertiary; Quantum Theory; Water

In the present study, the toxic effects of 1-octyl-3-methylimidazolium chloride ([Omim]Cl), 1-octyl-3-methylimidazolium bromide ([Omim]Br) and 1-octyl-3-methylimidazolium tetrafluoroborate ([Omim]BF4) in soil on Vicia faba (V. faba) seedlings at 0, 100, 200, 400, 600 and 800 mg kg(-1) were assessed for the first time at the cellular and molecular level. Moreover, the toxicity of these three ionic liquids (ILs) was evaluated, and the influence of anions on the toxicity of the ILs was assessed. The results showed that even at 100 mg kg(-1), the growth of V. faba seedlings was inhibited after exposure to the three ILs, and the inhibitory effect was enhanced with increasing concentrations of the three ILs. The level of reactive oxygen species (ROS) was increased after exposure to the three ILs, which resulted in lipid peroxidation, DNA damage and oxidative damage in the cells of the V. faba seedlings. In addition, the anion structure could influence the toxicity of ILs, and toxicity of the three tested ILs decreased in the following order: [Omim]BF4 > [Omim]Br > [Omim]Cl. Moreover, oxidative damage is the primary mechanism by which ILs exert toxic effects on crops, and ILs could reduce the agricultural productivity.

Topics: Anions; Ascorbic Acid; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Comet Assay; DNA Damage; Dose-Response Relationship, Drug; Glutathione; Hydrogen Peroxide; Imidazoles; Ionic Liquids; Lipid Peroxidation; Reactive Oxygen Species; Seedlings; Soil; Superoxide Dismutase; Superoxides; Vicia faba

In order to determine the adaptability of Adzuki beans as the interplanting crops in fruit yards, field and pot experimental treatments with full natural light and weak light (48% of full natural light) regimes were conducted to test the shade tolerance and physiological responses of three Adzuki bean varieties including Funan green Vigna angularis (FGVA), early-mature black V. angularis (EBVA) and late-mature black V. angularis (LBVA). The leaf photosynthetic characteristic parameters, photosynthetic pigment contents and the activity of RuBPCase were measured during the first bloom stage. The response of growth to weak light was likewise studied. The results showed that the photosynthetic characteristic parameters, i.e., the maximum net photosynthetic rate, light saturation point and light compensation point of the three Adzuki bean varieties under the weak light stress changed differently. The weak light stress induced the reduction of net photosynthetic rate, water use efficiency and RuBPCase activity of the three Adzuki bean varieties significantly. The contents of chlorophyll a and chlorophyll b in leaves of FGVA increased significantly, while Chl a/b and carotenoid content in the leaves decreased significantly after shading. But the other two varieties did not change obviously in photosynthetic pigments content after shading. The weak light changed the growth of the three Adzuki bean varieties, such as decreasing dry matter yield and dry matter accumulation efficiency, reducing root nodule and root-shoot ratio, debasing leaves quantity and leaf area index. The first bloom stage and maturing stage of FGVA advanced, while that of EBVA delayed under weak light. However, flowers were not strong enough to seed for LBVA under the weak light. In conclusion, according to the photosynthetic characteristics changes after shading, as well as the growth status, we concluded that the shade tolerance of the three Adzuki beans was ranked as FGVA>EBVA>LBVA.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Crops, Agricultural; Fabaceae; Light; Photosynthesis; Plant Leaves; Plant Roots

Greater exposure to Pb(Ⅱ) increases the likelihood of harmful effects in the environment. In this study, the aquatic unicellular alga Chlorella protothecoides (C. protothecoides) and Chlorella vulgaris (C. vulgaris) were chosen to assess the acute and chronic toxicity of Pb(Ⅱ) exposure. Results of the observations show dose-response relationships could be clearly observed between Pb(Ⅱ) concentration and percentage inhibition (PI). Exposure to Pb(Ⅱ) increased malondialdehyde (MDA) content by up to 4.22 times compared with the control, suggesting that there was some oxidative damage. ANOVA analysis shows that Pb(Ⅱ) decreased chlorophyll (chl) content, indicating marked concentration-dependent relationships, and the lowest levels of chl a, chl b, and total-chl were 14.53, 18.80, and 17.95% of the controls, respectively. A real-time PCR assay suggests the changes in transcript abundances of three photosynthetic-related genes. After 120 h exposure Pb(Ⅱ) reduced the transcript abundance of rbcL, psaB, and psbC, and the relative abundances of the three genes of C. protothecoides and C. vulgaris in response to Pb(Ⅱ) were 54.66-98.59, 51.68-95.59, 37.89-95.48, 36.04-94.94, 41.19-91.20, and 58.75-96.80% of those of the controls, respectively. As for 28 d treatments, the three genes displayed similar inhibitory trend. This research provides a basic understanding of Pb(Ⅱ) toxicity to aquatic organisms.

Topics: Chlorella; Chlorella vulgaris; Chlorophyll; Chlorophyll A; Environmental Pollutants; Lead; Malondialdehyde; Oxidation-Reduction; Photosynthesis; Species Specificity; Transcription, Genetic

This study investigated the effect of silicon (Si) on the photosynthetic gas exchange parameters (net CO2 assimilation rate [A], stomatal conductance to water vapor [gs], internal CO2 concentration [Ci], and transpiration rate [E]) and chlorophyll fluorescence a parameters (maximum quantum quenching [Fv/Fm and Fv'/Fm'], photochemical [qP] and nonphotochemical [NPQ] quenching coefficients, and electron transport rate [ETR]) in wheat plants grown in a nutrient solution containing 0 mM (-Si) or 2 mM (+Si) Si and noninoculated or inoculated with Pyricularia oryzae. Blast severity decreased due to higher foliar Si concentration. For the inoculated +Si plants, A, gs, and E were significantly higher in contrast to the inoculated -Si plants. For the inoculated +Si plants, significant differences of Fv/Fm between the -Si and +Si plants occurred at 48, 96, and 120 h after inoculation (hai) and at 72, 96, and 120 hai for Fv'/Fm'. The Fv/Fm and Fv'/Fm', in addition to total chlorophyll concentration (a + b) and the chlorophyll a/b ratio, significantly decreased in the -Si plants compared with the +Si plants. Significant differences between the -Si and +Si inoculated plants occurred for qP, NPQ, and ETR. The supply of Si contributed to decrease blast severity in addition to improving gas exchange performance and causing less dysfunction at the photochemical level.

Topics: Carbon Dioxide; Chlorophyll; Chlorophyll A; Magnaporthe; Photosynthesis; Photosystem II Protein Complex; Plant Diseases; Plant Leaves; Plant Stomata; Plant Transpiration; Silicon; Triticum

The antioxidant activity and individual bioactive compounds of lettuce, cultivated with 2.5-30% (v/v) of fresh or composted espresso spent coffee grounds, were assessed. A progressive enhancement of lettuce's antioxidant capacity, evaluated by radical scavenging effect and reducing power, was exhibited with the increment of fresh spent coffee amounts, while this pattern was not so clear with composted treatments. Total reducing capacity also improved, particularly for low spent coffee concentrations. Additionally, very significant positive correlations were observed for all carotenoids in plants from fresh spent coffee treatments, particularly for violaxanthin, evaluated by HPLC. Furthermore, chlorophyll a was a good discriminating factor between control group and all spent coffee treated samples, while vitamin E was not significantly affected. Espresso spent coffee grounds are a recognised and valuable source of bioactive compounds, proving herein, for the first time, to potentiate the antioxidant pool and quality of the vegetables produced.

Topics: Antioxidants; Carotenoids; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Coffee; Crops, Agricultural; Fertilizers; Lactuca; Lutein; Plant Extracts; Soil; Tocopherols; Xanthophylls

Bioaccumulation, subcellular distribution, and acute toxicity of yttrium (Y) were evaluated in Nymphoides peltata. The effects of Y concentrations of 1-5 mg L(-1) applied for 4 days were assessed by measuring changes in photosynthetic pigments, nutrient contents, enzymatic and non-enzymatic antioxidants, and ultrastructure. The accumulation of Y in subcellular fractions decreased in the order of cell wall > organelle > soluble fraction. Much more Y was located in cellulose and pectin than in other biomacromolecules. The content of some mineral elements (Mg, Ca, Fe, Mn, and Mo) increased in N. peltata, but there was an opposite effect for P and K. Meanwhile, ascorbate, and catalase activity decreased significantly for all Y concentrations. In contrast, peroxidase activity was induced, while initial rises in superoxide dismutase activity and glutathione content were followed by subsequent declines. Morphological symptoms of senescence, such as chlorosis and damage to chloroplasts and mitochondria, were observed even at the lowest Y concentration. Pigment content decreased as the Y concentration rose and the calculated EC50 and MPC of Y for N. peltata were 2 and 0.2 mg L(-1) after 4 days of exposure, respectively. The results showed that exogenous Y was highly available in water and that its high concentration in water bodies might produce harmful effects on aquatic organisms. N. peltata is proposed as a biomonitor for the assessment of metal pollution in aquatic ecosystems.

Topics: Ascorbic Acid; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Environmental Monitoring; Glutathione; Magnoliopsida; Plant Leaves; Superoxide Dismutase; Water Pollutants, Chemical; Yttrium

Environmental stress due to salt has been described to enhance lettuce processability, shelf life and consumer acceptability. Moderate salinity causes altered leaf carotenoid, lignin, phenolic and flavonol levels without noticeable changes in the green colour and morphology of lettuce. The aim of this study was to understand the improvements in processability, due to salt stress, related to textural properties and structural characteristics.. Physiological, phytochemical and structural changes were observed that were of significant relevance to salt stress (50, 100 and 150 mmol L(-1) NaCl). Fresh weight per plant, fresh weight per leaf, leaf area, water content, colour saturation, chlorophyll a and b and the area of the intercellular spaces decreased when the concentration of salt was increased. Solute concentration, elasticity, total and individual phenolic acids and the areas occupied by the palisade and spongy parenchyma cells increased when the concentration of salt was increased.. These data illustrate that salt stress can have a positive impact on certain structural parameters, especially tissue elasticity, that can be closely linked to a higher number of cells, of lower size and high leaf strength, explaining the postharvest longevity of lettuce. However, leaf growth and visual quality could be negatively affected by salt stress.

Topics: Carotenoids; Cell Wall; Chlorophyll; Chlorophyll A; Flavonols; Food Handling; Lactuca; Lignin; Phenols; Plant Leaves; Salinity; Sodium Chloride; Stress, Physiological; Water

The cultivation of higher plants in Space involves not only the development of new agro-technologies for the design of ecologically closed Space greenhouses, but also understanding of the effects of Space factors on biological systems. Among Space factors, ionising radiation is one of the main constraints to the growth of organisms. In this paper, we analyse the effect of low-LET radiation on leaf histology and cytology in Phaseolus vulgaris L. plants subjected to increasing doses of X-rays (0.3, 10, 50, 100 Gy). Leaves irradiated at tissue maturity were compared with not-irradiated controls. Semi-thin sections of leaves were analysed through light and epi-fluorescence microscopy. Digital image analysis was applied to quantify anatomical parameters, with a specific focus on the occurrence of signs of structural damage as well as alterations at subcellular level, such as the accumulation of phenolic compounds and chloroplast size. Results showed that even at high levels of radiation, general anatomical structure was not severely perturbed. Slight changes in mesophyll density and cell enlargement were detected at the highest level of radiation. However, at 100 Gy, higher levels of phenolic compounds accumulated along chloroplast membranes: this accompanied an increase in number of chloroplasts. The reduced content of chlorophylls at high levels of radiation was associated with reduced size of the chloroplasts. All data are discussed in terms of the possible role of cellular modifications in the maintenance of high radioresistance and photosynthetic efficiency.

Topics: Analysis of Variance; Chlorophyll; Chlorophyll A; Chloroplasts; Mesophyll Cells; Microscopy, Fluorescence; Phaseolus; Plant Epidermis; Plant Leaves; X-Rays

Silicon (Si) is an important structural element that can accumulate at high concentrations in grasses and sedges, and therefore Si structures might affect the optical properties of the leaves. To better understand the role of Si in light/leaf interactions in species rich in Si, we examined the total Si and silica phytoliths, the biochemical and morphological leaf properties, and the reflectance and transmittance spectra in grasses (Phragmites australis, Phalaris arundinacea, Molinia caerulea, Deschampsia cespitosa) and sedge (Carex elata). We show that these grasses contain >1% phytoliths per dry mass, while the sedge contains only 0.4%. The data reveal the variable leaf structures of these species and significant differences in the amount of Si and phytoliths between developing and mature leaves within each species and between grasses and sedge, with little difference seen among the grass species. Redundancy analysis shows the significant roles of the different near-surface silicified leaf structures (e.g., prickle hairs, cuticle, epidermis), phytoliths and Si contents, which explain the majority of the reflectance and transmittance spectra variability. The amount of explained variance differs between mature and developing leaves. The transmittance spectra are also significantly affected by chlorophyll a content and calcium levels in the leaf tissue.

Topics: Calcium; Carex Plant; Carotenoids; Chlorophyll; Chlorophyll A; Optical Phenomena; Plant Leaves; Poaceae; Silicon; Ultraviolet Rays

The extensive use of copper-bearing fungicides in vineyards is responsible for the accumulation of copper (Cu) in soils. Grass species able to accumulate Cu could be cultivated in the vineyard inter-rows for copper phytoextraction. In this study, the capacity of Festuca rubra cv Merlin and Sinapis alba to tolerate and accumulate copper (Cu) was first investigated in a hydroponic system without the interference of soil chemical-physical properties. After the amendment of Cu (5 or 10 mg Cu l-(1)) to nutrient solution, shoot Cu concentration in F. rubra increased up to 108.63 mg Cu kg(-1) DW, more than three times higher than in S. alba (31.56 mg Cu kg(-1) DW). The relationship between Cu concentration in plants and external Cu was dose-dependent and species specific. Results obtained from the hydroponic experiment were confirmed by growing plants in pots containing soil collected from six Italian vineyards. The content of soil organic matter was crucial to enhance Cu tolerance and accumulation in the shoot tissues of both plant species. Although S. alba produced more biomass than F. rubra in most soils, F. rubra accumulated significantly more Cu (up to threefold to fourfold) in the shoots. Given these results, we recommended that F. rubra cv Merlin could be cultivated in the vineyard rows to reduce excess Cu in vineyard soils.

Topics: Agriculture; Biodegradation, Environmental; Chlorophyll; Chlorophyll A; Copper; Festuca; Hydroponics; Plant Leaves; Plant Roots; Plant Shoots; Sinapis; Soil; Soil Pollutants

As the environment is inevitably exposed to ionizing radiation from natural and anthropogenic sources, it is important to evaluate gamma radiation induced stress responses in plants. The objective of this research is therefore to investigate radiation effects in Arabidopsis thaliana on individual and subcellular level by exposing 2-weeks-old seedlings for 7 days to total doses of 3.9 Gy, 6.7 Gy, 14.8 Gy and 58.8 Gy and evaluating growth, photosynthesis, chlorophyll a, chlorophyll b and carotenoid concentrations and antioxidative enzyme capacities. While the capacity of photosystem II (PSII measured as Fv/Fm) remained intact, plants started optimizing their photosynthetic process at the lower radiation doses by increasing the PSII efficiency (φPSII) and the maximal electron transport rate (ETRmax) and by decreasing the non-photochemical quenching (NPQ). At the highest radiation dose, photosynthetic parameters resembled those of control conditions. On subcellular level, roots showed increased superoxide dismutase (SOD) and ascorbate peroxidase (APX) capacities under gamma irradiation but catalase (CAT), syringaldazine peroxidase (SPX) and guaiacol peroxidase (GPX) activities, on the other hand, decreased. In the leaves no alterations were observed in SOD, CAT and SPX capacities, but GPX was highly affected. Based on these results it seems that roots are more sensitive for oxidative stress under gamma radiation exposure than leaves.

Topics: Arabidopsis; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Gamma Rays; Peroxidases; Photosynthesis; Plant Leaves; Plant Roots; Stress, Physiological; Superoxide Dismutase

Chlorophyll content, one of the most important physiological parameters related to plant photosynthesis, is usually used to predict yield potential. To map the quantitative trait loci (QTLs) underlying the chlorophyll content of rice leaves, a double haploid (DH) population was developed from an indica/japonica (Zhenshan 97/Wuyujing 2) crossing and two backcross populations were established subsequently by backcrossing DH lines with each of their parents. The contents of chlorophyll a and chlorophyll b were determined by using a spectrophotometer to directly measure the leaf chlorophyll extracts. To determine the leaf chlorophyll retention along with maturation, all measurements were performed on the day of heading and were repeated 30 days later. A total of 60 QTLs were resolved for all the traits using these three populations. These QTLs were distributed on 10 rice chromosomes, except chromosomes 5 and 10; the closer the traits, the more clustering of the QTLs residing on common rice chromosomal regions. In general, the majority of QTLs that specify chlorophyll a content also play a role in determining chlorophyll b content. Strangely, chlorophyll content in this study was found mostly to be lacking or to have a negative correlation with yield. In both backcross F1 populations, overdominant (or underdominant) loci were more important than complete or partially dominant loci for main-effect QTLs and epistatic QTLs, thereby supporting previous findings that overdominant effects are the primary genetic basis for depression in inbreeding and heterosis in rice.

Topics: Chlorophyll; Chlorophyll A; Chromosome Mapping; Crosses, Genetic; Genes, Plant; Genotype; Haploidy; Inbreeding; Oryza; Plant Leaves; Quantitative Trait Loci

Through its impact on morphogenesis, light is the key environmental factor that alters plant architectural development; however, the understanding that how light controls plant growth and developmental processes is still poor and needs further research. In this study, we monitored the effect of various monochromatic lights and plant growth regulators (PGRs) combinations on morphogenic and biochemical variation in wild grown-leaf derived callus cultures of Artemisia absinthium L. Combination of α-naphthalene acetic acid (NAA 1.0mg/l) and Thidiazuron (TDZ 2.0mg/l) resulted in optimum callogenic frequency (90%) when kept under fluorescent light for 4weeks (16/8h). In contrast to the control (white spectrum), red spectrum enhanced peroxidase activity, protease activity, total protein content and chlorophyll a/b ratio. Green spectrum was found to be more supportive for total phenolics, total flavonoids and antioxidant activity. Yellow light enhanced MDA content while white and green light improved total chlorophyll content and carotenoid content. A positive correlation among callogenic response, antioxidant activities and set of antioxidative enzyme activities was also observed in the current report. This study will help in understanding the influence of light on production of commercially important secondary metabolites and their optimization in the in vitro cultures of A. absinthium L.

Topics: Artemisia absinthium; Carotenoids; Chlorophyll; Chlorophyll A; Flavonoids; Light; Malondialdehyde; Peptide Hydrolases; Peroxidases; Phenols; Plant Leaves; Plant Proteins

Ionising radiation may have different effects on plant metabolism, growth and reproduction, depending on radiation dose, plant species, developmental stage and physiological traits. In this study, exposure of dwarf bean plants to different doses of X-rays (0.3, 10, 50, 100 Gy) was investigated with a multidisciplinary approach consisting of morphological, ecophysiological and biochemical analysis. Both mature and young leaves still growing during the X-rays exposure were compared with non-irradiated control leaves. In particular, leaf expansion, leaf anatomy and functional traits, as well as photosynthetic pigment content and Rubisco expression were analysed. Moreover, the activity of poly(ADP-ribose) polymerase (PARP) was also measured as an indicator of radiation-induced DNA damage. Our data showed that leaf growth is affected by high levels of radiation and demonstrate that mature leaves are more radio-resistant than young leaves, which experience severe dose-dependent changes in leaf functional traits. In particular, young leaves exhibited a reduction of area and an increase in specific mass and dry matter content, as well as a decline in Rubisco activity. Moreover, they showed elevated PARP activity and an increase in phenolic compounds in wall cells if compared with mature leaves. Both of these strategies have been interpreted as a way to help developing leaves withstand irradiation.

Topics: Biomass; Carotenoids; Chlorophyll; Chlorophyll A; Dose-Response Relationship, Radiation; Microscopy, Fluorescence; Phaseolus; Photosynthesis; Plant Leaves; Poly(ADP-ribose) Polymerases; Radiation, Ionizing; Ribulose-Bisphosphate Carboxylase; Water

The emergent hydrophyte Iris pseudacorus was constantly exposed over a 35-day period to atrazine in the laboratory. It could survive at an atrazine level up to 32 mg/L. Its relative growth rates were inhibited significantly when exposure dosage reached at or exceeded 2 mg/L (p < 0.05). No observed effect concentration and lowest observed effect concentration for growth were 1 and 2 mg/L, respectively. Chlorophyll a and b contents of the plant in all treatment groups were affected significantly, and chlorophyll a/b ratios of all atrazine treatment levels were pronouncedly higher than those of the control within 5 days of exposure (p < 0.05), but thereafter recovered to the level of the control. Differences of photosynthetic efficiency were significant between all atrazine treatments and the control; except for 1 mg/L on day 1 and 5, and 2 mg/L on day 1. I. pseudacorus did not show phytotoxicity symptoms after 35 days exposure to atrazine below 2 mg/L level, but photosynthetic efficiency had begun to decline.

Topics: Atrazine; Chlorophyll; Chlorophyll A; Herbicides; Iridaceae; Photosynthesis; Water Pollutants, Chemical

In this study, we have compared photosynthetic performance of barley leaves (Hordeum vulgare L.) grown under sun and shade light regimes during their entire growth period, under field conditions. Analyses were based on measurements of both slow and fast chlorophyll (Chl) a fluorescence kinetics, gas exchange, pigment composition; and of light incident on leaves during their growth. Both the shade and the sun barley leaves had similar Chl a/b and Chl/carotenoid ratios. The fluorescence induction analyses uncovered major functional differences between the sun and the shade leaves: lower connectivity among Photosystem II (PSII), decreased number of electron carriers, and limitations in electron transport between PSII and PSI in the shade leaves; but only low differences in the size of PSII antenna. We discuss the possible protective role of low connectivity between PSII units in shade leaves in keeping the excitation pressure at a lower, physiologically more acceptable level under high light conditions.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Fluorescence; Hordeum; Light; Photosynthesis; Photosystem II Protein Complex; Plant Leaves

Interactive effects of two heavy metal pollutants Cd and Pb in the growth medium were examined on their uptake, production of reactive oxygen species (ROS), induction of oxidative stress and antioxidative defence responses in Indica rice (Oryza sativa L.) seedlings. When rice seedlings in sand culture were exposed to 150 μM Cd (NO3)2 or 600 μM Pb (CH3COO)2 individually or in combination for 8-16 days, a significant reduction in root/shoot length, fresh weight, relative water content, photosynthetic pigments and increased production of ROS (O2˙- and H2O2) was observed. Both Cd and Pb were readily taken up by rice roots and localisation of absorbed metals was greater in roots than in shoots. When present together in the growth medium, uptake of both the metals Cd and Pb declined by 25-40%. Scanning electron microscope (SEM) imaging of leaf stomata revealed that Pb caused more distortion in the shape of guard cells than Cd. Dithizone staining of roots showed localisation of absorbed Cd on root hairs and epidermal cells. Both Cd and Pb caused increased lipid peroxidation, protein carbonylation, decline in protein thiol and increase in non-protein thiol. The level of reduced forms of non-enzymic antioxidants glutathione (GSH) and ascorbate (AsA) and their redox ratios (GSH/AsA) declined, whereas the activities of antioxidative enzymes superoxide dismutase (SOD) and guaiacol peroxidase (GPX) increased in metal treated seedlings compared to controls. In-gel activity staining also revealed increased intensities of SOD and GPX isoforms with metal treatments. Catalase (CAT) activity increased during early days (8 days) of metal exposure and declined by 16 days. Results suggest that oxidative stress is an important component in expression of Cd and Pb toxicities in rice, though uptake of both metals gets reduced considerably when present together in the medium.

Topics: Antioxidants; Ascorbic Acid; Biological Transport; Cadmium; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Glutathione; Hydrogen Peroxide; Lead; Lipid Peroxidation; Oryza; Oxidative Stress; Peroxidase; Plant Roots; Protein Carbonylation; Soil Pollutants; Sulfhydryl Compounds; Superoxide Dismutase; Superoxides

Under strong light conditions, long-lived chlorophyll triplets ((3)Chls) are formed, which can sensitize singlet oxygen, a species harmful to the photosynthetic apparatus of plants. Plants have developed multiple photoprotective mechanisms to quench (3)Chl and scavenge singlet oxygen in order to sustain the photosynthetic activities. The lumenal loop of light-harvesting chlorophyll a/b complex of photosystem II (LHCII) plays important roles in regulating the pigment conformation and energy dissipation. In this study, site-directed mutagenesis analysis was applied to investigate triplet-triplet energy transfer and quenching of (3)Chl in LHCII. We mutated the amino acid at site 123 located in this region to Gly, Pro, Gln, Thr and Tyr, respectively, and recorded fluorescence excitation spectra, triplet-minus-singlet (TmS) spectra and kinetics of carotenoid triplet decay for wild type and all the mutants. A red-shift was evident in the TmS spectra of the mutants S123T and S123P, and all of the mutants except S123Y showed a decrease in the triplet energy transfer efficiency. We propose, on the basis of the available structural information, that these phenomena are related to the involvement, due to conformational changes in the lumenal region, of a long-wavelength lutein (Lut2) involved in quenching (3)Chl.

Topics: Chlorophyll; Chlorophyll A; Circular Dichroism; Energy Transfer; Fluorescence; Light-Harvesting Protein Complexes; Lutein; Models, Molecular; Mutagenesis, Site-Directed; Mutation; Photobleaching; Photosynthesis; Photosystem II Protein Complex; Pisum sativum; Protein Binding

Dioon edule seedling mortality is mostly attributed to dehydration by prolonged drought, even when they present xeromorphic characteristics like the adult plants. The effect of germination date (GD) and soil water deficit on seedling tolerance to water stress was assessed. The seedlings germinated and grown from mature seeds every month from December to April GD were selected to evaluate the leaf area, photosynthetic pigment content, crassulacean acid metabolism (CAM) activity, stomatal conductance (gs) and leaflet anatomy at soil water potential (Ψs) of 0.0 MPa (day 1), -0.1 MPa (day 40), -1.0 MPa (day 90), -1.5 MPa (day 130), and a control (0.0 MPa at day 130) to recognize differences due to leaf development. The seedlings shifted from C3 to CAM cycling when exposed to water stress at Ψs of -1.0 MPa, like adult plants. The March-April GD seedlings with undeveloped sclerified hypodermis and stomata, presented reduced leaf area, lower Chlorophyll a/b ratio, higher CAM activity and midday partial stomatal closure when reached Ψs of -1.0 MPa. These have higher probability of dehydration during severe drought (February-April) than those of the December-February GD with similar Ψs. Plants used for restoration purposes must have full leaf development to increase the survival.

Topics: Adaptation, Physiological; Carotenoids; Chlorophyll; Chlorophyll A; Dehydration; Germination; Photosynthesis; Plant Stomata; Seedlings; Soil; Time Factors; Water; Zamiaceae

State transitions represent a photoacclimation process that regulates the light-driven photosynthetic reactions in response to changes in light quality/quantity. It balances the excitation between photosystem I (PSI) and II (PSII) by shuttling LHCII, the main light-harvesting complex of green algae and plants, between them. This process is particularly important in Chlamydomonas reinhardtii in which it is suggested to induce a large reorganization in the thylakoid membrane. Phosphorylation has been shown to be necessary for state transitions and the LHCII kinase has been identified. However, the consequences of state transitions on the structural organization and the functionality of the photosystems have not yet been elucidated. This situation is mainly because the purification of the supercomplexes has proved to be particularly difficult, thus preventing structural and functional studies. Here, we have purified and analysed PSI and PSII supercomplexes of C. reinhardtii in states 1 and 2, and have studied them using biochemical, spectroscopic and structural methods. It is shown that PSI in state 2 is able to bind two LHCII trimers that contain all four LHCII types, and one monomer, most likely CP29, in addition to its nine Lhcas. This structure is the largest PSI complex ever observed, having an antenna size of 340 Chls/P700. Moreover, all PSI-bound Lhcs are efficient in transferring energy to PSI. A projection map at 20 Å resolution reveals the structural organization of the complex. Surprisingly, only LHCII type I, II and IV are phosphorylated when associated with PSI, while LHCII type III and CP29 are not, but CP29 is phosphorylated when associated with PSII in state2.

Topics: Acclimatization; Arabidopsis; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Phosphorylation; Photosystem I Protein Complex; Photosystem II Protein Complex; Protein Stability; Thylakoids

Unicellular cyanobacterium Gloeobacter violaceus is an only known oxygenic photosynthetic organism that lacks thylakoid membrane. Molecular phylogenetic analyses indicate that G. violaceus is an early-branching cyanobacterium within cyanobacterial clade. Therefore, the photosynthetic system of G. violaceus is considered to be partly similar to that of the ancestral cyanobacteria that would lack thylakoid membrane. G. violaceus possesses chlorophyll (Chl) a as the only chlorophyll species like most cyanobacteria. It was proposed that the ancestral oxygenic photosynthetic organism had not only Chl a and phycobilins but also Chl b. However, no organism which contains both Chl a and Chl b and lacks thylakoid membrane has been found in nature. Therefore, we introduced the chlorophyllide a oxygenase gene responsible for Chl b biosynthesis into G. violaceus. In the resultant transformant, Chl b accumulated at approximately 11% of total Chl independent of growth phase. Photosystem I complexes isolated from the transformant contained Chl b at 9.9% of total Chl. The presence of Chl b in the photosystem I complexes did not inhibit trimer formation. Furthermore, time-resolved fluorescence spectrum demonstrated that Chl b transferred energy to Chl a in the photosystem I complexes and did not disturb the energy transfer among the Chl a molecules. These results show that G. violaceus is tolerant to artificially produced Chl b and suggest the flexibility of photosystem for Chl composition in the ancestral oxygenic photosynthetic organism.

Topics: Bacterial Proteins; Chlorophyll; Chlorophyll A; Cyanobacteria; Oxygenases; Photosystem I Protein Complex; Prochlorophytes; Time Factors

The transcript expression of a gene SrUGT85C2 has been documented for direct relation with steviol glycoside content in Stevia plant. Steviol glycoside and gibberellin biosynthetic routes are divergent branches of methyl erythritol-4 phosphate (MEP) pathway. So, SrUGT85C2 might be an influencing gibberellin content. Hence in the present study, transgenic Arabidopsis thaliana overexpressing SrUGT85C2 cDNA from Stevia rebaudiana was developed to check its effect on gibberellin accumulation and related plant growth parameters. The developed transgenics showed a noteworthy decrease of 78-83% in GA3 content. Moreover, the transgenics showed a gibberellin deficient phenotype comprising stunted hypocotyl length, reduced shoot growth and a significant fall in relative water content. Transgenics also showed 17-37 and 64-76% reduction in chlorophyll a and chlorophyll b contents, respectively. Reduction in photosynthetic pigments could be responsible for the noticed significant decrease in plant biomass. Like steviol glycoside and gibberellin biosynthesis, chlorophyll biosynthesis also occurs from the precursors isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) of MEP pathway in the plastids. The observed downregulated expression of genes encoding MEP pathway enzymes geranyl geranyl diphosphate synthase (GGDPS), copalyl diphosphate synthase (CDPS), kaurenoic acid oxidase (KAO), chlorophyll synthetase and chlorophyll a oxygenase in transgenics overexpressing SrUGT85C2 might be responsible for the reduction in gibberellins as well as chlorophyll. This study has documented for the first time the regulatory role of SrUGT85C2 in the biosynthesis of steviol glycoside, gibberellins and chlorophyll.

Topics: Arabidopsis; Chlorophyll; Chlorophyll A; Diterpenes, Kaurane; Erythritol; Gene Expression Regulation, Plant; Gibberellins; Glycosides; Plant Proteins; Plants, Genetically Modified; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Plant; Stevia; Sugar Phosphates; Transgenes

Five Malaysian rice (Oryza sativa L.) varieties, MR33, MR52, MR211, MR219, and MR232, were tested in pot culture under different salinity regimes for biochemical response, physiological activity, and grain yield. Three different levels of salt stresses, namely, 4, 8, and 12 dS m(-1), were used in a randomized complete block design with four replications under glass house conditions. The results revealed that the chlorophyll content, proline, sugar content, soluble protein, free amino acid, and yield per plant of all the genotypes were influenced by different salinity levels. The chlorophyll content was observed to decrease with salinity level but the proline increased with salinity levels in all varieties. Reducing sugar and total sugar increased up to 8 dS m(-1) and decreased up to 12 dS m(-1). Nonreducing sugar decreased with increasing the salinity levels in all varieties. Soluble protein and free amino acid also decreased with increasing salinity levels. Cortical cells of MR211 and MR232 did not show cell collapse up to 8 dS m(-1) salinity levels compared to susceptible checks (IR20 and BRRI dhan29). Therefore, considering all parameters, MR211 and MR232 showed better salinity tolerance among the tested varieties. Both cluster and principal component analyses depict the similar results.

Topics: Amino Acids; Carbohydrates; Chlorophyll; Chlorophyll A; Cluster Analysis; Oryza; Phylogeny; Plant Leaves; Plant Proteins; Plant Roots; Principal Component Analysis; Proline; Salinity; Seeds; Solubility

Salvia miltiorrhiza, which is commonly known as Danshen, is a traditional Chinese herbal medicine. To illustrate its physiological and biochemical responses to salt stress and to evaluate the feasibility of cultivating this plant in saline coastal soils, a factorial experiment under hydroponic conditions was arranged on the basis of a completely randomised design with three replications. Five salinity treatments (0, 25, 50, 75 and 100 mM NaCl) were employed in this experiment. The results showed that salinity treatments of <100 mM NaCl did not affect the growth of Salvia miltiorrhiza in a morphological sense, but significantly inhibit the accumulation of dry matter. Salinity treatments significantly decreased the Chl-b content but caused a negligible change in the Chl-a content, leading to a conspicuous overall decrease in the T-Chl content. The Na(+) content significantly increased with increasing hydroponic salinity but the K(+) and Ca(2+) contents were reversed, indicating that a high level of external Na(+) resulted in a decrease in both K(+) and Ca(2+) concentrations in the organs of Salvia miltiorrhiza. Salt stress significantly decreased the superoxide dismutase (SOD) activity of Salvia miltiorrhiza leaves in comparison with that of the control. On the contrary, the catalase (CAT) activity in the leaves markedly increased with the increasing salinity of the hydroponic solution. Moreover, the soluble sugar and protein contents in Salvia miltiorrhiza leaves dramatically increased with the increasing salinity of the hydroponic solution. These results suggested that antioxidant enzymes and osmolytes are partially involved in the adaptive response to salt stress in Salvia miltiorrhiza, thereby maintaining better plant growth under saline conditions.

Topics: Antioxidants; Calcium; Catalase; Chlorophyll; Chlorophyll A; Homeostasis; Plant Leaves; Plants, Medicinal; Potassium; Salinity; Salt Tolerance; Salt-Tolerant Plants; Salvia miltiorrhiza; Seedlings; Sodium Chloride; Stress, Physiological; Superoxide Dismutase

Albino tea has received increased attention due to its brisk flavour. To identify changes in the key chemical constituents conveying important qualities to albino tea, the metabolite profiles of four albino cultivars and one green tea cultivar were analysed. Compared to the green tea control, significantly decreased contents of chlorophyll (Chl) (p<0.01), total carotenoids (p<0.05), caffeine (p<0.01), and total catechins (p<0.05) were found in albino tea leaves with a few exceptions, whereas increases were noted in the Chl a/b ratio and the contents of both zeaxanthin and free amino acids, including theanine. Multivariate analysis identified catechins and carotenoids as the most important contributors to the metabolic profile variance between the albino and green tea cultivars. High levels of amino acids, along with low levels of chlorophylls, catechins and caffeine, contribute to the qualities of albino tea, which include reduced astringency and bitterness, along with a strong umami taste.

Topics: Amino Acids; Caffeine; Camellia sinensis; Catechin; Chlorophyll; Chlorophyll A; Discriminant Analysis; Humans; Plant Extracts; Plant Leaves; Quality Control; Taste; Tea; Xanthophylls; Zeaxanthins

The present study was designed to validate the applicability of photosynthetic performance using a PAM fluorometer and photosynthetic pigments in Euglena gracilis as endpoint parameters in toxicity assessment of liquid detergents using a dish washing liquid detergent during short- (0-72h) and long-term (7days) exposure. In short-term experiments, the detergent affected the photosynthetic efficiency with EC50 values (calculated for Fv/Fm) of 22.07%, 7.27%, 1.4% and 2.34%, after 0, 1, 24 and 72h, respectively. The relative electron transport rate (rETR) and quantum yield measured with increasing irradiances were also inhibited by the detergent. The most severe effect of the detergent on the light-harvesting pigments (μgmL(-1)) was observed after 72h where chlorophyll a and total carotenoids were decreased at concentrations above 0.1% and chlorophyll b was decreased at concentrations above 0.5%. In long-term experiments, the detergent reduced the photosynthetic efficiency of cultures giving an EC50 value of 0.867% for Fv/Fm. rETR and quantum yield with increasing irradiance were shown to be adversely affected at concentrations of 0.1% or above. A decrease in chlorophyll a and total carotenoids (μgmL(-1)) was observed at concentrations of 0.05% detergent or above. Chlorophyll b was shown to be comparatively less affected by detergent stress, and a significant decrease was observed at concentrations of 0.5% or above. However, there was no prominent decrease in per cell (Euglena) concentration of any pigment. It can be concluded that photosynthesis and light-harvesting pigments in E. gracilis were sensitive to detergent stress and can be used as sensitive parameters in toxicity assessment of detergents in aquatic environments.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Detergents; Electron Transport; Euglena gracilis; Photosynthesis; Photosystem II Protein Complex; Quantum Theory; Time Factors; Water Pollutants, Chemical

The present study was performed to screen out the extracts of algae and assess the seasonal variation in antimicrobial activity of Ulva pertusa against Gardnerella vaginalis. Seasonal variation in antibacterial activity was observed, with the extracts showing no activity during summer and autumn, and showing antibacterial activity from early winter (December) to middle spring (April). The maximum value of antimicrobial activity (6.5 mm inhibition zone at 5 mg disk(-1)) of U. pertusa against G. vaginalis was observed in April. Otherwise, for both chlorophyll a and b, the highest content (2.87 mg g(-1) and 1.37 mg g(-1)) was observed in March 2009. These results may reflect variation in cellular chemical compositions such as secondary metabolite(s) rather than chlorophyll and biological activities according to season.

Topics: Anti-Bacterial Agents; Chlorophyll; Chlorophyll A; Gardnerella vaginalis; Plant Extracts; Seasons; Ulva

The present study is aimed at assessing the extent of arsenic (As) toxicity under three different light intensities-optimum (400 μmole photon m(-2) s(-1)), sub-optimum (225 μmole photon m(-2) s(-1)), and low (75 μmole photon m(-2) s(-1))-exposed to Helianthus annuus L. var. DRSF-113 seedlings by examining various physiological and biochemical parameters. Irrespective of the light intensities under which H. annuus L. seedlings were grown, there was an As dose (low, i.e., 6 mg kg(-1) soil, As1; and high, i.e., 12 mg kg(-1) soil, As2)-dependent decrease in all the growth parameters, viz., fresh mass, shoot length, and root length. Optimum light-grown seedlings exhibited better growth performance than the sub-optimum and low light-grown seedlings; however, low light-grown plants had maximum root and shoot lengths. Accumulation of As in the plant tissues depended upon its concentration used, proximity of the plant tissue, and intensity of the light. Greater intensity of light allowed greater assimilation of photosynthates accompanied by more uptake of nutrients along with As from the medium. The levels of chlorophyll a, b, and carotenoids declined with increasing concentrations of As. Seedlings acquired maximum Chl a and b under optimum light which were more compatible to face As1 and As2 doses of As, also evident from the overall status of enzymatic (SOD, POD, CAT, and GST) and non-enzymatic antioxidant (Pro).

Topics: Arsenic; Biomass; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Dose-Response Relationship, Drug; Dose-Response Relationship, Radiation; Glutathione Transferase; Helianthus; Hydrogen Peroxide; Light; Malondialdehyde; Oxygen Consumption; Peroxidase; Photosynthesis; Plant Roots; Plant Stems; Proline; Seedlings; Superoxide Dismutase; Superoxides

Szechuan peppers are extensively used as a spice and in traditional medicine in Asia, primarily because of its active compounds, sanshools (S). However, there is only limited mention in agriculture, and there are no papers reporting its use as an herbicide safener. In this study, we provide the first evidence that S can effectively alleviate rice-seedling injury from metolachlor (M). We observed that the M-treated (0.25 μM) rice seedlings, which were 56.0%, 66.0%, and 57.0% of the non-treated control in shoot height, root length, and fresh biomass, respectively, were recovered by S to 93.1%, 97.6%, and 94.8%, respectively. The emergence rate was enhanced to over 80% in the M+S treatment, whereas it was below 60% in the M treatment. This M+S mixture elevated the rice-seedling root activity to higher than 87.0% of the value for the non-treated control. The activity of glutathione transferases in the combined treatments approximately doubles that of the M treatment and quadruples that of the non-treated controls. This effect was positively correlated with the induced expression of OsGSTU3. Our results suggest that S may represent a new group of safeners and enable the possibility of using these compounds for improving plant production or protecting rice from the phytotoxicity of metolachlor.

Topics: Acetamides; Amides; Chlorophyll; Chlorophyll A; Gene Expression Regulation, Plant; Glutathione Transferase; Herbicides; Oryza; Plant Roots; Plant Shoots; Protective Agents; Seedlings

The work explored sucrose metabolism in the leaves subtending the cotton boll (SBL) and its role in boll weight after waterlogging in cotton. Results showed that net photosynthesis rate (Pn), relative water content, contents of Chlorophyll a and Chlorophyll b, initial ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) activity and cytosolic fructose-1, 6-bisphosphatase (cy-FBPase) activity decreased with waterlogging in the SBL on fruiting branches 2-3 (FB2-3) and FB6-7. Activities of sucrose synthase (SuSy) and sucrose phosphate synthase (SPS) increased to the maximum up to 6 days of waterlogging then decreased with prolonged waterlogging. Rubisco activation and specific leaf weight increased and gene expressions of SuSy, SPS and rubisco activase (RCA) were all up-regulated with the duration of waterlogging, especially for the SBL on FB6-7. The induction of activity and gene expression of SuSy was most significant indicating its crucial role in sucrose metabolism after waterlogging. For the SBL in the later period of boll development on upper FB10-11 and FB14-15, the pattern seemed opposite to that of FB2-3 and FB6c7 as compensation effect in vegetative growth existed. Correlation analysis revealed that initial Rubisco activity and cy-FBPase activity were the main limitation to Pn reduction after waterlogging. Reduction in Pn, sucrose transformation rate and initial Rubisco activity directly decrease boll weight in waterlogged cotton. Besides the role in sucrose metabolism after waterlogging, SuSy also had a positive significant correlation with the duration of rapid-accumulation period for seed fiber weight (P<0.05). These findings elucidated mechanisms to waterlogging that affected seed fiber weight, which resulted from alteration in carbohydrates, enzymes and genes.

Topics: Carbohydrate Metabolism; Chlorophyll; Chlorophyll A; Cotton Fiber; Flowers; Fruit; Gene Expression Regulation, Plant; Gossypium; Organ Size; Photosynthesis; Plant Leaves; Plant Stems; Starch; Sucrose; Water; Weather

The present study explores the tolerance and metal removal response of a well-developed 2-week-old Phormidium mat after long-term exposure to Cu(2+)-enriched medium. Cu(2+) enrichment inhibited increase in mat biomass in a concentration-dependent manner. Mat area and the number of entrapped air bubbles decreased as Cu(2+) concentration increased in the medium. Decrease in number of air bubbles obviously reflects the adverse effect of Cu(2+) on photosynthetic performance of the mat. Metal enrichment did not substantially alter the amount of pigments, such as chlorophyll a, chlorophyll b, carotenoids, and phycocyanin, in the mat. Enhancement of Cu(2+) concentration in the medium led to changes in species composition of the test mat; however, Phormidium bigranulatum always remained the dominant organism. Relative share of green algae and some cyanobacterial taxa, namely, Lyngbya sp. and Oscillatoria tenuis, in the mat were increased by Cu(2+) enrichment. The mat successfully removed 80 to 94 % Cu(2+) from the growth medium containing 10 to 100 μM Cu(2+). Extracellular polysaccharides, whose share increased in the mat community after metal addition, seem to have contributed substantially to metal binding by the mat biomass.

Topics: Biodegradation, Environmental; Biomass; Chlorophyll; Chlorophyll A; Chlorophyta; Copper; Cyanobacteria; Metals; Photosynthesis; Toxicity Tests, Chronic

This study explored the effects of different light quality on photosynthetic pigment contents and photosynthetic characteristics of peanut (Qinhua 6) seedling leaves. The results showed that, compared with natural light, blue light (445-470 nm) could significantly improve the specific leaf area (SLA), chlorophyll a/b value and carotenoid content of peanut seedlings. Meanwhile, the net photosynthetic rate, stomatal conductance, and transpiration rate were higher, the intercellular CO2 content was lower, and the photosynthetic efficiency was improved significantly under blue light. Red light (610-660 nm) could improve the chlorophyll content significantly, and reduce SLA, chlorophyll a/b value and carotenoid content, with a lower photosynthetic efficiency than natural light. Green light (515-520 nm) and yellow light (590-595 nm) were not conducive to photosynthetic pigment accumulation of leaves, and significantly inhibited leaf photosynthesis of peanut seedlings.

Topics: Arachis; Chlorophyll; Chlorophyll A; Light; Photosynthesis; Plant Leaves; Plant Stomata; Plant Transpiration; Seedlings

Plants, in their life cycle, are usually exposed to various kinds of non-biological stresses including heavy metals. One of these heavy metals is nickel which affects many physiological processes of plants. Studies have shown that the changes in planting conditions can affect the qualitative and quantitative features of Catharanthus roseus; therefore, creating stressful conditions (e.g. NiCl2) can be an effective way to investigate the changes. In this research, we investigated the effect of 0, 2.5, 5, 10, 25 and 50 mM concentrations of NiCl2 on the degree of catalase enzyme activity, amount of proline aggregation and photosynthetic parameters on seeds of pink variety of C. roseus. The results indicated that the degree of catalase enzyme activity and the amount of proline aggregation increased in plants which were exposed to NiCl2 treatments, especially in high concentrations, while the total protein decreased. The stress of Ni also affected photosynthetic parameters, and decreased the amount of pigments, as well as the efficiency of photosystem II.

Topics: Catalase; Catharanthus; Chlorophyll; Chlorophyll A; Metals, Heavy; Nickel; Photosystem II Protein Complex; Plant Proteins; Proline; Stress, Physiological

The effect of pulsed electric field (PEF) treatments of different intensities on the electroporation of the cytoplasmatic membrane of Chlorella vulgaris, and on the extraction of carotenoids and chlorophylls were investigated. Staining the cells with propidium iodide before and after the PEF treatment revealed the existence of reversible and irreversible electroporation. Application of PEF treatments in the range of 20-25 kV cm(-1) caused most of the population of C. vulgaris to be irreversibly electroporated even at short treatment times (5 pulses of 3 µs). However, at lower electric field strengths (10 kV cm(-1)), cells that were reversibly electroporated were observed even after 50 pulses of 3 µs. The electroporation of C. vulgaris cells by PEF higher than 15 kV cm(-1) and duration is higher than 15 µs increased significantly the extraction yield of intracellular components of C. vulgaris. The application of a 20 kV cm(-1) for 75 μs increased the extraction yield just after the PEF treatment of the carotenoids, and chlorophylls a and b 0.5, 0.7, and 0.8 times, respectively. However, further increments in electric field strength and treatment time did not cause significant increments in the extraction yield. The extraction of carotenoids from PEF-treated C. vulgaris cells after 1 h of the application of the treatment significantly increased the extraction yield in comparison to the yield obtained from the cells extracted just after the PEF treatment. After PEF treatment at 20 kV cm(-1) for 75 µs, extraction yield for carotenoids, and chlorophylls a and b increased 1.2, 1.6, and 2.1 times, respectively. A high correlation was observed between irreversible electroporation and percentage of yield increase when the extraction was conducted after 1 h of the application of PEF treatment (R: 0.93), but not when the extraction was conducted just after PEF treatment (R: 0.67).

Topics: Carotenoids; Cell Membrane; Cell Membrane Permeability; Chlorella vulgaris; Chlorophyll; Chlorophyll A; Electricity; Electroporation; Propidium

Photochemical responses in the thylakoid membrane of Nymphoides peltatum to increasing lead (Pb) concentrations were investigated after 5 days of exposure. Pb accumulation increased in a concentration-dependent manner, with a maximum of 118.44 μg g(-1) at 100 μM. Nutrients (Zn, Mg, Mo, Ca, Fe and Mn), ATPase activity and pigment generally increased progressively at Pb concentrations of 12.5 and 25 μM, but then declined at concentrations of 50 and 100 μM. Moreover, Pb stress induced an increase in chlorophyll (Chl) a/b ratio in a different extent. No outstanding changes were observed in several Chl a fluorescence parameters at low Pb concentrations (12.5 and 25 μM), while significant changes (p < 0.05) were observed in these parameters at higher concentrations. The alterations of nutrients, ATPase activity and pigment content were associated with disturbances in the thylakoid membrane, indicated by the quenching of Chl a fluorescence. These results were indicative of a disarray in photochemical activities exerted by Pb phytotoxicity.

Topics: Chlorophyll; Chlorophyll A; Environmental Pollutants; Lead; Magnoliopsida; Thylakoids

As an excellent biological resource, Chlorella has wide applications for production of biofuel, bioactive substances and water environment restoration. Therefore, it is very important to understand the photosynthetic physiology characteristics of Chlorella. Magnesium ions play an important role in the growth of microalgae, not only the central atom of chlorophyll, but also the cofactor of some key enzyme in the metabolic pathway. A laboratory study was conducted to evaluate the effects of magnesium deficiency on several photosynthetic and physiological parameters and the triacylglyceride (TAG) accumulation of the green alga, Chlorella vulgaris, in the photoautotrophic culture process. Chlorella vulgaris biomass, protein, chlorophyll a and chlorophyll b contents decreased by 20%, 43.96%, 27.52% and 28.07% in response to magnesium deficiency, while the total oil content increased by 19.60%. Moreover, magnesium deficiency decreased the maximal photochemical efficiency F(v)/F(m) by 22.54%, but increased the non-photochemical quenching parameters qN. Our results indicated the decline of chlorophyll caused by magnesium, which affected the photosynthesis efficiency, lead to the growth inhibition of Chlorella vulgaris and affected the protein synthesis and increased the triacylglyceride (TAG) accumulation.

Topics: Biomass; Chlorella vulgaris; Chlorophyll; Chlorophyll A; Magnesium; Photosynthesis; Triglycerides

Chlorophyll plays a pivotal role in the plant physiology and its productivity. Cultivation of plants in crude oil contaminated soil has a great impact on the synthesis of chlorophyll pigment. Morpho-anatomy of the experimental plant also shows structural deformation in higher concentrations. Keeping this in mind, a laboratory investigation has been carried out to study the effect of crude oil on chlorophyll content and morpho-anatomy of Cyperus brevifolius plant. Fifteen-day-old seedling of the plant was planted in different concentrations of the crude oil mixed soil (i.e., 10,000, 20,000, 30,000, 40,000, and 50,000 ppm). A control setup was also maintained without adding crude oil. Results were recorded after 6 months of plantation. Investigation revealed that there is a great impact of crude oil contamination on chlorophyll content of the leaves of the experimental plant. It also showed that chlorophyll a, chlorophyll b, and total chlorophyll content of leaves grown in different concentrations of crude oil were found to be lower than those of the control plant. Further, results also demonstrated that chlorophyll content was lowest in the treatment that received maximum dose of crude oil. It also showed that chlorophyll content was decreased with increased concentration of crude oil. Results also demonstrated that there was a reduction in plant shoot and root biomass with the increase of crude oil concentration. Results also revealed that the shoot biomass is higher than root biomass. Morphology and anatomy of the experimental plant also show structural deformation in higher concentrations. Accumulation of crude oil on the cuticle of the transverse section of the leaves and shoot forms a thick dark layer. Estimation of the level of pollution in an environment due to oil spill is possible by the in-depth study of the harmful effects of oil on the morphology and anatomy and chlorophyll content of the plants grown in that particular environment.

Topics: Chlorophyll; Chlorophyll A; Cyperus; Petroleum; Petroleum Pollution; Plant Leaves; Plant Roots; Plant Shoots; Seedlings; Soil; Soil Pollutants; Stress, Physiological

The food industry and plant breeding programmes require fast, clean and low-cost screening techniques for nutritional compounds determination in food matrices. This is the first report on the study of the potential of near-infrared spectroscopy (NIRS) for the prediction of antioxidant compounds in summer squash tissues collected since 2009-2012. Modified partial least-squares (MPLS) regression was used to correlate spectral information and the different antioxidant compounds in the samples. The coefficients of determination in the external validation (r(2)ev) obtained were for ascorbic acid (0.77 and 0.86), chlorophyll a (0.79 and 0.66), chlorophyll b (0.86 and 0.79) and total phenolic compounds (0.65 and 0.68) in exocarp and mesocarp tissues, respectively, supporting that NIRS is able to predict in a rapid way these components for screening purposes. Major wavelengths influencing the calibration equations showed that chromophores as well as fibre components of the fruits highly participated in developing the NIR equations.

Topics: Antioxidants; Ascorbic Acid; Calibration; Chlorophyll; Chlorophyll A; Cucurbita; Food Analysis; Fruit; Least-Squares Analysis; Phenols; Predictive Value of Tests; Reproducibility of Results; Seasons; Spectroscopy, Near-Infrared; Vegetables

Eichhornia crassipes (Mart.) Solms. plantlets were grown in 0.1 and 1.0 ppm treatment solutions of Hoagland's hydroponic solutions modified with Hg(NO₃)2 in order to examine the specific cellular and biochemical mechanisms involved in the tolerance of this plant exposed to mercury. This study assessed the responses of chloroplast pigments, i.e., carotenoids and chlorophylls, and evaluated the enzymatic and nonenzymatic antioxidant systems. Inductively coupled plasma-atomic emission spectrometry (ICP-AES) revealed varying Hg(2+) levels in the young and mature leaf tissues, with greater amounts of Hg(2+) found in the tissues of the young leaves. Total chlorophyll levels, notably those of chlorophyll a, chlorophyll b, and carotenoids, showed significant elevation in young leaf tissues, while a decrease in their levels was observed in mature leaf tissues in comparison to those of the control plants. These results lend support to the protective role of increased chlorophyll and carotenoid levels in the photosynthetic apparatus of young E. crassipes leaves in the presence of Hg(2+). The antioxidant responses of Hg-treated E. crassipes plants were also measured, revealing a highly significant increase in catalase units, catalase and ascorbate peroxidase activities, and mercury-binding thiols in leaves from Hg-treated plants. Moreover, substantial differences in the degree of oxidative injury between the cells in leaves from the control and Hg-treated plants were evidenced by the lipid peroxidation activities monitored. The Hg-treatment-induced significant decrease in malondialdehyde (MDA) levels was observed in 0.1-ppm Hg(NO₃)2-exposed plants, while a highly significant increase in MDA levels was noted in 1.0-ppm Hg(NO₃)2-exposed plants. The high degree of lipid peroxidation at 1.0-ppm Hg treatment was evidently counteracted by the compensatory protective mechanism brought about by the increased levels in chloroplast pigments and the enhanced activities of the antioxidant systems. E. crassipes responded to mercury treatments by enhancing the synthesis of chlorophyll and carotenoid pigments, enzymatic, and nonenzymatic antioxidant substances, concomitantly increasing the antioxidative activities, thus rendering E. crassipes capable of tolerating Hg-induced stress. The potential of E. crassipes as a phytoremediator is evident.

Topics: Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Eichhornia; Environmental Pollutants; Lipid Peroxidation; Malondialdehyde; Mercury; Plant Leaves

Fipronil is an effective insecticide, but it presents highly toxic effects in nontarget aquatic organisms. The present study examined the enantioselective toxicity and degradation of fipronil enantiomers in a freshwater algae Scenedesmus obliguus suspension. There was a substantial difference in the acute toxicity of the enantiomers to S. obliguus, with 72-h median effective concentrations (EC50s) of 0.29 mg L(-1) and 1.50 mg L(-1) for the R-fipronil and S-fipronil, respectively. The influences on the concentration of chlorophyll a, chlorophyll b, and carotenoids were determined, and the effects of fipronil on the concentration of chlorophyll a and chlorophyll b were also enantioselective. The degradation of fipronil in algae suspension was enantioselective, with half-lives for R-fipronil and S-fipronil of 2.9 d and 3.2 d, respectively, and the enantiomer fraction reaching 0.65 at the day 17. The enantiomeric differences should be taken into consideration for fipronil risk assessment.

Topics: Biological Assay; Carotenoids; Chlorophyll; Chlorophyll A; Insecticides; Photosynthesis; Pyrazoles; Reproducibility of Results; Risk Assessment; Scenedesmus; Stereoisomerism; Suspensions; Time Factors

Cytokinins (CKs) are one of the main regulators of in vitro growth and development and might affect the developmental state and function of the photosynthetic apparatus of in vitro shoots. Effects of different cytokinin regimes including different types of aromatic cytokinins, such as benzyl-adenine, benzyl-adenine riboside and 3-hydroxy-benzyladenine alone or in combination were studied on the capacity of the photosynthetic apparatus and the pigment content of in vitro apple leaves after 3 weeks of culture. We found that the type of cytokinins affected both chlorophyll a and b contents and its ratio. Chlorophyll content of in vitro apple leaves was the highest when benzyl-adenine was applied as a single source of cytokinin in the medium (1846-2176 μg/1g fresh weight (FW) of the leaf). Increasing the concentration of benzyl-adenine riboside significantly decreased the chlorophyll content of the leaves (from 1923 to 1183 μg/1g FW). The highest chl a/chl b ratio was detected after application of meta-topolin (TOP) at concentrations of 2.0 and 6.0 μM (2.706 and 2.804). Chlorophyll fluorescence was measured both in dark-adapted (Fv/Fm test) and in light-adapted leaf samples (Yield test; Y(II)). The maximum quantum yield and efficiency of leaves depended on the cytokinin source of the medium varied between 0.683 and 0.861 (Fv/Fm) indicating a well-developed and functional photosynthetic apparatus. Our results indicate that the type and concentration of aromatic cytokinins applied in the medium affect the chlorophyll content of the leaves in in vitro apple shoots. Performance of the photosynthetic apparatus measured by chlorophyll fluorescence in the leaves was also modified by the cytokinin supply. This is the first ever study on the relationship between the cytokinin supply and the functionability of photosystem II in plant tissue culture and our findings might help to increase plantlet survival after transfer to ex vitro conditions.

Topics: Chlorophyll; Chlorophyll A; Cytokinins; Fluorometry; Malus; Photosynthesis; Photosystem II Protein Complex; Plant Growth Regulators; Plant Leaves; Spectrophotometry

The chemical structural differences distinguishing chlorophylls in oxygenic photosynthetic organisms are either formyl substitution (chlorophyll b, d, and f) or the degree of unsaturation (8-vinyl chlorophyll a and b) of a side chain of the macrocycle compared with chlorophyll a. We conducted an investigation of the conversion of vinyl to formyl groups among naturally occurring chlorophylls. We demonstrated the in vitro oxidative cleavage of vinyl side groups to yield formyl groups through the aid of a thiol-containing compound in aqueous reaction mixture at room temperature. Heme is required as a catalyst in aqueous solution but is not required in methanolic reaction mixture. The conversion of vinyl- to formyl- groups is independent of their position on the macrocycle, as we observed oxidative cleavages of both 3-vinyl and 8-vinyl side chains to yield formyl groups. Three new chlorophyll derivatives were synthesised using 8-vinyl chlorophyll a as substrate: 8-vinyl chlorophyll d, [8-formyl]-chlorophyll a, and [3,8-diformyl]-chlorophyll a. The structural and spectral properties will provide a signature that may aid in identification of the novel chlorophyll derivatives in natural systems. The ease of conversion of vinyl- to formyl- in chlorophylls demonstrated here has implications regarding the biosynthetic mechanism of chlorophyll d in vivo.

Topics: Catalysis; Chlorophyll; Chlorophyll A; Formates; Heme; Mercaptoethanol; Photosynthesis; Prochlorococcus; Protoporphyrins; Vinyl Compounds

We investigated the potentially different effects of one of the most commonly used glyphosate formulations in Argentina, Glifosato Atanor(®), and the technical-grade glyphosate on the pigment content, as biomass indicators of the algal fraction in a freshwater periphytic community. A laboratory bioassay was carried out in 250-ml beakers. Two treatments were used: technical-grade glyphosate acid and Glifosato Atanor(®) (isopropylamine salt of glyphosate 48 % w/v), which were at a concentration of 3 mg active ingredient per liter. Treatments and the control (without herbicide) were replicated in triplicate. The concentrations of chlorophyll a and b and carotenes were determined at 0, 2, 6, 10, 24, 48, 96 and 192 h after herbicide addition. A significant increase in pigment content was observed for both herbicides after a 2-day exposure. Moreover, the formulation had little or no effect compared to the active ingredient, suggesting that the additives of Glifosato Atanor(®) may not enhance glyphosate toxicity.

Topics: Analysis of Variance; Argentina; Chlorophyll; Chlorophyll A; Glycine; Glyphosate; Herbicides; Magnoliopsida; Microbiota; Pigments, Biological; Time Factors; Water Pollutants, Chemical

Ecophysiological traits of Prosopis juliflora (Sw.) DC. and a phylogenetically and ecologically similar native species, Anadenanthera colubrina (Vell.) Brenan, were studied to understand the invasive species' success in caatinga, a seasonally dry tropical forest ecosystem of the Brazilian Northeast. To determine if the invader exhibited a superior resource-capture or a resource-conservative strategy, we measured biophysical and biochemical parameters in both species during dry and wet months over the course of two years. The results show that P. juliflora benefits from a flexible strategy in which it frequently outperforms the native species in resource capture traits under favorable conditions (e.g., photosynthesis), while also showing better stress tolerance (e.g., antioxidant activity) and water-use efficiency in unfavorable conditions. In addition, across both seasons the invasive has the advantage over the native with higher chlorophyll/carotenoids and chlorophyll a/b ratios, percent N, and leaf protein. We conclude that Prosopis juliflora utilizes light, water and nutrients more efficiently than Anadenanthera colubrina, and suffers lower intensity oxidative stress in environments with reduced water availability and high light radiation.

Topics: Brazil; Carotenoids; Chlorophyll; Chlorophyll A; Ecosystem; Environment; Fabaceae; Forests; Introduced Species; Plant Leaves; Prosopis; Seasons; Stress, Physiological; Trees; Tropical Climate; Water

Photosynthetic light harvesting in plants is regulated by phosphorylation-driven state transitions: functional redistributions of the major trimeric light-harvesting complex II (LHCII) to balance the relative excitation of photosystem I and photosystem II. State transitions are driven by reversible LHCII phosphorylation by the STN7 kinase and PPH1/TAP38 phosphatase. LHCII trimers are composed of Lhcb1, Lhcb2, and Lhcb3 proteins in various trimeric configurations. Here, we show that despite their nearly identical amino acid composition, the functional roles of Lhcb1 and Lhcb2 are different but complementary. Arabidopsis thaliana plants lacking only Lhcb2 contain thylakoid protein complexes similar to wild-type plants, where Lhcb2 has been replaced by Lhcb1. However, these do not perform state transitions, so phosphorylation of Lhcb2 seems to be a critical step. In contrast, plants lacking Lhcb1 had a more profound antenna remodeling due to a decrease in the amount of LHCII trimers influencing thylakoid membrane structure and, more indirectly, state transitions. Although state transitions are also found in green algae, the detailed architecture of the extant seed plant light-harvesting antenna can now be dated back to a time after the divergence of the bryophyte and spermatophyte lineages, but before the split of the angiosperm and gymnosperm lineages more than 300 million years ago.

Topics: Arabidopsis; Arabidopsis Proteins; Chlorophyll; Chlorophyll A; Electrophoresis, Polyacrylamide Gel; Kinetics; Light-Harvesting Protein Complexes; MicroRNAs; Peptides; Phenotype; Phosphorylation; Photosynthesis; Photosystem I Protein Complex; Photosystem II Protein Complex; Protein Binding; Protein Multimerization; Thylakoids

In this study, the changes caused by chilling stress on some physiological parameters of pepino (Solanum muricatum Ait.) plant and the effects of ascorbic acid (100 mM) applied exogenously on these changes were examined. For this purpose, the photosynthetic pigments (chlorophyll a, chlorophyll b, total chlorophylls and carotenoids), ascorbic acid, total phenolic compounds, malondialdehyde and proline contents in leaves of pepino taken on 5th and 10th days were determined. As a result of chilling stress, it was found that while the photosynthetic pigments and proline contents decreased in pepino leaves, the ascorbic acid, total phenolic compounds and malondialdehyde contents increased. In plants which were subjected to pre-treatment of ascorbic acid on the 10th day of stress, ascorbic acid and proline contents increased while a decrease was observed in malondialdehyde content, compared to stress group without pre-treated. This study may be important for explaining resistance induced by treatment of exogenous ascorbic acid in pepino exposed to chilling stress.

Topics: Antioxidants; Ascorbic Acid; Carotenoids; Chlorophyll; Chlorophyll A; Cold Temperature; Cold-Shock Response; Malondialdehyde; Phenols; Plant Leaves; Proline; Solanum; Time Factors

Leaves of a pepper cultivar harboring the L(3) resistance gene were inoculated with Obuda pepper virus (ObPV), which led to the appearance of hypersensitive necrotic lesions approx. 72 h post-inoculation (hpi) (incompatible interaction), or with Pepper mild mottle virus (PMMoV) that caused no visible symptoms on the inoculated leaves (compatible interaction). ObPV inoculation of leaves resulted in ion leakage already 18 hpi, up-regulation of a pepper carotenoid cleavage dioxygenase (CCD) gene from 24 hpi, heat emission and declining chlorophyll a content from 48 hpi, and partial desiccation from 72 hpi. After the appearance of necrotic lesions a strong inhibition of photochemical energy conversion was observed, which led to photochemically inactive leaf areas 96 hpi. However, leaf tissues adjacent to these inactive areas showed elevated ΦPSII and Fv/Fm values proving the advantage of chlorophyll a imaging technique. PMMoV inoculation also led to a significant rise of ion leakage and heat emission, to the up-regulation of the pepper CCD gene as well as to decreased PSII efficiency, but these responses were much weaker than in the case of ObPV inoculation. Chlorophyll b and total carotenoid contents as measured by spectrophotometric methods were not significantly influenced by any virus inoculations when these pigment contents were calculated on leaf surface basis. On the other hand, near-infrared FT-Raman spectroscopy showed an increase of carotenoid content in ObPV-inoculated leaves suggesting that the two techniques detect different sets of compounds.

Topics: Capsicum; Chlorophyll; Chlorophyll A; Dioxygenases; Fluorescence; Plant Leaves; Plant Proteins; Spectrum Analysis, Raman; Tobamovirus

Hirschmanniella oryzae is the most common plant-parasitic nematode in flooded rice cultivation systems. These migratory animals penetrate the plant roots and feed on the root cells, creating large cavities, extensive root necrosis and rotting. The objective of this study was to investigate the systemic response of the rice plant upon root infection by this nematode. RNA sequencing was applied on the above-ground parts of the rice plants at 3 and 7 days post inoculation. The data revealed significant modifications in the primary metabolism of the plant shoot, with a general suppression of for instance chlorophyll biosynthesis, the brassinosteroid pathway, and amino acid production. In the secondary metabolism, we detected a repression of the isoprenoid and shikimate pathways. These molecular changes can have dramatic consequences for the growth and yield of the rice plants, and could potentially change their susceptibility to above-ground pathogens and pests.

Topics: Animals; Base Sequence; Chlorophyll; Chlorophyll A; Gene Expression Profiling; Gene Expression Regulation, Plant; Genes, Plant; Oryza; Plant Diseases; Plant Roots; Plant Shoots; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transcriptome; Tylenchoidea

The effects of spraying exogenous silicon (Si) (0, 1, 2, 3, 4 and 5 mmol L(-1)) on the growth, photosynthetic characteristics and activity of antioxidant enzymes in continuous-cropped 'Jinyan No. 4' cucumber seedlings were studied. The results showed that with the application of 1-3 mmol x L(-1) Si, electrolyte leakage (EL) and malondialdehyde (MDA) content in leaves were significantly decreased, while the contents of chlorophyll a (Chl a), chlorophyll b (Chl b), carotenoids (Car), chlorophyll (a+b) and photosynthetic rate (Pn) in leaves were significantly improved, the activities of superoxidase (SOD), peroxidase (POD), catalase (CAT) and ascorbic acid peroxidase (APX) were significantly increased, and the plant height, stem diameter and dry mass accumulation of cucumber seedlings were promoted. Compared with the low Si concentrations, excessive Si (4-5 mmol x L(-1)) resulted in higher EL and MDA, which were still lower than that in control, decreased the antioxidant enzymes activity and photosynthesis, and inhibited the growth of cucumber seedlings. These findings indicated that exogenous Si could enhance the capacity of scavenging active oxygen species and improve photosynthesis, protect cucumber seedlings from the lipid peroxidation, and increase the resistance to continuous-cropped cucumber obstacle. The optimal silicon concentration was 2 mmol x L(-1).

Topics: Antioxidants; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Cucumis sativus; Lipid Peroxidation; Malondialdehyde; Oxidation-Reduction; Peroxidases; Photosynthesis; Plant Leaves; Reactive Oxygen Species; Seedlings; Silicon

To study the effect of exogenous Ca2+ on photosynthetic parameters of Pinellia ternate and accumulations of active components under high temperature stress.. The pigment contents of P. ternata leaves, photosynthesis parameters and chlorophyll fluorescence parameters of P. ternata leaves, the contents of guanosine, adenosine and polysaccharide in P. ternata tubers were measured based on different concentrations of exogenous Ca2+ in heat stress when the plant height of P. ternata was around 10 cm.. The contents of total chlorophyll and ratio of chlorophyll a/b were relatively higher by spaying Ca2+. Compared with the control, spaying 6 mmol x L(-1) Ca2+ significantly enhanced the net photosynthetic rate (Pn), transpiration (Tr) and stomatal limitation (L8), but reduced intercellular CO2 concentration (C) in P. ternata leaves. With the increase of Ca2+ concentration, maximal PS II efficiency (Fv/Fm), actual photosynthetic efficiency (Yield) and photochemical quenching coefficient (qP) initially increased and then decreased, however, minimal fluorescence (Fo) and non-photochemical quenching coefficient (NPQ) went down first and then went up. The contents of guanosine and polysaccharide and dry weight of P. ternata tubers showed a tendency of increase after decrease, and the content of adenosine increased with the increase of Ca2+ concentration. The content of guanosine and polysaccharide in P. ternata tubers and its dry weight reached maximum when spaying 6 mmol x L(-1) Ca2+.. With the treatment of calcium ion, the inhibition of photosynthesis and the damage of PS II system were relieved in heat stress, which increased the production of P. ternata tubers.

Topics: Breeding; Calcium; Chlorophyll; Chlorophyll A; Dose-Response Relationship, Drug; Heat-Shock Response; Organ Size; Photosynthesis; Pinellia; Plant Leaves

LED lighting has several unique advantages over traditional lighting, including the ability to control spectral composition, the ability to produce very high light levels with low radiant heat output when cooled properly, and the ability to maintain useful light output for years without replacement. LED light sources have the capability of controlling true spectral composition, allowing wavelengths to match plant photoreceptors to provide more optimal production to regulate plant morphology and nutritional quality. In this paper, the effects of different light qualities on the growth, photosynthetic response and nutritional quality of savoy were studied. With 'Juhua-Xiaobaye' savoy as the test plant, full red light, full blue light, red/blue (3/1) light, red/blue (7/1) light and white/red/blue (3/2/1) light generated by light-emitting diodes were applied with white light generated by fluorescent lamps as control. The results showed that red light could increase biomass and stem diameter, but blue light showed the opposite effect. The chlorophyll (a+b) content was the highest in the red/blue (7/1) light treatment, and chlorophyll (a+b) content was correlated with the red/blue ratio positively. Blue light decreased chlorophyll (a + b) content of savoy, and increased chlorophyll a/b. The photosynthetic rate (Pn) and transpiration rate under red light were the highest, and increased by 43.8% and 55.1% compared with the control. Intercellular CO2 concentration and conductance were the highest under blue light. The fluorescence parameters of the plant were significantly affected by light quality. Values of the maximal photochemical efficiency of PS II (Fv/Fm), Fv/Fo and Phi(PS II) were the highest under white light. Soluble sugar, soluble protein and vitamin C contents were the highest under red, blue and white light, respectively. Comprehensive analysis indicated that red/blue (7/1) light was the best light combination to increase leaf chlorophyll (a+b) content and net photosynthetic rate, as well as plant growth and nutritional quality.

Topics: Biomass; Brassica; Chlorophyll; Chlorophyll A; Fluorescence; Light; Nutritive Value; Photosynthesis; Plant Development; Plant Leaves; Plant Stems

The effective region was segmented from the hyperspectral image of citrus leaf by threshold method with the average spectrum extracted and used to describe the corresponding leaf. Based on the different spectral pre-processing methods, the prediction models of three photosynthetic pigments (i. e., chlorophyll a, chlorophyll b, and carotenoid) were calibrated by partial least squares (PLS), BP neural network (BPNN) and least square support vector machine (LS-SVM). The LS-SVM model for chlorophyll a was established based on multiplicative scatter correction (MSC), and the correlation coefficient (Rp) and the root mean square error of prediction (RMSEP) were 0.898 3 and 0.140 4, respectively. The LS-SVM model for chlorophyll b with Rp = 0.912 3 and RMSEP = 0.042 6, was established based on standard normal variable (SNV). The PLS model for carotenoid was established with Rp = 0.712 8 and RMSEP = 0.062 4 based on moving average smoothing (MAS), but the result was no better than the other two. The results illustrated that these three photosynthetic pigments could be nondestructively and real time estimated by hyperspectral image.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Citrus; Least-Squares Analysis; Models, Theoretical; Neural Networks, Computer; Photosynthesis; Plant Leaves; Support Vector Machine

Effect of light quality, including red light, blue light, white light, red and blue mixing light with ratios of 8: 1, 8:2 and 8 : 3, on the growth characteristics and biochenmical composition of Chlorella pyrenoidosa was investigated based on light emitting diode (LED). Results showed that Chlorella pyrenoidosa grew best under blue light, and the optical density, specific growth rate and biomass of Chlorella pyrenoidosa was about 2.4, 0.10 d(-1) and 0.64 g x L(-1), respectively, while the optical density of Chlorella pyrenoidosa was between 1.0 and 1.7, the specific growth rate was between 0.07-0.10 d(-1) and the biomass was between 0.27 and 0.38 g x L(-1) under other light quality after 30 days of cultivation. Under blue light, the optical density, specific growth rate and biomass of Chlorella pyrenoidosa was approximately 2.05 times, 1.33 times and 2.06 times higher than red light, respectively. Moreover, red and blue mixing light was conducive to the synthesis of chlorophyll a and β-carotene of Chlorella pyrenoidosa, and blue light could promote the synthesis of chlorophyll b. Chlorophyll a and carotenoids content of Chlorella pyrenoidosa was 13.5 mg xg(-1) and 5.8 mg x g(-1) respectively under red and blue mixing light with a ratio of 8:1, while it was 8.4 mg x g(-1) and 3.6 mg x g(-1) respectively under blue light. Red and blue mixing light was more conducive to protein and total lipid content per dry cell of Chlorella pyrenoidosa. Protein and total lipid content was 489.3 mg x g(-1) and 311.2 mg x g(-1) under red and blue mixing light with a ratio of 8 : 3, while it was 400.9 mg x g(-1) and 231.9 mg x g(-1) respectively under blue light.

Topics: Biomass; Carotenoids; Chlorella; Chlorophyll; Chlorophyll A; Light

A quantitative method based on high performance liquid chromatography coupled with electrospray ionization tandem triple-quadrupole mass spectrometry (HPLC-ESI-QqQ-MS) has been established for five pigments in marine phytoplanktons. The HPLC method used ternary solvent systems and a reversed-phase C16-amide column. In addition, methanol, acetonitrile and aqueous ammonium acetate were used as mobile phases. Five pigments (chlorophyll a, chlorophyll b, β, β-carotene, lutein and fucoxanthin) were quantified in selective reaction mode. As results, good linear relationships were achieved between the concentrations and the peak areas of the five pigment standards. And their correlation coefficients (r2) were higher than 0.996. The recoveries of the pigment standards were between 82.77% and 99.83%. The inter-day and intra-day precisions were lower than 5% (n = 5). The detection limits of the pigments for this method were between 0.02 and 0.16 μg/L and the quantification limits were in the range from 0.06 to 0.54 μg/L. According to the above method, eleven algae (Heterosigma akashiwo (NMBRah03-2), Heterosigma akashiwo (NMBRah03-2-2), Karlodinium veneficum (NMBjah047-1), Prorocentrum minimum ( NMBjah042), Nannochloropsis oceanic (NMBluh014), Chlorella pyrenoidosa (NMBluh015-1), Pleurochrysis sp. (NMBjih026-1), Prymnesium sp. (NMBjih029), Skeletonema costatum (NMBguh004-1), Thalassiosira weiss- flogii (NMBguh021) and Thalassiosira pseudonana) (NMBguh005)) have been investigated for comparing the pigment distributions. The method is sensitive, accurate, reproducible, and useful for the study of alga compositions.

Topics: beta Carotene; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Lutein; Phytoplankton; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Xanthophylls

The effects of different dosages (0, 0.8%, 2%, 6% and 10%) of sewage sludge compost on the growth and photosynthetic characteristics of three turfgrass, i. e., Festuca arundinacea, Lolium perenne and Poa pratensis were studied. The results showed that the dosage of 6% had the optimal effects. The plant height, leaf area and dry biomass of three turfgrass increased significantly with the increasing sewage sludge compost dosage, while the root/shoot ratio decreased obviously. Compared with the control, the plant height of F. arundinacea, L. perenne and P. pratensis increased by 64.9%-180.8%, 97.3%-200.9% and 39.1%-156.5%, the leaf area per plant increased by 91.3%-417.4%, 186.4%-394.9% and 164.6%-508.3% and the dry biomass per plant increased by 333.4%-867.6%, 138.4%-445.1% and 316.3%-669.2%, respectively. With the increasing dosage of sewage sludge compost, the net photosynthetic rate (Pn), transpiration rate (Tr), intercellular CO2 concentration (Ci) and water use efficiency (WUE) of F. arundinacea increased first and decreased then, the Pn, Tr and WUE of P. pratensis showed a significantly increasing trend, while the Ci decreased after an initial increase, and the Pn and WUE of L. perenne increased gradually, while the Ci and Tr decreased after a gradual increase. With the in- creasing sewage sludge dosage, the contents of chlorophyll a and b of the three turfgrass species increased markedly, and then decreased when the dosage was more than 6%, but the ratio of chlorophyll a/b was little changed.

Topics: Biomass; Chlorophyll; Chlorophyll A; Festuca; Lolium; Photosynthesis; Plant Leaves; Poa; Sewage; Water

To provide the basis for improving ginseng production by exploring the difference of physio-biochemistry of Panax ginseng after increasing light intensity.. Activities of superoxide dismutase( SOD), hydrogen peroxidase (CAT), and peroxydase (POD), as well as contents of malondialdehyde (MDA), chlorophyll a and b under full-light condition were compared with those in the shade in spring and autumn.. Activity of three kinds of enzymes in the anti-oxidation system (SOD,CAT and POD) were 27.52%, 41.96% and 37.7% higher than the shade group, respectively. The content of MDA was 11.23% lower. There had no obvious difference on the content of chlorophyll a, but chlorophyll b was decreased slightly.. Increasing the light intensity properly can improve the activities of enzyme protection system, and lower the content of MDA, which indicates that the increasing of light will provide a theoretic guide for high-production technique of Panax ginseng.

Topics: Chemical Phenomena; Chlorophyll; Chlorophyll A; Light; Malondialdehyde; Oxidation-Reduction; Panax; Peroxidases; Superoxide Dismutase

Effect of nitric oxide donor (sodium nitroprusside, SNP, 500 μM) or hydrogen peroxide scavenger (dithiothreitol, DTT, 500 μM) on cadmium (Cd) or copper (Cu) uptake (150 μM solutions) and toxicity using Scenedesmus quadricauda was studied. Combined treatments (Cd or Cu + DTT or SNP) usually ameliorated metal-induced toxicity at the level of pigments, proteins, and mineral nutrients in comparison with respective metal alone. Viability tests (MTT and TTC) showed the lowest values preferentially in Cu treatments, indicating higher toxicity in comparison with Cd. Cd showed low impact on amino acids while strong Cu-induced depletion was mitigated by DTT and SNP. Amount of ROS and NO showed the most pronounced responses in SNP variants being rather reciprocal than parallel and regulated ascorbate peroxidase activity. Blot gel analyses of hsp70 protein did not reveal extensive changes after given exposure period. Phenols were elevated by DTT alone while all Cu treatments revealed depletion. Total Cu content decreased while total Cd content increased in metal + SNP or metal + DTT. Subsequent experiment using lower Cd, SNP or DTT doses (10 and 100 μM) revealed concentration-dependent impact on Cd uptake. Overall, DTT was found to be more suitable for the amelioration of metal-induced toxicity.

Topics: Amino Acids; Cadmium; Carotenoids; Chlorophyll; Chlorophyll A; Copper; Dithiothreitol; Free Radical Scavengers; Nitric Oxide; Nitric Oxide Donors; Nitroprusside; Peroxidase; Plant Proteins; Reactive Oxygen Species; Scenedesmus

The present study aims to assess the level and possible sources of organic and inorganic pollutants in Mahableshwar city in Western part of India and their effect on naturally growing foliose lichen Remototrachyana awasthii (Hale & Patw.) Divakar & A. Crespo. This lichen species growing abundantly in the area was collected from eight different sites on the basis of anthropogenic activities detected in the area. The concentration of inorganic heavy metals (Al, As, Cd, Cr, Fe, Pb, Mn, and Zn) and polycyclic aromatic hydrocarbons (PAHs) were analyzed and correlated with photosynthetic pigments (Chl a, Chl b, total chlorophyll, and carotenoid) together with chlorophyll degradation and protein contents. The concentration of most of the metals at different sites was significantly greater than at the control site (P<0.001). The highest metal content was found at Bus Stand and Panchgani, a tourist place that experiences heavy traffic activities. The concentration of PAHs, particularly of two-and three-ringed PAHs, was also found to be the highest in samples collected at Bus Stand area. The chlorophyll degradation and protein content were found to be the most sensitive parameters to assess the vitality of lichen thallus against wide range of air pollutants. The effectiveness of R. awasthii as a biomonitor will be investigated in the near future by comparing this species with other biomonitors.

Topics: Air Pollutants; Carotenoids; Chlorophyll; Chlorophyll A; Environmental Monitoring; India; Lichens; Metals, Heavy; Polycyclic Aromatic Hydrocarbons

Effects of gamma rays on the free polyamine (PA) levels were studied in Vigna radiata (L.) Wilczek. Seeds exposed to different doses of gamma rays were checked for damage on phenotype, germination frequency and alteration in photosynthetic pigments. Free polyamine levels were estimated from seeds irradiated in dry and water imbibed conditions. Polyamine levels of seedlings grown from irradiated seeds, and irradiated seedlings from unexposed seeds were also measured. Damage caused by gamma irradiation resulted in decrease in final germination percentage and seedling height. Photosynthetic pigments decreased in a dose dependent manner as marker of stress. Polyamines decreased in irradiated dry seeds and in seedlings grown from irradiated seeds. Radiation stress induced increase in free polyamines was seen in irradiated imbibed seeds and irradiated seedlings. Response of polyamines towards gamma rays is dependent on the stage of the life cycle of the plant.

Topics: Chlorophyll; Chlorophyll A; Fabaceae; Gamma Rays; Germination; Phenotype; Photosynthesis; Polyamines; Seedlings; Seeds

Both elevated ozone (O(3)) and limiting soil nitrogen (N) availability negatively affect crop performance. However, less is known about how the combination of elevated O(3) and limiting N affect crop growth and metabolism. In this study, we grew tobacco (Nicotiana sylvestris) in ambient and elevated O(3) at two N levels (limiting and sufficient). Results at the whole plant, leaf, and cellular level showed that primary metabolism was reduced by growth in limiting N, and that reduction was exacerbated by exposure to elevated O(3). Limiting N reduced the rates of photosynthetic CO(2) uptake by 40.8% in ambient O(3)-exposed plants, and by 58.6% in elevated O(3)-exposed plants, compared with plants grown with sufficient N. Reductions in photosynthesis compounded to cause large differences in leaf and whole plant parameters including leaf number, leaf area, and leaf and root biomass. These results were consistent with our meta-analysis of all published studies of plant responses to elevated O(3) and N availability. In tobacco, N uptake and allocation was also affected by growth in limiting N and elevated O(3), and there was an O(3)-induced compensatory response that resulted in increased N recycling from senescing leaves. In addition, transcript-based markers were used to track the progress through senescence, and indicated that limiting N and elevated O(3), separately and in combination, caused an acceleration of senescence. These results suggest that reductions in crop productivity in growing regions with poor soil fertility will be exacerbated by rising background O(3).

Topics: Air Pollutants; Carbon Dioxide; Chlorophyll; Chlorophyll A; Nicotiana; Nitrogen; Ozone; Photosynthesis; Plant Leaves; Plant Roots; Plant Stems

The changes in the structural characteristics of biomass residues during pyrolysis and composting were investigated. The biomass residues particles were prepared by pyrolysing at temperatures ranging from 350 to 400. For soilless production of the ornamental plant Anthurium andraeanum, pure sphagnum peat moss (P) has traditionally been used as the growing medium. This use of P must be reduced, however, because P is an expensive and nonrenewable resource. The current study investigated the use of biomass residues as substitutes for P in A. andraeanum production. Plants were grown for 15 months in 10 soilless media that contained different proportions of pyrolysed corn cobs (PC), composted corn cobs (C), pyrolysed garden wastes (PG), and P. Although the media altered the plant nutrient content, A. andraeanum growth, development, and yield were similar with media consisting of 50% P+50% PC, 50% P+35% PC+15% PG, and 100% P. This finding indicates that, when pyrolysed, organic wastes, which are otherwise an environmental problem, can be used to reduce the requirement for peat in the soilless culture of A. andraeanum.

Topics: Agriculture; Araceae; Biomass; Chlorophyll; Chlorophyll A; Culture Media; Elements; Micronutrients; Microscopy, Electron, Scanning; Plant Development; Plant Leaves; Soil; Temperature; Waste Products

Ozone (O3) has become one of the most toxic air pollutants to plants worldwide. However, investigations on O3 impacts on crops health and productivity in South America countries are still scarce. The present study analyzed the differences on the enzymatic and non-enzymatic antioxidant system in foliar tissue of two subtropical Phaseolus vulgaris varieties exposed to high O3 concentration. Both varieties were negatively impacted by the pollutant, but the responses between each variety were quite distinct. Results revealed that Irai has higher constitutive levels of reactive oxygen species (ROS) and ascorbate (AsA) concentration, but lower total thiol levels and catalase immunocontent. In this variety catalase protein concentration was increased after O3 exposure, indicating a better cellular capacity to reduce hydrogen peroxide. On the opposite, Fepagro 26-exposed plants increased ROS generation and AsA concentration, but had the levels of total thiol content and catalase protein unchanged. Furthermore, O3 treatment reduced the levels of chlorophylls a and b, and the relationship analysis between the chlorophyll ratio (a/b) and protein concentration were positively correlated indicating that photosynthetic apparatus is compromised, and thus probably is the biomass acquisition on Fepagro 26. Differently, O3 treatment of Irai did not affect chlorophylls a and b content, and loss on the protein content was lower. Altogether, these data suggest that early accumulation of ROS on Fepagro 26 are associated with an insufficient leaf antioxidant capacity, which leads to cell structure disruption and impairs the photosynthesis. Irai seems to be more tolerant to O3 toxic effects than Fepagro 26, and the observed differences on O3 sensitivity between the two varieties are apparently based on constitutive differences involved in the maintenance of intracellular redox homeostasis.

Topics: Antioxidants; Ascorbic Acid; Catalase; Chlorophyll; Chlorophyll A; Homeostasis; Oxidation-Reduction; Ozone; Phaseolus; Photosynthesis; Plant Proteins; Reactive Oxygen Species; Stress, Physiological; Sulfhydryl Compounds; Tropical Climate

Photosynthetic structures when imaged with nonlinear optical microscopy give rise to high third harmonic generation (THG) signal intensity due to the presence of chlorophylls and xanthophylls which have large second hyperpolarizabilitiy (γ) values. The γ value of trimers of the light-harvesting chlorophyll a/b pigment-protein complex of photosystem II (LHCII) isolated from pea (Pisum sativum) plants was investigated by the THG ratio technique at 1028 nm wavelength and found to have the value (-1600 ± 400) × 10(-41) m(2) V(-2). The large negative γ value of trimeric LHCII is due to the presence of chlorophyll a and chlorophyll b which have large negative γ values, while positive γ values of xanthophylls reduce the magnitude of the THG signal. Variation was observed between the measured γ value of LHCII and the approximated γ value of LHCII obtained by adding individual γ values of chlorophylls and xanthophylls. This difference can be attributed to the differing inter-pigment interactions of oriented chlorophylls and xanthophylls in the pigment-protein complex compared to randomly oriented non-interacting pigments in solution, as well as a differing dielectric environment of the pigments within LHCII versus the surrounding organic solvent.

Topics: Chlorophyll; Chlorophyll A; Octoxynol; Photosystem II Protein Complex; Pisum sativum; Plant Proteins; Protein Binding; Spectrophotometry

In this study, Secale cereale seedlings cultivated under 0 (control), 2 or 5mM Pb(NO3)2 concentrations were used to examine alterations in the organization and functionality of chlorophyll-protein complexes in thylakoid membranes under Pb ion stress. The studies were conducted on whole leaves of rye seedlings or thylakoid membranes isolated from Pb-treated and control plants. Using non-denaturing electrophoresis, it was assessed that increasing Pb concentrations resulted in an increase in the value of the ratio of the content of LHCII oligomers (mainly trimers) to the content of LHCII monomers. The parameters of chlorophyll fluorescence induction (q(p) and q(n)) indicated that the change in the LHCII supramolecular organization in the presence of Pb ions was connected with an increase in non-photochemical fluorescence quenching. Quantification of photosynthetic pigments showed that both Pb concentrations decreased the content of chlorophyll a, chlorophyll b, and carotenoids. The changes in the pigment content led to a significant reduction in light absorption by antenna complexes. However, the absorption spectra showed that red light was preferentially absorbed by antenna complexes in thylakoid membranes isolated from the Pb-treated plants. Examination of fluorescence emission spectra revealed that Pb ions decreased the fluorescence quantum yield of PSII. The emission spectra of thylakoids indicated a relative increase in the intensity of fluorescence emission from the trimeric and aggregated forms of the LHCII complexes in comparison to the intensity of fluorescence emission from PSI antenna complexes under excitation at 440 nm. Simultaneously, under excitation at 470 nm, we observed a rise in fluorescence intensity from the LHCII trimer after addition of 5mM Pb as well as a decrease in fluorescence intensity from the LHCII aggregates and PSI core and LHCI antenna complexes under both Pb concentrations. Pb treatments also reduced excitation energy absorbed by chlorophyll b and carotenoids within antenna complexes and transferred to chlorophyll a species emitting at 680 nm.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Environmental Pollutants; Lead; Photosystem II Protein Complex; Plant Leaves; Secale; Thylakoids

This study was conducted to compare the effects of foliar spray and rhizosphere irrigation with purple phototrophic bacteria (PPB) on growth and stevioside (ST) yield of Stevia. rebaudiana. The S. rebaudiana plants were treated by foliar spray, rhizosphere irrigation, and spray plus irrigation with PPB for 10 days, respectively. All treatments enhanced growth of S. rebaudiana, and the foliar method was more efficient than irrigation. Spraying combined with irrigation increased the ST yield plant (-1) by 69.2% as compared to the control. The soil dehydrogenase activity, S. rebaudiana shoot biomass, chlorophyll content in new leaves, and soluble sugar in old leaves were affected significantly by S+I treatment, too. The PPB probably works in the rhizosphere by activating the metabolic activity of soil bacteria, and on leaves by excreting phytohormones or enhancing the activity of phyllosphere microorganisms.

Topics: Chlorophyll; Chlorophyll A; Diterpenes, Kaurane; Glucosides; Oxidoreductases; Plant Leaves; Rhizosphere; Rhodopseudomonas; Soil; Solubility; Stevia

The influence of compost on the growth of bean plants irrigated with As-contaminated waters and its influence on the mobility of As in the soils and the uptake of As (as NaAs(III)O2) by plant components was studied at various compost application rates (3·10(4) and 6·10(4) kg ha(-1)) and at three As concentrations (1, 2 and 3 mg kg(-1)). The biomass and As and P concentrations of the roots, shoots and beans were determined at harvest time, as well as the chlorophyll content of the leaves and nonspecific and specifically bound As in the soil. The bean plants exposed to As showed typical phytotoxicity symptoms; no plants however died over the study. The biomass of the bean plants increased with the increasing amounts of compost added to the soil, attributed to the phytonutritive capacity of compost. Biomass decreased with increasing As concentrations, however, the reduction in the biomass was significantly lower with the addition of compost, indicating that the As phytotoxicity was alleviated by the compost. For the same As concentration, the As content of the roots, shoots and beans decreased with increasing compost added compared to the Control. This is due to partial immobilization of the As by the organic functional groups on the compost, either directly or through cation bridging. Most of the As adsorbed by the bean plants accumulated in the roots, while a scant allocation of As occurred in the beans. Hence, the addition of compost to soils could be used as an effective means to limit As accumulation in crops from As-contaminated waters.

Topics: Agricultural Irrigation; Arsenic; Arsenites; Biological Availability; Biomass; Chlorophyll; Chlorophyll A; Phaseolus; Phosphorus; Plant Leaves; Plant Roots; Soil; Soil Pollutants; Water Pollutants, Chemical

Nanoparticles (NPs) have emerged as one of the most innovative and promising application in agriculture. Since plants are recognized as essential component of all ecosystems, the effects of NPs on plants may pave a new insight to the ecosystems. Here, uptake and translocation of superparamagnetic iron oxide NPs (SPIONs), with various surface charges, on soybean has been probed; in addition, the effects of SPIONs on variations of chlorophyll, in hydroponic condition, together with their ability for reduction of iron deficiency chlorosis were explored. We find that SPIONs, which were entered and translocated in the soybean, increased chlorophyll levels, with no trace of toxicity. Furthermore, it was found that physicochemical characteristics of the SPIONs had a crucial role on the enhancement of chlorophyll content in subapical leaves of soybean. The equivalent ratio of chlorophyll a to b, in all treatments with conventional growth medium iron chelate and SPIONs (as iron source), indicated no significant difference on the photosynthesis efficiency. Finally, it was observed that the effect of SPIONs on the soybean chlorophyll content may have influence on both biochemical and enzymatic efficiency in different stages of the photosynthesis reactions.

Topics: Chlorophyll; Chlorophyll A; Germination; Glycine max; Magnetic Phenomena; Magnetite Nanoparticles; Microscopy, Electron, Transmission; Plant Components, Aerial; Plant Roots

Various plants possess non-photosynthetic, hydrophilic chlorophyll (Chl) proteins called water-soluble Chl-binding proteins (WSCPs). WSCPs are categorized into two classes; Class I (photoconvertible type) and Class II (non-photoconvertible type). Among Class II WSCPs, only Lepidium virginicum WSCP (LvWSCP) exhibits a low Chl a/b ratio compared with that found in the leaf. Although the physicochemical properties of LvWSCP have been characterized, its molecular properties have not yet been documented. Here, we report the characteristics of the LvWSCP gene, the biochemical properties of a recombinant LvWSCP, and the intracellular localization of LvWSCP. The cloned LvWSCP gene possesses a 669-bp open reading frame. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis revealed that the precursor of LvWSCP contains both N- and C-terminal extension peptides. RT-PCR analysis revealed that LvWSCP was transcribed in various tissues, with the levels being higher in developing tissues. A recombinant LvWSCP and hexa-histidine fusion protein (LvWSCP-His) could remove Chls from the thylakoid in aqueous solution and showed an absorption spectrum identical to that of native LvWSCP. Although LvWSCP-His could bind both Chl a and Chl b, it bound almost exclusively to Chl b when reconstituted in 40 % methanol. To clarify the intracellular targeting functions of the N- and C-terminal extension peptides, we constructed transgenic Arabidopsis thaliana lines expressing the Venus protein fused with the LvWSCP N- and/or C-terminal peptides, as well as Venus fused at the C-terminus of LvWSCP. The results showed that the N-terminal peptide functioned in ER body targeting, while the C-terminal sequence did not act as a trailer peptide.

Topics: Amino Acid Sequence; Arabidopsis; Base Sequence; Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Cloning, Molecular; DNA, Plant; Endoplasmic Reticulum; Genes, Plant; Lepidium; Phylogeny; Plants, Genetically Modified; Protein Binding; Recombinant Fusion Proteins; Sequence Homology, Amino Acid; Solubility

Heavy metal contamination, low pH and high substrate heterogeneity are multiple stress factors that often occur at the post-mining sites and make difficult the biological reclamation. Efficient tools for detection of the status of reclaimed vegetation at post-mining sites are needed. We tested the potential of visible to near-infrared (VNIR) spectroscopy to detect multiple stresses in Scots pine (Pinus sylvestris L.) at acidic substrates rich in As. The needle chemical traits (chlorophyll a + b - Cab; carotenoids - Car; Car/Cab; relative water content - RWC; soluble phenolics; lignin contents) were tested for sensitivity to different soil conditions of post-mining sites. For Scots pine growing on degraded substrates, at least three non-specific stress indicators (RWC, photosynthetic pigments and phenolics) are required to achieve good site separability corresponding to the stress load. We constructed and validated empirical models of selected needle chemical traits using VNIR spectroscopy: calibration of Cab (R(2) = 0.97, RMSE = 0.17 mg g(-1)), RWC (R(2) = 0.88, RMSE = 1.41 mg g(-1)), Car (R(2) = 0.66, RMSE = 0.08 mg g(-1)), phenolics (R(2) = 0.64, RMSE = 23.01 mg g(-1)) and lignin (R(2) = 0.45, RMSE = 3.32 mg g(-1)). The reflectance data yielded comparable site separability with the separability calculated from the laboratory data. The presented approach has potential for large-scale monitoring of Scots pine status, thus, assessment of reclamation quality in post-mining regions using air-born or satellite hyperspectral data.

Topics: Arsenic; Chlorophyll; Chlorophyll A; Lignin; Pinus sylvestris; Soil Pollutants; Spectroscopy, Near-Infrared; Stress, Physiological; Sulfur; Water

The effect of mixed inoculums of VAM (Vesicular Arbuscular Mycorrhizas) fungi on seed growth and photosynthetic apparatus in green house was monitored. The plants were watered daily with tap water. Plants were cultivated in natural environment in mid of March (2011). A direct relation between root length and water contents suggests a defense mechanism of MP (microrihzal plants) against the fungal stress. It was also supported by the fact that the leaf area of MP was much greater as compared to the NMP (non microrihzal plants) with elevated concentration of all chlorophyllus pigments in 30 days. An increase in the surface area of the leaf and concentration of the pigments, may be for an acceleration in absorption of CO₂ for reduction of it into glucose through oxidation of water molecule. The non-significant decline in glucose contents support the above hypothesis of rapid redox reaction mechanism which was established to overcome the stress. The positive effects of mycorrhizal which were already mentioned in the literature were reported in this article in relations of survival strategies of the plant, adapted in stress conditions. An increase in the chlorophyll contents (30 d) and leaf area of plants possibly attributed with absorption of solar radiation for the protection of plants. It was also supported by the higher concentration of carotenoids (30 d) that may have an additional function of regulation of certain developmental responses and screening of light to save the plants from stress conditions.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Glucose; Momordica charantia; Mycorrhizae; Photosynthesis; Plant Leaves; Plant Roots; Plant Shoots

Six cultivars of safflower which were (PI-387820, PI-251978, PI-170274, PI-387821, PI-386174 and Thori-78) grown in net house of NIAB under salinity (10 ds m⁻¹) and drought (60% field capacity) conditions and compared to their oxidative damage and antioxidative responses. Plant samples (leaves) were collected for the determination of malonidialdehyde (MDA), antioxidative enzymes (catalase, ascorbate peroxidase, glutathione reductase, and peroxidase), proline, and photosynthetic pigments. Salinity and drought decreased the chlorophyll a and b contents but a decrease in chlorophyll a and b was less in safflower variety (THORI-78) which could be a useful marker for selecting a stress tolerant variety. Both stresses considerable increases the accumulation of proline in PI-251978, PI-170274, PI-387821, PI-386174 and THORI-78 varieties of safflower whereas the proline accumulation did not appear to be an essential part of the protection mechanism against salinity and drought in variety PI-387820. Enzyme activity measurements revealed that THORI-78 can tolerate salinity and drought stress well by increasing the activity of catalase and APX enzymes whereas variety PI- 386174 showed increased activity of glutathione reductase enzyme under salinity and drought and appear to be very crucial antioxidative defenses during intense stress conditions. The results indicate that the photosynthetic pigments, proline and activities of the enzymes are important mechanism for the stress tolerance in safflower plant and can be considered as genetic improvement for the plant in salinity and drought soil conditions.

Topics: Ascorbate Peroxidases; Carthamus tinctorius; Catalase; Chlorophyll; Chlorophyll A; Droughts; Glutathione Reductase; Peroxidase; Plant Proteins; Proline; Salinity

The response of chickpea (Cicer arientinum L. cv. Giza 3) to treatment with two plant growth regulators putrescine (Put) and Indole-3-butyric acid (IBA) at 25, 50 and 100 mg L(-1) applied either alone or in combinations was studied. Spraying of Put and IBA either individually or in combination significantly increased the plant height, number and dry weight of branches, leaves and pods/plant and leaf area/plant at the two growth stages. Total photosynthetic pigments in fresh leaves were significantly promoted as a result of application of Put or IBA. Generally, application of Put and/or IBA at 100 mg L(-1) produced the highest numbers of pods which resulted in substantially the highest seed yield. Put and IBA increased the seed yield by 21.3 and 19.2%, respectively, while the combination of Put at 100 mgL(-1) and IBA at 50 mgL(-1) increased it by 27.4%. Greatest increases in straw and biological yield/fed (38.3 and 30.4%, respectively) were noted with the combination treatment of IBA 100 mg L(-1) plus Put at 100 mg L(-1). Put and IBA significantly increased the nitrogen, phosphorus, potassium, total soluble sugars and total free amino acids in chickpea seeds over control, but the effects were less marked than those of their combination. This response was greater following treatment with IBA than with Put. It could be conclude that spraying Put or/and IBA on chickpea plants have promotion effects on the seeds yield criteria which have promising potential as sources of low-cost protein and minerals for possible use as food/feed supplements.

Topics: Aerosols; Carotenoids; Chlorophyll; Chlorophyll A; Cicer; Crops, Agricultural; Dose-Response Relationship, Drug; Fruit; Indoles; Photosynthesis; Plant Growth Regulators; Plant Leaves; Putrescine

Fluazifop-p-butyl (FL) is one of the most popular graminicides from arylophenoxypropionate group. These herbicides act as inhibitors of acetyl-CoA carboxylase (ACCase) that catalyzes the formation of malonyl-CoA during metabolism of lipids and/or of some secondary compounds. On the other hand arylopropionates and cyclohexanediones cause phytotoxic effects by stimulating free-radicals generation and causing oxidative stress in susceptible plants. However, the importance of disturbances in plant pigments and polyamines accumulation for this effect is not clear. The aim of this work is to quantify the phytotoxicity of FL to non target maize plant and to explain how photosynthetic pigments, anthocyanins (ANC) and polyamines participate in this interaction. Obtained results showed reduction of chlorophyll a and b, but only in case of the highest herbicide dose. Lower FL concentrations caused increase of the photosynthetic pigments, or were not effective. A similar effect was stated for putrescine, while spermidine was reduced within epicotyl of leaf tissues. In case of 2-phenylethylamine (PEA), there was observed a lack of significant changes within leaves and an increase in epicotyl under the middle and the highest dose of the herbicide. Moreover, FL induced ANC accumulation in epicotyls of maize seedlings. The activity of such key enzymes of polyamine biosynthesis as: ornithine decarboxylase (ODC) and lysine decarboxylase (LDC), increased in leaves treated with herbicide at the lowest concentration and decreased under the highest. However, in case of epicotyls the decreasing tendency was observed with the exception of ODC under the highest FL dose. The activity of tyrosine decarboxylase (TyDC) was importantly elevated only within epicotyls under the lower FL concentrations. It was concluded that FL inhibits maize growth, and the intensity of the effect is positively correlated with the herbicide concentration. The phenomenon was related to changes in content of pigments, polyamines and activity of studied enzymes.

Topics: Anthocyanins; Carboxy-Lyases; Carotenoids; Chlorophyll; Chlorophyll A; Herbicides; Phenethylamines; Plant Leaves; Plant Proteins; Polyamines; Pyridines; Seedlings; Zea mays

Leaves of Lycium chinense L. are widely used in traditional Chinese medicine and as functional food in China and south-east Asia. Polyphenols are important active compounds in leaves of L. chinense L, but their compositions are still not very clear. Selenium has been reported to increase some components of plants. In this study, the composition of polyphenols of L. chinense leaves were analysed, and the effects of selenium on the polyphenols and other components were studied.. The main polyphenols were identified as rutin, chlorogenic acid, quercetin, kaempferol and apigenin-7-O-(6'-O-acetyl) glucose-rhamnose. Chlorogenic acid showed the highest average content of 78.36 g kg⁻¹. Rutin, quercetin and kaempferol were determined at low contents from 3.85 g kg⁻¹ to 7.08 g kg⁻¹. Correlation coefficients between selenium and chlorogenic acid, chlorophyll a, chlorophyll b and carotenoids were: R² = 0.963 (P = 0.002), 0.943 (P = 0.005), 0.957 (P = 0.003) and 0.943 (P = 0.005), respectively. Selenium (0.01-0.05 g kg⁻¹) significantly increased chlorogenic acid, chlorophyll and carotenoids by 200-400% and reduced rutin by 400-900%.. L. chinense can be an alternative source of chlorogenic acid. Selenium significantly increased chlorogenic acid, chlorophyll a, chlorophyll b and carotenoids, hence increasing the medicinal value of L. chinense leaves. Rutin, quercetin, kaempferol and apigenin-7-O-(6'-O-acetyl) glucose-rhamnose proved to be not significantly influenced by selenium.

Topics: Carotenoids; China; Chlorogenic Acid; Chlorophyll; Chlorophyll A; Down-Regulation; Fertilizers; Flavonoids; Functional Food; Hydroponics; Lycium; Nutritive Value; Plant Leaves; Rutin; Selenium; Sodium Selenite; Up-Regulation

Consumption of pulse crops, including field pea, is considered effective for a healthy diet. Hulls (seed coats) play an important role for protection of the cotyledon and embryo, but also as mediating positive effects on health outcomes. The biochemical attributes of field pea hulls were thus assessed to determine the occurrence of specific phytochemicals and their genotypic variability.. Sequestered bioproducts in mature hulls predominantly consisted of trans-lutein and chlorophylls a and b. Trace amounts of other carotenoid and pheophytin metabolites were identified. In developing hulls, violaxanthin, neoxanthin, lutein, zeaxanthin, chlorophylls a and b and β-carotene were detected. Genotypic differences in the accumulation of lutein and chlorophylls a and b were observed over years and locations. Polyphenolics and hydroxybenzoic acids were detected in the 'dun' and 'maple' field pea types-the only genotypes to have pigmented hulls. Unextractable patches of condensed tannin influenced the visual uniformity of the maple and dun genotypes, CDC Rocket and CDC Dundurn.. Within the yellow and green market classes, carotenoid and chlorophyll accumulation was consistent. Green cotyledon varieties sequestered higher concentrations of lutein than the yellow cotyledon varieties. Maple and dun types were more variable, reflective of different selection criteria. The occurrence of flavonoid-related compounds was correlated only with pigmented seed coat genotypes. The dietary potential of the chlorophylls and carotenoids that accumulated in the hulls split from the green and yellow field pea types is discussed as a value-added prospect in food supplements.

Topics: beta Carotene; Carotenoids; Chlorophyll; Chlorophyll A; Flavonoids; Genotype; Hydroxybenzoates; Lutein; Pisum sativum; Polyphenols; Seeds; Species Specificity; Xanthines

CN17 is a functional stay-green wheat variety that exhibits delayed leaf senescence and enhanced photosynthetic competence. To better understand these valuable traits, levels of chlorophyll a and b, soluble proteins, unsaturated fatty acids, and other components of CN17 were assayed. In addition, chloroplast ultrastructure, chloroplast number, and differences in gene expression between CN17 and a control variety, MY11, were examined. By 21 d post-anthesis (DPA), CN17 leaves exhibited a significantly higher maximal photochemical efficiency for photosystem II (PSII) (F(v) /F(m) ) and a significantly higher efficiency of excitation capture by open PSII reaction centres (F(v) '/F(m) '). In addition, chlorophyll degradation in CN17 was delayed by approximately 14 d, and was not blocked as observed in cosmetic stay-green phenotypes. The soluble protein content (Ps) of CN17 was higher than MY11 at all timepoints assayed, and the ratio of unsaturated to saturated fatty acids was significantly higher. CN17 also exhibited isolated granal lamellae associated with vesicles and diminished peroxidation, and between 35 and 42 DPA, a sharp decrease in chloroplast number was detected. Taken together, these results strongly support the hypothesis that chloroplast ultrastructure regeneration is responsible for the functional stay-green trait of CN17, and gene expression data provide insight into the mechanistic details.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Expressed Sequence Tags; Fatty Acids, Unsaturated; Gene Library; Photosynthesis; Photosystem II Protein Complex; Plant Leaves; Reactive Oxygen Species; Triticum

In the Ni-substituted chlorophylls, an ultrafast (<60 fs) deactivation channel is created, which is not present in Ni-porphyrins. This observation prompted us to investigate in detail the mechanism of excitation-to-heat conversion in Ni-substituted chlorophylls, experimentally, using time-resolved laser-induced optoacoustic spectroscopy, and theoretically, using group theory approach. The Ni-substituted chlorophylls show exceptional photostability and the optoacoustic measurements confirm the prompt and very efficient (100%) excitation-into-heat conversion in these complexes. Considering their excellent spectral properties and the loss-free excitation-into-heat conversion they are likely to become a new class of versatile photocalorimetric references. The curious features of the Ni-substituted chlorophylls originate from the symmetry of a ligand field created in the central cavity. The central N-Ni(2+) bonds, formed via the donation of two electrons from each of the sp(2) orbitals of two central nitrogens to an empty [Formula: see text] hybrid centered on Ni(2+), have a considerable covalent character. The extreme rate of excited state relaxation is then not due to a ladder of the metal centered d-states, often invoked in metalloporphyrins, but seems to result from a peculiar topology of the potential energy surface (a saddle-shaped crossing) due to the covalent character of the N-Ni(2+) bonds. This is confirmed by a strong 0→0 character of electronic transitions in these complexes indicating a similarity of their equilibrium geometries in the ground (S(0)) and the excited states (both Q(X) and Q(Y)). The excitation energy is very efficiently converted into molecular vibrations and dissipated as heat, involving the central Ni(2+). These Ni-substituted pigments pose a fine exemplification of symmetry control over properties of excited states of transition metal complexes.

Topics: Chlorophyll; Chlorophyll A; Energy Transfer; Hot Temperature; Light; Models, Chemical; Nickel; Photoacoustic Techniques; Photolysis; Plant Leaves; Solvents; Spectrum Analysis; Spinacia oleracea; Time Factors; Vibration

Chloroplast-located proteins which are encoded by the nuclear genome have to be imported from the cytosol into the organelle in a posttranslational manner. Among these nuclear-encoded chloroplast proteins are the light-harvesting chlorophyll a/b-binding proteins (LHCPs). After translation in the cytosol, precursor proteins of LHCPs are imported via the TOC/TIC translocase, processed to their mature size to insert into thylakoid membranes where they recruit chlorophylls a and b to form pigment-protein complexes. The translocation of proteins is a highly regulated process which employs several regulators. To analyze whether CAO (chlorophyll a oxigenase) which converts chlorophyll a to chlorophyll b at the inner chloroplast membrane, is one of these regulators, we performed import reactions utilizing a homozygous loss-of-function mutant (cao-1). We imported in vitro translated and (35)S-labeled precursor proteins of light-harvesting proteins of photosystem II LHCB1, LHCB4, and LHCB5 into chloroplasts isolated from cao-1 and show that import of precursor proteins and their processing to mature forms are not impaired in the mutant. Therefore, regulation of the import machinery cannot be responsible for the decreased steady-state levels of light-harvesting complex (LHC) proteins. Regulation does not take place at the transcriptional level either, because Lhcb mRNAs are not down-regulated. Additionally, reduced steady-state levels of LHCPs also do not occur due to posttranslational turnover of non-functional LHCPs in chloroplasts. Taken together, our data show that plants in the absence of CAO and therefore devoid of chlorophyll b are not influenced in their import behavior of LHC proteins.

Topics: Arabidopsis; Arabidopsis Proteins; Cell Nucleus; Chlorophyll; Chlorophyll A; Chloroplasts; Gene Expression Regulation, Plant; Light-Harvesting Protein Complexes; Mutation; Oxygenases; Plastocyanin; Protein Biosynthesis; Protein Stability; Protein Transport; RNA, Messenger

In this study, toxic effects of the cypermethrin in Allium cepa L. cells were investigated. For this aim, we investigated the changes in pigment contents, antioxidant enzymes, mitotic index and chromosomal abnormalities as indicators of toxicity. The seeds were treated with different doses (1.5, 3.0, 6.0 ppm) of cypermethrin for 72 h. The result showed that there was a significant alteration in the tested parameters depending on treatment dose in the seeds exposed to cypermethrin when compared to the control group. Cypermethrin exposure significantly reduced the carotenoid, chlorophyll a and b pigments in all treatment groups. The activity of superoxide dismutase showed a concentration-time dependent increase and the maximum increase was observed on day 15 of treatment at 6.0 ppm cypermethrin exposure. The activity of catalase increased gradually with increasing cypermethrin concentration, but a soft decrease in CAT activity was decreased after 15 days of 1.5 ppm and 3.0 ppm cypermethrin treatment. In the roots treated with 1.5, 3.0, and 6.0 ppm cypermethrin, the level of malondialdehyde was about 1.8, 2.4, and 3.4 times higher than the control group, respectively. It was also found that cypermethrin has a mitodepressive action on mitosis, and the MI was decreased depending on the dose of cyprmethrin. All of the concentrations of cypermethrin induced chromosomal abnormalities and the most common abnormality observed in the present study was chromosome bridges.

Topics: Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Chromosome Aberrations; Chromosomes, Plant; Lipid Peroxidation; Malondialdehyde; Micronucleus Tests; Mitosis; Mitotic Index; Onions; Plant Cells; Plant Leaves; Plant Roots; Pyrethrins; Superoxide Dismutase

The temperature-dependent disassembly process of three monomeric isoforms, namely Lhcb1, Lhcb2, and Lhcb3, of the major light-harvesting chlorophyll (Chl) a/b complexes of photosystem II (LHCIIb) were characterized by observing the changes of absorption spectra, circular dichroism (CD), and dissociation processes of the bound pigments to the in vitro reconstituted complexes subjected to high temperatures. Our results suggest that the three isoforms of LHCIIb undergo conformational rearrangements, structural changes, and dissociations of the bound pigments when the ambient temperature increases from 20 to 90°C. The conformation of the complexes changed sensitively to the changing temperatures because the absorption peaks in the Soret region (436 and 471 nm) and the Qy region (650-660 and 680 nm) decreased immediately upon elevating the ambient temperatures. Analyzing temperature-dependent denaturing and pigment dissociation process, we can divide the disassembly process into three stages: The first stage, appeared from 20°C to around 50-60°C, was characterized by the diminishment of the absorption around 650-660 and 680 nm, accompanied by the blue-shift of the peak at 471 nm and disappearance of the absorbance at 436 nm, which is related to changes in the transition energy of the Chl b cluster, and the red-most Chl a cluster in the LHCIIb. The second stage, beginning at about 50-60°C, was signified by the diminishment of the CD signal between (+)483 nm and (-)490 nm, which implied the disturbance of dipole-dipole interaction of pigments, and the onset of the pigment dissociation. The last stage, beginning at about 70-80°C, indicates the complete dissociation of the pigments from the complex. The physiological aspects of the three stages in the denaturing process are also discussed.

Topics: Chlorophyll; Chlorophyll A; Chromatography, Thin Layer; Circular Dichroism; Energy Transfer; Hot Temperature; Kinetics; Light; Light-Harvesting Protein Complexes; Photosystem II Protein Complex; Pigments, Biological; Pisum sativum; Plant Proteins; Protein Isoforms; Recombinant Proteins; RNA, Plant; Spectrum Analysis

The Poisson-TrEsp method (where TrEsp stands for transition charges from electrostatic potentials) has been successfully applied to calculate excitonic couplings in a variety of pigment-protein complexes. It relies on an isomorphism that allows for relating the excitonic coupling between transition densities in dielectric media to their Coulomb coupling. This isomorphism was derived by Hsu et al. (J. Chem. Phys. 114, 3065, (2001)) using time-dependent density functional response theory. In this article, we provide an alternative and simple derivation by first-order perturbation theory. An application of Poisson-TrEsp to photosystem I trimers reveals that the local field correction/screening factor depends on the mutual orientation of the pigments and on the dielectric boundaries rather than on distance. A mean correction factor of f = 0.69 is determined for this system.

Topics: Chlorophyll; Chlorophyll A; Electric Conductivity; Linear Models; Models, Theoretical; Photosystem I Protein Complex; Poisson Distribution; Protein Multimerization; Static Electricity

Zygophyllum xanthoxylum, a C(3) woody species, is a succulent xerophyte that is well adapted to arid environments. Our previous investigations showed that Na(+) has a positive effect on the growth of Z. xanthoxylum under drought conditions, which was closely related to high Na(+) accumulation in leaves. To reveal the physiological mechanisms underlying how Na(+) accumulation improves the drought resistance of Z. xanthoxylum, 3-week-old seedlings were treated with a series of additional external NaCl concentrations (5-150 mM) in sand culture experiments. Seedlings were also subjected to water deficit (30% of field water capacity) in the presence or absence of additional NaCl (50 mM) in pot experiments. The results indicated that 50 mM NaCl could mitigate deleterious impacts of water deficit on the growth of Z. xanthoxylum, by improving the relative water content, inducing a significant drop in leaf water potential and, concomitantly, increasing leaf turgor pressure and chlorophyll concentrations resulting in an enhancement of overall plant photosynthetic activity (i.e., photosynthetic rate and water use efficiency). Furthermore, NaCl (50 mM) could alleviate the inhibitory effect of water deficit on the activity of photosystem II in Z. xanthoxylum. The contribution of Na(+) to the total osmotic potential varied from 8% in the control to 13% in plants subjected to water deficit and, surprisingly, to 28% in plants grown in the presence of 50 mM NaCl under water deficit; however, the contribution of K(+) significantly decreased from 13 to 8%. These findings suggest that, under arid environments, Z. xanthoxylum is able to accumulate a high concentration of Na(+) in its leaves and use it directly for osmotic adjustment, which was coupled with an improvement in leaf hydration and photosynthetic activity.

Topics: Chlorophyll; Chlorophyll A; Osmosis; Photosynthesis; Photosystem II Protein Complex; Plant Stomata; Potassium; Quantum Theory; Sodium; Sodium Chloride; Water; Zygophyllum

In view of the folklore use of green leaves to treat inflammation, the anti-inflammatory property of chlorophylls and their degradation products were studied. Chlorophyll a and pheophytin a (magnesium-free chlorophyll a) from fresh leaves showed potent anti-inflammatory activity against carrageenan-induced paw edema in mice and formalin-induced paw edema in rats. Chlorophyll a inhibited bacterial lipopolysaccharide-induced TNF-α (a pro-inflammatory cytokine) gene expression in HEK293 cells, but it did not influence the expression of inducible nitric acid synthase and cyclooxygenase-2 genes. Chlorophyll b only marginally inhibited both inflammation and TNF-α gene expression. But both chlorophyll a and chlorophyll b showed the same level of marginal inhibition on 12-O-tetradecanoyl-phorbol-13-acetate-induced NF-κB activation. Chlorophylls and pheophytins showed in vitro anti-oxidant activity. The study shows that chlorophyll a and its degradation products are valuable and abundantly available anti-inflammatory agents and promising for the development of phytomedicine or conventional medicine to treat inflammation and related diseases.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Carrageenan; Chlorophyll; Chlorophyll A; Chromolaena; Cyclooxygenase 2; Edema; Eupatorium; Formaldehyde; HEK293 Cells; Humans; Inflammation; Lipopolysaccharides; Mice; Moraceae; NF-kappa B; Nitric Oxide Synthase Type II; Pheophytins; Plant Extracts; Rats; Rats, Wistar; Tetradecanoylphorbol Acetate; Tumor Necrosis Factor-alpha

Changes in function and composition of the photosynthetic apparatus as well as the ultrastructure of chloroplasts in mesophyll cells were analyzed in flag leaves of the high-yield barley (Hordeum vulgare) variety cv. Lomerit during senescence under field conditions in two successive years. In contrast to previous results obtained with the elder variety cv. Carina photosystem II efficiency measured by F(v)/F(m) was found to be rather stable until a very late stage of senescence. Chlorophyll a fluorescence and P700 absorbance measurements revealed that the activities of the two photosystems declined in parallel. An increase in the chlorophyll a/b ratio at a late stage of senescence was observed to coincide with a decline in the level of the Lhcb1 apoprotein of the light harvesting complex (LHC) and the level of the corresponding transcript. Ultrastructural investigations revealed the presence of gerontoplasts that have long, single or pairwise thylakoids and lack large grana stacks. It is hypothesized that the early degradation of grana thylakoids harboring the major LHC reduced the risk of photoinhibition and might be causally related to the high yield of the barley variety cv. Lomerit.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Fluorescence; Gene Expression Regulation, Plant; Hordeum; Immunoblotting; Light-Harvesting Protein Complexes; Mesophyll Cells; Photosynthesis; Plant Leaves; Protein Subunits; Quantum Theory; Real-Time Polymerase Chain Reaction; Ribulose-Bisphosphate Carboxylase; RNA, Messenger

The structure of the major light-harvesting chlorophyll a/b complex (LHCII) was analyzed by pulsed EPR measurements and compared with the crystal structure. Site-specific spin labeling of the recombinant protein allowed the measurement of distance distributions over several intra- and intermolecular distances in monomeric and trimeric LHCII, yielding information on the protein structure and its local flexibility. A spin label rotamer library based on a molecular dynamics simulation was used to take the local mobility of spin labels into account. The core of LHCII in solution adopts a structure very similar or identical to the one seen in crystallized LHCII trimers with little motional freedom as indicated by narrow distance distributions along and between α helices. However, distances comprising the lumenal loop domain show broader distance distributions, indicating some mobility of this loop structure. Positions in the hydrophilic N-terminal domain, upstream of the first trans-membrane α helix, exhibit more and more mobility the closer they are to the N terminus. The nine amino acids at the very N terminus that have not been resolved in any of the crystal structure analyses give rise to very broad and possibly bimodal distance distributions, which may represent two families of preferred conformations.

Topics: Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Pisum sativum; Protein Structure, Quaternary; Protein Structure, Secondary; Protein Structure, Tertiary; Spin Labels

In shoot cultures of Thymus daenensis, hyperhydricity syndrome promoted by benzyladenine (BA) is characterised by the development of chlorophyll-deficient shoots with a high water content and reduced growth that is less differentiated. By removing the BA from the culture medium, the hyperhydricity was reversed, and the reversion toward a normal growth in vitro was more efficient in shoots treated with 5 μM of salicylic acid (SA), showing a significant increase in chlorophyll b after 4 weeks of culture. In the present study, the effect of salicylic acid on the reversion of shoot hyperhydricity was investigated at the level of the free, soluble and insoluble conjugated polyamine content. In T. daenensis micropropagated shoots, the level of polyamines was high, with a predominance of putrescine. BA, which triggered hyperhydricity, caused a reduction of the polyamine (PA) content by one-half due to a decrease in the putrescine content and insoluble conjugated PAs that were not detected in the hyperhydric shoots. In the reverted shoots, changes of the free polyamines, spermidine and, more notably, spermine, were shown. The spermine content doubled after 4 weeks of culture, and its amount was the same as that found in normal shoots, suggesting that free spermine could be particularly involved in the reversion of hyperhydricity. In the SA-reverted tissues, the PA pattern was marked with a transient increase of free putrescine, spermidine and spermine and an enhancement of soluble conjugated spermine. This transitory SA-dependent amplification of PAs was concomitant with a remarkable transient increase of H(2)O(2), suggesting that SA may be implicated in PA signalling pathways for tissue differentiation during the reversion of hyperhydricity in T. daenensis.

Topics: Benzyl Compounds; Carotenoids; Cell Differentiation; Chlorophyll; Chlorophyll A; Culture Media; Culture Techniques; Hydrogen Peroxide; Plant Diseases; Plant Shoots; Polyamines; Putrescine; Salicylic Acid; Solubility; Spermine; Thymus Plant; Time Factors

The effects of changes in the chlorophyll (chl) content on the kinetics of the OJIP fluorescence transient were studied using two different approaches. An extensive chl loss (up to 5-fold decrease) occurs in leaves suffering from either an Mg(2+) or SO(4)(2-) deficiency. The effects of these treatments on the chl a/b ratio, which is related to antenna size, were very limited. This observation was confirmed by the identical light intensity dependencies of the K, J and I-steps of the fluorescence rise for three of the four treatments and by the absence of changes in the F(685 nm)/F(695 nm)-ratio of fluorescence emission spectra measured at 77K. Under these conditions, the F(0) and F(M)-values were essentially insensitive to the chl content. A second experimental approach consisted of the treatment of wheat leaves with specifically designed antisense oligodeoxynucleotides that interfered with the translation of mRNA of the genes coding for chl a/b binding proteins. This way, leaves with a wide range of chl a/b ratios were created. Under these conditions, an inverse proportional relationship between the F(M) values and the chl a/b ratio was observed. A strong effect of the chl a/b ratio on the fluorescence intensity was also observed for barley Chlorina f2 plants that lack chl b. The data suggest that the chl a/b ratio (antenna size) is a more important determinant of the maximum fluorescence intensity than the chl content of the leaf.

Topics: Beta vulgaris; Chlorophyll; Chlorophyll A; Fluorescence; Hordeum; Magnesium; Oligonucleotides, Antisense; Plant Leaves; Plant Proteins; Sulfates; Triticum

The fate of chloroplast DNA (cpDNA) during plastid development and conversion between various plastid types is still not very well understood. This is especially true for the cpDNA found in plastids of naturally senescing leaves. Here, we describe changes in plastid nucleoid structure accompanied with cpDNA degradation occurring during natural senescence of the free-growing deciduous woody species Acer pseudoplatanus L. Natural senescence was investigated using three types of senescing leaves: green (G), yellow-green (YG) and yellow (Y). The extent of senescence was evaluated at the level of photosynthetic pigment degradation, accumulation of starch and plastid ultrastructure. Determination of cpDNA amount was carried out by in planta visualization with 4,6-diamidino-2-phenylindole, by Southern hybridization, and by dot-blot using an rbcL gene probe. During natural senescence, plastid nucleoids undergo structural rearrangements accompanied by an almost complete loss of cpDNA. Furthermore, senescence-associated protein components exhibiting strong binding to an ∼10kbp rbcL-containg cpDNA fragment were identified. This interaction might be important for rbcL expression and Rubisco degradation during the course of natural senescence in trees.

Topics: Acer; Carotenoids; Cellular Senescence; Chlorophyll; Chlorophyll A; Chloroplasts; Climate; DNA, Chloroplast; Plant Leaves; Ribulose-Bisphosphate Carboxylase; Starch

The spectral reflectance signature of living organisms provides information that closely reflects their physiological status. Because of its high potential for the estimation of geomorphic biological parameters, particularly of gross photosynthesis of plants, two-dimensional spectroscopy, via the use of hyperspectral instruments, has been widely used in remote sensing applications. In genetics research, in contrast, the reflectance phenotype has rarely been the subject of quantitative analysis; its potential for illuminating the pathway leading from the gene to phenotype remains largely unexplored. In this study, we employed hyperspectral imaging techniques to identify Arabidopsis mutants with altered leaf pigment status. The techniques are comprised of two modes; the first is referred to as the 'targeted mode' and the second as the 'non-targeted mode'. The 'targeted' mode is aimed at visualizing individual concentrations and compositional parameters of leaf pigments based on reflectance indices (RIs) developed for Chls a and b, carotenoids and anthocyanins. The 'non-targeted' mode highlights differences in reflectance spectra of leaf samples relative to reference spectra from the wild-type leaves. Through the latter approach, three mutant lines with weak irregular reflectance phenotypes, that are hardly identifiable by simple observation, were isolated. Analysis of these and other mutants revealed that the RI-based targeted pigment estimation was robust at least against changes in trichome density, but was confounded by genetic defects in chloroplast photorelocation movement. Notwithstanding such a limitation, the techniques presented here provide rapid and high-sensitive means to identify genetic mechanisms that coordinate leaf pigment status with developmental stages and/or environmental stress conditions.

Topics: Agrobacterium; Anthocyanins; Arabidopsis; Carotenoids; Chlorophyll; Chlorophyll A; Chloroplasts; Chromosome Mapping; Chromosomes, Plant; Genetic Complementation Test; Genetic Loci; Image Processing, Computer-Assisted; Phenotype; Plant Leaves; Spectrum Analysis

The possible use of municipal solid waste compost (MSWC) in the production of marigold and basil seedlings examined. Six medium prepared from commercial peat (CP) and MSWC (0, 15, 30, 45, 60, and 100% v/v). There was not any plant growth when MSWC used alone (100%). The addition of MSWC in low content (15% and 30%) improved seed emergence for marigold and basil respectively, while greater content revealed opposed impacts. Mean emergence time delayed as MSWC content increased into substrates. Addition of MSWC (especially in content greater than 30%) into CP reduced (from 34 to 64%) plant height, leaf number and stem diameter as a consequence reduced plant fresh weight (plant biomass) for both species. The number of lateral stems decreased (up to 81%) in basil when MSWC added into substrate mixtures. Chlorophyll b content decreased (up to 58%) in substrates with MSWC content greater than 15% or 30% while similar reduction observed in content of Chlorophyll a and total carotenoids for basil with MSWC > 60%. However, Chlorophyll a and total carotenoids content increased as MSWC content increased for marigold. K and Na leaf content increased but P equivalent decreased as MSWC content increased. Nursery-produced basil and marigold seedlings grown in 15% MSWC; displayed quality indices similar to those recorded for conventional mixtures of peat and may act as component substitute.

Topics: Asteraceae; Biomass; Chlorophyll; Chlorophyll A; Ocimum basilicum; Plant Leaves; Plant Stems; Refuse Disposal; Seedlings; Soil

Taraxacum formosanum, a well-known Chinese herb shown to be protective against hepatic cancer as well as liver and lung damage, may be attributed to the presence of abundant carotenoids and chlorophylls. However, the variety and content of chlorophylls remain uncertain. The objectives of this study were to develop an high-performance liquid chromatography-diode array detection-mass spectrometry method for determination of chlorophylls in T. formosanum and preparation by column chromatography. An HyPURITY C18 column and a gradient mobile phase of water (A), methanol (B), acetonitrile (C), and acetone (D) could resolve 10 chlorophylls and an internal standard Fast Green FCF within 30 min with a flow rate at 1 mL/min and detection at 660 nm. Both chlorophylls a and a' were present in the largest amount (1389.6 μg/g), followed by chlorophylls b and b' (561.2 μg/g), pheophytins a and a' (31.7 μg/g), hydroxychlorophyll b (26.5 μg/g), hydroxychlorophylls a and a' (9.8 μg/g), and chlorophyllides a and a' (0.35 μg/g). A glass column containing 52 g of magnesium oxide-diatomaceous earth (1:3, w/w) could elute chlorophylls with 800 mL of acetone containing 50% ethanol at a flow rate of 10 mL/min. Some new chlorophyll derivatives including chlorophyllide b, pyropheophorbide b, hydroxypheophytin a, and hydroxypheophytin a' were generated during column chromatography but accompanied by a 63% loss in total chlorophylls. Thus, the possibility of chlorophyll fraction prepared from T. formosanum as a raw material for future production of functional food needs further investigation.

Topics: Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Mass Spectrometry; Pheophytins; Taraxacum

Enantioselectivity in ecotoxicity of chiral pesticides in the aquatic environment has been a subject of growing interest. In this study, the toxicological impacts of hexaconazole enantiomers were investigated with freshwater algae Scenedesmus obliquus. After 96 h of exposure, the EC(50) values for rac-hexaconazole, (+)-hexaconazole, and (-)-hexaconazole were 0.178, 0.355, and 0.065 mg l(-1) , respectively. Therefore, the acute toxicities of hexaconazole enantiomers were enantioselective. In addition, the different toxic effects were evaluated when S. obliquus were exposed to 0.2, 0.5, and 1.0 mg l(-1) of rac-hexaconazole, (+)-hexaconazole, and (-)-hexaconazole during 96 h, respectively. The chlorophyll a and chlorophyll b contents of S. obliquus treated by (-)-hexaconazole were lower than those exposed to (+)-hexaconazole, whereas the malondialdehyde contents of S. obliquus treated by (-)-form were higher than those exposed to (+)-form at higher concentrations. In general, catalase activities were significantly upregulated by exposure to (-)-enantiomer than (+)-enantiomer at all three concentrations. However, superoxide dismutase activities exposed to (-)-hexaconazole were lower than that exposed to (+)-hexaconazole at 0.2 mg l(-1) and 0.5 mg l(-1) . On the basis of these data, the acute toxicity and toxic effects of hexaconazole against S. obliquus were enantioselective, and such enantiomeric differences must be taken into consideration in pesticide risk assessment.

Topics: Antioxidants; Biomarkers; Chlorophyll; Chlorophyll A; Chlorophyta; Ecotoxicology; Environmental Pollutants; Malondialdehyde; Oxidative Stress; Stereoisomerism; Time Factors; Triazoles

Chlorophyll is a deleterious molecule that generates reactive oxygen species and must be converted to nontoxic molecules during plant senescence. The degradation pathway of chlorophyll a has been determined; however, that of chlorophyll b is poorly understood, and multiple pathways of chlorophyll b degradation have been proposed. In this study, we found that chlorophyll b is degraded by a single pathway, and elucidated the importance of this pathway in avoiding cell death. In order to determine the chlorophyll degradation pathway, we first examined the substrate specificity of 7-hydroxymethyl chlorophyll a reductase. 7-hydroxymethyl chlorophyll a reductase reduces 7-hydroxymethyl chlorophyll a but not 7-hydroxymethyl pheophytin a or 7-hydroxymethyl pheophorbide a. These results indicate that the first step of chlorophyll b degradation is its conversion to 7-hydroxymethyl chlorophyll a by chlorophyll b reductase, although chlorophyll b reductase has broad substrate specificity. In vitro experiments showed that chlorophyll b reductase converted all of the chlorophyll b in the light-harvesting chlorophyll a/b protein complex to 7-hydroxymethyl chlorophyll a, but did not completely convert chlorophyll b in the core antenna complexes. When plants whose core antennae contained chlorophyll b were incubated in the dark, chlorophyll b was not properly degraded, and the accumulation of 7-hydroxymethyl pheophorbide a and pheophorbide b resulted in cell death. This result indicates that chlorophyll b is not properly degraded when it exists in core antenna complexes. Based on these results, we discuss the importance of the proper degradation of chlorophyll b.

Topics: Alcohol Oxidoreductases; Arabidopsis; Arabidopsis Proteins; Cell Death; Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Magnesium; Phenotype; Photosystem I Protein Complex; Substrate Specificity

The chlorophyll content is an important experimental parameter in agronomy and plant biology research. In this report, we explore the feasibility of determining total concentration of extracts containing chlorophyll a and chlorophyll b by chlorophyll fluorescence. We found that an excitation at 457 nm results in the same integrated fluorescence emission for a molecule of chlorophyll a and a molecule of chlorophyll b. The fluorescence yield induced by 457 nm is therefore proportional to total molar chlorophyll concentration. Based on this observation, we designed an instrument to determine total chlorophyll concentrations. A single light emitting diode (LED) is used to excite chlorophyll extracts. After passing through a long-pass filter, the fluorescence emission is assessed by a photodiode. We demonstrate that this instrument facilitates the determination of total chlorophyll concentrations. We further extended the functionality of the instrument by including LEDs emitting at 435 and 470 nm wavelengths, thereby preferentially exciting chlorophyll a and chlorophyll b. This instrument can be used to determine chlorophyll a and chlorophyll b concentrations in a variety of organisms containing different ratios of chlorophylls. Monte-Carlo simulations are in agreement with experimental data such that a precise determination of chlorophyll concentrations in carotenoid-containing biological samples containing a concentration of less than 5 nmol/mL total chlorophyll can be achieved.

Topics: Arabidopsis; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Chromatography, Thin Layer; Fluorescence; Kinetics; Light; Liriodendron; Lolium; Monte Carlo Method; Nicotiana; Spectrometry, Fluorescence; Synechocystis

A field study was conducted to investigate the impact of ambient solar UV on the various growth, physiological and yield parameters of four sorghum (Sorghum bicolor L.) varieties-Indore-12, Indore-26, CSV-23 and Indore-27 by excluding either UV-B (<315 nm) or UV-A/B (<400 nm) components of solar spectrum. Exclusion of UV significantly enhanced plant height, area and specific leaf weight of flag leaf, biomass accumulation, yield parameters and harvest index in all the sorghum varieties. Chlorophyll b was significantly enhanced and chlorophyll a increased to a lesser extent, UV-B absorbing substances and chlorophyll a/b ratio were significantly decreased by the exclusion of solar UV. The enhancement in the vegetative growth and yield by UV exclusion might be linked to the remarkable increase in rate of photosynthesis in sorghum varieties. The magnitude of the response was high in I-26 and I-27 as compared to CSV-23 and I-12 after exclusion of solar UV. All the varieties of sorghum had a negative cumulative stress response index (CSRI), the sensitivity of the sorghum varieties was in the following sequence I-12>CSV-23>I-26>I-27. Thus I-27 was the most sensitive and I-12 the least sensitive variety to present level of solar UV radiation. The differences in UV sensitivity identified among sorghum varieties might be useful in breeding programs for increased tolerance to UV-B radiation.

Topics: Adaptation, Physiological; Biomass; Chlorophyll; Chlorophyll A; Photosynthesis; Plant Leaves; Sorghum; Species Specificity; Ultraviolet Rays

This study investigated the removal of nitrogen and phosphorus from the effluent of a submerged anaerobic membrane bioreactor (SAnMBR) by means of a lab-scale photobioreactor in which algae biomass was cultured in a semi-continuous mode for a period of 42 days. Solids retention time was 2 days and a stable pH value in the system was maintained by adding CO(2). Nitrogen and phosphorus concentrations in the SAnMBR effluent fluctuated according to the operating performance of the bioreactor and the properties of its actual wastewater load. Despite these variations, the anaerobic effluent proved to be a suitable growth medium for microalgae (mean biomass productivity was 234 mg l(-1)d(-1)), achieving a nutrient removal efficiency of 67.2% for ammonium (NH(4)(+)-N) and 97.8% for phosphate (PO(4)(-3)-P). When conditions were optimum, excellent water quality with very low ammonium and phosphate concentrations was obtained.

Topics: Anaerobiosis; Biomass; Bioreactors; Cell Count; Chlorophyll; Chlorophyll A; Membranes, Artificial; Microalgae; Nitrogen; Phosphorus; Photobioreactors; Pilot Projects; Solubility; Waste Disposal, Fluid; Wastewater

A central goal of plant ecophysiological studies is to generate patterns of physiological behavior that are applicable to a species, which can be complicated when plant size is considered. Studies indicate that plant size can influence numerous ecophysiological parameters, especially in vascular epiphytes. The few studies that have included ferns in their analyses suggest that plant size is less important in ferns than angiosperms. This study investigates this apparent disparity by examining the relationship between plant size and an array of ecophysiological parameters in the epiphytic fern Asplenium auritum, especially the role of plant size in determining responses to water stress.. Plants were classified according to size and measured for a variety of functional traits, including maximum photosynthetic rate, stomatal conductance, water-use efficiency, stomatal density, chlorophyll content, chlorophyll a/b ratio, specific leaf area, whole plant drying rates, and desiccation tolerance. Results for all traits were compared across size classes to determine size-related differences.. Plant size significantly influenced most traits examined, most notably photosynthetic rate, stomatal density, stomatal conductance, whole plant drying rates, and recovery from desiccation. We report the first evidence of size-mediated shifts in desiccation tolerance in plants: small individuals tolerated water loss, whereas larger individuals avoided desiccation.. Our findings indicate that size-mediated ecophysiological shifts are more important than previously thought in ferns, particularly in regard to water relations. Desiccation tolerance may allow young fern sporophytes to establish in stressful environments and is subsequently lost in older individuals.

Topics: Analysis of Variance; Chlorophyll; Chlorophyll A; Costa Rica; Ferns; Oxygen Consumption; Photosynthesis; Plant Leaves; Plant Stomata; Water

By using a free-air controlled enrichment (FACE) system, this paper studied the effects of elevated atmospheric ozone (O3) concentration (150% of ambient O3) on the flag leaf photosynthetic pigment contents of wheat varieties Yannong 19, Yangmai 16, Jiaxin 002, Yangmai 15, and Yangfumai 2. For the test varieties, no significant differences were observed in the flag leaf chlorophyll a, chlorophyll b, chlorophyll (a+b), and carotenoid contents between treatments elevated O3 concentration and ambient O3 at booting and anthesis stages, but the photosynthetic pigment contents in treatment elevated O3 concentration all decreased after anthesis, with a significant decrease of chlorophyll a, chlorophyll b, and chlorophyll (a+b) contents, which indicated that elevated O3 had minor effects on the synthesis of photosynthetic pigments but accelerated their decline process. Different wheat varieties had genetic difference in the responses of flag leaf photosynthetic pigment contents to elevated O3, among which, Yangmai 15 and Jiaxin 002 had better tolerance to ozone stress. The flag leaf chlorophyll a, chlorophyll b, and chlorophyll (a+b) contents at grain-filling stage (about 21 days after anthesis) had a significant positive correlation with the 1000-grain mass.

Topics: Atmosphere; Chlorophyll; Chlorophyll A; Ozone; Photosynthesis; Plant Leaves; Triticum

Sexual dimorphisms of dioecious plants are important in controlling and maintaining sex ratios under changing climate environments. Yet, little is known about sex-specific responses to elevated CO(2) with soil nitrogen (N) deposition. To investigate sex-related physiological and biochemical responses to elevated CO(2) with N deposition, Populus cathayana Rehd. was employed as a model species. The cuttings were subjected to two CO(2) regimes (350 and 700 μmol mol(-1)) with two N levels (0 and 5 g N m(-2) year(-1)). Our results showed that elevated CO(2) and N deposition separately increased the total number of leaves, leaf area (LA), leaf mass, net photosynthetic rate (P(n)), light saturated photosynthetic rate (P(max)), chlorophyll a (Chl a), and chlorophyll a to chlorophyll b ratio (Chl a/b) in both males and females of P. cathayana. However, the effects on LA, leaf mass, P(n), P(max), Chl a and Chl a/b were weakened under the combined treatment of elevated CO(2) and N deposition. Males had higher leaf mass, P(n), P(max), apparent quantum yield (Φ), carboxylation efficiency (CE), Chl a, Chl a/b, leaf N, and root carbon to N ratio (C/N) than did females under elevated CO(2) with N deposition. In contrast to males, females had significantly higher levels of soluble sugars in leaves and greater starch accumulation in roots and stems under the same condition. The results of the present work imply that P. cathayana females are more responsive and suffer from greater negative effects on growth and photosynthetic capacity than do males when grown under elevated CO(2) with soil N deposition.

Topics: Carbohydrate Metabolism; Carbon; Carbon Dioxide; Chlorophyll; Chlorophyll A; Nitrogen; Photosynthesis; Populus; Starch

A broad regional understanding of tropical forest leaf photosynthesis has long been a goal for tropical forest ecologists, but it has remained elusive due to difficult canopy access and high species diversity. Here we develop an empirical model to predict sunlit, light-saturated, tropical leaf photosynthesis using leaf and simulated canopy spectra. To develop this model, we used partial least squares (PLS) analysis on three tropical forest datasets (159 species), two in Hawaii and one at the biosphere 2 laboratory (B2L). For each species, we measured light-saturated photosynthesis (A), light and CO(2) saturated photosynthesis (A(max)), respiration (R), leaf transmittance and reflectance spectra (400-2,500 nm), leaf nitrogen, chlorophyll a and b, carotenoids, and leaf mass per area (LMA). The model best predicted A [r(2) = 0.74, root mean square error (RMSE) = 2.9 μmol m(-2) s(-1))] followed by R (r(2) = 0.48), and A(max) (r(2) = 0.47). We combined leaf reflectance and transmittance with a canopy radiative transfer model to simulate top-of-canopy reflectance and found that canopy spectra are a better predictor of A (RMSE = 2.5 ± 0.07 μmol m(-2) s(-1)) than are leaf spectra. The results indicate the potential for this technique to be used with high-fidelity imaging spectrometers to remotely sense tropical forest canopy photosynthesis.

Topics: Carbon Dioxide; Carotenoids; Cell Respiration; Chlorophyll; Chlorophyll A; Ecosystem; Hawaii; Light; Nitrogen; Photosynthesis; Plant Leaves; Plant Physiological Phenomena; Regression Analysis; Trees; Tropical Climate

Oryza rufipogon L. (O. rufipogon) or a common wild rice, showed considerable aluminum (Al) tolerance. In this study, we examined the physiologic and genetic response of wild rice short term and long term to Al toxicity, respectively. In the short term study, morin staining, DAPI staining and aniline blue staining were used to detect Al distribution, cell division and callose production in the roots of O. rufipogon. The results indicated cell division could be enhanced by Al within low concentration range. In the long term study, we chose Oryza sativa L (O. sativa) (the close sib of O. rufipogon) as a reference. It showed that O. rufipogon grew better than O. sativa when treated with Al of 1.4 mmol/l concentration and also experienced a short period of root growth stimulation. This study gave some basic data to explain the mechanisms Oryza rufipogon L. developed to deal with Al and lay a good foundation to further study. SSH (suppression subtractive hybridization) proved that transcripts of the small subunit of Rubisco and a Photosystem I P700 apoprotein were enhanced under long term Al treatment in wild rice. Further investigation via the assessment of the content of chlorophyll a, b indicated that the content of chlorophyll a, b in the leaves of O. rufipogon generally rose after Al treatment for 15 days. This indicated that intake of Al can affect photosynthesis of plant.

Topics: Aluminum; Aniline Compounds; Blotting, Northern; Cell Division; Chlorophyll; Chlorophyll A; Electrophoresis, Polyacrylamide Gel; Gene Expression Regulation, Plant; Nucleic Acid Hybridization; Oryza; Photosynthesis; Plant Leaves; Plant Roots; Ribulose-Bisphosphate Carboxylase; Time Factors

To evaluate the physiological response of Tillandsia capillaris Ruiz & Pav. f. capillaris, T. recurvata L., and T. tricholepis Baker to different air pollution sources, epiphyte samples were collected from a noncontaminated area in the province of Córdoba (Argentina) and transplanted to a control site as well as three areas categorized according to the presence of agricultural, urban, and industrial (metallurgical and metal-mechanical) emission sources. A foliar damage index (FDI) was calculated with the physiological parameters chlorophyll a, chlorophyll b, malondialdehyde (MDA), hydroperoxyconjugated dienes, sulfur (S) content, and dry weight-to-fresh weight ratio. In addition, electrical conductivity (E-cond), relative water content (RWC), dehydration kinetics (Kin-H(2)O), total phenols (T-phen), soluble proteins (S-prot), and activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase were determined. The parameters E-cond, FDI, SOD, RWC, and Kin-H(2)O can serve as suitable indicators of agricultural air pollution for T. tricholepis and T. capillaris, and CAT, Kin-H(2)O, and SOD can do the same for T. recurvata. In addition, MDA, T-phen, and S-prot proved to be appropriate indicators of urban pollution for T. recurvata. Moreover, FDI, E-cond, and SOD for T. recurvata and MDA for T. tricholepis, respectively, could be used to detect deleterious effects of industrial air pollution.

Topics: Air Pollutants; Air Pollution; Antioxidants; Argentina; Ascorbate Peroxidases; Catalase; Chlorophyll; Chlorophyll A; Environmental Monitoring; Malondialdehyde; Plant Leaves; Sulfur; Superoxide Dismutase; Tillandsia

Excitation energy transfer in the light-harvesting complex II of higher plants is modeled using excitonic couplings and local transition energies determined from structure-based calculations recently (Müh et al., 2010). A theory is introduced that implicitly takes into account protein induced dynamic localization effects of the exciton wavefunction between weakly coupled optical and vibronic transitions of different pigments. Linear and non-linear optical spectra are calculated and compared with experimental data reaching qualitative agreement. High-frequency intramolecular vibrational degrees of freedom are found important for ultrafast subpicosecond excitation energy transfer between chlorophyll (Chl) b and Chla, since they allow for fast dissipation of the excess energy. The slower ps component of this transfer is due to the monomeric excited state of Chlb 605. The majority of exciton relaxation in the Chla spectral region is characterized by slow ps exciton equilibration between the Chla domains within one layer and between the lumenal and stromal layers in the 10-20ps time range. Subpicosecond exciton relaxation in the Chla region is only found within the terminal emitter domain (Chls a 610/611/612) and within the Chla 613/614 dimer. Deviations between measured and calculated exciton state life times are obtained for the intermediate spectral region between the main absorbance bands of Chla and Chlb that indicate that besides Chlb 608 another pigment should absorb there. Possible candidates, so far not identified by structure-based calculations, but by fitting of optical spectra and mutagenesis studies, are discussed. Additional mutagenesis studies are suggested to resolve this issue.

Topics: Chlorophyll; Chlorophyll A; Circular Dichroism; Energy Transfer; Kinetics; Light-Harvesting Protein Complexes; Models, Molecular; Photosystem II Protein Complex; Protein Multimerization; Spectrometry, Fluorescence; Temperature; Vibration

In deep lakes, water column stratification isolates the surface water from the deeper bottom layers, creating a three dimensional differentiation of the chemical, physical, biological and optical characteristics of the waters. Chromophoric dissolved organic matter (CDOM) and total suspended solids (TSS) play an important role in the attenuation of ultraviolet and photosynthetically active radiation. In the present analysis of spectral irradiance, we show that the wavelength composition of the metalimnetic visible irradiance was influenced by epilimnetic spatial distribution of CDOM. We found a low occurrence of blue-green photons in the metalimnion where epilimnetic concentrations of CDOM are high. In this field study, the spatial variation of the spectral irradiance in the metalimnion correlates with the observed metalimnetic concentrations of chlorophyll a as well as chlorophyll a : chlorophyll b/c ratios. Dissolved oxygen, pH, and nutrients trends suggest that chlorophyll a concentrations were representative of the phytoplankton biomass and primary production. Thus, metalimnetic changes of spectral irradiance may have a direct impact on primary production and an indirect effect on the spatial trends of pH, dissolved oxygen, and inorganic nutrients in the metalimnion.

Topics: Biomass; Chlorophyll; Chlorophyll A; Hydrogen-Ion Concentration; Oxygen; Phytoplankton; Seasons; Ultraviolet Rays; Water

The host plant Helianthemum sessiliflorum was inoculated with the mycorrhizal desert truffle Terfezia boudieri Chatin, and the subsequent effects of the ectomycorrhizal relationship on host physiology were determined. Diurnal measurements revealed that mycorrhizal (M) plants had higher rates of photosynthesis (35%), transpiration (18%), and night respiration (49%) than non-mycorrhizal (NM) plants. Consequently, M plants exhibited higher biomass accumulation, higher shoot-to-root ratios, and improved water use efficiency compared to NM plants. Total chlorophyll content was higher in M plants, and the ratio between chlorophyll a to chlorophyll b was altered in M plants. The increase in chlorophyll b content was significantly higher than the increase in chlorophyll a content (2.58- and 1.52-fold, respectively) compared to control. Calculation of the photosynthetic activation energy indicated lower energy requirements for CO(2) assimilation in M plants than in NM plants (48.62 and 61.56 kJ mol(-1), respectively). Continuous measurements of CO(2) exchange and transpiration in M plants versus NM plants provided a complete picture of the daily physiological differences brought on by the ectomycorrhizal relationships. The enhanced competence of M plants to withstand the harsh environmental conditions of the desert is discussed in view of the mycorrhizal-derived alterations in host physiology.

Topics: Ascomycota; Biomass; Carbon Dioxide; Chlorophyll; Chlorophyll A; Cistaceae; Energy Metabolism; Photosynthesis; Plant Roots; Plant Shoots; Plant Transpiration; Symbiosis

Difference fluorescence line-narrowing spectroscopy at 4.5 K was employed to investigate electron-phonon and electron-vibrational coupling strengths of the lower exciton level of water-soluble chlorophyll-binding protein (WSCP) from cauliflower reconstituted with chlorophyll a or chlorophyll b, respectively. The electron-phonon coupling is found to be moderate with integral Huang-Rhys factors S in the order of 0.81-0.85. A weak dependence of S on excitation wavelength within the inhomogeneously broadened fluorescence origin band is attributed to a sizable contribution of nonresonant excitation that varies with excitation wavelength. The strongly asymmetric and highly structured one-phonon profile is characterized by a peak phonon frequency (ω(m)) of ~24 cm(-1) and further discernible peaks at 48 and 88 cm(-1), respectively. A structural assignment of this unusual one-phonon profile is proposed. As will be shown in the accompanying paper (part II) (DOI 10.1021/jp111457t), the parameters of electron-phonon coupling readily account for shape and position of the fluorescence origin bands at 666.1 and 683.8 nm for chlorophyll b- and chlorophyll a-WSCP, respectively. A rich structure of S(1)→S(0) vibrational frequencies was resolved in the wavenumber range between 180 and 1665 cm(-1) for both chlorophyll a- and chlorophyll b-WSCP. The corresponding individual Huang-Rhys factors fall in the range between 0.0011 and 0.0500. To the best of our knowledge, this is the first report of S-factors for vibrational modes of chlorophyll b. Most remarkable is the presence of two additional modes at 228 and 327 cm(-1) compared with the vibrational spectrum of chlorophyll in solution. The additional modes can most likely be attributed to H-bond formation in the vicinity of the chlorophyll molecule bound by WSCP.

Topics: Brassica; Chlorophyll; Chlorophyll A; Electrons; Light-Harvesting Protein Complexes; Recombinant Proteins; Spectrometry, Fluorescence; Thermodynamics; Water

Persistent spectral hole burning at 4.5 K has been used to investigate the excitonic energy level structure and the excited state dynamics of the recombinant class-IIa water-soluble chlorophyll-binding protein (WSCP) from cauliflower. The hole-burned spectra are composed of four main features: (i) a narrow zero-phonon hole (ZPH) at the burn wavelength, (ii) a number of vibrational ZPHs, (iii) a broad low-energy hole at ~665 and ~683 nm for chlorophyll b- and chlorophyll a-WSCP, respectively, and (iv) a second satellite hole at ~658 and ~673 nm for chlorophyll b- and chlorophyll a-WSCP, respectively. The doublet of broad satellite holes is assigned to an excitonically coupled chlorophyll dimer. The lower-energy holes at ~665 and ~683 nm for chlorophyll b- and chlorophyll a-WSCP, respectively, represent the lower exciton states. Taking into account the parameters of electron-phonon coupling, the lower exciton state can be assigned as the fluorescence origin. The lower exciton state is populated by two processes: (i) exciton relaxation from the higher exciton state and (ii) vibrational relaxation within the lower exciton state. Assuming identical site energies for the two excitonically coupled chlorophyll molecules, the dipole-dipole interaction energy J is directly determined to be 85 and 100 cm(-1) for chlorophyll b- and chlorophyll a-WSCP, respectively, based on the positions of the satellite holes. The Gaussian low-energy absorption band identified by constant fluence hole burning at 4.5 K has a width of ~150 cm(-1) and peaks at 664.9 and 682.7 nm for chlorophyll b- and chlorophyll a-WSCP, respectively. The action spectrum is broader and blue-shifted compared to the fluorescent lower exciton state. This finding can be explained by a slow protein relaxation between energetically inequivalent conformational substates within the lowest exciton state in agreement with the results of Schmitt et al. (J. Phys. Chem. B2008, 112, 13951).

Topics: Brassica; Chlorophyll; Chlorophyll A; Electrons; Light-Harvesting Protein Complexes; Recombinant Proteins; Spectrometry, Fluorescence; Thermodynamics; Vibration; Water

The generation of ROS (reactive oxygen species) in plant galls may induce the degradation of the membrane systems of a plant cell and increase the number of plastoglobules. This numerical increase has been related to the prevention of damage to the thylakoid systems, and to the maintenance of photosynthesis rates. To investigate this hypothesis in gall systems, a comparative study of the ultrastructure of chloroplasts in non-galled leaves and in leaf galls of A. australe and A. spruceanum was conducted. Also, the pigment composition and the photosynthetic performance as estimated by chlorophyll fluorescence measurements were evaluated. The ultrastructural analyses revealed an increase in the number and size of plastoglobules in galls of both species studied. The levels of total chlorophylls and carotenoids were lower in galls than in non-galled tissues. The chlorophyll a/b ratio did not differ between the non-galled tissues and both kinds of galls. The values of maximum electron transport rate (ETR(MAX)) were similar for all the samples. The occurrence of numerous large plastoglobules in the galled tissues seemed to be related to oxidative stress and to the recovery of the thylakoid membrane systems. The maintenance of the ETR(MAX) values indicated the existence of an efficient strategy to maintain similar photosynthetic rates in galled and non-galled tissues.

Topics: Aspidosperma; Carotenoids; Chlorophyll; Chlorophyll A; Chloroplasts; Electron Transport; Fluorescence; Lipoproteins; Oxidative Stress; Photosynthesis; Plant Leaves; Plant Proteins; Plant Tumors; Reactive Oxygen Species; Thylakoids

Modifications in growing techniques can affect the yield and nutritional quality of various cultivated plant species. Owing to its high nutritional value, pepper (Capsicum annuum L.) was used in this study as a model plant to investigate the effect of natural biostimulants on yield and fruit quality parameters under conditions of reduced fertilisation.. A positive influence of biostimulant treatment on yield parameters was observed. The overall increase in the pigment content of leaves after biostimulant application agreed well with the higher total and commercial yields of treated pepper cultivars compared with their controls. The results showed that natural biostimulants had a positive effect on the vitamin C and total phenolic contents in pepper fruits during the hot summer season. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) antioxidant activities were also significantly higher (P < 0.05) in treated plants and correlated strongly with all measured quality parameters except total phenolic content.. Generally, biostimulants improved the antioxidant activity, vitamin C and phenolic contents in fruits as well as the pigment content in leaves of treated compared with non-treated pepper plants grown hydroponically. Thus the application of biostimulants could be considered as a good production strategy for obtaining high yields of nutritionally valuable vegetables with lower impact on the environment.

Topics: Antioxidants; Ascorbic Acid; Capsicum; Carotenoids; Chlorophyll; Chlorophyll A; Fruit; Hydroponics; Nutritive Value; Phenols; Plant Diseases; Plant Growth Regulators; Plant Leaves; Quality Control; Species Specificity

The impact of UV-B radiation on photosynthetic related parameters was studied in Oryza sativa L. cv. Safari plants, after an UV-B irradiation performed 1h per day for 7days (between 8 and 14days after germination) with a ten narrow-band (λ 311nm) that resulted in a total biological effective UV-B (UVB(BE)) of 2.975kJm(-2)day(-1) and a total of 20.825kJm(-2). Gas exchange measurements were severely affected, showing reductions higher than 80% in net photosynthesis (P(n)), stomatal conductance and photosynthetic capacity (A(max)), 1day after the end of the 7-days UV-B treatment. Similarly, several fluorescence parameters (F(o), F(v)/F(m), Fv'/Fm', ϕ(e), q(P) and q(E)) and thylakoid electron transport (involving both photosystems) were also severely reduced. Concomitantly, a decline of xanthophylls, carotenes, Chl a, Chl (a+b) and Chl (a/b) values was accompanied by the increase of the lipoperoxidation level in chloroplast membranes, altogether reflecting a loss of protection against oxidative stress. Seven days after of the end of UV-B treatment, most fluorescence parameters recovered, but in P(n), A(max), thylakoid electron transport rates, Chl a and lipid classes, as well as the level of lipoperoxidation, the impacts were even stronger than immediately after the end of stress, denoting a clear loss of performance of photosynthetic structures. However, only a moderate impact on total lipids was observed, accompanied by some changes in the relative weight of the major chloroplast membrane lipid classes, with emphasis on the decrease of MGDG and the increase of phospholipids. That suggested an ability to de novo lipid synthesis allowing qualitative changes in the lipid matrix. Notably, the leaves developed after the end of UV-B irradiation showed a much lower impact, with significantly decreased values only in P(n) and g(s), rises in several fluorescence parameters, thylakoid electron transport, photosynthetic pigments (xanthophylls and chls) and DEPS, while lipid classes presented values close to control. The results showed a global impact of UV-B in the photosynthetic structures and performance in irradiated leaves, but revealed also a low impairment extent in the leaves entirely developed after the end of the irradiation, reflecting a remarkable recovery of the plant after the end of stress, what could constitute an advantage under occasional UV-B exposure events in this vital worldwide staple food crop.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Chloroplasts; Electron Transport; Gases; Lipid Metabolism; Lipid Peroxidation; Oryza; Photosynthesis; Plant Leaves; Thylakoids; Ultraviolet Rays; Xanthophylls

The uptake of Se (VI) by two aquatic plants, Myriophyllum spicatum L. and Ceratophyllum demersum L., and its effects on their physiological characteristics have been studied. Plants were cultivated outdoors under semi-controlled conditions and in two concentrations of Na selenate solution (20 μg Se L(-1) and 10 mg Se L(-1)). The higher dose of Se reduced the photochemical efficiency of PSII in both species, while the lower dose had no effect on PSII. Addition of Se had no effect on the amounts of chlorophyll a and b. The concentration of Se in plants grown in 10 mg Se L(-1), averaged 212 ± 12 μg Seg(-1) DM in M. spicatum (grown from 8-13 d), and 492 ± 85 μg Se g(-1) DM in C. demersum (grown for 31 d). Both species could take up a large amount of Se. The amount of soluble Se compounds in enzyme extracts ranged from 16% to 26% in control, and in high Se solution from 48% to 36% in M. spicatum and C. demersum, respectively. Se-species were determined using HPLC-ICP-MS. The main soluble species in both plants was selenate (∼37%), while SeMet and SeMeSeCys were detected at trace levels.

Topics: Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Cysteine; Cystine; Magnoliopsida; Mass Spectrometry; Organoselenium Compounds; Photosystem II Protein Complex; Selenic Acid; Selenium Compounds; Selenocysteine; Selenomethionine; Water

Two-month old seedlings of perennial ryegrass (Lolium perenne L.) were subjected to four different levels of salinity for 7 days. The NaCl treatments reduced turf quality and normalized transpiration rates. Both chlorophyll (Chl) a and Chl b contents decreased in the grass exposed to 255 mM relative to the control. An increase in the lipid peroxidationin was observed. The activity of leaf superoxide dismutase increased while, peroxidase and catalase activities decreased in response to NaCl treatments. The expression of Chl Cu/ZnSOD, Cyt Cu/ZnSOD, FeSOD, CAT, POD, GPX and GR was up-regulated for NaCl-treated grass. Salt stress increased accumulation of Na(+) and decreased K(+)/Na(+) ratio, Mg(2+) and P content in both shoots and roots of perennial ryegrass. The findings of this study suggest that salt stress may cause toxicity to perennial ryegrass through oxidative injury and damage to Chl and cell membrane integrity.

Topics: Catalase; Chlorophyll; Chlorophyll A; Gene Expression Regulation, Plant; Glutathione Reductase; Ions; Lipid Peroxidation; Lolium; Peroxidase; Peroxidases; Plant Leaves; Plant Roots; Plant Transpiration; Salinity; Seedlings; Sodium Chloride; Stress, Physiological; Superoxide Dismutase

Since the complication of monitoring and evaluating the problems about the transgenic expression and its impacts on the receptor in the transgenic crop breeding and other relevant evaluated works, the authors in the present work tried to assess the differences of spectral parameters of the transgenic rice in contrast with its parent group quantitatively and qualitatively, fulfilling the growth monitoring of the transgenic samples. The spectral parameters (spectral morphological characteristics and indices) chosen are highly related to internal or external stresses to the receipts, and thus could be applied as indicators of biophysical or biochemical processes changes of plant. By ASD portable field spectroradiometer with high-density probe, fine foliar spectra of 8 groups were obtained. By analyzing spectral angle and continuum removal, the spectral morphological differences and their locations of sample spectra were found which could be as auxiliary priori knowledge for quantitative analysis. By investigating spectral indices of the samples, the quantitative differences of spectra were revealed about foliar chlorophyll a+b and carotenoid content. In this study both the spectral differences between transgenic and parent groups and among transgenic groups were investigated. The results show that hyperspectral technique is promising and a helpful auxiliary tool in the study of monitoring the transgenic crop and other relevant researches. By this technique, quantitative and qualitative results of sample spectra could be provided as prior knowledge, as certain orientation, for laboratory professional advanced transgenic breeding study.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Oryza; Plants, Genetically Modified; Remote Sensing Technology; Spectrum Analysis; Transgenes

Time-dependent density functional theory (TDDFT) calculations have been used to understand the excited-state properties of modified chlorophyll f (Chlide f), Chlide a, Chlide b, and axial ligated (with imidazole, H(2)O, CH(3)OH, CH(3)COOH, C(6)H(5)OH) Chlide f molecules. The computed differences among the Q(x), Q(y), B(x), and B(y) band absorbance wavelengths of Chlide a, b, and f molecules are found to be comparable with the experimentally observed shifts for these bands in chlorophyll a (chl a), chl b, and chl f molecules. Our computations provide evidence that the red shift in the Q(y) band of chl f is due to the extended delocalization of macrocycle chlorin ring because of the presence of the -CHO group. The local contribution from the -CHO substituent to the macrocycle chlorin ring stabilizes the corresponding molecular orbitals (lowest unoccupied molecular orbital (LUMO) of the Chlide f and LUMO-1 of the Chlide b). All the absorption bands of Chlide f shift to higher wavelengths on the addition of axial ligands. Computed redox potentials show that, among the axial ligated Chlide f molecules, Chlide f-imidazole acts as a good electron donor and Chlide f-CH(3)COOH acts as a good electron acceptor.

Topics: Chlorophyll; Chlorophyll A; Chlorophyllides; Electrons; Ligands; Models, Molecular; Spectrum Analysis

The reversible associations between the light-harvesting complexes (LHCs) and the core complexes of PSI and PSII are essential for the photoacclimation mechanisms in higher plants. Two types of Chls, Chl a and Chl b, both function in light harvesting and are required for the biogenesis of the photosystems. Chl b-less plants have been studied to determine the function of the LHCs because the Chl b deficiency has severe effects specific to the LHCs. Previous studies have shown that the amounts of the LHCs, especially the LHCII trimer, were decreased in the mutants; however, it is still unclear whether Chl b is required for the assembly of the LHCs and for the association of the LHCs with PSI and PSII. Here, to reveal the function of Chl b in the LHCs, we investigated the oligomeric states of the LHCs, PSI and PSII in the Arabidopsis Chl b-less mutant. A two-dimensional blue native-PAGE/SDS-PAGE demonstrated that the PSI-LHCI supercomplex was fully assembled in the absence of Chl b, whereas the trimeric LHCII and PSII-LHCII supercomplexes were not detected. The PSI-NAD(P)H dehydrogenase (NDH) supercomplexes were also assembled in the mutant. Furthermore, we detected two forms of monomeric LHC proteins. The faster migrating forms, which were detected primarily in the mutant, were probably apo-LHC proteins, whereas the slower migrating forms were probably the LHC proteins that contained Chl a. These findings increase our understanding of the Chl b function in the assembly of LHCs and the association of the LHCs with PSI, PSII and NDH.

Topics: Arabidopsis; Chlorophyll; Chlorophyll A; Electrophoresis, Polyacrylamide Gel; Fluorescence; Hot Temperature; Immunoblotting; Intracellular Membranes; Light-Harvesting Protein Complexes; Mutation; Phenotype; Photosystem I Protein Complex; Photosystem II Protein Complex; Protein Structure, Quaternary; Protein Subunits; Proteomics; Thylakoids

In the present study the effect of inorganic nutrients (CuSO₄ & ZnSO₄) on morphogenic and biochemical responses from nodal explants in Withania somnifera L. was investigated. Incorporation of either Copper sulphate (25-200 μM) or Zinc sulphate (50-500 μM) in the optimized Murashige and Skoog (MS) medium highly influenced the shoot bud formation and subsequent elongation, which induced maximum percentage (95%) regeneration, number (61.7 ± 0.25) of shoots with shoot length (5.46 ± 0.16 cm) on CuSO₄ (100 μM) and maximum percentage regeneration (100%), number of shoots (66.1 ± 0.96) with shoot length (6.24 ± 0.21 cm) on ZnSO₄ (300 μM) after 12 weeks of culture. Healthy growing in vitro microshoots rooted efficiently on ½ MS medium supplemented with NAA (0.5 μM), which induced (16.2 ± 0.12) roots with root length (3.30 ± 0.12 cm) after 4 weeks. Pigment content increased with increasing concentration of Cu and Zn and the maximum Chl. a (0.47), (0.41); Chl. b (0.52), (0.42); total Chl. (0.99), (0.83) and Carotenoid (0.16), (0.16) mg/g FW contents in regenerants were found on CuSO₄ (100 μM) and ZnSO₄ (300 μM), respectively. Maximum proline content (0.17), (0.16) μg/g FW was observed on high concentrations of CuSO₄ (200 μM) and ZnSO₄ (500 μM) respectively, in the basal medium. Regenerated plantlets were acclimatized successfully in soilrite with a survival rate of 95%. No morphological variations were detected among the micropropagated plants when compared with seedling raised plants of the same age.

Topics: Acclimatization; Biomass; Carotenoids; Chlorophyll; Chlorophyll A; Copper; Copper Sulfate; Environment; Ions; Meristem; Photosynthesis; Pigments, Biological; Plant Roots; Plant Shoots; Proline; Soil; Withania; Zinc; Zinc Sulfate

The effects of Mn(2+) deficiency on light absorption, transmission, and oxygen evolution of maize chloroplasts were investigated by spectral methods. Several effects of Mn(2+) deficiency were observed: (1) the skeleton of pigment protein complexes and oxygen-evolving center and the combination between pigment and protein were damaged; (2) the light absorption of chloroplasts was obviously decreased; (3) the energy transfer among amino acids within PS II protein-pigment complex and decreased energy transport from tyrosine residue to chlorophyll a and from chlorophyll b and carotenoid to chlorophyll a were inhibited; (4) the oxygen-evolving of chloroplast was significantly inhibited. However, Mn(2+) addition decreased the damage of light absorption, transmission, and oxygen evolution of maize chloroplasts caused by Mn(2+) deficiency.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Electron Transport; Manganese; Oxygen; Photosystem II Protein Complex; Spectrometry, Fluorescence; Spectrophotometry, Ultraviolet; Tyrosine; Zea mays

The present study aimed at investigating effects of zinc and aluminum (0.15 and 0.3mM) in duckweed (Lemna minor L.) over a 15-day period. High bioaccumulation of both metals was accompanied by an increase in dry weight under higher metal treatments. Antioxidant response was observed under both metal stresses, with large increases in superoxide dismutase and peroxidases. Catalase activity declined only in duckweed exposed to Zn while lipid peroxidation as well as H(2)O(2), proline and ascorbate levels increased. The results suggest induction of oxidative stress under both aluminum and zinc toxicity, and also demonstrate duckweed's capacity to upregulate its antioxidative defense. Additionally, Zn was found to be more toxic than Al to duckweed for the concentrations applied. Due to its high bioaccumulation potential and tolerance via increased antioxidant capacity, duckweed has a potential for phytoremediation of water bodies polluted by low levels of zinc and aluminum.

Topics: Aluminum; Antioxidants; Araceae; Catalase; Chlorophyll; Chlorophyll A; Ecosystem; Environmental Monitoring; Hydrogen Peroxide; Lipid Peroxidation; Malondialdehyde; Oxidants; Oxidative Stress; Peroxidases; Superoxide Dismutase; Water Pollutants, Chemical; Zinc

To understand further the role of deoxynivalenol (DON) in development of Fusarium head blight (FHB), we investigated effects of the toxin on uninfected barley tissues. Leaf segments, 1 to 1.2 cm long, partially stripped of epidermis were floated with exposed mesophyll in contact with DON solutions. In initial experiments with the leaf segments incubated in light, DON at 30 to 90 ppm turned portions of stripped tissues white after 48 to 96 h. The bleaching effect was greatly enhanced by addition of 1 to 10 mM Ca(2+), so that DON at 10 to 30 ppm turned virtually all stripped tissues white within 48 h. Content of chlorophylls a and b and of total carotenoid pigment was reduced. Loss of electrolytes and uptake of Evans blue indicated that DON had a toxic effect, damaging plasmalemmas in treated tissues before chloroplasts began to lose pigment. When incubated in the dark, leaf segments also lost electrolytes, indicating DON was toxic although the tissues remained green. Thus, loss of chlorophyll in light was due to photobleaching and was a secondary effect of DON, not required for toxicity. In contrast to bleaching effects, some DON treatments that were not toxic kept tissues green without bleaching or other signs of injury, indicating senescence was delayed compared with slow yellowing of untreated leaf segments. Cycloheximide, which like DON, inhibits protein synthesis, also bleached some tissues and delayed senescence of others. Thus, the effects of DON probably relate to its ability to inhibit protein synthesis. With respect to FHB, the results suggest DON may have multiple roles in host cells of infected head tissues, including delayed senescence in early stages of infection and contributing to bleaching and death of cells in later stages.

Topics: Antifungal Agents; Carotenoids; Cellular Senescence; Chlorophyll; Chlorophyll A; Cycloheximide; Fusarium; Gene Expression Regulation, Plant; Hordeum; Plant Leaves; Trichothecenes

Cherry tomato plants (Lycopersicon esculentum Mill) were sprayed with fluoranthene and mixture of fluoranthene and mannitol solutions for 30d. The exposure was carried out in growth chambers in field conditions, and the air was filtered through charcoal filters to remove atmospheric contaminants. Plants were sprayed with 10microM fluoranthene as mist until they reached the fruiting stage, and the eco-physiological parameters were measured to determine the effects of the treatments. We measured CO(2) uptake and water vapour exchange, chlorophyll fluorescence, leaf pigment contents, visual symptoms and biomass allocation. Fluoranthene which was deposited as mist onto leaves negatively affected both growth and the quality of tomato plants, while other treatments did not. The photosynthetic rate measured at saturated irradiance was approximately 37% lower in fluoranthene-treated plants compared with the control group. Other variables, such as stomata conductance, the photochemical efficiency of PSII in the dark, Chl a, Chl b, and the total chlorophyll contents of the tomato leaves were significantly reduced in the fluoranthene-treated plants. Tomato plants treated with fluoranthene showed severe visible injury symptoms on the foliage during the exposure period. Mannitol (a reactive oxygen scavenger) mitigated effects of fluoranthene; thus, reactive oxygen species generated through fluoranthene may be responsible for the damaged tomato plants. It is possible for fluoranthene to decrease the aesthetic and hence the economic value of this valuable crop plant.

Topics: Air Pollutants; Biomass; Carbon Dioxide; Chlorophyll; Chlorophyll A; Enzyme Inhibitors; Fluorenes; Mannitol; Photosystem II Protein Complex; Pigments, Biological; Reactive Oxygen Species; Solanum lycopersicum

A number of spectroscopic characteristics of three almost isoenergetic, red-shifted chlorophylls (chls) in the PS II antenna complex CP29 are investigated with the aim of (i) determining whether their band shapes are substantially identical or not, (ii) addressing the topical problem of whether they are involved in excitonic interactions with other chls, and (iii) establishing whether their binding sites may be defined as "mixed" with respect to their capacity to bind chls a and b. The three chls A2-CHL612, A3-CHL613, and B3-CHL614 were analyzed after in vitro apoprotein-pigment reconstitution using the CP29 coding sequence from Arabidopsis thaliana for both the wild-type and mutant complexes. Difference spectra thermal broadening analyses indicated that the half-bandwidths varied between 12 and 15 nm (at room temperature), due mainly to differences in the optical reorganization energy (25-40 cm(-1)). Moreover, only the A2 chl displayed an intense vibrational band in the 300-600 cm(-1) interval from the 0-0 transition. We conclude that within the red absorbing (approximately 680 nm) antenna chls of a single chl-protein complex a marked spectral band shape heterogeneity exists. By analysis of the absorption and circular dichroism spectra no evidence was found of significantly strong excitonic interactions. The single gene mutation of the A3 and B3 binding sites causes absorption changes in both the long wavelength chl a absorbing region and in the chl b spectral region. This has previously been observed and was attributed to "mixed" chl a/b binding sites [Bassi, R., Croce, R., Cugini, D., and Sandona, D. (1999) Proc. Natl. Acad. Sci. U.S.A. 96,10056-10061]. This interpretation, while in principle not being unreasonable, is shown to be incorrect for these two chls.

Topics: Apoproteins; Arabidopsis; Arabidopsis Proteins; Binding Sites; Chlorophyll; Chlorophyll A; Chloroplast Proteins; Energy Transfer; Light-Harvesting Protein Complexes; Mutagenesis, Site-Directed; Photosystem II Protein Complex; Plant Proteins; Recombinant Proteins; Ribonucleoproteins; Spectrometry, Fluorescence

Diurnal courses of photosynthetic gas exchange parameters, chlorophyll a fluorescence characteristics and the de-epoxidation state of the xanthophyll cycle pigments (DEPS) were measured during the gradual acclimation of 4-year-old Norway spruce seedlings to different photosynthetic photon flux density (PPFD) and air temperature (T(air)) regimes, simulating cloudy days with moderate T(air) (LI, maximum PPFD 300 micromol m(-2)s(-1), T(air) range 15-25 degrees C), sunny days with moderate T(air) (HI, maximum PPFD 1000 micromol m(-2)s(-1), T(air) range 15-25 degrees C) and hot sunny days (HI-HT, maximum PPFD 1000 micromol m(-2)s(-1), T(air) range 20-35 degrees C). The plants were acclimated inside a growth chamber and each acclimation regime lasted for 13d. Acclimation to HI conditions led to a strong depression of the net CO(2) assimilation rates (A(N)), particularly during noon and afternoon periods. Exposure to the HI-HT regime led to a further decrease of A(N) even during the morning period. Insufficient stomatal conductance was found to be the main reason for depressed A(N) under HI and HI-HT conditions. Only slight changes of the maximum photosystem II (PSII) photochemical efficiency (F(V)/F(M)), in the range of 0.78-0.82, supported the resistance of the Norway spruce photosynthetic apparatus against PSII photoinhibition during acclimation to both HI and HI-HT conditions. The HI plants showed increased content of xanthophyll cycle pigments (VAZ) and enhanced efficiency of thermal energy dissipation within PSII (D) that closely correlated with the increased DEPS. In contrast, acclimation to the HI-HT regime resulted in a slight reduction of VAZ content and significantly diminished D and DEPS values during the entire day in comparison with HI plants. These results indicate a minor role of the xanthophyll cycle-mediated thermal dissipation in PSII photoprotection under elevated temperatures. The different contributions of the thermal dissipation and non-assimilatory electron transport pathways in PSII photoprotection during acclimation of the Norway spruce photosynthetic apparatus to excess irradiance and heat stresses are discussed.

Topics: Acclimatization; Carbon Dioxide; Chlorophyll; Chlorophyll A; Circadian Rhythm; Gases; Light; Norway; Photons; Photosynthesis; Photosystem II Protein Complex; Picea; Plant Stomata; Temperature; Xanthophylls

The hypothesis that CO(2) gas exchange and chlorophyll a fluorescence (ChlaF) of lichens vary according to the light regimes of their original habitat, as observed in vascular plants, was tested by analysing the photosynthetic performance of 12 populations of seven dorsoventral, foliose lichens collected from open, south-exposed rocks to densely shaded forests. Light response curves were induced at optimum thallus water content and ChlaF emission curves at the species-specific photon flux at which the quantum yield of CO(2) assimilation is the highest and is saturating the photosynthetic process. Photosynthetic pigments were quantified in crude extracts. The results confirm that the maximum rate of gross photosynthesis is correlated with the chlorophyll content of lichens, which is influenced by light as well as by nitrogen availability. Like leaves, shade tolerant lichens emit more ChlaF than sun-loving ones, whereas the photosynthetic quantum conversion is higher in the latter.

Topics: Altitude; Carbon Dioxide; Carotenoids; Chlorophyll; Chlorophyll A; Ecosystem; Electron Transport; Fluorescence; Lichens; Light; Organ Size; Photosynthesis

To assess the aquatic ecosystem safety for silica (SiO2) nanoparticles (NPs), the growth inhibition and photosynthetic pigment contents of Scenedesmus obliquus in logarithm growth phase exposed to SiO2 NPs and SiO2 bulk particles (BPs) suspensions were measured. SiO2 NPs with 10-20 nm diameters were found to be toxic. The 20% effective concentration (EC20) values for 72 and 96 hr were 388.1 and 216.5 mg/L, respectively. The contents of chlorophyll decreased significantly under moderate and high concentration (50, 100, and 200 mg/L) of SiO2 NPs after 96-hr exposure, but the carotenoids did not. SiO2 BPs were found to be nontoxic up to 200 mg/L. The toxicity of SiO2 NPs probablely due to their sorption to algal cells surface. The results imply that there is potential harm to aquatic environment by using SiO2 NPs, and it should deserve special concern.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Microscopy, Electron, Scanning; Quantum Dots; Scenedesmus; Silicon Dioxide; Spectrophotometry, Ultraviolet; Water Pollutants, Chemical

From time integration of the electron dynamics under a density functional tight binding Hamiltonian in the presence of external time varying electric fields, we obtain the absorption spectra of a series of chlorophylls and bacteriochlorophylls. We obtain good agreement with the observed experimental energies as well as with fully ab initio results in the literature for the main absorption bands. As a first step towards an atomistic description of energy transfer between chromophores in photosynthetic antenna systems we calculate the coupling energy between the excitations of two chlorophyll a molecules as a function of the distance as well as the transfer of energy between these when one of them is subjected to laser illumination.

Topics: Absorption; Bacteriochlorophylls; Chlorophyll; Chlorophyll A; Energy Transfer; Photosynthesis; Thermodynamics

Chlorophyll (Chl) a/b-binding proteins from Prochlorothrix hollandica known as Pcb antennae were studied by femtosecond transient absorption technique to identify energy transfer rates and pathways in Pcb and Pcb-PS I complexes. Carotenoids transfer energy to Chl with low efficiency of approximately 25% in Pcb complexes. Interestingly, analysis of transient absorption spectra identified a pathway from the hot S(1) state of zeaxanthin and/or beta-carotene as the major energy transfer channel between carotenoids and chlorophylls in Pcb whereas the S(2) state contributes only marginally to energy transfer. Due to energetic reasons, no energy transfer is possible via the relaxed S(1) state of carotenoids. The low overall energy transfer efficiency of carotenoids recognizes chlorophylls as the main light-harvesting pigments. Besides Chl a, presence of Chl b, which transfers energy to Chl a with nearly 100% efficiency, significantly broadens the spectral range accessible for light-harvesting and improves cross section of Pcb complexes. The major role of carotenoids in Pcb is photoprotection.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Energy Transfer; Photosystem I Protein Complex; Prochlorothrix; Spectrometry, Fluorescence

We report a new device for the estimation of the content of chlorophyll a pigment in water samples as an indicator of water quality. The extraction of the pigment from water was also optimized. 10 mL of water was filtered through a nylon filter (45 microm pore size and 13 mm of diameter), after the chlorophylls were dissolved by immersing the filter in 1 mL of a low non-hazardous solvent as ethanol. An in-valve in-tube SPME device coupled to capillary liquid chromatography with diode array detection was designed. A capillary column of 70 cm in length (0.32 mm i.d. coated with 5% diphenyl-95% polydimethylsiloxane, 3 microm coating thickness) was used as the loop of the injection valve for preconcentration and a Zorbax SB C(18) (SiO(2)-based) 150 mm x 0.5 mm i.d., 5 microm column (Agilent) was used as analytical column. The achieved detection limit was 0.05 microg L(-1) and the working range of concentrations was 0.1-1 microg L(-1). % RSD values between 2 and 11 were obtained. Chlorophyll a in several water matrices was determined with good results in presence of other pigments such as chlorophyll b, pheophytin a and pheophytin b.

Topics: Chlorophyll; Filtration; Limit of Detection; Pheophytins; Quality Control; Solid Phase Extraction; Solvents; Water Pollutants, Chemical

Light-harvesting antenna system possesses an inherent property of photoprotection. The single-helix proteins found in cyanobacteria play role in photoprotection and/or pigment metabolism. The photoprotective functions are also manifested by the two- and four-helix proteins. The photoprotection mechanism evolved earlier to the mechanism of light-harvesting of the antenna complex. Here, the light-harvesting complex genes of photosystems I and II from Arabidopsis are enlisted, and almost similar set of genes are identified in rice. Also, the three-helix early light-inducible proteins (ELIPs), two-helix stress-enhanced proteins (SEPs), and one-helix high light-inducible proteins [one-helix proteins (OHPs)] are identified in rice. Interestingly, two independent genomic loci encoding PsbS protein are also identified with implications on additional mode of non-photochemical quenching (NPQ) mechanism in rice. A few additional LHC-related genes are also identified in rice (LOC_Os09g12540, LOC_Os02g03330). This is the first report of identification of light-harvesting complex genes and light-inducible genes in rice.

Topics: Amino Acid Sequence; Arabidopsis; Chlorophyll; Chlorophyll A; Evolution, Molecular; Genome, Plant; Light-Harvesting Protein Complexes; Molecular Sequence Data; Oryza; Protein Binding; Sequence Homology, Amino Acid

Active ground optical remote sensing (AGORS) devices mounted on overhead irrigation booms could help to improve seedling quality by autonomously monitoring seedling stress. In contrast to traditionally used passive optical sensors, AGORS devices operate independently of ambient light conditions and do not require spectral reference readings. Besides measuring red (590-670 nm) and near-infrared (>760 nm) reflectance AGORS devices have recently become available that also measure red-edge (730 nm) reflectance. We tested the hypothesis that the additional availability of red-edge reflectance information would improve AGORS of plant stress induced chlorophyll breakdown in Scots pine (Pinus sylvestris). Our results showed that the availability of red-edge reflectance information improved AGORS estimates of stress induced variation in chlorophyll concentration (r2>0.73, RMSE<1.69) when compared to those without (r2=0.57, RMSE=2.11).

Topics: Biosensing Techniques; Chlorophyll; Chlorophyll A; Linear Models; Pinus sylvestris; Reproducibility of Results; Seedlings; Stress, Physiological

Phytopigment estimation has a considerable interest in the evaluation of freshwater bodies' quality, because it takes into account the synergistic effect of nutrients like phosphorus or nitrogen on algal growth producing eutrophication. Furthermore, their increasing concentration constitutes the first step in the formation of biofilms on the surface sediments, adding a new and very important element to the dynamic nature of the surface sediments. In this study the distribution of phytoplankton--in terms of chlorophyll-a, chlorophyll-b, phaeophytin-a, phaeophytin-b, total carotenoids, total chlorophyll, and total phaeophytin--was evaluated in river bed sediments. Samples collected at sites with low levels of nutrients (P,N) and metal concentrations showed lower phytopigment concentrations than those collected at the sampling sites affected by sources of pollution. Phytoplankton concentrations were directly and highly related to the organic matter concentrations--in particular to the humic fraction--as well as to the total nitrogen (N), total phosphorus (P(T)) and available phosphorus (P(A)) concentrations in sediments. In addition, phytoplankton also correlates positively with Cu, Zn, Fe and Al extracted in oxalate, being Cu the variable that most influences the phytopigment growth. These are essential metals for the metabolism of the phytoplankton, so therefore the increase in metal concentrations can increase algal growth, unless they reach toxic levels.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Environmental Monitoring; Geologic Sediments; Metals; Nitrogen; Phosphorus; Pigments, Biological; Rivers; Spain

Salicylic acid-deficient NahG transgenic lines and sid2 mutants were used to evaluate the role of this compound in the development of the short-lived, annual plant Arabidopsis thaliana, with a particular focus on the interplay between salicylic acid and other phytohormones. Low salicylic acid levels led to increased growth, as well as to smaller abscisic acid levels and reduced damage to PSII (as indicated by F(v)/F(m) ratios) during the reproductive stages in rosette leaves of NahG transgenic lines and sid2 mutants, compared with wild-type plants. Furthermore, salicylic acid deficiency highly influenced seed yield and composition. Seed production increased by 4.4-fold and 3.5-fold in NahG transgenic lines and sid2 mutants, respectively, compared to the wild type. Salicylic acid deficiency also improved seed composition in terms of antioxidant vitamin concentrations, seeds of salicylic acid-deficient plants showing higher levels of alpha- and gamma-tocopherol (vitamin E) and beta-carotene (pro-vitamin A) than seeds of wild-type plants. Seeds of salicylic acid-deficient plants also showed higher nitrogen concentrations than seeds of wild-type plants. It is concluded that (i) the sid2 gene, which encodes for isochorismate synthase, plays a central role in salicylic acid biosynthesis during plant development in A. thaliana, (ii) salicylic acid plays a role in the regulation of growth, senescence, and seed production, (iii) there is a cross-talk between salicylic acid and other phytohormones during plant development, and (iv) the concentrations of antioxidant vitamins in seeds may be influenced by the endogenous levels of salicylic acid in plants.

Topics: Arabidopsis; Biomass; Carbon; Carotenoids; Chlorophyll; Chlorophyll A; Fruit; Intramolecular Transferases; Mixed Function Oxygenases; Mutation; Nitrogen; Photosystem II Protein Complex; Plant Growth Regulators; Plant Leaves; Plants, Genetically Modified; Salicylic Acid; Seeds

The light-harvesting chlorophyll a/b-protein complex of photosystem II (LHCII) is the most abundant membrane protein in green plants, and its degradation is a crucial process for the acclimation to high light conditions and for the recovery of nitrogen (N) and carbon (C) during senescence. However, the molecular mechanism of LHCII degradation is largely unknown. Here, we report that chlorophyll b reductase, which catalyzes the first step of chlorophyll b degradation, plays a central role in LHCII degradation. When the genes for chlorophyll b reductases NOL and NYC1 were disrupted in Arabidopsis thaliana, chlorophyll b and LHCII were not degraded during senescence, whereas other pigment complexes completely disappeared. When purified trimeric LHCII was incubated with recombinant chlorophyll b reductase (NOL), expressed in Escherichia coli, the chlorophyll b in LHCII was converted to 7-hydroxymethyl chlorophyll a. Accompanying this conversion, chlorophylls were released from LHCII apoproteins until all the chlorophyll molecules in LHCII dissociated from the complexes. Chlorophyll-depleted LHCII apoproteins did not dissociate into monomeric forms but remained in the trimeric form. Based on these results, we propose the novel hypothesis that chlorophyll b reductase catalyzes the initial step of LHCII degradation, and that trimeric LHCII is a substrate of LHCII degradation.

Topics: Alcohol Oxidoreductases; Arabidopsis; Cellular Senescence; Chlorophyll; Chlorophyll A; Electrophoresis, Gel, Two-Dimensional; Escherichia coli; Light; Light-Harvesting Protein Complexes

The toxicity of the chiral herbicides rac-metolachlor and S-metolachlor to Chlorella pyrenoidosa was determined and compared in this study, based on four different test endpoints: the growth inhibition rate, the chlorophyll a and chlorophyll b concentration, the catalase activity, and the ultrastructural morphology of cells. The 24, 48, 72, and 96h EC(50) values of rac-metolachlor were 0.196, 0.241, 0.177 and 0.152mgL(-1), respectively; these values were higher than those of S-metolachlor, which were 0.116, 0.106, 0.081 and 0.068mgL(-1), respectively. This indicates that S-metolachlor was more toxic to C. pyrenoidosa than rac-metolachlor. The Chla and Chlb concentration of C. pyrenoidosa treated by rac-metolachlor was higher than that treated by S-metolachlor. In general, the catalase activity of C. pyrenoidosa treated by S-metolachlor was higher than that exposed to rac-metolachlor, and catalase activity was inhibited at high concentrations of both herbicides. The ultrastructural morphology of cells grown in the two herbicides was observed by transmission electron microscopy. The cell wall separated from the cell membrane, accumulated starch granules were observed in the chloroplast, and some lipid droplets and unknown electron-opaque deposits were also observed in the cytoplasm. The mechanism of the toxicity of rac- and S-metolachlor toxicity to C. pyrenoidosa was explored, and the enantioselective toxicity of rac- and S-metolachlor to C. pyrenoidosa was determined. These results will help to develop an understanding of the biologically mediated environmental processes of rac- and S-metolachlor.

Topics: Acetamides; Catalase; Cell Growth Processes; Chlorella; Chlorophyll; Chlorophyll A; Herbicides; Stereoisomerism

Effects of Pb accumulation on the contents of chlorophylls (a and b), carotenoid, ascorbic acid (AsA), non-protein SH groups and protein were investigated in aquatic macrophyte Elodea canadensis. Pb accumulation in E. canadensis tissues increased with increasing metal concentrations. The increases at 1, 10 and 100 mg/L Pb are about 12.0, 44.6 and 71.1 times greater than control, respectively. Contents of chlorophylls, carotenoid and protein were adversely affected by Pb accumulation. Induction of non-protein SH groups and AsA showed that Pb accumulation caused oxidative stress. It is also possible that increased non-protein SH groups by Pb accumulation may be due to their role in Pb detoxification.

Topics: Ascorbic Acid; Carotenoids; Chlorophyll; Chlorophyll A; Hydrocharitaceae; Lead; Oxidative Stress; Plant Proteins; Water Pollutants, Chemical

The effects of high Zn concentration were investigated in sugar beet (Beta vulgaris L.) plants grown in a controlled environment in hydroponics. High concentrations of Zn sulphate in the nutrient solution (50, 100 and 300 microm) decreased root and shoot fresh and dry mass, and increased root/shoot ratios, when compared to control conditions (1.2 microm Zn). Plants grown with excess Zn had inward-rolled leaf edges and a damaged and brownish root system, with short lateral roots. High Zn decreased N, Mg, K and Mn concentrations in all plant parts, whereas P and Ca concentrations increased, but only in shoots. Leaves of plants treated with 50 and 100 microm Zn developed symptoms of Fe deficiency, including decreases in Fe, chlorophyll and carotenoid concentrations, increases in carotenoid/chlorophyll and chlorophyll a/b ratios and de-epoxidation of violaxanthin cycle pigments. Plants grown with 300 microm Zn had decreased photosystem II efficiency and further growth decreases but did not have leaf Fe deficiency symptoms. Leaf Zn concentrations of plants grown with excess Zn were high but fairly constant (230-260 microg.g(-1) dry weight), whereas total Zn uptake per plant decreased markedly with high Zn supply. These data indicate that sugar beet could be a good model to investigate Zn homeostasis mechanisms in plants, but is not an efficient species for Zn phytoremediation.

Topics: Beta vulgaris; Biological Transport; Carbon Dioxide; Chlorophyll; Chlorophyll A; FMN Reductase; Hydroponics; Minerals; Nitrogen; Oxygen; Photosystem II Protein Complex; Plant Structures; Trace Elements; Xanthophylls; Zinc; Zinc Sulfate

Accumulation of carbon, nitrogen and sulphur content in Rinodina sophodes, crustose poleotolerent lichen growing naturally in and around six sites of Kanpur city was estimated, and their influence on the photosynthetic pigments of the lichen was studied. Maximum carbon concentration was recorded at highly polluted area while higher accumulation of nitrogen was recorded near village in outskirt of the city having higher ammonia emission. The concentration of sulphur was not detected in most of the sites except a single site where it had a quite lower value (0.22%). Photosynthetic pigments (chlorophyll a and b) increased parallel to the level of traffic density. Multiple correlation analysis revealed that chlorophyll a had highly significant correlation (1%) with chlorophyll b (r=0.9986) and total chlorophyll (r=0.9307). Carbon is directly correlated with nitrogen (r=0.3035), sulphur (r=0.1743) and chlorophyll degradation (r=0.2685) while negatively correlated with chlorophyll a (-0.3323), chlorophyll b (r=-0.3429) and total chlorophyll (r=-0.0824). Nitrogen showed negative correlation between all photosynthetic pigments and chlorophyll degradation, while in case of sulphur, it was high positive correlation at 1% with chlorophyll degradation (0.9445).

Topics: Carbon; Chlorophyll; Chlorophyll A; India; Lichens; Nitrogen; Pigments, Biological; Sulfur

A study was conducted over 2 years to determine whether growth under elevated CO(2) (700 μmol mol(-1) ) and temperature (ambient + 4 °C) conditions modifies photochemical efficiency or only the use of electron transport products in spring wheat grown in field chambers. Elevated atmospheric CO(2) concentrations increased crop dry matter at maturity by 12-17%, while above-ambient temperatures did not significantly affect dry matter yield. In measurements with ambient CO(2) at ear emergence and after anthesis, growth at elevated CO(2) concentrations decreased flag leaf light-saturated carbon assimilation. The quantum yield of electron transport (Φ(PSII) ) measured at ambient CO(2) and higher irradiances increased at ear emergence and decreased after anthesis in plants grown at elevated CO(2) . At higher light intensities, but not in low light, photochemical quenching (qP) decreased after growth in elevated CO(2) conditions. Growth under CO(2) enrichment increased dark- (Fv:Fm) and light-adapted (Fv':Fm') photochemical efficiencies, and decreased the chlorophyll a:b ratio, suggesting an increase in light-harvesting complexes relative to PSII reaction centres. A relatively higher decrease in carbon assimilation than the decrease in Φ(PSII) pointed to a sink other than CO(2) assimilation for electron transport products at defined growth stages. With higher light intensities, warmer temperatures increased Φ(PSII) and Fv':Fm' at ear emergence and decreased Φ(PSII) after anthesis; in ambient-but not elevated-CO(2) , warmer temperatures also decreased qP after anthesis. CO(2) fixation increased or did not change with temperature, depending on the growth stage and year. We conclude that elevated CO(2) decreases the carbon assimilation capacity, but increases photochemistry and resource allocation to light harvesting, and that elevated levels of CO(2) can mitigate photochemistry inhibition as a result of warm temperatures.

Topics: Acclimatization; Atmosphere; Carbon; Carbon Dioxide; Chlorophyll; Chlorophyll A; Fluorescence; Global Warming; Hot Temperature; Photochemical Processes; Photosynthesis; Triticum

Two lichen species (Peltigera rufescens and Cladina arbuscula subsp. mitis) and one moss species (Racomitrium lanuginosum) growing on a copper mine heaps (probably 200-300yr old) in the village of Spania dolina (Slovak Republic) were assessed for selected physiological parameters, including composition of assimilation pigments, chlorophyll a fluorescence, soluble proteins and free amino acid content. The lichen C. arbuscula subsp. mitis was collected also at a control locality where total copper concentration in the soil was approximately 3% that of the waste heaps. Concentrations of Al, Co, Cu, Ni, Sb and Zn were highest in thalli of Peltigera, while the moss Racomitrium contained the highest content of Fe and Pb. Thalli of Cladina contained less metals than the cyanolichen Peltigera, and except for Zn metal concentrations in Cladina from the control locality were lower than in thalli of the same species from copper mine heaps. Regardless of the species or locality, the composition of assimilation pigments and chlorophyll a fluorescence showed that the tested lichens and moss were in good physiological condition and adapted to increased copper levels in the soil. There were significantly different amounts of total free amino acids in Peltigera, Cladina and Racomitrium from the Cu-polluted field. However, differences in amount of free amino acids in control, as well as Cu-polluted thalli of Cladina were less pronounced.

Topics: Adaptation, Physiological; Amino Acids; Ascomycota; Bryopsida; Chlorophyll; Chlorophyll A; Copper; Environmental Monitoring; Lichens; Metals; Proteins; Soil Pollutants

The major light-harvesting chlorophyll a/b complex (LHCII) greatly enhances the efficiency of photosynthesis in green plants. Recombinant LHCII can be assembled in vitro from its denatured, bacterially expressed apoprotein and plant pigments. This makes it an interesting candidate for biomimetic light-harvesting in photovoltaic applications. Due to its almost 20 pigments bound per apoprotein, LHCII absorbs efficiently in the blue and red spectral domains of visible light but less efficiently in the green domain, the so-called "green gap" in its absorption spectrum. Here we present a hybrid complex of recombinant LHCII with organic dyes that add to LHCII absorption in the green spectral region. One or three Rhodamine Red dye molecules were site-specifically attached to cysteine side chains in the apoprotein and did not interfere with LHCII assembly, function and stability. The dyes transferred their excitation energy virtually completely to the chlorophylls in LHCII, partially filling in the green gap. Thus, organic dyes can be used to increase the absorption cross section and, thus, the light-harvesting efficiency of recombinant LHCII.

Topics: Chlorophyll; Chlorophyll A; Fluorescent Dyes; Protein Folding; Rhodamines

In order to reveal the induced resistance mechanism of tobacco treated with copper solution to potato virus Y-vein necrosis strain (PVY(N)), disease indexes, contents of virus and some physiological and biochemical indexes in tobacco were studied. The results showed that when treated at the copper concentration of 0.8 mg x L(-1), the symptom displayed and vein necrosis on tobacco were postponed, the disease index and content of virus sharply decreased , and the content of chlorophyll a, chlorophyll b and phenylalanine ammonia lyase (PAL) activity remarkably increased. Furthermore, vein necrosis closely linked to contents of total phenol and flavonoid. In this study, the contents of total phenol and flavonoid were promoted when treated with a solution at the copper concentration of 0.8 mg x L(-1). But the contents of total phenol and flavonoid reached to the first peak at the 3rd day after inoculation, and then decreased to the lowest levels which even were lower than those of the control after inoculating PVY(N). Then the contents of total phenol and flavonoid increased slowly from the 6td but still lower than those of the control. The result implied that spraying copper solution might play an important role in induced resistance of tobacco to vein necrosis disease and strengthen the antiviral capability to PVY(N).

Topics: Chlorophyll; Chlorophyll A; Copper; Immunity, Innate; Nicotiana; Phenylalanine Ammonia-Lyase; Potyvirus

To develop a better understanding of compositional changes occurring during the production of commercial teas, we determined by high-performance liquid chromatography (HPLC) changes in ingredient levels during each of several manufacturing steps used to produce Kamairi-cha, a premium green tea. Kamairi-cha uses pan-frying instead of the usual blanching technique to inactivate the enzymes responsible for producing traditional black tea. The resulting tea lacks the characteristic bitterness of green tea, producing a green tea that is described as sweet tasting. The processing steps used to produce this pan-fried tea were as follows: 1st roasting, 1st rolling, 2nd roasting, 2nd rolling, 1st firing, and 2nd firing. The results show that during production at temperatures up to 300 degrees C, raw leaves lost (in percent) 97.3 water, 94 two chlorophylls, 14.3 seven catechins, and 2.75 caffeine. A separate analysis showed that the final product contained 21.67 mg/g dry wt of the biologically active amino acid theanine. The results of this 1st report on changes in individual catechins and other tea ingredients in tea leaves during pan-frying make it possible to select production conditions that maximize levels of beneficial tea ingredients. The possible significance of the results for the human diet is discussed.

Topics: Alkaloids; Caffeine; Catechin; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Food Handling; Glutamates; Hot Temperature; Korea; Plant Leaves; Tea; Water

Persistent nonphotochemical hole burning and delta-FLN spectra obtained at 4.5 K are reported for monomeric chlorophyll (Chl) a/b light-harvesting complexes of photosystem II (LHC II) of green plants. The hole burned spectra of monomeric LHC II appear to be similar to those obtained before for trimeric LHC II (Pieper et al. J. Phys. Chem. B 1999, 103, 2412). They are composed of three main features: (i) a homogeneously broadened zero-phonon hole coincident with the burn wavelength, (ii) an intense, broad hole in the vicinity of approximately 680 nm as a result of efficient excitation energy transfer to a low-energy trap state, and (iii) a satellite hole at approximately 649 nm which is correlated with the low-energy 680 nm hole. Zero-phonon hole action spectroscopy reveals that the low-energy absorption band is located at 679.6 nm and possesses a width of approximately 110 cm(-1) which is predominantly due to inhomogeneous broadening at 4.5 K. The electron-phonon coupling of the above-mentioned low-energy state(s) is weak with a Huang-Rhys factor S in the order of 0.6 and a peak phonon frequency (omega(m)) of approximately 22 cm(-1) within a broad and strongly asymmetric one-phonon profile. The resulting Stokes shift 2S omega(m) of approximately 26.4 cm(-1) readily explains the position of the fluorescence origin band at 680.8 nm. Thus, we conclude that the 679.6 nm state(s) is (are) the fluorescent state(s) of monomeric LHC II at 4.5 K. The absorption intensity of the lowest Q(y) state is shown to roughly correspond to that of one out of the eight Chl a molecules bound in the monomeric subunit. In addition, the satellite hole structure produced by hole burning within the 679.6 nm state is weak with only one shallow satellite hole observed in the Chl b spectral range at 648.8 nm. These results suggest that the 679.6 nm state is widely localized on a Chl a molecule, which may belong to a Chl a/b heterodimer. These characteristics are different from those expected for Chl a612, which has been associated with the fluorescent state at room temperature. Alternatively, the 679.6 nm state may be assigned to Chl a604, which is located in a cluster with several Chl b molecules resulting in a relatively weak excitonic coupling.

Topics: Chlorophyll; Chlorophyll A; Photosystem II Protein Complex; Protein Multimerization; Spectrometry, Fluorescence; Spinacia oleracea

Chlorophyll b is one of the major photosynthetic pigments of plants. The regulation of chlorophyll b biosynthesis is important for plants in order to acclimate to changing environmental conditions. In the chloroplast, chlorophyll b is synthesized from chlorophyll a by chlorophyllide a oxygenase (CAO), a Rieske-type monooxygenase. The activity of this enzyme is regulated at the level of protein stability via a feedback mechanism through chlorophyll b. The Clp protease and the N-terminal domain (designated the A domain) of CAO are essential for the regulatory mechanism. In this study, we aimed to identify the specific amino acid residue or the sequence within the A domain that is essential for this regulation. To accomplish this goal, we randomly introduced base substitutions into the A domain and searched for potentially important residues by analyzing 1,000 transformants of Arabidopsis thaliana. However, none of the single amino acid substitutions significantly stabilized CAO. Therefore, we generated serial deletions in the A domain and expressed these deletions in the background of CAO-deficient Arabidopsis mutant. We found that the amino acid sequence (97)QDLLTIMILH(106) is essential for the regulation of the protein stability. We furthermore determined that this sequence induces the destabilization of green fluorescent protein. These results suggest that this sequence serves as a degradation signal that is recognized by proteases functioning in the chloroplast.

Topics: Amino Acid Sequence; Arabidopsis; Arabidopsis Proteins; Chlorophyll; Chlorophyll A; Chloroplasts; Endopeptidase Clp; Enzyme Stability; Oxygenases; Protein Structure, Tertiary; Sequence Deletion

Light harvesting complex II (LHCII) is the most abundant protein in the thylakoid membrane of higher plants and green algae. LHCII acts to collect solar radiation, transferring this energy mainly toward photosystem II, with a smaller amount going to photosystem I; it is then converted into a chemical, storable form. We performed time-resolved femtosecond visible pump/mid-infrared probe and visible pump/visible probe absorption difference spectroscopy on purified LHCII to gain insight into the energy transfer in this complex occurring in the femto-picosecond time regime. We find that information derived from mid-infrared spectra, together with structural and modeling information, provides a unique visualization of the flow of energy via the bottleneck pigment chlorophyll a604.

Topics: Absorption; Chlorophyll; Chlorophyll A; Hydrogen Bonding; Infrared Rays; Light; Light-Harvesting Protein Complexes; Models, Molecular; Protein Conformation; Spectrophotometry, Infrared; Thermodynamics; Time Factors

Demetalation kinetics of natural chlorophyll (Chl) d purified from Acaryochloris marina was first studied and compared with those of Chls a and b. The demetalation rate constant of Chl d, which possessed a formyl group at the 3-position, was five-fold smaller than that of Chl a possessing a vinyl group at the same position in aqueous acetone at the proton concentration of 1.2 x 10(-3) M at 25 degrees C. In contrast, the demetalation rate constant of Chl b possessing a formyl group at the 7-position was 26 times smaller than that of Chl a. The activation energy of demetalation reaction of Chl d was larger than that of Chl a, but smaller than that of Chl b. These indicate that the substitution effect of 3-formyl group on the acidic removal of central magnesium in Chls was smaller than that of 7-formyl group.

Topics: Chlorophyll; Chlorophyll A; Cyanobacteria; Kinetics; Magnesium; Photosynthesis; Porphyrins; Spectrophotometry

The present work aimed to study the physiological effects of cadmium (Cd) and copper (Cu) in pea (Pisum sativum). Pea plants were exposed to increasing doses of cadmium chloride (CdCl2) and copper chloride (CuCl2) for 20 d. The examined parameters, namely root and shoot lengths, the concentration of photosynthetic pigments and the rate of photosynthesis were affected by the treatments especially with high metals concentrations. The analysis of heavy metals accumulation shows that leaves significantly accumulate cadmium for all the tested concentrations. However, copper was significantly accumulated only with the highest tested dose. This may explain the higher inhibitory effects of cadmium on photosynthesis and growth in pea plants. These results are valuable for understanding the biological consequences of heavy metals contamination particularly in soils devoted to organic agriculture.

Topics: beta Carotene; Cadmium Chloride; Chlorophyll; Chlorophyll A; Copper; Photosynthesis; Pisum sativum; Plant Leaves; Soil Pollutants

Characteristics of the monsoonal bloom of phytoplankton at Orissa Coast in the Bay of Bengal were studied through bimonthly observation from April 2001 to December 2002. Three photosynthetic pigments chlorophyll-a (Chl a), chlorophyll-b (Chl b) and carotenoid (Car) were analyzed by absorption spectroscopic method. The seasonal variation of Chl a included phytoplankton bloom in the coastal area during monsoon period. The water column integrated Chl a reached to 68 mg m(-2) at the station-1(St1), and amounted to 20 mg m(-2) at 30 km off the river mouth during August 2001. In contrast the same amount was found at 15 km off the Mahanadi river mouth during August 2002. Salinity during this period varied from 5 psu at the St1 to 27 psu at the edge of the bloom area. The total amount of river discharge in the monsoon period calculated from daily river discharge data reported by Water Resources Department in India was 84 x 10(9) m(3) during 2001 and 20 x 10(9) m(3) during 2002. Both nitrate and phosphate concentrations showed negative quadratic relationship with salinity throughout the observation period. Extrapolated nitrate and phosphate concentration discharge from the Mahanadi river were 10.8 and 4 microg-at l(-1), respectively. Microscopic identification revealed dominance of fluvial Chlorophyceae and diatoms during the monsoon period showing influence of the freshwater discharge.

Topics: Animals; Carotenoids; Chlorophyll; Chlorophyll A; Environmental Monitoring; Eutrophication; India; Nitrates; Oceans and Seas; Phosphates; Phytoplankton; Rain; Rivers; Salinity; Seasons; Seawater; Tropical Climate

The objectives of this study were to develop a high performance liquid chromatography-mass spectrometry (HPLC-MS) method for determination of chlorophylls and their derivatives in Gynostemma pentaphyllum Makino, a traditional Chinese herb possessing vital biological activities. Chlorophylls were extracted with a quaternary solvent system of hexane-acetone-ethanol-toluene (10:7:6:7, v/v/v/v), followed by separation of a total of 15 chlorophylls and their derivatives within 32 min using a gradient mobile phase of acetone, acetonitrile and methanol and a HyPURITY C18 column, with detection at 660 nm and flow rate at 1 mL/min. Identification was carried out on the basis of retention behavior, absorption spectra and mass spectra using atmospheric pressure chemical ionization (APCI) in positive ion mode for detection. Of the 15 analytes, chlorophyll a, chlorophyll b, pheophytin a and pheophytin b were quantified by using standard calibration curves, with the other 11 being quantified with an internal standard Fast Green FCF. Chlorophyll extracts in G. pentaphyllum were found to contain pheophytin a (2508.3 microg/g), pheophytin a' (111.2 microg/g), chlorophyll a (113.8 microg/g), chlorophyll a' (11.0 microg/g), hydroxypheophytin a (88.6 microg/g), hydroxypheophytin a' (66.5 microg/g), pyropheophytin a (76.0 microg/g), hydroxychlorophyll a (23.8 microg/g), pheophytin b (319.6 microg/g), pheophytin b' (13.2 microg/g), chlorophyll b (287.9 microg/g), chlorophyll b' (11.1 microg/g), hydroxychlorophyll b (15.0 microg/g), hydroxypheophytin b (11.2 microg/g) and hydroxypheophytin b' (8.5 microg/g).

Topics: Calibration; Chlorophyll; Chlorophyll A; Chromatography, Liquid; Gynostemma; Mass Spectrometry; Molecular Structure; Pheophytins; Reference Standards; Reproducibility of Results

In a two-year field experiment with wheat cultivars under different application rates of fertilizer N, the wheat leaf pigment concentrations were monitored with hyper-spectral remote sensing, and quantitative monitoring models were established. The results showed that the pigment concentrations in wheat leaves increased with increasing N application rate, and differed significantly among test cultivars. With the growth of wheat, the relative concentration of chlorophyll a + b varied more obviously than those of chlorophyll b and carotenoid (Car), and the sensitive bands of the pigments occurred mostly within visible light range, especially in red-edge district. The analyses on the relationships between eight existing vegetation indices and leaf pigment concentrations indicated that the concentrations of chlorophyll a, chlorophyll b, and chlorophyll a + b were highly correlated with red edge position, and the relationships to REP(LE) were better than to REP(IG), giving the determination coefficient R2 as 0.835, 0.841 and 0.840, and standard error SE as 0.264, 0.095 and 0.353, respectively. However, the R2 values between Car and different spectral indices decreased significantly, and the differences among the spectrum indices were very small. The tests of the monitoring models with independent datasets indicated that REP(LE) and REP(IG) were the best to predict leaf pigment concentrations. The R2 of chlorophyll a, chlorophyll a + b, and Car for REP(LE) were 0.805, 0.744 and 0.588, with the RE being 9.0%, 9.7% and 14.6%, respectively, and the R2 and RE of chlorophyll b for REP(IG) were 0.632 and 18.2%, respectively. It was suggested that the red-edge parameters of hyper-spectral reflectance had stable relationships with the pigment concentrations in wheat leaves, and especially, REP(LE) could be used to reliably estimate the concentrations of leaf chlorophyll a and chlorophyll a + b.

Topics: Chlorophyll; Chlorophyll A; Models, Theoretical; Plant Leaves; Reproducibility of Results; Spectrum Analysis; Triticum

The major light-harvesting chlorophyll a/ b complex (LHCIIb) of the photosynthetic apparatus in green plants can be viewed as a protein scaffold binding and positioning a large number of pigment molecules that engage in rapid excitation energy transfer. This property makes LHCIIb potentially interesting as a light harvester (or a model thereof) in photoelectronic applications. Such applications would require the immobilization of LHCIIb (or similar dye-protein complexes) on a solid surface. In this work, the immobilization of recombinant LHCIIb is tested and optimized on functionalized gold surfaces via a histidine 6 tag (His tag) in the protein moiety. Immobilization efficiency and kinetics are analyzed by using surface plasmon resonance (SPR) and surface plasmon field-enhanced fluorescence spectroscopy (SPFS). The latter was also used to assess the integrity of immobilized LHCIIb by recording Chl b-sensitized Chl a emission spectra. Since His tags have been included in a substantial number of recombinant proteins, the immobilization technique developed here for LHCIIb presumably can be extended to a large range of other membrane and water-soluble proteins.

Topics: Biotinylation; Chlorophyll; Chlorophyll A; Gold; Histidine; Kinetics; Light; Models, Chemical; Molecular Conformation; Photochemistry; Photosystem II Protein Complex; Spectrometry, Fluorescence; Streptavidin; Surface Plasmon Resonance; Time Factors

Many fruit-tree species, including relevant Citrus spp varieties exhibit a reproductive biology that impairs breeding and strongly constrains genetic improvements. In citrus, juvenility increases the generation time while sexual sterility, inbreeding depression and self-incompatibility prevent the production of homozygous cultivars. Genomic technology may provide citrus researchers with a new set of tools to address these various restrictions. In this work, we report a valuable genomics-based protocol for the structural analysis of deletion mutations on an heterozygous background.. Two independent fast neutron mutants of self-incompatible clementine (Citrus clementina Hort. Ex Tan. cv. Clemenules) were the subject of the study. Both mutants, named 39B3 and 39E7, were expected to carry DNA deletions in hemizygous dosage. Array-based Comparative Genomic Hybridization (array-CGH) using a Citrus cDNA microarray allowed the identification of underrepresented genes in these two mutants. Subsequent comparison of citrus deleted genes with annotated plant genomes, especially poplar, made possible to predict the presence of a large deletion in 39B3 of about 700 kb and at least two deletions of approximately 100 and 500 kb in 39E7. The deletion in 39B3 was further characterized by PCR on available Citrus BACs, which helped us to build a partial physical map of the deletion. Among the deleted genes, ClpC-like gene coding for a putative subunit of a multifunctional chloroplastic protease involved in the regulation of chlorophyll b synthesis was directly related to the mutated phenotype since the mutant showed a reduced chlorophyll a/b ratio in green tissues.. In this work, we report the use of array-CGH for the successful identification of genes included in a hemizygous deletion induced by fast neutron irradiation on Citrus clementina. The study of gene content and order into the 39B3 deletion also led to the unexpected conclusion that microsynteny and local gene colinearity in this species were higher with Populus trichocarpa than with the phylogenetically closer Arabidopsis thaliana. This work corroborates the potential of Citrus genomic resources to assist mutagenesis-based approaches for functional genetics, structural studies and comparative genomics, and hence to facilitate citrus variety improvement.

Topics: Alleles; Arabidopsis; Chlorophyll; Chlorophyll A; Chromosome Mapping; Chromosomes, Artificial, Bacterial; Chromosomes, Plant; Citrus; Fast Neutrons; Gene Dosage; Genome, Plant; Genomics; Multigene Family; Mutagenesis; Oligonucleotide Array Sequence Analysis; Phenotype; Polymorphism, Single Nucleotide; Populus; Sequence Deletion; Species Specificity; Vitis

Chloroplast biogenesis involves careful coordination of both plastid and nuclear gene expression, which is achieved in part by retrograde signaling from the chloroplast to the nucleus. This can be demonstrated by the fact that the herbicide, Norflurazon (NF), which causes bleaching of chloroplasts, prevents the light induction of photosynthesis-related genes in the nucleus. It has been proposed that the tetrapyrrole pathway intermediate Mg-protoporphyrin IX acts as the signaling molecule in this pathway and accumulates in the chloroplasts and cytosol of the cell after NF treatment. Here we present data that demonstrate that this model is too simplistic. We have developed a sensitive liquid chromatography-mass spectrometry (LC/MS) method to measure tetrapyrrole intermediates and have shown that no Mg-protoporphyrin IX, nor indeed any other chlorophyll-biosynthesis intermediate, can be detected in NF-treated plants under conditions in which nuclear gene expression is repressed. Conversely when endogenous Mg-protoporphyrin IX levels are artificially increased by supplementation with the tetrapyrrole precursor, 5-aminolevulinic acid, the expression of nuclear-encoded photosynthetic genes is induced, not repressed. We also demonstrate that NF-treatment leads to a strong down-regulation of tetrapyrrole biosynthesis genes, consistent with the absence of an accumulation of tetrapyrrole intermediates. Finally, there is no correlation between nuclear-gene expression and any of the chlorophyll biosynthetic intermediates over a range of growth conditions and treatments. Instead, it is possible that a perturbation of tetrapyrrole synthesis may lead to localized ROS production or an altered redox state of the plastid, which could mediate retrograde signaling.

Topics: Arabidopsis; Cell Nucleus; Chlorophyll; Chlorophyll A; Chromatography, Liquid; Gene Expression Regulation, Plant; Herbicides; Mass Spectrometry; Plastids; Protoporphyrins; Pyridazines; Seedlings; Signal Transduction

Comparisons were made between the content of Al, Cd, Cu, Fe, Mn, Pb, Sb and Zn in thalli of the lichen Xanthoria parietina and selected physiological processes. Assimilation pigments, activity of photosystem II, parietin, reactive substances produced using thiobarbituric acid (TBA) and CO(2) gas exchange were measured in order to assess tolerance to atmospheric sources of pollution. As expected, lichen thalli accumulated high amounts of the measured elements in relation to distance from pollution sources in Kosice, Slovak Republic (US Steel factory and vehicular traffic in the city center). However, except for TBA reactive substances production and to some extent CO(2) gas exchange at the most polluted station, none of the tested physiological parameters showed a clear correlation between accumulation of elements and physiological damage. This reflected a high degree of pollution tolerance in this lichen and corresponded with its high abundance in Kosice.

Topics: Carbon Dioxide; Carotenoids; Chlorophyll; Chlorophyll A; Emodin; Environmental Monitoring; Environmental Pollutants; Lichens; Metals; Photosystem II Protein Complex; Slovakia; Thiobarbituric Acid Reactive Substances

There are four LhcII genes of Dunaliella salina have been submitted to the database of GenBank. However, little is known about Lhca genes of this green alga, although this knowledge might be available to study the composition and phylogenesis of Lhc gene family. Recently, one Lhca gene was been cloned from the green alga D. salina by PCR amplification using degenerate primers. This cDNA, designated as DsLhca1, contains an open reading frame encoded a protein of 222 amino acids with a calculated molecular mass of 27.8 kDa. DsLhca1 is predicted to contain three transmembrane domains and a N-terminal chloroplast transit peptide (cTP) with length of 33 amino acids. The genomic sequence of DsLhca1 is composed of five introns. The deduced polypeptide sequence of this gene showed a lower degree of identity (less than 30%) with LHCII proteins from D. salina. But its homology to Lhca proteins of other algae (Volvox carteri Lhca_AF110786) was higher with pairwise identities of up to 67.1%. Phylogenetic analysis indicated that DsLhcal protein cannot be assigned to any types of Lhca proteins in higher plants or in Chlamydomonas reinhardtii.

Topics: Algal Proteins; Amino Acid Sequence; Base Sequence; Chlorophyll; Chlorophyll A; Chlorophyta; Light-Harvesting Protein Complexes; Molecular Sequence Data; Photosystem I Protein Complex; Phylogeny

The light-harvesting chlorophyll a/b-binding protein of photosystem II (LHCIIb) fulfills multiple functions, such as light harvesting and energy dissipation under different illuminations. The crystal structure of LHCIIb at the near atomic resolution reveals an antiparallel strands structure in the lumenal loop between the transmembrane helices B/C. To study the structural and functional significances of this structure, three amino acids (Val-119, His-120, and Ser-123) in this region have been exchanged to Phe, Leu, and Gly, respectively, and the influence of the mutagenesis on the structure and function of LHCIIb has been investigated. The results are as follows. 1) Circular dichroism spectra of the mutations reveals that the antiparallel strands in the lumenal region are very important for adjusting pigment conformation in the neoxanthin domain of LHCIIb. Although the mutagenesis causes only a slight loss of the Neo binding in the complexes (V119F, 0.09; S123G, 0.19; and H120L, 0.27), it imparts remarkable changes to the pigment conformation. 2) Substituting Ser-123 with Gly results in a higher susceptibility to photodamage, an increased tendency to aggregate, and enhanced fluorescence quenching induced by the medium acidification. These results demonstrate that this antiparallel strands domain plays an important role in regulating the pigment conformation and in adjusting the aggregation and the fluorescence yield of LHCIIb.

Topics: Chlorophyll; Chlorophyll A; Circular Dichroism; Fluorescence; Light-Harvesting Protein Complexes; Models, Molecular; Mutagenesis, Site-Directed; Mutation; Photobleaching; Photosystem II Protein Complex; Pisum sativum; Plant Proteins; Protein Binding; Recombinant Fusion Proteins

The effects of additional nitrogen on the toxicity and removal of ferrocyanide and ferricyanide by cyanogenic plants were investigated. Maize (Zea mays L. var. ZN 304) seedlings were grown in the hydroponic solutions with or without additional nitrogen, and amended with either potassium ferrocyanide or potassium ferricyanide at 25.0 +/- 0.5 degrees C for 144 h. Various physiological parameters were monitored to determine the responses of the plant seedlings to the exposure of these two chemicals. A remarkable decrease in transpiration rate, biomass, shoot length, chlorophyll contents, and soluble proteins was evident for maize seedlings grown in the N-free hydroponic solutions spiked with either ferrocyanide or ferricyanide (P < 0.01), but slight changes were observed in the selective parameters in the N-containing hydroponic solutions spiked with either of these chemicals (P > 0.05). A higher removal of ferrocyanide than ferricyanide was registered in the N-free hydroponic solutions, but more ferricyanide than ferrocyanide was removed by maize grown in the N-containing nutrient solutions (P < 0.01). Although roots of maize accumulated iron cyanides, more cyanide was recovered in plant materials of those grown in the N-containing hydroponic solutions than the N-free nutrient solutions (P < 0.05). Mass balance analysis indicated that the majority of the iron cyanides removed from solution was assimilated by maize and additional nitrogen had a significantly negative impact on the uptake of both chemicals (P < 0.01). Results of this study suggest that uptake and assimilation mechanisms for ferrocyanide and ferricyanide might be quite different in maize and the application of the external nitrogen has a substantial influence on the removal of both iron cyanides by plants. None of the iron cyanide complexes can serve as a sole nitrogen source to support maize growth.

Topics: Analysis of Variance; Chlorophyll; Chlorophyll A; Ferricyanides; Ferrocyanides; Hydroponics; Nitrogen; Plant Roots; Plant Shoots; Plant Transpiration; Seedlings; Volatilization; Zea mays

* Over 6 d of dark-induced senescence, leaf segments of wild-type Lolium temulentum lost > 96% chlorophyll a + b; leaves from plants containing a staygreen mutation introgressed from Festuca pratensis, which has a lesion in the senescence-associated fragmentation of pigment-proteolipid complexes, retained over 43% of total chlorophyll over the same period. * Mutant segments preferentially retained thylakoid membrane proteins (exemplified by LHCP II) but lost other cellular proteins at the same rate as wild-type tissue. The protein synthesis inhibitor D-MDMP inhibited chlorophyll degradation and partially prevented protein loss in both genotypes, but tissues treated with the ineffective L-stereoisomer were indistinguishable from water controls. * Principal-components analysis of leaf reflectance spectra distinguished between genotypes, time points and D-MDMP treatments, showing the disruption of pigment metabolism during senescence brought about by the staygreen mutation, by inhibition of protein synthesis and by combinations of the two factors. * The build-up of oxidized, dephytylated and phaeo-derivatives of chl a during senescence of staygreen tissue was prevented by D-MDMP and associated with characteristic difference spectra when senescent mutant tissue was compared with wild-type or inhibitor-treated samples. The suitability of senescence as a subject for systems biology approaches is discussed.

Topics: Cellular Senescence; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Festuca; Genotype; Lolium; Pigments, Biological; Plant Proteins; Principal Component Analysis; Propionates

The interaction between physical and biological factors responsible for the cessation of ripple migration on a sandy intertidal flat was examined during a microalgal bloom period in late winter/early spring, as part of a wider study into the biostabilisation of intertidal sediments. Ripple positions and ripple geometry were monitored, and surface sediment was sampled, at weekly intervals over a 5-week period. Ripples remained in the same position for at least 4 weeks, during which time there was a progressive reduction in bedform height (smoothing) and deposition of some 1.5 cm sediment, mainly in the ripple troughs (surface levelling). The mean chlorophyll a (chl a) sediment content was 6.0 microg gDW(-1) (DW: dry weight) (0-1 mm depth fraction), with a maximum value of 7.4 microg gDW(-1) half way through the bloom. Mean colloidal-S carbohydrate (S: saline extraction) content was 131 microg GE gDW(-1) (GE: glucose equivalent) (0-1 mm), with a maximum of 261 microg GE gDW(-1 )towards the end of the bloom. Important accessory pigments were peridinin (indicative of dinophytes) and fucoxanthin (diatoms). Stepwise multiple regression showed that peridinin was the best predictor of chl a. For the first time, in situ evidence for the mediation of (wave) ripple migration by microalgae is provided. Results indicate that diatoms, and quite possibly dinophytes, can have a significant effect on intertidal flat ripple mobility on a temporal scale of weeks. In addition, microalgal effects appear capable of effecting a reduction in bed roughness on a spatial scale of up to 10(-2 )m, with a subsequent reduction in bottom stress and bed erodability. It is suggested that a unique combination of environmental conditions, in conjunction with the microalgal bloom(s), promoted the initial cessation of ripple movement, and that stationary-phase, diatom-derived extracellular polymeric substances (EPS) (and possibly dinophyte-derived EPS) may have prolonged the condition. It is reasonable to suppose that ripple stabilisation by similar processes may have contributed to ripple mark preservation in the geological record. A conceptual model of sandy intertidal flat processes is presented, illustrating two conditions: (i) a low EPS/microalgae sediment content with low ripple stabilisation and preservation potential; and (ii) a high EPS/microalgae content with higher preservation potential.

Topics: Carbohydrates; Carotenoids; Chlorophyll; Chlorophyll A; England; Environmental Microbiology; Eukaryota; Geologic Sediments; Water Movements; Xanthophylls; Zeaxanthins

Three isoforms of the major light-harvesting chlorophyll (Chl) a/b complexs of photosystem II (LHCIIb) in the pea, namely, Lhcb1, Lhcb2, and Lhcb3, were obtained by overexpression of apoprotein in Escherichia coli and by successfully refolding these isoforms with thylakoid pigments in vitro. The sequences of the protein, pigment stoichiometries, spectroscopic characteristics, thermo- and photostabilities of different isoforms were analysed. Comparison of their spectroscopic properties and structural stabilities revealed that Lhcb3 differed strongly from Lhcb1 and Lhcb2 in both respects. It showed the lowest Qy transition energy, with its reddest absorption about 2 nm red-shifted, and the highest photostability under strong illuminations. Among the three isoforms, Lhcb 2 showed lowest thermal stability regarding energy transfer from Chl b to Chl a in the complexes, which implies that the main function of Lhcb 2 under high temperature stress is not the energy transfer.

Topics: Chlorophyll; Chlorophyll A; Escherichia coli; Isoenzymes; Light-Harvesting Protein Complexes; Photosystem II Protein Complex; Pisum sativum; Plant Proteins; Protein Folding; Protein Structure, Quaternary; Recombinant Proteins; Thylakoids

Energy-dependent quenching of excess absorbed light energy (qE) is a vital mechanism for regulating photosynthetic light harvesting in higher plants. All of the physiological characteristics of qE have been positively correlated with charge transfer between coupled chlorophyll and zeaxanthin molecules in the light-harvesting antenna of photosystem II (PSII). We found evidence for charge-transfer quenching in all three of the individual minor antenna complexes of PSII (CP29, CP26, and CP24), and we conclude that charge-transfer quenching in CP29 involves a delocalized state of an excitonically coupled chlorophyll dimer. We propose that reversible conformational changes in CP29 can "tune" the electronic coupling between the chlorophylls in this dimer, thereby modulating the energy of the chlorophyll-zeaxanthin charge-transfer state and switching on and off the charge-transfer quenching during qE.

Topics: Arabidopsis Proteins; Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Chloroplast Proteins; Electron Transport; Electrophysiology; Light; Light-Harvesting Protein Complexes; Lutein; Models, Molecular; Photosystem II Protein Complex; Protein Conformation; Recombinant Proteins; Ribonucleoproteins; Structure-Activity Relationship; Xanthophylls; Zeaxanthins

Water stress is one of the most important environmental factors that reduce growth, development and production of plants. Stress was applied with polyethyleneglycol (PEG) 6000 and water potentials were: zero (control), -0.15 (PEG 10%), -0.49 (PEG 20%), -1.03 (PEG 30%) and -1.76 (PEG 40%) MPa. The roots and leaves respiration of two maize (Zea mays L.) cultivars -704 and 301- were determined in various concentrations of PEG 6000. Oxygen uptake declined in leaves and roots with increasing PEG concentrations. Decrease of oxygen uptake in roots and leaves of 704 variety were higher than 301 variety. Chlorophyll a, b and total chlorophyll content were significantly decreased (p < 0.05), but carotenoids content increased (p < 0.05) under water stress. Decrease of chlorophyll content in 704 var. was higher than 301 var., but carotenoids content in 301 var. was higher than 704 var. Relative Water Content (RWC) was used to indicate the degree of stress. RWC decreased with increasing PEG concentrations. Lowering of RWC reduced growth and increased shoot/root ratio. Decrease of water content in 704 plants was higher than 301 plants. Shoot/root ratio in 704 var. was higher than 301 var.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Oxygen; Oxygen Consumption; Photosynthesis; Pigmentation; Polyethylene Glycols; Pressure; Water; Zea mays

Growth parameters and biochemical changes were studied in roots and leaves of 15 day old maize grown in a nutrient solution containing various copper concentrations (0, 25, 50, 75 and 100 microM). An accumulation of H2O2 was observed in roots and shoots. The leaf chlorophyll a, b and carotenoid contents decreased with increasing Cu concentration. The results demonstrated adverse effects of Cu on N metabolism and plant growth. Cu exposure elevated Cu concentration and decreased Nitrate Reductase (NR) activity in the roots and shoots. However, Cu exposure increased total free amino acid content in the leaves.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Copper; Dose-Response Relationship, Drug; Electrons; Nitrate Reductase; Photochemistry; Plant Roots; Plant Shoots; Seeds; Time Factors; Zea mays

The saline pond microalga, Dunaliella salina (Dunal) Teod. maintained in De Walne's (basal) medium under laboratory conditions was confirmed by amplifying the chromosomal DNA of the microalga by PCR with specific primers MA1 and MA2. Seaweed extracts obtained from Sargassum wightii and Ulva lactuca were amended separately at 1.0%, 1.5%, 2.0% and 2.5% levels to the basal medium in order to assess their potential on the growth and concentration of pigments, viz. Chl a, Chl b and beta-carotene of the alga. beta-Carotene was isolated and visible absorption spectrum was taken at 443 and 475 nm confirmed the presence of 9-cis-beta-carotene and all-trans-beta-carotene isomers. Maximum yield, highest division rate (mu) and highest pigment concentrations were observed in the cells grown in 1.5% S. wightii and 2.0% U. lactuca amended medium and these cells were subjected to DAPI staining. The results of epifluorescence microscopy and image analysis revealed a significant enhancement of the cell and nuclear area of the microalgae.

Topics: beta Carotene; Chlorophyll; Chlorophyll A; Chlorophyta; DNA; India; Industrial Microbiology; Microscopy, Confocal; Microscopy, Fluorescence; Plant Extracts; Polymerase Chain Reaction; Seaweed; Water Microbiology

A method for the isolation of chlorophylls from spinach by counter-current chromatography was developed. An initial extraction protocol was devised to avoid the notorious sensitivity of chlorophylls to degradation by light, heat, oxygen, acids and bases. Further purification and separation of chlorophylls a and b were achieved using counter-current chromatography. Chlorophyll structures and purities were established by HPLC, fast atom bombardment mass spectrometry and nuclear magnetic resonance. Purity was estimated to be >95% (100% by HPLC). Typical yields from 30g of freeze-dried spinach were 300mg of chlorophyll a and 100mg of chlorophyll b.

Topics: Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Countercurrent Distribution; Nuclear Magnetic Resonance, Biomolecular; Spectrometry, Mass, Fast Atom Bombardment; Spinacia oleracea

Action spectra for photosystem II (PSII)-driven oxygen evolution and of photosystem I (PSI)-mediated H(2) photoproduction and photoinhibition of respiration were used to determine the participation of chlorophyll (Chl) a/b-binding Pcb proteins in the functions of pigment apparatus of Prochlorothrix hollandica. Comparison of the in situ action spectra with absorption spectra of PSII and PSI complexes isolated from the cyanobacterium Synechocystis 6803 revealed a shoulder at 650 nm that indicated presence of Chl b in the both photosystems of P. hollandica. Fitting of two action spectra to absorption spectrum of the cells showed a chlorophyll ratio of 4:1 in favor of PSI. Effective antenna sizes estimated from photochemical cross-sections of the relevant photoreactions were found to be 192+/-28 and 139+/-15 chlorophyll molecules for the competent PSI and PSII reaction centers, respectively. The value for PSI is in a quite good agreement with previous electron microscopy data for isolated Pcb-PSI supercomplexes from P. hollandica that show a trimeric PSI core surrounded by a ring of 18 Pcb subunits. The antenna size of PSII implies that the PSII core dimers are associated with approximately 14 Pcb light-harvesting proteins, and form the largest known Pcb-PSII supercomplexes.

Topics: Bacterial Proteins; Carrier Proteins; Chlorophyll; Chlorophyll A; Models, Biological; Photosystem I Protein Complex; Photosystem II Protein Complex; Prochlorothrix; Synechocystis

Among the herbicides used in vineyards, the pre-emergence soil-applied flumioxazin (fmx) is a recently used molecule that inhibits chlorophyll biosynthesis in weed species. The aim of this work is to further characterize the effects of fmx on the non-target grapevine (Vitis vinifera L. cv. Chardonnay) using cutting as a model. Several photosynthesis parameters were estimated during 25 days after treatment with various fmx concentrations (from 0.5mM to 50mM). Measuring chlorophyll fluorescence it appeared that fmx or a by-product penetrated the plant throughout roots and spread throughout vessels. Besides the initial target, protox, fmx affected other functions related to photosynthesis. Fmx induced a simultaneous drop of both P(n), g(s) and T. Fmx caused stomatal closure, which partially explains the decrease of the net photosynthesis. The decline in F(v)/F(m) indicates that the photochemistry of PSII and its ability to reduce the primary acceptor Q(A) are also affected by fmx in grapevine. Fmx leads to a decrease in the coefficients of both photochemical and non-photochemical quenching. Simultaneous stomatal closure and decrease in the quantum yield of CO(2) assimilation indicate a change in energy metabolism following fmx stress. After ten days of fmx treatment, analyses of the response of net carbon assimilation in leaves to different intercellular CO(2) concentrations have shown a decrease of the maximum carboxylation velocity of RuBP. Stomatal closure, PSII photochemistry, change in energy metabolism and RuBP activity were affected by fmx treatment.

Topics: Benzoxazines; Chlorophyll; Chlorophyll A; Herbicides; Photosynthesis; Phthalimides; Spectrometry, Fluorescence; Vitis

The apoprotein of the major light-harvesting chlorophyll a/b complex (LHCIIb) is post-translationally imported into the chloroplast, where membrane insertion, protein folding, and pigment binding take place. The sequence and molecular mechanism of the latter steps is largely unknown. The complex spontaneously self-organises in vitro to form structurally authentic LHCIIb upon reconstituting the unfolded recombinant protein with the pigments chlorophyll a, b, and carotenoids in detergent micelles. Former measurements of LHCIIb assembly had revealed two apparent kinetic phases, a faster one (tau1) in the range of 10 s to 1 min, and a slower one (tau2) in the range of several min. To unravel the sequence of events we analysed the binding of chlorophylls into the complex by using time-resolved fluorescence measurements of resonance energy transfer from chlorophylls to an acceptor dye attached to the apoprotein. Chlorophyll a, offered in the absence of chlorophyll b, bound with the faster kinetics (tau1) exclusively whereas chlorophyll b, in the absence of chlorophyll a, bound predominantly with the slower kinetics (tau2). In double-jump experiments, LHCIIb assembly could be dissected into a faster chlorophyll a and a subsequent, predominantly slower chlorophyll b-binding step. The assignment of the faster and the slower kinetic phase to predominantly chlorophyll a and exclusively chlorophyll b binding, respectively, was verified by analysing the assembly kinetics with a circular dichroism signal in the visible domain presumably reflecting the establishment of pigment-pigment interactions. We propose that slow chlorophyll binding is confined to the exclusively chlorophyll b binding sites whereas faster binding occurs to the chlorophyll a binding sites. The latter sites can bind both chlorophylls a and b but in a reversible fashion as long as the complex is not stabilised by proper occupation of the chlorophyll b sites. The resulting two-step model of LHCIIb assembly is able to reconcile the highly specific binding sites containing either chlorophyll a or b, as seen in the recent crystal structures of LHCIIb, with the observation of promiscuous binding sites able to bind both chlorophyll a and b in numerous reconstitution analyses of LHCIIb assembly.

Topics: Animals; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Circular Dichroism; Energy Transfer; Fluorescence; Kinetics; Light-Harvesting Protein Complexes; Models, Biological; Protein Binding; Protein Structure, Quaternary; Time Factors

The light-harvesting complexes (LHCs) of land plants and green algae have essential roles in light capture and photoprotection. Though the functional diversity of the individual LHC proteins are well described in many land plants, the extent of this family in the majority of green algal groups is unknown. To examine the evolution of the chlorophyll a/b antennae system and to infer its ancestral state, we initiated several expressed sequence tag projects from a taxonomically broad range of chlorophyll a/b-containing protists. This included representatives from the Ulvophyceae (Acetabularia acetabulum), the Mesostigmatophyceae (Mesostigma viride), and the Prasinophyceae (Micromonas sp.), as well as one representative from each of the Euglenozoa (Euglena gracilis) and Chlorarachniophyta (Bigelowiella natans), whose plastids evolved secondarily from a green alga. It is clear that the core antenna system was well developed prior to green algal diversification and likely consisted of the CP29 (Lhcb4) and CP26 (Lhcb5) proteins associated with photosystem II plus a photosystem I antenna composed of proteins encoded by at least Lhca3 and two green algal-specific proteins encoded by the Lhca2 and 9 genes. In organisms containing secondary plastids, we found no evidence for orthologs to the plant/algal antennae with the exception of CP29. We also identified PsbS homologs in the Ulvophyceae and the Prasinophyceae, indicating that this distinctive protein appeared prior to green algal diversification. This analysis provides a snapshot of the antenna systems in diverse green algae, and allows us to infer the changing complexity of the antenna system during green algal evolution.

Topics: Amino Acid Sequence; Biological Evolution; Chlorophyll; Chlorophyll A; Chlorophyta; DNA, Complementary; Light-Harvesting Protein Complexes; Molecular Sequence Data; Plants; Sequence Homology, Amino Acid; Species Specificity

Eelgrass beds are important habitats for many organisms, but there has been a decline in the area covered by eelgrass during the last decades due to increased eutrophication resulting in increased shading from phytoplankton. The use of herbicides in terrestrial agriculture has also increased over the last century, and while the effects of herbicides on non-target organisms have been well studied in freshwater they are overlooked in coastal waters. It is not known if herbicides have any effect on the distribution of eelgrass (Zostera marina), or how natural phytoplankton communities respond to the same herbicides. Direct and indirect effects of the herbicides Glyphosate, Bentazone and MCPA both as single toxicants and as mixtures, on the eelgrass plants were investigated in this study. The direct effects on eelgrass were examined by measuring the four different endpoints; the relatively growth rate as length and weight, the chlorophyll a and b ratio, as well as the RNA-DNA ratio, at the end of a 3 days exposure period. The indirect effect was investigated by measuring the effect on a natural phytoplankton production from Roskilde Fjord six times during 1 year. The results showed that the chlorophyll a-b and RNA-DNA ratios were the most sensitive endpoints in single herbicide experiments. The effects of herbicide mixtures on eelgrass were much larger compared to the single substances. Nearly a halving was found for both the relatively growth rate in length and weight, and the RNA-DNA as well as the chlorophyll a-b ratios were also significant reduced. This indicates a possible synergistic effect, and calculations based on the concentration addition model indicate that the low concentrations mixture has a synergistic effect, whereas the high concentration mixture has an antagonistic effect on eelgrass (Z. marina). The low concentrations mixture is the one with the highest relevance for coastal areas. The effect on phytoplankton showed some variation over the year but mainly in a stimulation in primary production. This means that herbicides affect eelgrass both directly and indirectly.

Topics: 2-Methyl-4-chlorophenoxyacetic Acid; Chlorophyll; Chlorophyll A; Denmark; DNA, Plant; Glycine; Glyphosate; Herbicides; Phytoplankton; RNA, Plant; Seasons; Seawater; Water Pollutants, Chemical; Zosteraceae

The plastid-encoded psaJ gene encodes a hydrophobic low-molecular-mass subunit of photosystem I (PSI) containing one transmembrane helix. Homoplastomic transformants with an inactivated psaJ gene were devoid of PSI-J protein. The mutant plants were slightly smaller and paler than wild-type because of a 13% reduction in chlorophyll content per leaf area caused by an approximately 20% reduction in PSI. The amount of the peripheral antenna proteins, Lhca2 and Lhca3, was decreased to the same level as the core subunits, but Lhca1 and Lhca4 were present in relative excess. The functional size of the PSI antenna was not affected, suggesting that PSI-J is not involved in binding of light-harvesting complex I. The specific PSI activity, measured as NADP(+) photoreduction in vitro, revealed a 55% reduction in electron transport through PSI in the mutant. No significant difference in the second-order rate constant for electron transfer from reduced plastocyanin to oxidized P700 was observed in the absence of PSI-J. Instead, a large fraction of PSI was found to be inactive. Immunoblotting analysis revealed a secondary loss of the luminal PSI-N subunit in PSI particles devoid of PSI-J. Presumably PSI-J affects the conformation of PSI-F, which in turn affects the binding of PSI-N. This together renders a fraction of the PSI particles inactive. Thus, PSI-J is an important subunit that, together with PSI-F and PSI-N, is required for formation of the plastocyanin-binding domain of PSI. PSI-J is furthermore important for stability or assembly of the PSI complex.

Topics: Chlorophyll; Chlorophyll A; Chloroplast Proton-Translocating ATPases; Chloroplasts; Electron Transport; Gene Silencing; Kinetics; Light-Harvesting Protein Complexes; Mutation; NADP; Nicotiana; Oxidation-Reduction; Phenotype; Photosystem I Protein Complex; Photosystem II Protein Complex; Plant Leaves; Plant Proteins; Plants, Genetically Modified; Plastocyanin; Protein Subunits; Spectrometry, Fluorescence; Thylakoids

Chromium is a highly toxic non-essential metal for microorganisms and plants. Due to its widespread industrial use, chromium (Cr) has become a serious pollutant in diverse environmental settings. The presence of Cr leads to the selection of algal populations able to tolerate high levels of Cr compounds. The diverse Cr-resistance mechanisms displayed by microorganisms include biosorption, diminished accumulation, precipitation, reduction of Cr(6+) to Cr(3+), and chromate efflux. In this paper we describe the effects of Cr(6+) (the more toxic species) on the photosynthetic and photoreceptive apparatus of the fresh water unicellular alga Chlamydomonas reinhardtii. We measured the effect of the heavy metal by means of in vivo absorption microspectroscopy of both the thylakoid compartments and the eyespot. The decomposition of the overall absorption spectra in pigment constituents indicates that Cr(6+) induced a complete pheophinitization of the chrorophylls and a modification of the carotenoids present in the eyespot only when its concentration is equal or greater than 10 microM. Due to this low tolerance level, C. reinhardtii could be used as indicator of Cr pollution, but it is not feasible for bioremediation purposes.

Topics: Animals; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Chromium; Photosynthesis; Thylakoids; Water Pollutants, Chemical

An open top chamber experiment was carried out in the summer of 2003 to examine the effect of nitrous acid (HONO) gas on the physiological status of Scots pine saplings (Pinus sylvestris). Four-year-old pine trees were exposed to two different levels of HONO gas (at ca. 2.5 ppb and 5.0 ppb) and a control (filtered air) from early evening to early morning (18:00-6:00), in duplicate open top chambers. Significant decreases in the ratios of chlorophylls a to b, an increase in the carbon to nitrogen (C/N) ratio, and a reduction of maximum yield of PS II (F(v)/F(m)) in pine needles were also observed after the 2 months' fumigation. Cation contents of pine needles were also decreased by the fumigation with HONO gas. The results could be explained by the harmful effects of OH radicals, generated from photolysis of HONO gas, and/or aqueous phase HONO (NO(2)(-)/HONO), on the photosynthetic capacity of pine needles.

Topics: Air Pollutants; Carbon; Chlorophyll; Chlorophyll A; Environmental Exposure; Environmental Monitoring; Free Radicals; Metals; Nitrogen; Nitrous Acid; Photosynthesis; Pinus sylvestris

In this work we modeled the circular dichroism (CD) spectrum of LHCII, the main light harvesting antenna of photosystem II of higher plants. Excitonic calculations are performed for a monomeric subunit, taken from the crystal structure of trimeric LHCII from spinach [Liu, Z. F., Yan, H. C., Wang, K. B., Kuang, T. Y., Zhang, J. P., Gui, L. L., An, X. M., and Chang, W. R. (2004) Nature 428, 287-292]. All of the major features of the CD spectrum above 450 nm are satisfactorily reproduced, and possible orientations of the Chl and carotenoid transition dipole moments are identified. The obtained modeling parameters are used to simulate the CD spectra of two complexes with altered pigment composition: a mutant lacking Chls a 611-612 and a complex lacking the carotenoid neoxanthin. By removing the relevant pigment(s) from the structure, we are able to reproduce their spectra, which implies that the alteration does not disturb the overall structure. The CD spectrum of trimeric LHCII shows a reversed relative intensity of the two negative bands around 470 and 490 nm as compared to monomeric LHCII. The simulations reproduce this reversal, indicating that it is mainly due to interactions between chromophores in different monomeric subunits, and the trimerization does not induce observable changes in the monomeric structure. Our simulated spectrum resembles one of two different trimeric CD spectra reported in literature. We argue that the differences in the experimental trimeric CD spectra are caused by changes in the strength of the monomer-monomer interactions due to the differences in detergents used for the purification of the complexes.

Topics: Chlorophyll; Chlorophyll A; Circular Dichroism; Light-Harvesting Protein Complexes; Models, Chemical; Mutation; Photosystem II Protein Complex

Chlorophyll (Chl) d is a major chlorophyll in a novel oxygenic prokaryote Acaryochloris marina. Here we first report the redox potential of Chl d in vitro. The oxidation potential of Chl d was +0.88 V vs. SHE in acetonitrile; the value was higher than that of Chl a (+0.81 V) and lower than that of Chl b (+0.94 V). The oxidation potential order, Chl b>Chl d>Chl a, can be explained by inductive effect of substituent groups on the conjugated pi-electron system on the macrocycle. Corresponding pheophytins showed the same order; Phe b (+1.25 V)>Phe d (+1.21 V)>Phe a (+1.14 V), but the values were significantly higher than those of Chls, which are rationalized in terms of an electron density decrease in the pi-system by the replacement of magnesium with more electronegative hydrogen. Consequently, oxidation potential of Chl a was found to be the lowest among Chls and Phes. The results will help us to broaden our views on photosystems in A. marina.

Topics: Acetonitriles; Chlorophyll; Chlorophyll A; Cyanobacteria; Dimethylformamide; Electrochemistry; In Vitro Techniques; Models, Chemical; Molecular Structure; Oxidation-Reduction; Petroselinum; Pheophytins; Solvents

The possibility that the chlorophyll (chl) ring distortions observed in the crystal structures of chl-protein complexes are involved in the transition energy modulation, giving rise to the spectral forms, is investigated. The out-of-plane chl-macrocycle distortions are described using an orthonormal set of deformations, defined by the displacements along the six lowest-frequency, out-of-plane normal coordinates. The total chl-ring deformation is the linear combination of these six deformations. The two higher occupied and the two lower unoccupied chl molecular orbitals, which define the Q(y) electronic transition, have the same symmetry as four of the six out-of-plane lowest frequency modes. We assume that a deformation along the normal-coordinate having the same symmetry as a given molecular orbital will perturb that orbital and modify its energy. The changes in the chl Q(y) transition energies are evaluated in the Peridinin-Chl-Protein complex and in light harvesting complex II (LHCII), using crystallographic data. The macrocycle deformations induce a distribution of the chl Q(y) electronic energy transitions which, for LHCII, is broader for chla than for chlb. This provides the physical mechanism to explain the long-held view that the chla spectral forms in LHCII are both more numerous and cover a wider energy range than those of chlb.

Topics: Animals; Biophysical Phenomena; Biophysics; Carotenoids; Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Molecular Conformation; Protozoan Proteins; Static Electricity; Thermodynamics

Single molecule spectroscopy experiments are reported for native peridinin-chlorophyll a-protein (PCP) complexes, and three reconstituted light-harvesting systems, where an N-terminal construct of native PCP from Amphidinium carterae has been reconstituted with chlorophyll (Chl) mixtures: with Chl a, with Chl b and with both Chl a and Chl b. Using laser excitation into peridinin (Per) absorption band we take advantage of sub-picosecond energy transfer from Per to Chl that is order of magnitude faster than the Förster energy transfer between the Chl molecules to independently populate each Chl in the complex. The results indicate that reconstituted PCP complexes contain only two Chl molecules, so that they are spectroscopically equivalent to monomers of native-trimeric-PCP and do not aggregate further. Through removal of ensemble averaging we are able to observe for single reconstituted PCP complexes two clear steps in fluorescence intensity timetraces attributed to subsequent bleaching of the two Chl molecules. Importantly, the bleaching of the first Chl affects neither the energy nor the intensity of the emission of the second one. Since in strongly interacting systems Chl is a very efficient quencher of the fluorescence, this behavior implies that the two fluorescing Chls within a PCP monomer interact very weakly with each other which makes it possible to independently monitor the fluorescence of each individual chromophore in the complex. We apply this property, which distinguishes PCP from other light-harvesting systems, to measure the distribution of the energy splitting between two chemically identical Chl a molecules contained in the PCP monomer that reaches 280 cm(-1). In agreement with this interpretation, stepwise bleaching of fluorescence is also observed for native PCP complexes, which contain six Chls. Most PCP complexes reconstituted with both Chl a and Chl b show two emission lines, whose wavelengths correspond to the fluorescence of Chl a and Chl b. This is a clear proof that these two different chromophores are present in a single PCP monomer. Single molecule fluorescence studies of PCP complexes, both native and artificially reconstituted with chlorophyll mixtures, provide new and detailed information necessary to fully understand the energy transfer in this unique light-harvesting system.

Topics: Animals; Carotenoids; Chlorophyll; Chlorophyll A; Dinoflagellida; Fluorescence; Light-Harvesting Protein Complexes; Protein Conformation; Protozoan Proteins; Spectrometry, Fluorescence

Topics: Chlorophyll; Chlorophyll A; Isotopes; Magnesium; Plant Physiological Phenomena

The differential pigment composition and photosynthetic activity of sun and shade leaves of deciduous (Acer pseudoplatanus, Fagus sylvatica, Tilia cordata) and coniferous (Abies alba) trees was comparatively determined by studying the photosynthetic rates via CO(2) measurements and also by imaging the Chl fluorescence decrease ratio (R(Fd)), which is an in vivo indicator of the net CO(2) assimilation rates. The thicker sun leaves and needles in all tree species were characterized by a lower specific leaf area, lower water content, higher total chlorophyll (Chl) a+b and total carotenoid (Cars) content per leaf area unit, as well as higher values for the ratio Chl a/b compared to the much thinner shade leaves and needles that possess a higher Chl a+b and Cars content on a dry matter basis and higher values for the weight ratio Chls/Cars. Sun leaves and needles exhibited higher rates of maximum net photosynthetic CO(2) assimilation (P(Nmax)) measured at saturating irradiance associated with higher maximum stomatal conductance for water vapor efflux. The differences in photosynthetic activity between sun and shade leaves and needles could also be sensed via imaging the Chl fluorescence decrease ratio R(Fd), since it linearly correlated to the P(Nmax) rates at saturating irradiance. Chl fluorescence imaging not only provided the possibility to screen the differences in P(N) rates between sun and shade leaves, but in addition permitted detection and quantification of the large gradients in photosynthetic rates across the leaf area existing in sun and shade leaves.

Topics: Carbon Dioxide; Carotenoids; Chlorophyll; Chlorophyll A; Fluorescence; Gases; Light; Photosynthesis; Pigmentation; Plant Leaves; Spectrometry, Fluorescence; Sunlight; Trees

Most Chl a in PSI complexes was removed without any loss of P700 by ether treatment, yielding antenna-depleted P700-Chl a protein complexes (CP1s) with a Chl a/P700 ratio of 12. On addition of about 60 molecules of Chl b per P700 with phosphatidylglycerol, about 20 molecules of Chl b per P700 were bound to the complexes. The ratio of the bound Chl b to the added Chl b was about one-third, irrespective of the amount of Chl b added. The same partition ratio was obtained on reconstitution with Chl a, suggesting that the binding affinity of Chl b for the Chl a-binding sites is similar to that of Chl a. The relative quantum efficiency of P700 photooxidation, determined by the increase in its initial rate, increased in proportion to the increase in number of bound Chl b molecules. The degree of the increase was the same as expected if the bound Chl b had the same antenna activity as the bound Chl a. The bound Chl b emitted fluorescence with a peak at 660 nm, and its yield was as high as the Chl a remaining in the complexes. However, the excitation spectrum of the Chl a fluorescence, detected at 680 nm, was almost the same as the absorption spectrum of the Chl b-bound complexes, indicating efficient energy transfer of the bound Chl b to Chl a. These results suggest that Chl b primarily occupies the Chl a-binding sites close to the reaction center region, acting as an efficient antenna for P700.

Topics: Binding Sites; Binding, Competitive; Chlorophyll; Chlorophyll A; Photosystem I Protein Complex; Protein Binding; Spectrophotometry; Spinacia oleracea

Solution culture experiments were conducted to investigate the effects of excessive Cu on the seed germination and growth of three plant species of the genus Elsholtzia (Elsholtzia haichowensis, Elsholtzia aypriani and Elsholtzia ciliata), and to compare Cu uptake and tolerance mechanisms of the three plants. The results showed that E. haichowensis had higher tolerance to excessive Cu than E. aypriani and E. ciliata, and that the adaptive Cu tolerance mechanism in E. haichowensis might involve the active participation of proteins.

Topics: Biodegradation, Environmental; China; Chlorophyll; Chlorophyll A; Germination; Industrial Waste; Lamiaceae; Metals; Mining; Plant Proteins; Plant Roots; Plant Shoots; Seeds; Soil Pollutants

Time-local and time-nonlocal theories are used in combination with optical spectroscopy to characterize the water-soluble chlorophyll binding protein complex (WSCP) from cauliflower. The recombinant cauliflower WSCP complexes reconstituted with either chlorophyll b (Chl b) or Chl a/Chl b mixtures are characterized by absorption spectroscopy at 77 and 298 K and circular dichroism at 298 K. On the basis of the analysis of these spectra and spectra reported for recombinant WSCP reconstituted with Chl a only (Hughes, J. L.; Razeghifard, R.; Logue, M.; Oakley, A.; Wydrzynski, T.; Krausz, E. J. Am. Chem. Soc. U.S.A. 2006, 128, 3649), the "open-sandwich" model proposed for the structure of the pigment dimer is refined. Our calculations show that, for a reasonable description of the data, a reduction of the angle between pigment planes from 60 degrees of the original model to about 30 degrees is required when exciton relaxation-induced lifetime broadening is included in the analysis of optical spectra. The temperature dependence of the absorption spectrum is found to provide a unique test for the two non-Markovian theories of optical spectra. Based on our data and the 1.7 K spectra of Hughes et al. (2006), the time-local partial ordering prescription theory is shown to describe the experimental results over the whole temperature range between 1.7 K and room temperature, whereas the alternative time-nonlocal chronological ordering prescription theory fails at high temperatures. Modified-Redfield theory predicts sub-100 fs exciton relaxation times for the homodimers and a 450 fs time constant in the heterodimers. Whereas the simpler Redfield theory gives a similar time constant for the homodimers, the one for the heterodimers deviates strongly in the two theories. The difference is explained by multivibrational quanta transitions in the protein which are neglected in Redfield theory.

Topics: Brassica; Chlorophyll; Chlorophyll A; Circular Dichroism; Kinetics; Light-Harvesting Protein Complexes; Models, Chemical; Models, Molecular; Optics and Photonics; Spectrum Analysis; Static Electricity

Recent progress in resolution of the structure of the light harvesting complex II provides the basis for theoretical predictions on nonlinear optical properties from microscopic calculations. An approach to absorption and fluorescence is presented within the framework of Bloch equations using a correlation expansion of relevant many particle interactions. The equations derived within the framework of this theory are applied to describe fluorescence saturation phenomena. The experimentally observed decrease of the normalized fluorescence quantum yield from 1 to 0.0001 upon increasing the intensity of laser pulse excitation at 645 nm by five orders of magnitude [R Schödel et al., Biophys. J. 71, 3370 (1996)] is explained by Pauli blocking effects of optical excitation and excitation energy transfer.

Topics: Chlorophyll; Chlorophyll A; Light; Light-Harvesting Protein Complexes; Photosystem II Protein Complex; Quantum Theory; Spectrometry, Fluorescence

The green alga Botryococcus protuberans was isolated from its natural environment and its morphology under different cultural conditions was examined. The alga was characterized by a high starch content and reddish oil drops as the assimilatory products. Photosynthetic pigments, Chl a, Chl b, carotenoids and xanthophylls are present. Modification of environmental conditions in modified Chu-10 medium resulted in optimum growth of the alga. Fatty acid composition revealed palmitic acid being the major component, while lauric acid, myristic acid and stearic acid were found in less quantity.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Chlorophyta; Energy-Generating Resources; Fatty Acids; Light; Reproduction, Asexual; Xanthophylls

The effect of mercury (Hg) on the biochemical parameters of Lycopersicon esculentum Mill leaf was studied. Application of mercuric chloride in varying concentrations (0.5, 1.0, 1.5 and 2.0 mM HgCl2 kg(-1) sand) caused significant reduction that went up to 89% and 72% chlorophyll a and chlorophyll b contents respectively (at flowering stage), 69% in carotenoid content, 64% in total soluble protein content and 91% in nitrate reductase activity (all at post-flowering stage). The amounts of nitrate and proline increased maximally (151% and 143% respectively) at the flowering stage, whereas total soluble sugar enhanced by 57% at the post-flowering stage. Changes observed in most of the parameters, were concentration dependent. Such studies seem to be able to discover suitable bioindicators of heavy metal pollution.

Topics: Carbohydrate Metabolism; Carotenoids; Chlorophyll; Chlorophyll A; Mercury; Nitrate Reductase; Nitrates; Plant Leaves; Plant Proteins; Proline; Soil Pollutants; Solanum lycopersicum

The canopy reflectance of winter wheat infected with stripe rust was measured in the field through artificial inoculation, and the pigment contents of the wheat leaves were determined indoor. The correlation between pigment contents and canopy hyperspectra data and the first derivative data of the disease wheat were analyzed respectively. Using linear and non-linear regression methods, and choosing a part of samples, the estimation models about pigment contents of disease wheat were built. Through the test of the other part samples, the result shows that the model containing the normalized value of the sum of first derivative within green edge (SD(g)) and the sum of first derivative within red edge (SD(r)) is the best one. The model was used to estimate the contents of chlorophyll a and chlorophyll b and carotenoid of the disease wheat, and the relative errors were 17.0%, 16.3% and 12.4%, respectively. This study shows that canopy hyperspectra data can be used to estimate the pigment contents of crops leaves and the estimation precision is high. This conclusion has great practice and application value to monitor the grow-ing way of and disease influence on crops by using hyperspectral remote sensing.

Topics: Basidiomycota; Carotenoids; Chlorophyll; Chlorophyll A; Host-Pathogen Interactions; Pigments, Biological; Plant Diseases; Plant Leaves; Seasons; Spectrophotometry; Triticum

With strawberry ' French 3' (Fragaria x ananassa Duch cv. French 3) as test material, this paper studied the effects of foliar spraying DA-6 at the rates of 10, 20 and 30 mg x L(-1) in late autumn on its seedlings net photosynthetic rate, metabolism of reactive oxygen species, and plant growth. Compared with the control, foliar spraying DA-6 of 20 and 30 mg x L(-1) increased the net photosynthetic rate, chlorophyll a and b contents, and SOD and CAT activities significantly, and decreased the reactive oxygen and MDA contents. Moreover, foliar spraying DA-6 of 20 and 30 mg x L(-1) increased the average dry weight per leaf, and the dry mass of shoot and root significantly. The ratio of root to shoot after spraying DA-6 of 20 and 30 mg x L(-1) was 29.9% and 29.3% higher than the control, respectively. Foliar spraying appropriate concentration of DA-6 in late autumn could improve the plant growth of strawberry seedlings.

Topics: Biomass; Catalase; Chlorophyll; Chlorophyll A; Fragaria; Hydrogen Peroxide; Photosynthesis; Plant Growth Regulators; Plant Leaves; Seasons; Superoxide Dismutase

To explore the possible regulatory role of chlorophyllase (Chlase) in chlorophyll (Chl) degradation during leaf senescence, RNAi Arabidopsis (Arabidopsis thaliana) plants were constructed to repress the expressions of AtCLH1 and/or AtCLH2. Transcript levels of AtCLH1 and/or AtCLH2 were dramatically lowered and Chlase activity was correspondingly inhibited, but the Chl degradation kinetics was not affected in the RNAi plants. Results of further analysis indicated that the Chl a/b ratio decreased in AtCLH1 RNAi lines, in comparison with the increasing Chl a/b ratio in the wide type during leaf senescence. In addition, an induced Chlase activity was consistently detected at the initial stage of senescence in all the plants examined. In contrast, transcript levels of both AtCLH1 and AtCLH2 decreased dramatically upon the initiation of senescence in both the wide-type and the RNAi plants. Interestingly, compared with the wide type, lower but still significant transcript levels of the RNAi targeted Chlase gene(s) were sustained during the whole period of dark incubation in all the three RNAi lines examined, indicating the functioning of some compensatively regulating mechanism. Based on these results, along with related reports, we conclude that Chlase might be required at the initial stage of leaf senescence, quite likely playing a role in converting Chl b to a.

Topics: Arabidopsis Proteins; Carboxylic Ester Hydrolases; Chlorophyll; Chlorophyll A; Gene Expression Regulation, Plant; Genetic Vectors; Plant Leaves; Plants, Genetically Modified; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference

The effect of copperchloride (CuCl2) on the level of chlorophyll (a+b), proline, protein and abscisic acid in sunflower (Helianthus annuus L.) seedlings were investigated Control and copper treated (0.4, 0.5 and 0.6 mM) seedlings were grown for ten days in Hoagland solution. Abscisic acid content was determined in root, shoot and leaf tissues of seedlings by HPLC. Copper stress caused significant increase of the abscisic acid contents in roots, shoots and leaves of seedlings. The increase was dependent on the copper salt concentration. Enhanced accumulation of proline in the leaves of seedlings exposed to copper was determined, as well as a decrease of chlorophyll (a+b) and total protein (p < 0.05 or p < 0.01). It was observed that the level of chlorophyll (a+b) and total protein (p < 0.05 or p < 0.01) remarkably decreased as copper concentration increased to 0.6 mM, although the levels of proline and abscisic acid in the leaves of plants were increased--a dose-depended behavior The same trends were also observed with the level of abscisic acid of stems and roots. Copper has dose- depended effects on chlorophyll, proline, protein and abscisic acid level of sunflower (Helianthus annuus L.) seedlings. Thus, we assumed that copper levels increase above some critical points seedling growth get negative effects. This assumption is in line with previous findings.

Topics: Abscisic Acid; Chlorophyll; Chlorophyll A; Copper; Helianthus; Plant Leaves; Plant Proteins; Proline; Seedlings

Marine pollutants induce changes in microalgal metabolism. In this study effects of tributyl-tin chloride (TBTCl) on a marine microalga Tetraselmis suecica was studied. The changes induced by TBTCI on growth rate, viability and biochemicals were assessed. In acute exposure to TBTCl, EC50 estimated for 24 hr was 2.02 microg ml(-1), whereas total lethality was observed at 4 microg ml(-1). In chronic exposure to TBTCl, at higher concentrations (0.5-1 microg ml(-1)) growth rate, chlorophyll pigments, carbohydrate and protein contents were reduced. The results of this study indicate that TBTCl toxicity made drastic changes in growth and biochemical composition of T. suecica.

Topics: Carbohydrate Metabolism; Chlorophyll; Chlorophyll A; Chlorophyta; Plant Proteins; Trialkyltin Compounds; Water Pollutants, Chemical

The Madagascar Periwinkle, Catharanthus roseus (L.) G. Don (a valued medicinal plant) was exposed to different concentrations ofheavymetals like, CdCl, and PbCl, with a view to observe their bioaccumulation efficiency. Germination was inhibited by both the heavy metals in the seeds previously imbibed in GA, and KNO, for 24 hr. EC50 (the effective concentration which inhibits root length by 50%) was recorded as 180 microM for CdCl2, and 50 microM for PbCl2. Both alpha-amylase and protease activity were reduced substantially on treatment of seeds with increasing concentrations of CdCl2, and PbCl2. Malondialdehyde (MDA) a product of lipoxigenase (LOX) activity also increased due to the treatment of both CdCl, and PbCl2. When two-months-old plants grown in normal soil were transferred to soils containing increasing amounts of these two heavy metals, senescence of lower leaves and extensive chlorosis were noticed after four days of transfer However, plants gradually acclimatized and after 20 days the chlorophyll content was almost comparable to normal. Plants receiving CdCl2 treatment (250 microg g(-1) and less) became acclimatized after two weeks and started normal growth. But PbCl2 of 432 microg g(-1) and less could not affect the plant growth throughout, after a preliminary shock was erased. In case of CdCl2 treatment, a stunted growth with reduced leaf area, reduced biomass and sterility were recorded after six months, while plants show normal growth and flowering in case of PbCl2 treatment. Total alkaloid was also found to be decreased in the roots of CdCl2 treated plants. No change was observed in case of PbCl2. GA3 treatments to the CdCl2 treated plants show internode elongation and increase in leaf area with relatively elongated leaves and thinning of stem diameter AAS analyses of leaves of treated plants exhibited 5-10% accumulation of cadmium, but there was no accumulation of lead at all.

Topics: Alkaloids; alpha-Amylases; Biodegradation, Environmental; Cadmium; Catharanthus; Chlorophyll; Chlorophyll A; Gibberellins; Lead; Lipid Peroxidation; Malondialdehyde; Peptide Hydrolases; Plant Growth Regulators; Plant Leaves; Plant Roots; Soil Pollutants

In the Champagne vineyard, most of the areas treated in early 2000 with the newly approved herbicide flazasulfuron had vines with altered growth and yellow leaves throughout the growing season. In order to clarify the physiological perturbations caused on the non-target grapevine and their potential consequences, C nutrition of grape plants grown in vineyards treated or not with flazasulfuron in 2000 was characterized during the following season. Vines from treated areas exhibited yellow leaves and an alteration of photosynthetic activity, characterized by declines in leaf gas exchanges (by 85%) and photosynthetic pigment concentrations (by 88%), and a marked disorganization of the leaf plastids. The herbicide also caused a decrease in leaf starch and soluble carbohydrate levels (-74% and -90%, respectively). Surprisingly, some vines re-greened after bloom, then exhibiting similar carbohydrate physiology to those grown in a non-treated area. Thus, recovery of CO(2) fixation rates, plastid ultra-structure, pigment concentrations and carbohydrate levels was found in re-greening leaves. Unlike the informations available in the literature, our results showed that flazasulfuron may be phytotoxic for grapevine. However, this toxicity was overcome the following year, indicating that vines have the potential to recover from this herbicide stress after one season.

Topics: Carbohydrate Metabolism; Carbon Dioxide; Carotenoids; Chlorophyll; Chlorophyll A; Herbicides; Microscopy, Electron, Transmission; Photosynthesis; Plant Leaves; Plastids; Starch; Sulfonylurea Compounds; Vitis

The results of a survey aimed at investigating whether NO2 and NH3 emitted by road traffic can influence lichen diversity, lichen vitality and the accumulation of nitrogen in lichen thalli are reported. For this purpose, distance from a highway in a rural environment of central Italy was regarded as the main parameter to check this hypothesis. The results of the present survey indicated that road traffic is not a relevant source of NH3. On the other hand, NO2 concentrations, although rather low, were negatively correlated with distance from the highway according to a typical logarithmic function. No association between NO2 concentrations and the diversity of epiphytic lichens was found, probably because of the low NO2 values measured. Also bark properties were not influenced by distance from the highway. Accumulation of nitrogen, reduction in the content of chlorophyll a, chlorophyll b and total carotenoids were found in transplanted thalli of Evernia prunastri, but NO2 was not responsible for these changes, which were probably caused by applications of N-based fertilizers.

Topics: Air Pollutants; Ammonia; Biodiversity; Carotenoids; Chlorophyll; Chlorophyll A; Environmental Monitoring; Geography; Italy; Lichens; Nitrogen Dioxide; Plant Bark; Trees; Vehicle Emissions

We studied the effects of hyperhydricity on subcellular ultrastructure and physiology of leaves during in vitro regeneration of apple plants. Morphological, anatomical and ultrastructural differences between healthy leaf tissues obtained from greenhouse-grown plants and healthy and hyperhydric leaves obtained from shoots raised from nodal shoot explants in a bioreactor were investigated by electron microscopy and confocal laser scanning microscopy. Compared with healthy leaves, hyperhydric leaves showed abnormal, often discontinuous development of the epidermis and cuticle. Stomata were malformed. The leaf lamina appeared thickened and was characterized by poor differentiation between the palisade and spongy mesophyll tissue. Hyperhydric leaves had a significantly lower chloroplast number per cell and chloroplasts showed reduced thylakoid stacking compared with healthy leaves. Hyperhydricity resulted in a general decrease in concentrations of reduced and oxidized pyridine nucleotides, reflecting a reduction in metabolic activity. The activities of antioxidant enzymes, such as superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase were higher in hyperhydric leaves than in healthy leaves, indicating that hyperhydricity was associated with oxidative stress. Chlorophyll fluorescence measurements provided evidence of oxidative damage to the photosynthetic machinery in hyperhydric leaves: photochemical efficiency of photosystem II, effective quantum efficiency and photochemical quenching were all lower in hyperhydric leaves compared with healthy leaves.

Topics: Antioxidants; Ascorbate Peroxidases; Carotenoids; Catalase; Chlorophyll; Chlorophyll A; Energy Metabolism; Fluorescence; Malus; Peroxidases; Photosynthesis; Plant Epidermis; Plant Leaves; Pyridines; Reactive Oxygen Species; Superoxide Dismutase; Water

This work was aimed at finding materials that could be used as alternatives to lichens as air quality monitors since the high natural variability and the large amount of lichen collected are two drawbacks of the use of these organisms. Lichen Flavoparmelia caperata(L.) Hale was exposed in three different forms (transplant, detached from the substratum and as a biomass-ground and homogeneized) and compared to the planetree bark (Platanus hybrida Brot.), exposed as a biomass, and two organic synthetic materials (Chelex(R) 100 resin and cellulose acetate). Materials were exposed for two months in the winter, spring and summer, at three Portuguese coastal cities. The results showed that the airborne accumulations of Cu, Ni, Pb and Sr were partially dependent on the meteorological conditions but mainly dependent on the nature of the exposed material. The standard deviations of the synthetic materials or homogenized biomass were the same or greater than lichen transplants or detached. The accumulation by biological materials, of the four studied elements, was comparable to the lichen transplant accumulation. The replacement of the traditional transplants by the biomass was not considered advantageous, since their preparation is time-consuming. Therefore lichens remain the most suitable in biomonitoring studies. The exposure of detached lichen allows the accurate measurement of the exposed area/volume so it can be useful to relate atmospheric deposition rates with the lichen metal content. The synthetic materials accumulated Cu and Ni and should only be used as an alternative to traditional transplants when these are the elements of interest.

Topics: Air Pollutants; Cellulose; Chelating Agents; Chlorophyll; Chlorophyll A; Environmental Monitoring; Lichens; Magnoliopsida; Metals, Heavy; Plant Bark; Resins, Synthetic

Linear alkylbenzene sulfonate (LAS) is the most commonly used anionic surfactant in laundry detergents and cleaning agents. LAS compounds are found in surface waters and soils. The short-term acute toxicity of LAS to weeping willows (Salix babylonica L.) was investigated. Willow cuttings were grown in hydroponic solution spiked with LAS at 24.0+/-1 degrees C for 192 h. The normalized relative transpiration of plants was used to determine toxicity. Severe reduction of the transpiration was only found for high doses of LAS (240 mgl(-1)). Chlorophyll contents in leaves of treated plants varied with the dose of LAS, but there was no significant linear correlation. The activities of the enzymes superoxide dismutases (SOD), catalase (CAT), and peroxidase (POD) were quantified at the end of experiments. At higher concentrations of LAS (240 mgl(-1)), the activities of SOD and CAT were decreased. The correlation between the dose of LAS and the POD activity in leaf cells was the highest of all enzyme assays (R(2)=0.5). EC(50) values for a 50% inhibition of the transpiration of the trees were estimated to 374 mgl(-1) (72 h) and 166 mgl(-1) (192 h). Results from this experiment indicated that phytotoxic effects of LAS on willow trees are not expected for normal environmental conditions.

Topics: Alkanesulfonic Acids; Benzenesulfonates; Catalase; Chlorophyll; Chlorophyll A; Peroxidase; Plant Leaves; Salix; Superoxide Dismutase; Surface-Active Agents

Adaptation of the halotolerant alga Dunaliella salina to iron deprivation involves extensive changes of chloroplast morphology, photosynthetic activities, and induction of a major 45-kDa chloroplast protein termed Tidi. Partial amino acid sequencing of proteolytic peptides suggested that Tidi resembles chlorophyll a/b-binding proteins which compose light-harvesting antenna complexes (LHC) (Varsano, T., Kaftan, D., and Pick, U. (2003) J. Plant Nutr. 26, 2197-2210). Here we show that Tidi shares the highest amino acid sequence similarity with light-harvesting I chlorophyll a/b-binding proteins from higher plants but has an extended proline-rich N-terminal domain. The accumulation of Tidi is reversed by iron supplementation, and its level is inversely correlated with photosystem I (PS-I) reaction center proteins. In native gel electrophoresis, Tidi co-migrates with enlarged PS-I-LHC-I super-complexes. Single particle electron microscopy analysis revealed that PS-I units from iron-deficient cells are larger (31 and 37 nm in diameter) than PS-I units from control cells (22 nm). The 77 K chlorophyll fluorescence emission spectra of isolated complexes suggest that the Tidi-LHC-I antenna are functionally coupled to the reaction centers of PS-I. These findings indicate that Tidi acts as an accessory antenna of PS-I. The enlargement of PS-I antenna in algae and in cyanobacteria under iron deprivation suggests a common limitation that requires rebalancing of the energy distribution between the two photosystems.

Topics: Amino Acid Sequence; Blotting, Northern; Chlorophyll; Chlorophyll A; Cloning, Molecular; Cyanobacteria; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Eukaryota; Immunohistochemistry; Iron; Iron Deficiencies; Light; Light-Harvesting Protein Complexes; Microscopy, Electron; Molecular Sequence Data; Photosystem I Protein Complex; Proline; Protein Structure, Tertiary; Proteins; RNA, Messenger; Sequence Analysis, Protein; Sequence Homology, Amino Acid; Spectrometry, Fluorescence; Temperature; Thylakoids; Time Factors; Transcription, Genetic

Protein composition and native state of chlorophylls were analyzed in two wheat (Triticum durum L.) genotypes with different tolerance to drought, Barakatli-95 (drought-tolerant) and Garagylchyg-2 (drought-sensitive), during water deficit. It is shown that the plants subjected to water deficit appear to have a slight increase in alpha- and beta-subunits of CF1 ATP-synthase complex (57.5 and 55 kD, respectively) in Barakatli-95 and their lower content in Garagylchyg-2. Steady-state levels of the core antenna of PS II (CP47 and CP43) and light-harvesting Chl a/b-apoproteins (LHC) II in the 29.5-24 kD region remained more or less unchanged in both wheat genotypes. The synthesis of 36 kD protein and content of low-molecular-weight polypeptides (21.5, 16.5, and 14 kD) were noticeably increased in the tolerant genotype Barakatli-95. Drought caused significant changes in the carotenoid region of the spectrum (400-500 nm) in drought-sensitive genotype Garagylchyg-2 (especially in the content of pigments of the violaxanthin cycle). A shift of the main band from 740-742 to 738 nm is observed in the fluorescence spectra (77 K) of chloroplasts from both genotypes under water deficiency, and there is a stimulation of the ratio of fluorescence band intensity F687/F740.

Topics: Adaptation, Biological; Chlorophyll; Chlorophyll A; Dehydration; Electrophoresis, Polyacrylamide Gel; Genotype; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Plant Proteins; Rosaniline Dyes; Spectrometry, Fluorescence; Spectrophotometry; Thylakoids; Triticum

The organization of pigment molecules in photosystems is strictly determined. The peripheral antennae have both chlorophyll a and b, but the core antennae consist of only chlorophyll a in green plants. Furthermore, according to the recent model obtained from the crystal structure of light-harvesting chlorophyll a/b-protein complexes II (LHCII), individual chlorophyll-binding sites are occupied by either chlorophyll a or chlorophyll b. In this study, we succeeded in altering these pigment organizations by introducing a prokaryotic chlorophyll b synthesis gene (chlorophyllide a oxygenase (CAO)) into Arabidopsis. In these transgenic plants (Prochlirothrix hollandica CAO plants), approximately 40% of chlorophyll a of the core antenna complexes was replaced by chlorophyll b in both photosystems. Chlorophyll a/b ratios of LHCII also decreased from 1.3 to 0.8 in PhCAO plants. Surprisingly, these transgenic plants were capable of photosynthetic growth similar to wild type under low light conditions. These results indicate that chlorophyll organizations are not solely determined by the binding affinities, but they are also controlled by CAO. These data also suggest that strict organizations of chlorophyll molecules are not essential for photosynthesis under low light conditions.

Topics: Agrobacterium tumefaciens; Arabidopsis; Bacterial Proteins; Chlorophyll; Chlorophyll A; Chloroplasts; Genes, Bacterial; Genes, Plant; Microscopy, Electron; Mutation; Oxygenases; Phenotype; Photosynthesis; Plants, Genetically Modified; Prochlorothrix; Recombinant Proteins; Transformation, Genetic

In this paper, field studies were conducted to determine the potential of alterations in physiology, and the intraspecific differences in sensitivity of 10 wheat (Triticum aestivum) cultivars to enhanced UV-B radiation. The supplemental UV-B radiation was 5.00 kJ x m(-2), simulating a depletion of 20% stratospheric ozone in Kunming area. The results showed that six of 10 tested wheat cultivars showed significant changes in total chlorophyll content, among which, chlorophyll a was strongly reduced, but chlorophyll b was reduced to a lesser extent, leading to a decrease in chlorophyll a/b ratio. The effects of UV-B on MDA and flavonoid contents also showed intraspecific difference. The MDA content of 5 cultivars was significant increased, while that of 2 cultivars was decreased. Four cultivars had significantly increased flavonoid content, but the flavonoid content in 2 cultivars was significantly decreased. The changes of chlorophyll and flavonoid contents were significantly correlated with the content of MDA(r = -0.78, P < 0.01 and r = -0.75, P < 0.05 respectively). There was a correlation between flavonoid and anti-UV-B radiation of wheat.

Topics: Chlorophyll; Chlorophyll A; Flavonoids; Species Specificity; Triticum; Ultraviolet Rays

A set of equations for determining chlorophyll a (Chl a) and accessory chlorophylls b, c2, c1 + c2 and the special case of Acaryochloris marina, which uses Chl d as its primary photosynthetic pigment and also has Chl a, have been developed for 90% acetone, methanol and ethanol solvents. These equations for different solvents give chlorophyll assays that are consistent with each other. No algorithms for Chl c compounds (c2, c1 + c2) in the presence of Chl a have previously been published for methanol or ethanol. The limits of detection (and inherent error, +/- 95% confidence limit), for chlorophylls in all organisms tested, was generally less than 0.1 microg/ml. The Chl a and b algorithms for green algae and land plants have very small inherent errors (< 0.01 microg/ml). Chl a and d algorithms for Acaryochloris marina are consistent with each other, giving estimates of Chl d/a ratios which are consistent with previously published estimates using HPLC and a rarely used algorithm originally published for diethyl ether in 1955. The statistical error structure of chlorophyll algorithms is discussed. The relative error of measurements of chlorophylls increases hyperbolically in diluted chlorophyll extracts because the inherent errors of the chlorophyll algorithms are constants independent of the magnitude of absorbance readings. For safety reasons, efficient extraction of chlorophylls and the convenience of being able to use polystyrene cuvettes, the algorithms for ethanol are recommended for routine assays of chlorophylls. The methanol algorithms would be convenient for assays associated with HPLC work.

Topics: Acetone; Animals; Chlorophyll; Chlorophyll A; Diatoms; Dinoflagellida; Ethanol; Methanol; Solvents; Species Specificity; Spectrophotometry; Spinacia oleracea; Synechococcus

Trididemnum miniatum is a colonial ascidian harboring the photosymbiotic prokaryote Prochloron sp. These bacterial cells are located in the tunic of the host animal. The present study revealed, by ultrastructural analysis, that the Prochloron cells were exclusively distributed and proliferated in the tunic. They were shown to be embedded in the tunic matrix and to have no direct contact with ascidian cells. Some tunic cells of the ascidians, however, did phagocytize and digest the symbiont. Round cell masses were sometimes found in the tunic and appeared to consist of disintegrating cyanobacterial cells. The thoracic epidermis of ascidian zooids was often digitated, and the epidermal cells extended microvilli into the tunic. Since there were no Prochloron cells in the alimentary tract of the ascidian zooids, the photosymbionts would not be considered part of the typical diet of the host ascidians. Thin layer chromatography showed that the symbionts possessed both chlorophyll a and b, while a 16S rRNA gene phylogeny supported the identification of the photosymbiont of T. miniatum as Prochloron sp.

Topics: Animals; Base Sequence; Chlorophyll; Chlorophyll A; Chromatography, Thin Layer; Microscopy, Electron, Transmission; Molecular Sequence Data; Photobiology; Phylogeny; Prochloron; RNA, Ribosomal, 16S; Symbiosis; Urochordata

The application of wood ash from biofuel sources to the forest has been suggested as a source of nutrients for trees and for restoration of acidified soils and surface waters. Studies on the effects of wood ash on aquatic organisms and ecosystems are, however, few. This study investigated the effects of wood ash (0.1-10 g l(-1)) on the freshwater moss Fontinalis antipyretica Hedw., which has previously been shown to be a sensitive test organism for assessing environmental pollution. After nine weeks of treatment with wood ash, a significant effect of enhanced stem growth was observed at higher concentrations (1-10 g l(-1)). The concentration of wood ash was also correlated with the growth of secondary branches. Photosynthesis (oxygen evolution after 4 h of exposure to wood ash) was significantly lower in the alkaline treatments (no pH adjustment) compared to the treatments with neutral wood ash solutions (pH adjusted to 7.5). Furthermore, photosynthesis in the alkaline wood ash treatments was significantly lower compared to the control. The ratio between the optical density value before and after acidification (OD(665/665a)) was higher for all wood ash concentrations compared to the control. The OD(665/665a) values ranged from 1.52 to 1.61 and there was a difference, however not significant, between the alkaline and the neutral treatment at 10 g l(-1) wood ash. This study clearly demonstrated that wood ash may be beneficial for F. antipyretica at moderate concentrations (0.5-5 g l(-1)), particularly when a sudden increase in pH is avoided.

Topics: Bryopsida; Chlorophyll; Chlorophyll A; Hydrogen-Ion Concentration; Oxygen; Photosynthesis; Plant Shoots; Plant Stems; Water Pollutants; Wood

The aim of this study was to identify, in the lichen Ramalina lacera, antioxidants that could provide indications of air pollution stress, and respond earlier than traditionally used structural/physiological parameters. The pollution-sensitive lichen R. lacera was transplanted from its relatively unpolluted natural habitat to two air-polluted sites for a period of up to 6 months. The superoxide dismutase and catalase activities, total water- and lipid-soluble low-molecular-weight antioxidant capacities and chlorophyll b/chlorophyll a ratios were assessed every 6 weeks. The earliest signs of oxidative stress were detected in the activities of fungal copper/zinc-superoxide dismutase, algal iron-superoxide dismutase and water-soluble low-molecular-weight antioxidants, which increased significantly as early as 42 days after exposure to pollution. Catalase activity increased in lichens transplanted to the polluted sites after 90 days. All activities decreased towards the end of the experiment. The impact of air pollution on R. lacera, using the traditionally employed parameter of chlorophyll b/chlorophyll a ratio, was only detected after 6 months of exposure to air pollution. Our results indicate that antioxidant parameters may serve as improved early-warning indicators of air pollution stress in lichens.

Topics: Air Pollutants; Air Pollution; Antioxidants; Catalase; Chlorophyll; Chlorophyll A; Environmental Monitoring; Enzyme Activation; Geography; Israel; Lichens; Oxidation-Reduction; Superoxide Dismutase

Reconstitution of the 16 kDa N-terminal domain of the peridinin-chlorophyll-protein, N-PCP, with mixtures of chlorophyll a (Chl a) and Chl b, resulted in 32 kDa complexes containing two pigment clusters, each bound to one N-PCP. Besides homo-chlorophyllous complexes, hetero-chlorophyllous ones were obtained that contain Chl a in one pigment cluster, and Chl b in the other. Binding of Chl b is stronger than that of the native pigment, Chl a. Energy transfer from Chl b to Chl a is efficient, but there are only weak interactions between the two pigments. Individual homo- and hetero-chlorophyllous complexes were investigated by single molecule spectroscopy using excitation into the peridinin absorption band and scanning of the Chl fluorescence, the latter show frequently well resolved emissions of the two pigments.

Topics: Animals; Carotenoids; Chlorophyll; Chlorophyll A; Eukaryota; Protozoan Proteins; Spectrophotometry

Light-induced pigment oxidation and its relation to excited state quenching in photosystems antennae have been investigated in isolated thylakoids. The results indicate that (i) chlorophyll oxidation takes place in two sequential steps. A slow initial phase is followed by a steep increase in the bleaching rate when more than one quarter of the chromophores are oxidised. (ii) During the initial slow phase, the carotenoid pool is bleached with an apparent rate which is about three times faster than that found for chlorophyll a and more than six times faster than that of chlorophyll b. (iii) Pigment bleaching has been observed both in photosystem I and photosystem II, and it has been possible to estimate a similar carotenoid bleaching rate in the two photosystems. (iv) The protection conferred by singlet state quenchers in the initial slow phase of pigment oxidation is modest. Taking into consideration that both the photosystems are subjected to the oxidative treatment, a somewhat larger protective effect than those estimated for photo-inhibition in thylakoids [S. Santabarbara, F.M. Garlaschi, G. Zucchelli, R.C. Jennings, Biochim. Biophys. Acta 1409 (1999) 165-170] can be computed, although it is less than 50% of the expected level on the basis of the observed reciprocity to the number of incident photons. (v) Pigment oxidation is associated with the loss of membrane ultra-structure, which is interpreted as originating from a decrease in grana stacking. The dynamics of loss of membrane ultra-structure parallel the phases observed for chlorophyll photo-bleaching.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Circular Dichroism; Kinetics; Light; Oxidation-Reduction; Photochemistry; Photosystem II Protein Complex; Plant Proteins; Spectrometry, Fluorescence; Thylakoids; Time Factors

Topics: Cadmium; Carotenoids; Chlorophyll; Chlorophyll A; Dose-Response Relationship, Drug; Matricaria; Photosynthesis; Plant Structures; Soil Pollutants

In living organisms the photosynthesis involves the absorption of light by the light-harvesting (LH) antenna complexes. By mimicking the photosynthesis process the artificial photonic device composed of bio-molecular hetero structure is developed. The proposed photonic device is composed of Chl a, Chl b, phycoerythrin and phycocyanin. To fabricate the highly ordered structure of biomolecules, the deposition of ordered Chl a/Chl b molecules onto solid substrates was done by the Langmuir-Blodgett (LB) technique, and the self-assembly technique was used for the deposition of phycocyanin molecule. To optimize the photocurrent generation, the photoelectric response characteristics of Chl a and Chl b LB films were measured according to the number of deposition layers, and effects of phycocyanin layer and various phycoerythrin concentrations on the photocurrent generation were analyzed. The biophotonic device was fabricated by the combination of the hetero biofilms and the photocurrent generation of the proposed device was observed. It was observed that the photocurrent generation of the proposed biodevice was improved by using the appropriate biopigments to be selected. The proposed artificial biophotonic device consisting of biopigments can be used to generate photocurrent by mimikng the photosynthesis in living system.

Topics: Chlorophyll; Chlorophyll A; Cyanobacteria; Electron Transport; Light; Light-Harvesting Protein Complexes; Photons; Phycocyanin; Phycoerythrin; Pigmentation; Pigments, Biological; Spectrophotometry; Surface Plasmon Resonance; Time Factors

Dunaliella salina is a useful model for studying the respective roles of each LHCII protein at the molecular level in extreme environmental conditions. However, information about the LhcII genes in D. salina is very limited. In order to identify more LhcII genes in D. salina, two additional LhcII cDNAs were obtained by screening a cDNA library. The genomic DNA was amplified by PCR using a specific primer set corresponding to the 5' and 3' untranslated regions of each transcript. The untranslated regions of the two additional genes are obviously different from each other; therefore they are two genes. Each gene contains an open reading frame for a protein of 253 amino acids. The two deduced proteins in D. salina are 99% identical at the amino acid sequence level to the previously reported LHCII protein in the same genus D. tertiolecta. Unrooted phylogenetic tree showed that types of LHCII proteins in D. salina did not correspond to any types in C. reinhardtii.

Topics: Amino Acid Sequence; Chlorophyll; Chlorophyll A; Chlorophyta; DNA, Complementary; Molecular Sequence Data; Photosystem II Protein Complex; Sequence Analysis, DNA

Chlorophyllide a oxygenase (CAO) that converts chlorophyllide a to chlorophyllide b was overexpressed in tobacco to increase chlorophyll (Chl) b biosynthesis and alter the Chl a/b ratio. Transgenic plants along with their wild-type cultivars were grown in low and high light intensities. In low light there was 20% increase in chlorophyll b contents in transgenic plants, which resulted in 16% reduction in the Chl a/b ratio. In high light, total Chl contents were 31% higher in transgenic plants than those of wild type. The increase in Chl a was 19% and that of Chl b was 72% leading to 31% decline of Chl a/b ratio. The increase in Chl b contents was accompanied by enhanced CAO expression that was highly pronounced in low light. As compared to low light, in high light Lhcb1 and Chl a/b transcripts abundance was significantly increased in transgenic plants suggesting a close relationship between Chl b synthesis and cab gene expression. However, there was a small increase in expression of LHCII proteins, which did not correspond to 72% increase in Chl b content in transgenic line, implying that LHCPII has the ability to bind more Chl b molecules.

Topics: Arabidopsis; Blotting, Northern; Blotting, Western; Chlorophyll; Chlorophyll A; Gene Expression; Genetic Vectors; Light; Light-Harvesting Protein Complexes; Nicotiana; Oxygenases; Polymerase Chain Reaction; RNA, Plant; Transformation, Genetic

Several mechanisms have been proposed to explain how algae can tolerate heavy metals. In order to better understand the mechanisms determining metal tolerance, we examined the interaction of copper with two strains of the copper-tolerant green algae Oocystis nephrocytioides, isolated from algal communities differing only in copper exposure. The strains were cultured in chemically-defined media containing 0.04 microM Cu(total) (pCu 12.4) or 2 microM Cu(total) (pCu 10.6). Growth, photosynthesis rate, content of chlorophyll a and b, copper accumulation, its cellular distribution and ultrastructural localization, as well as the influence of algal growth on extracellular copper complexation were determined. Both strains had comparable growth and photosynthesis rates. The cellular content of both chlorophyll a and b was reduced, by roughly the same extent, at pCu 10.6 compared to pCu 12.4. Copper titration of the media indicated the production of copper-complexing ligands by O. nephrocytioides cultured at pCu 12.4 that increased with increased algal density during cell growth. No additional ligands were detected at pCu 10.6. Copper-complexing ligands had a conditional stability constant of K = 10(13) at pH 7.3. The intracellular concentration of copper in O. nephrocytioides was 80 microM at pCu 12.4 and increased to 7.5mM at pCu 10.6. The proportion of intracellular Cu accumulated increased from 8% of total Cu content at pCu 12.4 to 60% at pCu 10.6. By electron spectroscopic imaging, intracellular Cu was detected in the thylakoids and the pyrenoid of O. nephrocytioides cells. The results indicate that the tolerance of O. nephrocytioides to Cu is constitutive and does not need to be induced by previous exposure to Cu. We propose that accumulation and sequestration of Cu in thylakoids and, to a lesser extent, adsorption of copper to the algal cell surface represent the most important tolerance mechanism, for O. nephrocytioides.

Topics: Chlorophyll; Chlorophyll A; Chlorophyta; Copper; Culture Media; Edetic Acid; Ligands; Microscopy, Energy-Filtering Transmission Electron; Photosynthesis; Thylakoids

Perfluorooctane sulfonic acid (PFOS) is an anthropogenic contaminant detected in various environmental and biologic matrices. This compound is a fluorinated surfactant, a class of molecules renowned for their persistence and their global distribution but for which few ecotoxicological data are currently available, especially under field conditions. The toxicity of PFOS to the aquatic macrophytes Myriophyllum sibiricum and M. spicatum was investigated using 12,000 L outdoor microcosms. Replicate microcosms (n = 3) were treated with 0.3, 3, 10, and 30 mg/L PFOS as the potassium salt and assessed at regular intervals during a period of 42 days. M. sibiricum was more sensitive to PFOS under these simulated field conditions than M. spicatum. Toxicity was observed in the evaluated end points at > 3 mg/L PFOS for EC10s and > 12 mg/L PFOS for EC50s for M. spicatum and in M. sibiricum at > 0.1 mg/L PFOS for EC10s and > 1.6 mg/L PFOS for EC50s. The no observed-effect concentration (NOEC) for M. spicatum was consistently > or = 11.4 mg/L PFOS, whereas the NOEC for M. sibiricum was > or = 0.3 mg/L PFOS. A risk assessment for these plants estimated a negligible probability of toxicity being observed in these plants from PFOS exposure at current environmental concentrations.

Topics: Alkanesulfonic Acids; Carotenoids; Chlorophyll; Chlorophyll A; Fluorocarbons; Magnoliopsida; No-Observed-Adverse-Effect Level; Plant Roots; Risk Assessment; Water Pollutants, Chemical

The conformational distribution of the N-terminal domain of the major light-harvesting chlorophyll a/b protein (LHCIIb) has been characterized by electron-electron double resonance yielding distances between spin labels placed in various domains of the protein. Distance distributions involving residue 3 near the N terminus turned out to be bimodal, revealing that this domain, which is involved in regulatory functions such as balancing the energy flow through photosystems (PS) I and II, exists in at least two conformational states. Models of the conformational sub-ensembles were generated on the basis of experimental distance restraints from measurements on LHCIIb monomers and then checked for consistency with the experimental distance distribution between residues 3 in trimers. Only models where residue 3 is located above the core of the protein and extends into the aqueous phase on the stromal side fit the trimer data. In the other state, which consequently is populated only in monomers, the N-terminal domain extends sideways from the protein core. The two conformational states may correspond to two functional states of LHCIIb, namely trimeric LHCIIb associated with PSII in stacked thylakoid membranes and presumably monomeric LHCIIb associated with PSI in nonstacked thylakoids. The switch between these two is known to be triggered by phosphorylation of Thr-6. A similar phosphorylation-induced conformational change of the N-terminal domain has been observed by others in bovine annexin IV which, due to the conformational switch, also loses its membrane-aggregating property.

Topics: Animals; Annexin A4; Cattle; Chlorophyll; Chlorophyll A; Crystallography, X-Ray; Dimerization; Electron Spin Resonance Spectroscopy; Electrons; Escherichia coli; Light; Light-Harvesting Protein Complexes; Macromolecular Substances; Models, Molecular; Mutation; Oxygen; Phosphorylation; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Pisum sativum; Protein Conformation; Protein Structure, Secondary; Protein Structure, Tertiary; Recombinant Proteins; Spin Labels; Threonine; Thylakoids; Time Factors

The red-most fluorescence emission of photosystem I (733 nm at 4 K) is associated with the Lhca4 subunit of the antenna complex. It has been proposed that this unique spectral feature originates from the low energy absorption band of an excitonic interaction involving chlorophyll A5 and a second chlorophyll a molecule, probably B5 (Morosinotto, T., Breton, J., Bassi, R., and Croce, R. (2003) J. Biol. Chem. 278, 49223-49229). Because of the short distances between chromophores in Lhc proteins, the possibility that other pigments are involved in the red-shifted spectral forms could not be ruled out. In this study, we have analyzed the pigment-pigment interactions between nearest neighboring chromophores in Lhca4. This was done by deleting individual chlorophyll binding sites by mutagenesis, and analyzing the changes in the spectroscopic properties of recombinant proteins refolded in vitro. The red-shifted (733 nm) fluorescence peak, the major target of this analysis, was lost upon mutations affecting sites A4, A5, and B5 and was modified by mutating site B6. In agreement with the shorter distance between chlorophylls A5 and B5 (7.9 A) versus A4 and A5 (12.2 A) in Lhca4 (Ben-Shem, A., Frolow, F., and Nelson, N. (2003) Nature 426, 630-635), we conclude that the low energy spectral form originates from an interaction involving pigments in sites A5 and B5. Mutation at site B6, although inducing a 15-nm blue-shift of the emission peak, maintains the red-shifted emission. This implies that chromophores responsible for the interaction are conserved and suggests a modification in the pigment organization. Besides the A5-B5 pair, evidence for additional pigment-pigment interactions between chlorophylls in sites B3-A3 and B6-A6 was obtained. However, these features do not affect the red-most spectral form responsible for the 733-nm fluorescence emission band.

Topics: Arabidopsis; Binding Sites; Chlorophyll; Chlorophyll A; Circular Dichroism; Light-Harvesting Protein Complexes; Models, Molecular; Mutagenesis, Site-Directed; Photosystem I Protein Complex; Pigments, Biological; Recombinant Proteins; Spectrometry, Fluorescence; Spectrophotometry

Chlorophylls were quantitatively studied in the leaves of black locust (Robinia pseudoacacia L.) seedlings exposed to electromagnetic fields of high frequency. Exposure system was designed and built up to make possible simultaneous exposure of seedling lots (3 months old) to low power density electromagnetic fields corresponding to a frequency of 400 MHz. After three weeks of daily exposures (1, 2, 3 and 8 hours), chlorophyll levels were measured using adequate spectral device. Statistical analysis of experimental results was performed by means of t-test to identify significant modifications induced by electromagnetic treatment in exposed samples in comparison to the control. Chlorophyll-a as well as chlorophyll-b level was found to decrease except the exposure time of two hours, where a considerable enhancement was noticed. It was revealed that the ratio of the two main types of chlorophyll was decreasing logarithmically to the increase of daily exposure time.

Topics: Chlorophyll; Chlorophyll A; Dose-Response Relationship, Radiation; Electromagnetic Fields; Environmental Exposure; Plants; Radiation Tolerance; Seeds; Spectrophotometry; Time Factors

We determined hydrogen, carbon and nitrogen isotopic compositions of chlorophylls a and b isolated from leaves of five C3 higher plant species (Benthamidia japonica, Prunus japonica, Acer carpinifolium, Acer argutum and Querus mongloica), and hydrogen and carbon isotopic compositions of phytol and chlorophyllides in the chlorophylls to understand isotopic fractionations associated with chlorophyll biosynthesis in these species. Chlorophylls are depleted in D relative to ambient water by approximately 189 per thousand and enriched in (13)C relative to bulk tissue by approximately 1.6 per thousand. These data can be explained by the contribution of isotopic fractionations during phytol and chlorophyllide biosyntheses. Phytol is more depleted in both D (by approximately 308 per thousand) and (13)C (by approximately 4.3 per thousand), while chlorophyllides are less depleted in D (by approximately 44 per thousand) and enriched in (13)C (by approximately 4.8 per thousand). Such inhomogeneous distribution of isotopes in chlorophylls suggests that (1) the phytol in chlorophylls reflects strong D- and (13)C-depletions due to the isotopic fractionations during the methylerythritol phosphate pathway followed by hydrogenation, and (2) the chlorophyllides reflect D- and (13)C-enrichments in tricarboxylic acid cycle. On the other hand, chlorophylls are slightly ( approximately 1.2 per thousand) depleted in (15)N relative to the bulk tissue, indicating that net isotopic fractionation of nitrogen during chlorophyll biosynthesis is small compared with those of hydrogen and carbon.

Topics: Carbon Isotopes; Chlorophyll; Chlorophyll A; Chlorophyllides; Hydrogen; Magnoliopsida; Models, Chemical; Molecular Structure; Nitrogen Isotopes; Phytol; Water

With the availability of structural models for photosystem I (PSI) in cyanobacteria and plants it is possible to compare the excitation transfer networks in this ubiquitous photosystem from two domains of life separated by over one billion years of divergent evolution, thus providing an insight into the physical constraints that shape the networks' evolution. Structure-based modeling methods are used to examine the excitation transfer kinetics of the plant PSI-LHCI supercomplex. For this purpose an effective Hamiltonian is constructed that combines an existing cyanobacterial model for structurally conserved chlorophylls with spectral information for chlorophylls in the Lhca subunits. The plant PSI excitation migration network thus characterized is compared to its cyanobacterial counterpart investigated earlier. In agreement with observations, an average excitation transfer lifetime of approximately 49 ps is computed for the plant PSI-LHCI supercomplex with a corresponding quantum yield of 95%. The sensitivity of the results to chlorophyll site energy assignments is discussed. Lhca subunits are efficiently coupled to the PSI core via gap chlorophylls. In contrast to the chlorophylls in the vicinity of the reaction center, previously shown to optimize the quantum yield of the excitation transfer process, the orientational ordering of peripheral chlorophylls does not show such optimality. The finding suggests that after close packing of chlorophylls was achieved, constraints other than efficiency of the overall excitation transfer process precluded further evolution of pigment ordering.

Topics: Algorithms; Biophysics; Chlorophyll; Chlorophyll A; Cyanobacteria; Dimerization; Kinetics; Light-Harvesting Protein Complexes; Macromolecular Substances; Models, Biological; Models, Molecular; Models, Statistical; Normal Distribution; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Plant Proteins; Protein Binding

Susceptibility to photoinhibition of the evergreen conifers Abies alba Mill., Picea abies (L.) Karst. and Pinus mugo Turra was investigated in an unheated greenhouse during winter and spring 2003. Photosynthetic performance of the seedlings was assessed by chlorophyll a fluorescence and analyses of chlorophyll and total carotenoid concentrations in needles. During winter months, maximum quantum yield of PSII photochemistry (ratio of variable to maximum fluorescence, Fv/Fm) was significantly greater in A. alba than in P. abies and P. mugo. Abies alba also sustained higher maximum apparent electron transport rate (ETRmax) than P. abies and P. mugo. Total concentrations of chlorophyll and carotenoids in needles decreased during the winter in P. mugo and P. abies, but remained stable in A. alba. For all species, Fv/Fm decreased from December until February and then increased to a maximum in April. Photoinhibition was greatest (Fv/Fm < 0.80) in all seedlings in February, the month with the lowest mean temperature. Saturating photosynthetic photon flux (PPFsat) and ETRmax were positively related to air temperature. All species had lower values of ETRmax and PPFsat in winter than in spring. Non-photochemical quenching of chlorophyll fluorescence (NPQ) was highest at low air temperatures. Differences among species in susceptibility to winter photoinhibition resulted from their specific light preferences and led to different mechanisms to cope with photoinhibitory stress. The more shade-tolerant A. alba sustained a higher photosynthetic capacity in winter than P. abies and P. mugo. Winter photoinhibition in P. abies, P. mugo and, to a lesser extent, in A. alba may reflect adaptive photoprotection of the photosynthetic apparatus in winter.

Topics: Abies; Carotenoids; Chlorophyll; Chlorophyll A; Cold Temperature; Ecosystem; Light; Photosynthesis; Picea; Pinus; Plant Leaves; Seasons; Species Specificity; Time Factors

Net fluxes of H+, K+ and Ca2+ ions from maize (Zea mays L.) isolated leaf segments were measured non-invasively using ion-selective vibrating microelectrodes (the MIFE technique). Leaf segments were isolated from the blade base, containing actively elongating cells (basal segments), and from non-growing tip regions (tip segments). Ion fluxes were measured in response to bright white light (2600 micromoles m-2 s-1) from either the leaf segments or the underlying mesophyll (after stripping the epidermis). Fluxes measured from the mesophyll showed no significant difference between basal and tip regions. In leaf segments (epidermis attached), light-induced flux kinetics of all ions measured (H+, Ca2+ and K+) were strikingly different between the two regions. It appears that epidermal K+ fluxes are required to drive leaf expansion growth, whereas in the mesophyll light-induced K+ flux changes are likely to play a charge balancing role. Light-stimulated Ca2+ influx was not directly attributable either to leaf photosynthetic performance or to leaf expansion growth. It is concluded that light-induced ion flux changes are associated with both leaf growth and photosynthesis.

Topics: Calcium; Carbon Dioxide; Chlorophyll; Chlorophyll A; Ethanol; Fluorescence; Hydrogen; Ion Transport; Light; Microelectrodes; Oxygen; Photosynthesis; Pigments, Biological; Plant Epidermis; Plant Leaves; Potassium; Zea mays

The presence of actinorhizas and arbuscular mycorrhizas may reduce plant stresses caused by adverse soil conditions. A greenhouse experiment was conducted using a sediment with a high pH, resulting from the disposal of waste originated at an acetylene and polyvinylchloride factory, in which Black alder (Alnus glutinosa) seedlings were inoculated either with Glomus intraradices BEG163 (originally isolated from the same sediment), Frankia spp. or both symbionts. After a 6-month growth period, plants inoculated with both symbionts had significantly greater leaf area, shoot height and total biomass when compared with the uninoculated control, the Frankia spp. and the G. intraradices treatments alone. In dual inoculated plants the N and P leaf content was significantly increased. A defoliation experiment was performed to evaluate the stress recovery of A. glutinosa and plants inoculated with both symbionts had a faster leaf regrowth and produced greater numbers of leaves. The dual inoculation resulted in greater numbers of and larger root nodules than when inoculated with Frankia spp. alone. The length and NADH diaphorase activity of the extraradical mycelium of G. intraradices was also significantly greater when dual inoculation was performed. The inoculation with Frankia spp. alone was shown to improve A. glutinosa growth, whereas G. intraradices alone had no positive effect under these environmental conditions. However, when the two symbionts were inoculated together a synergistic effect was observed resulting in a greater benefit for the plants and for both symbionts. The relevance of these findings for the phytorestoration of anthropogenic stressed sediments with high pH is discussed.

Topics: Alnus; Biodegradation, Environmental; Biomass; Carbon; Chlorophyll; Chlorophyll A; Frankia; Fungi; Geologic Sediments; Hydrogen-Ion Concentration; Industrial Waste; Nitrogen; Phosphorus; Plant Leaves; Plant Shoots; Potassium; Refuse Disposal; Soil Pollutants; Symbiosis

Changes in photosynthetic pigment and protein contents, growth and the spectral characteristics in Chlorella vulgaris in response to selenium stress were investigated. Carotenoids (beta-carotene and xanthophylls) and chlorophyll (Chl a and Chl b) contents in cells exposed to Se 50 mg/L increased primarily and decreased afterwards, while photosynthetic pigments in cells exposed to Se 800 mg/L decreased significantly. Chlorophyll (Chl) absorption peak at 693 nm and prominent Chl emission peak at 700 nm, weakened significantly after Se stress. The excitation spectra showed a decrease in excitation energy transfer efficiency in Se-stressed cells. Total soluble protein decreased after Se stress. The changes in total Se, Mg(2+), Ca(2+), K(+) and Na(+) concentrations in culture medium and cells were also determined by ICP-AES.

Topics: beta Carotene; Calcium; Carotenoids; Chlorella vulgaris; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Magnesium; Potassium; Selenium; Sodium; Xanthophylls

Different dilution levels of tannery treated effluent and their corresponding concentration of chromium (Cr6+) were studied in a petridish culture experiment on seed germination and seedling growth in radish (Raphanus sativus L). The different concentrations of Cr6+ (2, 5 and 10 ppm) and treated tannery effluent (10, 25 and 50%) showed reduction in seedling growth and related enzymatic activities with increase in concentration of Cr6+ in treatments and effluent both. The low concentration of chromium (2 ppm) and effluent dilution (10%) showed significant growth reduction separately. At this concentration of chromium and effluent dilution chlorophyll content, amylase, catalase and protein contents remained unchanged while with increase in Cr6+ concentration (>2ppm) and effluent dilution (> 10%) in treatments showed growth inhibitory effects.

Topics: alpha-Amylases; Amylases; beta-Amylase; Biomass; Catalase; Chlorophyll; Chlorophyll A; Chromium; Germination; Industrial Waste; Plant Roots; Raphanus; Seeds; Tanning; Waste Disposal, Fluid; Water Pollutants, Chemical

Land plants change the compositions of light-harvesting complexes (LHC) and chlorophyll (Chl) a/b ratios in response to the variable light environments which they encounter. In this study, we attempted to determine the mechanism which regulates Chl a/b ratios and whether the changes in Chl a/b ratios are essential in regulation of LHC accumulation during light acclimation. We hypothesized that changes in the mRNA levels for chlorophyll a oxygenase (CAO) involved in Chl b biosynthesis are an essential part of light response of Chl a/b ratios and LHC accumulation. We also examined the light-intensity dependent response of CAO-overexpression and wild-type Arabidopsis thaliana plants. When wild-type plants were acclimated from low-light (LL) to high-light (HL) conditions, CAO mRNA levels decreased and the Chl a/b ratio increased. In transgenic plants overexpressing CAO, the Chl a/b ratio remained low under HL conditions; thereby suggesting that changes in the CAO mRNA levels are necessary for those in Chl a/b ratios upon light acclimation. Under HL conditions, the accumulation of Lhcb1, Lhcb3 and Lhcb6 was enhanced in plants overexpressing CAO. On the contrary, in a CAO-deficient mutant, chlorina 1-1, theaccumulation of Lhcb1, Lhcb2, Lhcb3, Lhcb6 and Lhca4 was reduced. In comparison to wild-type, beta-carotene levels were reduced in CAO-overexpressing plants, while they were elevated in chlorina 1-1 mutants. These results imply that the transcriptional control of CAO is a part of the regulatory mechanism for the accumulation of a distinct set of LHC proteins upon light acclimation.

Topics: Acclimatization; Arabidopsis; beta Carotene; Chlorophyll; Chlorophyll A; Gene Expression Regulation, Plant; Light; Light-Harvesting Protein Complexes; Oxygenases; Plants, Genetically Modified; RNA, Messenger; Time Factors

Pesticides, firmly attached to the topsoil, might enter nearby watercourses at periods with high erosive loss of sediments. Therefore, exposure of aquatic organisms to these low mobility pesticides, in many cases, will coincide with a high sediment concentration. In this study, both individual and combined effects of propiconazole and runoff sediment on the aquatic model bryophyte Vesicularia dubyana are studied. Individual exposure to propiconazole induced responses in V. dubyana at rather low concentration levels (approximately 1 microg/L), showing that harmful effects of propiconazole potentially may occur in watercourses draining propiconazole-treated fields. Individual exposure to the sediment size fractions S1 (0.16-2 microm) and S2 (0.03-0.16 microm) caused plant stress at a concentration of 100 mg/L. The coarser fraction S1 showed strong inhibition effects on photosynthesis, probably due to light attenuation. Compared to S1, the suspension with the finer fraction S2 showed lower turbidity, higher nutrient content, and a higher proportion of sediment-bound propiconazole. The combined effects of propiconazole and suspended sediment are dependent on concentrations of sediment and propiconazole. At low sediment concentration (e.g., 100 mg/L), neither S1 nor S2 reduce the toxicity of propiconazole, as only 2% of propiconazole are bound to particles. An increase in sediment concentration decreases the bioavailable concentration of propiconazole; however, at the same time, this increases the turbidity, thereby inhibiting plant photosynthesis.

Topics: Adsorption; Agriculture; Bryophyta; Carbon; Chlorophyll; Chlorophyll A; Dose-Response Relationship, Drug; Environmental Monitoring; Fungicides, Industrial; Geologic Sediments; Kinetics; Nitrogen; Peroxidases; Photosynthesis; Plants; Rain; Soil; Triazoles; Water; Water Pollutants, Chemical

The authentication of virgin olive oil samples requires usually the use of sophisticated and very expensive analytical techniques, so there is a need for fast and inexpensive analytical techniques for use in a quality control methodology. Virgin olive oils present an intense fluorescence spectra. Synchronous excitation-emission fluorescence spectroscopy (SEEFS) was assessed for origin determination of virgin olive oil samples from five French registered designation of origins (RDOs) (Nyons, Vallée des Baux, Aix-en-Provence, Haute-Provence, and Nice). The spectra present bands between 600 and 700 nm in emission due to chlorophylls a and b and pheophytins a and b. The bands between 275 and 400 nm in emission were attributed to alpha-, beta-, and gamma-tocopherols and to phenolic compounds, which characterize the virgin olive oils compared to other edible oils. The chemometric treatment (PLS1) of synchronous excitation-emission fluorescence spectra allows one to determine the origin of the oils from five French RDOs (Baux, Aix, Haute-Provence, Nice, and Nyons). Results were quite satisfactory, despite the similarity between two denominations of origin (Baux and Aix) that are composed by some common cultivars (Aglandau and Salonenque). The interpretation of the regression coefficients shows that RDOs are correlated to chlorophylls, pheophytins, tocopherols, and phenols compounds, which are different for each origin. SEEFS is part of a global analytic methodology that associates spectroscopic and chromatographic techniques. This approach can be used for traceability and vindicates the RDOs.

Topics: Chlorophyll; Chlorophyll A; France; Olive Oil; Phenols; Pheophytins; Plant Oils; Spectrometry, Fluorescence; Tocopherols

Mg(II)-porphyrin-ligand and (bacterio)chlorophyl-ligand coordination interactions have been studied by solution and solid-state MAS NMR spectroscopy. (1)H, (13)C and (15)N coordination shifts due to ring currents, electronic perturbations and structural effects are resolved for imidazole (Im) and 1-methylimidazole (1-MeIm) coordinated axially to Mg(II)-OEP and (B)Chl a. As a consequence of a single axial coordination of Im or 1-MeIm to the Mg(II) ion, 0.9-5.2 ppm (1)H, 0.2-5.5 ppm (13)C and 2.1-27.2 ppm (15)N coordination shifts were measured for selectively labeled [1,3-(15)N]-Im, [1,3-(15)N,2-(13)C]-Im and [1,3-(15)N,1,2-(13)C]-1-MeIm. The coordination shifts depend on the distance of the nuclei to the porphyrin plane and the perturbation of the electronic structure. The signal intensities in the (1)H NMR spectrum reveal a five-coordinated complex, and the isotropic chemical shift analysis shows a close analogy with the electronic structure of the BChl a-histidine in natural light harvesting 2 complexes. The line broadening of the ligand responses support the complementary IR data and provide evidence for a dynamic coordination bond in the complex.

Topics: Bacteriochlorophyll A; Bacteriochlorophylls; Carbon Isotopes; Chlorophyll; Chlorophyll A; Histidine; Imidazoles; Isotope Labeling; Light-Harvesting Protein Complexes; Magnesium; Magnetic Resonance Spectroscopy; Molecular Structure; Nitrogen Isotopes; Photosynthesis; Rhodobacter sphaeroides; Spectrophotometry, Infrared; Spinacia oleracea

Photosystem I contains two potential electron transfer pathways between P(700) and F(X). These branches are made up of the electron transfer chain components A, A(0), and A(1). The primary electron acceptor A(0) is a chlorophyll a monomer that could be one or both of the two chlorophyll molecules, eC-A(3)/eC-B(3), identified in the 2.5 A resolution structure. The eC-A(3)/eC-B(3) chlorophylls are both coordinated by the sulfur atom of a methionine. This coordination is highly unusual, as interactions between the acid Mg(2+) and the soft base sulfur are weak. The eC-A(3)/eC-B(3) chlorophylls also are located close to one of the connecting chlorophylls that may link the antenna and the electron transfer chain chlorophylls. Due to their location in the structure, the eC-A(3)/eC-B(3) chlorophylls may play a role in both excitation energy transfer and electron transfer. To test the role of the eC-A(3)/eC-B(3) chlorophylls in electron transfer, Met-684 of PsaA and Met-664 of PsaB have been changed to His, Ser, and Leu. Replacement of either M(A684) or M(B664) results in a significant alteration in growth phenotype. The His and Leu mutants are very light sensitive in the presence of oxygen. Growth is impaired to a greater extent in the B-side mutants. However, all of the mutants are able to grow anaerobically at comparable rates. The His and Ser mutants all accumulate PSI at a level similar to that of wild type, whereas the Leu mutants have reduced amounts of PSI. Ultrafast transient absorbance measurements show that the (A(0)(-) - A(0)) difference signal accumulates in the MH(A684) and MH(B664) mutants under neutral conditions, demonstrating that electron transfer between A(0)(-) and A(1) is blocked or significantly slowed. The results show that both the A-branch and the B-branch of the ETC are active in PSI from Chlamydomonas reinhardtii.

Topics: Animals; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Electron Spin Resonance Spectroscopy; Electron Transport; Ligands; Mutagenesis, Site-Directed; Oxidation-Reduction; Photosystem I Protein Complex; Pigments, Biological; Protein Subunits; Spectrophotometry; Thylakoids

All chlorophyll (Chl)-binding proteins involved in photosynthesis of higher plants are hydrophobic membrane proteins integrated into the thylakoids. However, a different category of Chl-binding proteins, the so-called water-soluble Chl proteins (WSCPs), was found in members of the Brassicaceae, Polygonaceae, Chenopodiaceae, and Amaranthaceae families. WSCPs from different plant species bind Chl a and Chl b in different ratios. Some members of the WSCP family are induced after drought and heat stress as well as leaf detachment. It has been proposed that this group of proteins might have a physiological function in the Chl degradation pathway. We demonstrate here that a protein that shared sequence homology to WSCPs accumulated in etiolated barley (Hordeum vulgare) seedlings exposed to light for 2 h. The novel 22-kD protein was attached to the outer envelope of barley etiochloroplasts, and import of the 27-kD precursor was light dependent and induced after feeding the isolated plastids the tetrapyrrole precursor 5-aminolevulinic acid. HPLC analyses and spectroscopic pigment measurements of acetone-extracted pigments showed that the 22-kD protein is complexed with chlorophyllide. We propose a novel role of WSCPs as pigment carriers operating during light-induced chloroplast development.

Topics: Adaptation, Physiological; Aminolevulinic Acid; Chlorophyll; Chlorophyll A; Chlorophyllides; Chloroplasts; Darkness; Disasters; Hordeum; Hot Temperature; Light; Light-Harvesting Protein Complexes; Photosynthesis; Plant Proteins

Tests during the last few years have confirmed that 4-nonylphenol (4-NP) can have oestrogen-like effects (xeno-hormone) on animal organisms. The objective was to firstly evaluate the ecotoxicological effects of 4-NP on plants such as hydrocultures. To clarify how this substance interferes with the photosynthetic system of plants, various tests were carried out using the basil plant (Ocimum basilicum).. The effect of the pollutant 4-NP on the pigment content in the leaves of the basil plant was analysed with the use of High-Performance-Liquid-Chromatography (HPLC).. A general assessment of the HPLC data revealed that plants that came in contact with the 4-nonylphenol showed a change in pigmentation. More chlorophyll a and b was produced, although at the same time a higher production of degradation products and by-products of the chlorophylls was observed. These occurrences can therefore be seen as an impairment of the photosynthetic process. The contaminated plants produced less xanthophylls than the non-contaminated ones, though these differences were statistically not significant.. The variations on the pigment content in the leaves of the basil plant can be interpreted as a consequence of the 4-NP application. It was, however, not investigated whether the plants absorbed the pollutants directly. The effect could have been caused by adsorption of the oily substance to the roots, and this could have led to a hindrance of the uptake of nutrients and possibly water. In order to clarify this further, biochemical experiments are being conducted.

Topics: Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Ocimum basilicum; Phenols; Photosynthesis; Plant Leaves

Plant and aphid biomass, photosynthetic pigment (chlorophylls a and b and carotenoids) concentrations, and chlorophyll a/b and chlorophyll/carotenoid ratios were quantified in aphid-infested 'Tugela' near-isogenic lines (Tugela, Tugela-Dn1, Tugela-Dn2, and Tugela-Dn5). The objectives were to quantify changes of photosynthetic pigments (chlorophylls a and b, and carotenoids) caused by aphid feeding and assess resistance of wheat isolines through aphid and plant biomass analysis. Biomass of bird cherry-oat aphid, Rhopalosiphum padi (L.) (Hemiptera: Aphididae)-infested plants was lower than Russian wheat aphid, Diuraphis noxia (Mordvilko) (Hemiptera: Aphididae),- infested plants. When infested by D. noxia, all lines showed increased biomass over time, except Tugela where biomass decreased on day 12. No difference in plant biomass was detected among R. padi-infested and uninfested wheat lines. Biomass of D. noxia from Tugela (D. noxia-susceptible) was significantly higher than from plants with Diuraphis noxia-resistant Dn genes. Diuraphis noxia biomass from Tugela-Dn1 and Dn2 lines was not different from each other, but they were lower than from Tugela-Dn5. In contrast, there was no difference in R. padi biomass among wheat lines. Concentrations of chlorophylls a and b and carotenoids were significantly lower in D. noxia-infested plants compared with R. padi-infested and uninfested plants. When infested by D. noxia, chlorophyll a and b concentrations were not different among wheat lines on day 3, but they were lower in Tugela and Tugela-Dn1 than in Tugela-Dn2 and -Dn5 plants on days 6 and 12. However, no difference was detected in chlorophyll a/b or chlorophyll/carotenoid ratio among Tugela lines. The study demonstrated that Dn genes in the Tugela isolines conferred resistance to D. noxia but not to R. padi. Tugela-Dn1 was antibiotic, Tugela-Dn2 was tolerant and antibiotic, and Tugela-Dn5 was moderately antibiotic.

Topics: Animals; Aphids; Carotenoids; Chlorophyll; Chlorophyll A; Genotype; Plant Diseases; Triticum

Chlorophyll a (Chl a) and chlorophyll b (Chl b) plant pigments, which are important in the food industry and are beneficial as environmental pollution indicators, have been extracted with a novel solvent mixture (1:1 v/v acetone-propanol) not containing chloroform and simultaneously determined by first-derivative spectrophotometry. The results were statistically compared to those obtained by the ordinary absorption spectrophotometric reference utilizing the principle of additivity of absorbances. The testing of the developed method in synthetic mixtures of Chl a and Chl b and in real plant material samples (grass, spinach, chard, purslane, black cabbage, crisp lettuce, rocket, dill and seaweed) proved successful in that the developed extractive derivative spectrophotometric method was both rapid and precise, and was not dependent on the Chl a/b ratio in contrast to the reference method which was adversely affected by the latter parameter.

Topics: Calibration; Chlorophyll; Chlorophyll A; Plant Extracts; Solvents; Species Specificity; Spectrophotometry; Vegetables

A peroxidative activity was found in solubilized thylakoid membranes of olives (Olea europaea) cv. hojiblanca that catalyses degradation of chloroplast pigments in the presence of H2O2 and 2,4-dichlorophenol (DCP). The intermediate products of this degradation were analyzed using HPLC with diode array detection and the results indicated that 13(2)-OH-chlorophyll a and 13(2)-OH-chlorophyll b were the primary catabolites. The peroxidative activity assosiated with the thylakoid membranes affected, not only chlorophyll a and chlorophyll b, but also other accessory pigments in the photosynthetic process, such as the carotenoids. Quantitatively, the progressive decrease of the ratios Chl a/b and total Chls a+b/carotenoids indicated a more rapid disappearance of Chl a than of Chl b and a faster degradation of Chls a+b than of carotenoids.

Topics: Carotenoids; Chlorophenols; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Hydrogen Peroxide; Olea; Thylakoids

A Petri dish system in which development of oak (Quercus robur L.) microcuttings is stimulated by the late stage ectomycorrhizal (EM) fungus Piloderma croceum J. Erikss. & Hjortst. in a long pre-symbiotic stage was optimised to allow synchronous, rhythmic plant growth. Addition of indole-3-acetic acid or activated charcoal to the medium caused an early and more intensive EM formation coupled with suppression of most developmental effects of P. croceum. Leaf area, chlorophyll fluorescence, and content were compared in inoculated and uninoculated plants grown at two relative humidity levels (45 and 95%) and under consideration of three possible answers to inoculation, that is, no or EM formation after the 1st or the 2nd shoot flush. The culture conditions for uninoculated plants were suboptimal, leading toward photochemical stress reflected by a non photochemical quenching (qE) increase and a reduced Chl content at the end of the assay. Prior to EM formation, inoculation itself enhanced the optimal (Fv/Fm) and effective (phiPSII) quantum yield in leaves of the 1st shoot flush under reduced relative humidity. It also fully protected the plants against stress during the complete assays. The results indicate that inoculated plants only form EM once they have acquired a sufficient development level and C-providing capacity. However, the fungus actively improves the development and photosynthesis of plants up to the pre-mycorrhizal stage, helping them to reach this capacity.

Topics: Charcoal; Chlorophyll; Chlorophyll A; Culture Media; Fungi; Humidity; Indoleacetic Acids; Mycorrhizae; Photosynthesis; Plant Roots; Quercus

Electric field-induced absorption changes (electrochromism or Stark effect) of the light-harvesting PSII pigment-protein complexes LHCIIb, CP29, CP26 and CP24 were investigated. The results indicate the lack of strong intermolecular interactions in the chlorophyll a (Chl a) pools of all complexes. Characteristic features occur in the electronic spectrum of Chl b, which reflect the increased values of dipole moment and polarizability differences between the ground and excited states of interacting pigment systems. The strong Stark signal recorded for LHCIIb at 650-655 nm is much weaker in CP29, where it is replaced by a unique Stark band at 639 nm. Electrochromism of Chl b in CP26 and CP24 is significantly weaker but increased electrochromic parameters were also noticed for the Chl b transition at 650 nm. The spectra in the blue region are dominated by xanthophylls. The differences in Stark spectra of Chl b are linked to differences in pigment content and organization in individual complexes and point to the possibility of electron exchange interactions between energetically similar and closely spaced Chl b molecules.

Topics: Chlorophyll; Chlorophyll A; Electrochemistry; Electromagnetic Fields; Light-Harvesting Protein Complexes; Photosystem II Protein Complex; Protein Binding; Spectrum Analysis

Acclimation of the photosynthetic apparatus of chlorophyll b-less barley mutant chlorina f2 to low light (100 micromolm(-2)s(-1); LL) and extremely high light level (1000 micromolm(-2)s(-1); HL) was examined using techniques of pigment analysis and chlorophyll a fluorescence measurements at room temperature and at 77 K. The absence of chlorophyll b in LL-grown chlorina f2 resulted in the reduction of functional antenna size of both photosystem II (by 67%) and photosystem I (by 21%). Chlorophyll fluorescence characteristics of the LL-grown mutant indicated no impairment of the utilization of absorbed light energy in photosystem II photochemistry. Thermal dissipation of excitation energy estimated as non-photochemical quenching of minimal fluorescence (SV(0)) was significantly higher as compared to the wild-type barley grown under LL. Despite impaired assembly of pigment-protein complexes, chlorina f2 was able to efficiently acclimate to HL. In comparison with chlorina f2 grown under LL, HL-grown chlorina f2 was characterized by unaffected maximal photochemical efficiency of photosystem II (F(V)/F(M), doubled content of both beta-carotene and the xanthophyll cycle pigments and considerably reduced efficiency of excitation energy transfer from carotenoids to chlorophyll a. The enormous xanthophyll cycle pool size was however associated with reduced SV(0) capacity. We suggest that the substantial part of the xanthophyll cycle pigments is not bound to the remaining pigment-protein complexes and acts as filter for excitation energy, thereby contributing to the efficient photoprotection of chlorina f2 grown under HL.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Energy Metabolism; Fluorescence; Hordeum; Light; Light-Harvesting Protein Complexes; Photosynthesis; Photosystem II Protein Complex; Plant Leaves; Plants, Genetically Modified; Temperature

Growth and photosynthesis responses were measured for Scots pine (Pinus sylvestris L. cv.) inoculated with ectomycorrhizal fungi (Suillus bovinus) under 6.5 and 25 mg/L Cu treatments to evaluate ectomycorrhizal seedlings' tolerance to heavy metal stress. Results showed that excessive Cu can significantly impair the growth and photosynthesis of pine seedlings, but such impairment is much smaller to the ectomycorrhizal seedlings. Under 25 mg/L Cu treatment, the dry weight of ectomycorrhizal seedlings is 25% lower than the control in contrary to 53% of the non-mycorrhizal seedlings, and the fresh weight of ectomycorrhizal roots was significantly higher than those of non-mycorrhizal roots, about 25% and 42% higher at 6.5 and 25 mg/L Cu treatments respectively. Furthermore, ectomycorrhizal fungi induced remarkable difference in the growth rate and pigment content of seedlings under excessive Cu stress. At 25 mg/L Cu, the contents of total chlorophyll, chlorophyll-a and chlorophyll-b were 30% higher in ectomycorrhizal plants than those in non-mycorrhizal plants. O2 evolution and electron transport of PSI and PSII were restrained by elevated Cu stress. However, no significant improvement was observed in reducing the physiological restraining in ectomycorrhizal seedlings over the non-mycorrhizal ones.

Topics: Chlorophyll; Chlorophyll A; Copper; Mycorrhizae; Photosynthesis; Pinus; Plant Roots; Seedlings; Soil Pollutants

The light-harvesting chlorophyll a/b complex (LHCIIb) spontaneously assembles from its pigment and protein components in detergent solution. The formation of functional LHCIIb can be detected in time-resolved experiments by monitoring the establishment of excitation energy transfer from protein-bound chlorophyll b to chlorophyll a. To detect the possible initial steps of chlorophyll binding that may not yet give rise to chlorophyll b-to-a energy transfer, we have monitored LHCIIb assembly by measuring excitation energy transfer from a fluorescent dye, covalently bound to the protein, to the chlorophylls. In order to exclude interference of the dye with protein folding or pigment binding, the experiments were repeated with the dye bound to four different positions in the protein. Initial chlorophyll binding occurs at roughly the same rate as the establishment of chlorophyll b-to-a energy transfer, in the range of 10 s. However, under limiting chlorophyll concentrations, the binding of chlorophyll a clearly precedes that of chlorophyll b. The complex containing the apoprotein, carotenoids, and chlorophyll a but no chlorophyll b is biochemically unstable and therefore cannot be isolated. However, chlorophyll a binding into this weak complex is specific, as it does not occur with a C-terminal deletion mutant of Lhcb1 which still contains most chlorophyll-ligating amino acids but is unable to fold and assemble into functional LHCIIb. As a scenario for LHCIIb assembly in the thylakoid, we propose the initial formation of a labile Lhcb1-chlorophyll a-carotenoid complex that then becomes stabilized by the binding (or formation in situ) of chlorophyll b.

Topics: Animals; Binding Sites; Boron Compounds; Carotenoids; Chlamydomonas; Chlorophyll; Chlorophyll A; Cysteine; Energy Transfer; Fluorescence Resonance Energy Transfer; Fluorescent Dyes; Gene Deletion; Kinetics; Light-Harvesting Protein Complexes; Peptide Hydrolases; Photosynthetic Reaction Center Complex Proteins; Pigments, Biological; Protein Binding; Protein Folding; Protein Structure, Tertiary; Spectrometry, Fluorescence; Time Factors

Data are presented from 39 species of mosses and 16 liverworts for ratios of chlorophylls and total carotenoids, and light saturation of photosynthetic electron flow or photosynthetic CO2 uptake, in relation to the postulate that bryophyte cells in general show shade-plant characteristics.. Pigment concentrations were measured by spectrophotometer in 80 % acetone extracts. Light-saturation curves were constructed by (modulated) chlorophyll florescence and for some species by infra-red gas analysis.. The pigment measurements were widely variable but broadly in line with the findings of previous authors. Median values (mosses/liverworts) were: total chlorophyll, 1.64/3.76 mg g(-1); chlorophyll a : b, 2.29/1.99; chlorophylls : carotenoids, 4.74/6.75). The PPFD values at 95 % saturation (estimated from fitted curves) also ranged widely, but were almost all <1000 micromol m(-2) s(-1); the median for mosses was 583 and for liverworts 214 micromol m(-2) s(-1). The two highest PPFD95% values were from Polytrichum species with lamella systems forming a ventilated photosynthetic tissue. Total chlorophyll, chlorophyll a : b and chlorophylls : carotenoids all correlated significantly with PPFD95%.. Bryophytes include but are not inherently shade plants. Light-saturation levels for species of open sun-exposed habitats are lower than for vascular sun plants and are probably limited by CO2 diffusion into unistratose leaves; this limit can only be exceeded by bryophytes with ventilated photosynthetic tissues which provide increased area for CO2 uptake.

Topics: Bryophyta; Carotenoids; Chlorophyll; Chlorophyll A; England; Light; Photosynthesis; Pigments, Biological; Plant Leaves; Regression Analysis; Species Specificity

Topics: Araceae; Cadmium; Carcinogens, Environmental; Chlorophyll; Chlorophyll A; Chromium; Copper; Water Pollutants

The excitation energy transfer between photosystem (PS) II complexes was studied in isolated grana disks and thylakoids using chlorophyll a fluorescence induction measurements in the presence of DCMU under stacked and destacked conditions. Destacking of grana was achieved using a sonication protocol in a buffer without MgCl(2). The degree of stacking was controlled and quantified by atomic force microscopy and by the concomitant absorption changes. As expected from the literature, intact thylakoids showed a strong dependency of the connectivity of PSII centers, the F(m)/F(o) ratio as well as the fraction of PSIIbeta centers on the MgCl(2) concentration. In contrast, these parameters did not change in isolated grana disks. In particular, the connectivity remained constantly high irrespective of the degree of destacking. These differences were explained by the high protein density in grana disks, which hinders separation and mixing of proteins sufficiently to change energy transfer properties. Due to the occurrence of stroma lamella in intact thylakoids, intermixing of PSII and PSI is possible and allows for changes in F(m)/F(o) ratio as is the separation of LHCII from PSII, thus leading to an increase in the fraction of PSIIbeta. Even if mixing and separation of proteins are impaired in isolated grana disks, destacking should lead to a decrease in connectivity if transversal excitation energy transfer between two opposite membranes is significant. Because the connectivity is constant over all degrees of destacking employed, we conclude that the energy transfer in granas is mainly lateral.

Topics: Chlorophyll; Chlorophyll A; Energy Transfer; Fluorometry; Lipid Bilayers; Magnesium Chloride; Microscopy, Atomic Force; Photosystem II Protein Complex; Sonication; Spectrophotometry; Spinacia oleracea; Thylakoids

The major light-harvesting protein of photosystem II (LHCIIb) is the most abundant chlorophyll-binding protein in the thylakoid membrane. It contains three membrane-spanning alpha helices; the first and third one closely interact with each other to form a super helix, and all three helices bind most of the pigment cofactors. The protein loop domains connecting the alpha helices also play an important role in stabilizing the LHCIIb structure. Single amino acid exchanges in either loop were found to be sufficient to significantly destabilize the complex assembled in vitro [Heinemann, B., and Paulsen, H. (1999) Biochemistry 38, 14088-14093. Mick, V., Eggert, K., Heinemann, B., Geister, S., and Paulsen, H (2004) Biochemistry 43, 5467-5473]. This work presents an analysis of such point mutations in the lumenal loop with regard to the extent and nature of their effect on LHCIIb stability to obtain detailed information on the contribution of this loop to stabilizing the complex. Most of the mutant proteins yielded pigment-protein complexes if their reconstitution and/or isolation was performed under mild conditions; however, the yields were significantly different. Several mutations in the vicinity of W97 in the N-proximal section of the loop gave low reconstitution yields even under very mild conditions. This confirms our earlier notion that W97 may be of particular relevance in stabilizing LHCIIb. The same amino acid exchanges accelerated thermal complex dissociation in the absence of lithium dodecyl sulfate (LDS) and raised the accessibility of the lumenal loop to protease; both effects were well correlated with the reduction in reconstitution yields. We conclude that a detachment of the lumenal loop is a possible first step in the dissociation of LHCIIb. Dramatically reduced complex yields in the presence but not in the absence of LDS were observed for some but not all mutants, particularly those near the C-proximal end of the loop. We conclude that complex stabilities in the absence and in the presence of LDS do not correlate and most likely are determined by different structural characteristics, at least in LHCIIb but maybe also in other membrane proteins.

Topics: Amino Acid Substitution; Chlorophyll; Chlorophyll A; Electrophoresis, Polyacrylamide Gel; Hydrolysis; Kinetics; Light-Harvesting Protein Complexes; Photosystem II Protein Complex; Pisum sativum; Plant Proteins; Protein Denaturation; Protein Folding; Protein Structure, Tertiary; Sodium Dodecyl Sulfate; Temperature; Trypsin

We investigated adaptive responses of the photosynthetic electron transport to a decline in the carbon assimilation capacity. Leaves of different ages from wild-type tobacco (Nicotiana tabacum) L. var Samsun NN and young mature leaves of tobacco transformants with impaired photoassimilate export were used. The assimilation rate decreased from 280 in young mature wild-type leaves to below 50 mmol electrons mol chlorophyll(-1) s(-1) in older wild-type leaves or in transformants. The electron transport capacity, measured in thylakoids isolated from the different leaves, closely matched the leaf assimilation rate. The numbers of cytochrome (cyt)-bf complexes and plastocyanin (PC) decreased with the electron transport and assimilation capacity, while the numbers of photosystem I (PSI), photosystem II, and plastoquinone remained constant. The PC to PSI ratio decreased from five in leaves with high assimilation rates, to values below one in leaves with low assimilation rates, and the PC versus flux correlation was strictly proportional. Redox kinetics of cyt-f, PC, and P700 suggest that in leaves with low electron fluxes, PC is out of the equilibrium with P700 and cyt-f and the cyt-f reoxidation rate is restricted. It is concluded that the electron flux is sensitive to variations in the number of PC, relative to PSI and cyt-bf, and PC, in concert with cyt-bf, is a key component that adjusts to control the electron transport rate. PC dependent flux control may serve to adjust the electron transport rate under conditions where the carbon assimilation is diminished and thereby protects PSI against over-reduction and reactive oxygen production.

Topics: Carbon; Chlorophyll; Chlorophyll A; Electron Transport; Light; Logistic Models; Nicotiana; Oxidation-Reduction; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Plant Leaves; Plastocyanin; Thylakoids; Time Factors

The effects of light intensity on photosynthetic characteristics and possible mechanisms of preventing photodamage for two Ficus species seedlings grown under different irradiance were studied in rainy season. Ficus tinctoria Corner. is a sun species. Ficus subulata Bl. is a shade-tolerant species. With the increase of growth light intensity, maximum photosynthetic rate (P(max)), midrib angle (MA), chlorophyll a to b ratio (Chl a/b), carotenoid to chlorophyll ratio (Car/Chl), and thermal dissipation (NPQ) increased, while chlorophyll content and the amount of light harvesting complex of photosystem II decreased in both Ficus species. Those changes could reduce the existence of excessive light energy and ameliorated the extent of photoinhibition of photosynthesis. Under strong irradiance, photosynthetic rate was higher in F. tinctoria grown under 100% and 36% relative irradiance (RI) than in F. subulata, but their thermal dissipation capacity did not differ significantly. The results suggested that under strong irradiance seedlings could reduce excessive light energy through increasing both light energy utilization by photosynthesis and thermal dissipation in F. tinctoria, but through increasing thermal dissipation only in F. subulata. Reducing excessive light energy could protect photosynthetic apparatus from photoinhibition or photodamage. Under low irradiance, excessive light energy was reduced mainly through increasing light energy utilization by photosynthesis in F. tinctoria when grown under 100% and 36% RI and through increasing both photosynthetic utilization and thermal dissipation when grown under 12% and 4% RI, and through increasing thermal dissipation only in F. subulata. The extent of photoinhibition was greater in F. subulata than in F. tinctoria under the same irradiance of leaf surface, while the diurnal changes in photoinhibition was similar between the two species when grown under 100% RI. Under 100% RI, MA is larger in F. subulata than in F. tinctoria, which resulted in a lower leaf surface irradiance in F. subulata than in F. tinctoria. This was the main reason that the two Ficus species showed similar diurnal photoinhibition pattern under 100% RI. Acclimation of MA to irradiance was important to Ficus plants, especially to F. subulata.

Topics: Chlorophyll; Chlorophyll A; Circadian Rhythm; Ficus; Light; Photosynthesis

Asynchronous cultures of wild-type Euglena gracilis were tested for their morphophysiological response to 10 mM MnSO4. Growth was only moderately slowed (15%), while oxygen evolution was never compromised. Inductively coupled plasma analyses indicated that the Mn cell content doubled with respect to controls, but no signs of localised accumulation were detected with X-ray microanalysis. Evident morphological alterations were found at the plastid level with transmission electron microscopy and confocal laser scanning microscopy. An increase in the plastid mass, accompanied by frequent aberrations of chloroplast shape and of the organisation of the thylakoid system, was observed. These aspects paralleled a decrease in the molar ratio of chlorophyll a to b and an increase in the fluorescence emission ratio of light-harvesting complex II to photosystem II, the latter evaluated by in vivo single-cell microspectrofluorimetry. These changes were observed between 24 and 72 h of treatment. However, the alterations in the pigment pattern and photosystem II fluorescence were no longer observed after 96 h of Mn exposure, notwithstanding the maintenance of the large plastid mass. The response of the photosynthetic apparatus probably allows the alga to limit the photooxidative damage linked to the inappropriately large peripheral antennae of photosystem II. On the whole, the resistance of Euglena gracilis to Mn may be due to an exclusion-tolerance mechanism since most Mn is excluded from the cell, and the small amount entering the organism is tolerated by means of morphophysiological adaptation strategies, mainly acting at the plastid level.

Topics: Adaptation, Physiological; Animals; Chlorophyll; Chlorophyll A; Chloroplasts; Drug Resistance; Euglena gracilis; Light-Harvesting Protein Complexes; Manganese; Manganese Compounds; Microscopy, Confocal; Microscopy, Electron, Transmission; Oxidative Stress; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plastids; Sulfates; Thylakoids

The effects of clinostating on physiological processes and biochemical characteristics of wheat plants (Triticum aestivum L.) both healthy and infected by the wheat streak mosaic virus (WSMV) were studied. In six experiments, each lasting over 30 days, healthy and infected plants of the dwarf Apogee variety were grown under conditions of continuous horizontal and vertical clinostating with 2 rpm at 21 +/- 2 degrees C and 6000 1x (the optimal moisture of a substrate being maintained). The control variants (healthy and infected) were simultaneously grown under the same conditions of temperature and illumination in stationary containers and in open pots. During the experiment, visual observations were carried out over the state of tested plants. After completing the experiment, biometric indices, pigment, carbohydrate and dry matter contents were determined in all the plants. It was shown that clinostating sharply reduced the reproductive function of healthy plants and considerably affected their biomass (productivity) and concentration of chlorophylls and sugars. The viral infection resulted in further reduction of these characteristics. In control variants the viral effect was more significant. We speculate that clinostating reduced the rate of reproduction and spread of the virus.

Topics: Biomass; Carbohydrates; Chlorophyll; Chlorophyll A; Gravitation; Mosaic Viruses; Pigments, Biological; Plant Leaves; Plant Roots; Rotation; Seeds; Triticum; Weightlessness Simulation

The photosynthetic apparatus contains several protein complexes, many of which are regulated by environmental conditions. In this study, the influences of microgravity on PSI and PSII in Brassica rapa plants grown aboard the space shuttle were examined. We found that Brassica plants grown in space had a normal level of growth relative to controls under similar conditions on Earth. Upon return to Earth, cotyledons were harvested and thylakoid membranes were isolated. Analysis of chlorophyll contents showed that the Chl a/b ratio (3.5) in flight cotyledons was much higher than a ratio of 2.42 in the ground controls. The flight samples also had a reduction of PSI complexes and a corresponding 30% decrease of PSI photochemical activity. Immunoblotting showed that the reaction centre polypeptides of PSI were more apparently decreased (e.g. by 24-33% for PsaA and PsaB, and 57% for PsaC) than the light-harvesting complexes. In comparison, the accumulation of PSII complex was less affected in microgravity, thus only a slight reduction in D1, D2 and LHCII was observed in protein blots. However, there was a 32% decrease of OEC1 in the flight samples, indicating a defective OEC subcomplex. In addition, an average 54% increase of the 54 kDa CF1-beta isoform was found in the flight samples, suggesting that space-grown plants suffered from certain stresses, consistent with implications of the increased Chl a/b ratio. Taken together, the results demonstrated that Brassica plants can adapt to spaceflight microgravity, but with significant alterations in chloroplast structures and photosynthetic complexes, and especially reduction of PSI and its activity.

Topics: Biomass; Brassica rapa; Chlorophyll; Chlorophyll A; Chloroplasts; Cotyledon; Electron Transport; Immunologic Techniques; Light-Harvesting Protein Complexes; Microscopy, Electron, Transmission; Photosynthesis; Photosystem I Protein Complex; Photosystem II Protein Complex; Pigments, Biological; Plant Leaves; Space Flight; Thylakoids; Weightlessness

Properties of mesophyll cells and photosynthetic membranes of Arabidopsis thaliana (L.) Heynh. and Pisum sativum (L.) plants grown in a horizontal clinostat and in control conditions were compared. Obtained data have show that under clinorotation conditions seedlings have experienced the following cell morphology changes structural chloroplast rearrangement in palisade cells, pigment content alteration, and cell aging acceleration.

Topics: Arabidopsis; Carotenoids; Chlorophyll; Chlorophyll A; Chloroplasts; Gravitation; Microscopy, Electron; Photosynthesis; Pigments, Biological; Pisum sativum; Plant Leaves; Rotation; Thylakoids; Weightlessness Simulation

We recently put forth a model of a protochlorophyllide (Pchlide) light-harvesting complex operative during angiosperm seedling de-etiolation (Reinbothe, C., Lebedev, N., and Reinbothe, S. (1999) Nature 397, 80-84). This model, which was based on in vitro reconstitution experiments with zinc analogs of Pchlide a and Pchlide b and the two NADPH:protochlorophyllide oxidoreductases (PORs), PORA and PORB, of barley, predicted a 5-fold excess of Pchlide b, relative to Pchlide a, in the prolamellar body of etioplasts. Recent work (Scheumann, V., Klement, H., Helfrich, M., Oster, U., Schoch, S., and Rüdiger, W. (1999) FEBS Lett. 445, 445-448), however, contradicted this model and reported that Pchlide b would not be present in etiolated plants. Here we demonstrate that Pchlide b is an abundant pigment in barley etioplasts but is rather metabolically unstable. It is rapidly converted to Pchlide a by virtue of 7-formyl reductase activity, an enzyme that had previously been implicated in the chlorophyll (Chl) b to Chl a reaction cycle. Our findings suggest that etiolated plants make use of 7-formyl reductase to fine tune the levels of Pchlide b and Pchlide a and thereby may regulate the steady-state level of light-harvesting POR-Pchlide complex.

Topics: Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Hordeum; Oxidoreductases; Oxidoreductases Acting on CH-CH Group Donors; Protochlorophyllide

Cytochrome b(6)f complexes contain a molecule of chlorophyll a (Chla), which, in Chlamydomonas reinhardtii, can be exchanged for extraneous chlorophyll during protracted incubation of the purified complex in detergent solution. The specificity of the site and its location in the complex have been studied by photochemical coupling and circular dichroism spectroscopy. Following substitution of the original chlorophyll with [(3)H]Chla, the complex was irradiated in the Soret absorption band of Chla to complete bleaching and the amount of radioactivity covalently bound to each b(6)f subunit determined. Strong labeling was found to be associated with cytochrome f. The labeling originates from [(3)H]Chla molecules bound to a slowly exchanging site and showing the properties of the endogenous Chl, not from molecules dissolved in the detergent belt surrounding the complex. Chlorophyll b (Chlb) can compete with Chla, albeit with a lower affinity. Irradiation of [(3)H]Chlb introduced into the slowly exchanging site yielded the same labeling pattern that was observed with [(3)H]Chla. Proteolytic cleavage showed [(3)H]Chla labeling to be strictly restricted to the C-terminal region of cytochrome f. Circular dichroism spectra of the native complex revealed a bilobed signal characteristic of excitonic interaction between chlorophylls. The structural and evolutionary implications of these findings are discussed.

Topics: Animals; Centrifugation, Density Gradient; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Circular Dichroism; Cytochrome b Group; Cytochrome b6f Complex; Dimerization; Hydrolysis; Light; Papain; Photochemistry; Protein Subunits; Spectrophotometry, Ultraviolet; Thylakoids; Tritium

Using a Synechocystis sp. PCC 6803 mutant strain that lacks photosystem (PS) I and that synthesizes chlorophyll (Chl) b, a pigment that is not naturally present in the wild-type cyanobacterium, the functional consequences of incorporation of this pigment into the PS II core complex were investigated. Despite substitution of up to 75% of the Chl a in the PS II core complex by Chl b, the modified PS II centers remained essentially functional and were able to oxidize water and reduce Q(A), even upon selective excitation of Chl b at 460 nm. Time-resolved fluorescence decay measurements upon Chl excitation showed a significant reduction in the amplitude of the 60-70 ps component of fluorescence decay in open Chl b-containing PS II centers. This may indicate slower energy transfer from the PS II core antenna to the reaction center pigments or slower energy trapping. Chl b and pheophytin b were present in isolated PS II reaction centers. Pheophytin b can be reversibly photoreduced, as evidenced from the absorption bleaching at approximately 440 and 650 nm upon illumination in the presence of dithionite. Upon excitation at 685 nm, transient absorption measurements using PS II particles showed some bleaching at 650 nm together with a major decrease in absorption around 678 nm. The 650 nm bleaching that developed within approximately 10 ps after the flash and then remained virtually unchanged for up to 1 ns was attributed to formation of reduced pheophytin b and oxidized Chl b in some PS II reaction centers. Chl b-containing PS II had a lower rate of charge recombination of Q(A)(-) with the donor side and a significantly decreased yield of delayed luminescence in the presence of DCMU. Taken together, the data suggest that Chl b and pheophytin b participate in electron-transfer reactions in PS II reaction centers of Chl b-containing mutant of Synechocystis without significant impairment of PS II function.

Topics: Chlorophyll; Cyanobacteria; Electron Transport; Energy Metabolism; Kinetics; Light-Harvesting Protein Complexes; Luminescent Measurements; Oxidation-Reduction; Oxygen; Pheophytins; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Proteins; Spectrometry, Fluorescence; Temperature

A supercomplex containing the photosystem I (PSI) and chlorophyll a/b light-harvesting complex I (LHCI) has been isolated using a His-tagged mutant of Chlamydomonas reinhardtii. This LHCI-PSI supercomplex contained approximately 215 chlorophyll molecules of which 175 were estimated to be chlorophyll a and 40 to be chlorophyll b, based on P700 oxidation and chlorophyll a/b ratio measurements. Its room temperature long wavelength absorption peak was at 680 nm, and it emitted chlorophyll fluorescence maximally at 715 nm (77 K). The LHCI was composed of four or more different types of Lhca polypeptides including Lhca3. No LHCII proteins or other phosphoproteins were detected in the LHCI-PSI supercomplexes suggesting that the cells from which they were isolated were in State 1. Electron microscopy of negatively stained samples followed by image analysis revealed the LHCI-PSI supercomplex to have maximal dimensions of 220 A by 180 A and to be approximately 105 A thick. An averaged top view was used to model in x-ray and electron crystallographic data for PSI and Lhca proteins respectively. We conclude that the supercomplex consists of a PSI reaction center monomer with 11 Lhca proteins arranged along the side where the PSI proteins, PsaK, PsaJ, PsaF, and PsaG are located. The estimated molecular mass for the complex is 700 kDa including the bound chlorophyll molecules. The assignment of 11 Lhca proteins is consistent with a total chlorophyll level of 215 assuming that the PSI reaction center core binds approximately 100 chlorophylls and that each Lhca subunit binds 10 chlorophylls. There was no evidence for oligomerization of Chlamydomonas PSI in contrast to the trimerization of PSI in cyanobacteria.

Topics: Animals; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Light; Light-Harvesting Protein Complexes; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex

The energy transfer processes between Chls b and Chls a have been studied in the minor antenna complex CP29 by femtosecond transient absorption spectroscopy. Two samples were analyzed: the native CP29, purified from higher plants, and the recombinant one, reconstituted in vitro with the full pigment complement. The measurements indicate that the transfer kinetics in the two samples are virtually identical, confirming that the reconstituted CP29 has the same spectroscopic properties as the native one. In particular, three lifetimes (150 fs, 1.2 ps, and 5-6 ps) were identified for Chl b-652 nm to Chl a energy transfer and at least one for Chl b-640 nm (600-800 fs). Considering that the complexes bind two Chls b per polypeptide, the observation of more than two lifetimes for the Chl b to Chl a energy transfer, in both samples, clearly indicates the presence of the so-called mixed Chl binding sites--sites which are not selective for Chl a or Chl b, but can accommodate either species. The kinetic components and spectra are assigned to specific Chl binding sites in the complex, which provides further information on the structural organization.

Topics: Chlorophyll; Chlorophyll A; Dose-Response Relationship, Radiation; Energy Transfer; Macromolecular Substances; Mutation; Protein Structure, Tertiary; Radiation Dosage; Recombinant Proteins; Zea mays

Integrity of chloroplast membranes is essential to photosynthesis. Loss of thylakoid membrane integrity has been proposed as a consequence of ozone (O(3)) exposure and therefore may be a mechanistic basis for decreased photosynthetic rates commonly associated with ozone exposure. To investigate this hypothesis, Pinus ponderosa seedlings were exposed to ambient air or ozone concentrations maintained at 0.15 or 0.30 microliter l(-1) for 10 h day(-1) for 51 days during their second growing season. Over the course of the study, foliage samples were periodically collected for thylakoid membrane, chlorophyll and protein analyses. Additionally, gas-exchange measurements were made in conjunction with foliage sampling to verify that observed chloroplastic responses were associated with ozone-induced changes in photosynthesis. Needles exposed to elevated ozone exhibited decreases in chlorophyll a and b content. The decreases were dependent on the duration and intensity of ozone exposure. When based on equal amounts of chlorophyll, ozone-exposed sample tissue exhibited an increase in total protein. When based on equal amounts of protein, ozone-exposed samples exhibited an increase in 37 kDa proteins, possibly consisting of breakdown products, and a possible decrease in 68 kDa proteins, Rubisco small subunit. There was also a change in the ratio of Photosystem I protein complexes CPI and CPII that may have contributed to decreased photosynthesis. Net photosynthetic rates were decreased in the high ozone treatment suggesting that observed structural and biochemical changes in the chloroplast were associated with alterations of the photosynthetic process.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Environmental Exposure; Oxidants, Photochemical; Ozone; Photosynthesis; Pinus; Pinus ponderosa; Seedlings

Structure and function of chloroplasts are known to after during senescence. The senescence-induced specific changes in light harvesting antenna of photosystem II (PSII) and photosystem I (PSI) were investigated in Cucumis cotyledons. Purified light harvesting complex II (LHCII) and photosystem I complex were isolated from 6-day non-senescing and 27-day senescing Cucumis cotyledons. The chlorophyll a/b ratio of LHCII obtained from 6-day-old control cotyledons and their absorption, chlorophyll a fluorescence emission and the circular dichroism (CD) spectral properties were comparable to the LHCII preparations from other plants such as pea and spinach. The purified LHCII obtained from 27-day senescing cotyledons had a Chl a/b ratio of 1.25 instead of 1.2 as with 6-day LHCII and also exhibited significant changes in the visible CD spectrum compared to that of 6-day LHCII, indicating some specific alterations in the organisation of chlorophylls of LHCII. The light harvesting antenna of photosystems are likely to be altered due to aging. The room temperature absorption spectrum of LHCII obtained from 27-day senescing cotyledons showed changes in the peak positions. Similarly, comparison of 77K chlorophyll a fluorescence emission characteristics of LHCII preparation from senescing cotyledons with that of control showed a small shift in the peak position and the alteration in the emission profile, which is suggestive of possible changes in energy transfer within LHCII chlorophylls. Further, the salt induced aggregation of LHCII samples was lower, resulting in lower yields of LHCII from 27-day cotyledons than from normal cotyledons. Moreover, the PSI preparations of 6-day cotyledons showed Chl a/b ratios of 5 to 5.5, where as the PSI sample of 27-day cotyledons had a Chl a/b ratio of 2.9 suggesting LHCII association with PSI. The absorption, fluorescence emission and visible CD spectral measurements as well as the polypeptide profiles of 27-day cotyledon-PSI complexes indicated age-induced association of LHCII of PSII with PSI obtained from 27-day cotyledons. We modified our isolation protocols by increasing the duration of detergent Triton X-100 treatment for preparing the PSI and LHCII complexes from 27-day cotyledons. However, the PSI complexes isolated from senescing samples invariably proved to have significantly low Chl a/b ratio suggesting an age induced lateral movement and possible association of LHCII with PSI complexes. The analyses of polypeptide comp

Topics: Chlorophyll; Chlorophyll A; Circular Dichroism; Cucumis sativus; Energy Transfer; Light-Harvesting Protein Complexes; Peptides; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Spectrometry, Fluorescence; Thylakoids

The major light-harvesting chlorophyll a/b protein (LHCIIb) of higher plants is one of the few membrane proteins that can be refolded in vitro. During folding, the apoprotein is assembled with pigments to form a structurally authentic and functional pigment--protein complex. All reconstitution procedures used so far include solubilization of the apoprotein in sodium dodecyl sulfate (SDS) where the protein adopts approximately half of its alpha-helical folding present in the native structure. This paper shows that this preformed alpha-helix is not a prerequisite for LHCIIb folding in vitro. The apoprotein can also be reconstituted starting from a solution in guanidinium hydrochloride (Gnd) where the protein contains no detectable helical structure. Reconstitution yields are somewhat lower in the Gnd than in the SDS procedure, but the reconstitution products exhibit very similar biochemical and spectroscopic properties. The kinetics of LHCIIb assembly, as assessed by time-resolved fluorescence measurements, are virtually the same in both reconstitution procedures. This demonstrates that the initiation of alpha-helix formation is not a rate-limiting step in LHCIIb apoprotein folding.

Topics: Apoproteins; Chlorophyll; Chlorophyll A; Circular Dichroism; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Protein Folding; Spectrometry, Fluorescence

A monolithic C18-bonded silica rod column (Merck Chromolith) was applied to the separation of mixtures of chlorophylls a and b and their derivatives originated by hydrolysis of the phytyl ester linkage (chlorophyllides), loss of the central Mg atom (pheophytins), or both processes (pheophorbides). Mobile phases containing two different ion-pair reagents, ammonium acetate and buffered pyridine, were tested. Both eluents achieved the resolution of the eight pigments in less than 5 min. The method based on the pyridine-containing mobile phase was applied to the separation of chlorophylls and their green coloured degradation products in senescing leaves of deciduous trees, green fruits and a marine microalga.

Topics: Chlorophyll; Chlorophyll A; Esters; Metals; Phytic Acid; Pyridines; Silicon Dioxide

Measurements of dipole strengths of chlorophylls in solution are reviewed and correlated. The refractive index dependence is found to be expressible in a simple empirical fashion that does not rely on the concept of vacuum dipole strength. The index dependence in some respects contradicts the dependence expected on the basis of effective field theories.

Topics: Bacterial Proteins; Bacteriochlorophylls; Chlorophyll; Chlorophyll A; Refractometry; Solutions; Static Electricity

The lichens Ramalina celastri (Spreng.) Krog & Swinsc., Punctelia microsticta (Müll. Arg.) Krog and Canomaculina pilosa (Stizenb.) Elix & Hale were transplanted simultaneously to 17 urban-industrial sites in a northwestern area of Córdoba city, Argentina. The transplantation sites were set according to different environmental conditions: traffic, industries, tree cover, building height, topographic level, position in the block and distances from the river and from the power plant. Three months later, chlorophyll a, chlorophyll b, phaeophytin a, soluble proteins, hydroperoxy conjugated dienes, malondialdehyde concentration and sulfur accumulation were determined, and a pollution index was calculated for each sampling site. Redundancy analysis was applied to detect the variation pattern of the lichen variables that can be 'best' explained by the environmental variables considered. The present study provides information about both the specific pattern response of each species to atmospheric pollution, and environmental conditions that determine it. As regards pollutants emission sources R. celastri showed a chemical response associated mainly with pollutant released by the power plant and traffic. P. microsticta and C. pilosa responded mainly to industrial sources. Regarding environmental conditions that affect the spreading of air pollutants and their incidence on the bioindicator, the topographic level and tree cover surrounding the sampling site were found to be important for R. celastri, tree cover surrounding the sampling site and the building height affected P. microsticta, while building height did so for C. pilosa.

Topics: Air Pollutants; Biomarkers; Chlorophyll; Chlorophyll A; Environment; Environmental Monitoring; Lichens; Power Plants; Proteins; Sensitivity and Specificity; Vehicle Emissions

Peripheral chlorophyll a/b binding antenna of photosystem I (LHCI) from green algae and higher plants binds specific low energy absorbing chlorophylls (red pigments) that give rise to a unique red-shifted emission. A three-dimensional structural model of the Lhca4 polypeptide from the LHCI from higher plants was constructed on the basis of comparative sequence analysis, secondary structure prediction, and homology modeling using LHCII as a template. The obtained model of Lhca4 helps to visualize protein ligands to nine chlorophylls (Chls) and three potential His residues to extra Chls. Central domain of the Lhca4 comprising the first (A) and the third (C) transmembrane (TM) helices that binds 6 Chl molecules and two carotenoids is conserved structurally, whereas the interface between the first and the second TM helices and the outer surface of the second TM helix differ significantly among the LHCI and LHCII polypeptides. The model of Lhca4 predicts a histidine residue in the second TM helix, a potential binding site for extra Chl in close proximity to Chls a5 and b5 (labeling by Kühlbrandt). The interpigment interactions in the formed pigment cluster are suggested to cause a red spectral shift in absorption and emission. Modeling of the LHCI-730 heterodimer based on the model structures of Lhca1 and Lhca4 allowed us to suggest potential sites of pigment-pigment interactions that might be formed upon heterodimerization or docking of the LHCI dimers to the surface of PSI.

Topics: Amino Acid Sequence; Arginine; Binding Sites; Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Computer Simulation; Consensus Sequence; Dimerization; Light-Harvesting Protein Complexes; Models, Molecular; Molecular Sequence Data; Molecular Structure; Photosystem I Protein Complex; Photosystem II Protein Complex; Pigments, Biological; Protein Conformation; Protein Structure, Secondary; Protein Subunits; Sequence Alignment

Topics: Ascorbate Peroxidases; Carbon Radioisotopes; Chlorophyll; Chlorophyll A; Chlorophyllides; Hordeum; Hydrogen Peroxide; Light; Oxidoreductases Acting on CH-CH Group Donors; Peroxidases; Plant Leaves; Protochlorophyllide; Salicylic Acid

We conducted two experiments to determine the usefulness of a chlorophyll content meter (CCM) for the measurement of foliar chlorophyll concentration in sugar maple (Acer saccharum Marsh.) in the fall color period. In Experiment 1, four sugar maple trees were visually assigned to each of four fall foliage color categories in October 1998. On four dates in the fall of 1999, leaves were taken from the trees and analyzed for chlorophyll concentration by absorbance of pigment extracts and by determination of the chlorophyll content index (CCI) with a CCM. The two measures of chlorophyll concentration were strongly correlated (P < 0.001, r2 = 0.72). In Experiment 2, the CCI of leaves from sugar maple trees subjected to one of four fertilization treatments (lime, lime + manure, lime + 10:10:10 N,P,K fertilizer and an untreated control) were determined with a CCM. Treatment effects were distinguishable between all pairwise comparisons (P < 0.001), except for the lime versus lime + NPK fertilizer treatments.

Topics: Acer; Chlorophyll; Chlorophyll A; Plant Leaves; Trees

Nicotiana tabacum L. plantlets were cultured in vitro photoautotrophically (0% sucrose) and photomixotrophically (3% or 5% sucrose) at two irradiances (80 or 380 mumol m-2 s-1) with the aim of investigating the effect of these culture conditions on photosynthetic parameters and on protective systems against excess excitation energy. In plantlets grown photoautotrophically under higher irradiance photoinhibition was demonstrated. These plantlets had a decreased chlorophyll (Chl) a + b content and Chl a/b ratio, an increased content of xanthrophyll cycle pigments and a higher deepoxidation state, a decreased maximum photochemical efficiency of photosystem II (PS II) and actual photochemical efficiency of PS II, and an increased non-photochemical quenching. In the photoautotrophically grown plantlets and those photomixotrophically grown with 3% sucrose, the increase of growth irradiance from 80 to 380 mumol m-2 s-1 stimulated the activities of ascorbate-glutathione cycle enzymes with the exception of ascorbate peroxidase. Ascorbate peroxidase activity was not affected by the increase in growth irradiance but a significant decrease with increasing sucrose concentration was evident. The higher concentration of sucrose in the medium (5%) in combination with the higher irradiance inhibited photosynthesis (decrease in Chl a + b content and net photosynthetic rate) but no significant changes in activities of ascorbate-glutathione cycle enzymes were found. These results suggest that exogenous sucrose added to the medium improved high irradiance and oxidative stress resistance of the plantlets but the effect of sucrose is concentration dependent.

Topics: Antioxidants; Ascorbic Acid; Chlorophyll; Chlorophyll A; Glutathione; Nicotiana; Photobiology; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Sucrose; Xanthophylls

Changes in the chloroplast ultra-structure and photochemical function were studied in detached barley (Hordeum vulgare L. cv. Akcent) leaf segments senescing in darkness or in continuous white light of moderate intensity (90 mumol m-2 s-1) for 5 days. A rate of senescence-induced chlorophyll degradation was similar in the dark- and light-senescing segments. The Chl a/b ratio was almost unchanged in the dark-senescing segments, whereas in the light-senescing segments an increase in this ratio was observed indicating a preferential degradation of light-harvesting complexes of photosystem II. A higher level of thylakoid disorganisation (especially of granal membranes) and a very high lipid peroxidation were observed in the light-senescing segments. In spite of these findings, both the maximal and actual photochemical quantum yields of the photosystem II were highly maintained in comparison with the dark-senescing segments.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Darkness; Hordeum; Light; Lipid Peroxidation; Microscopy, Electron; Photosystem II Protein Complex; Plant Leaves

The resolution of quaternary mixtures of chlorophylls a and b and pheophytins a and b has been accomplished by partial least-squares (PLS) multivariate calibration, applied to the fluorescence signals of these pigments. The total luminescence information of the compounds has been used to optimize the spectral data set to perform the calibration. After preliminary studies, a method is described in acetone media, to avoid emulsions with the olive oil samples. Different scanning paths have been selected for each method. For the simultaneous determination of the pigments in olive oil samples, a comparative study of the results found by using excitation, emission, and synchronous spectral data, as analytical signal, was performed. The excitation spectra were selected as the better analytical signals for the determination of the pigments in olive oil samples. The optimum wavelength range to record the excitation spectra (lambda(em) = 662 nm) was selected to minimize the contribution of pheophytin a and to maximize the contribution of the other pigments, which are the minor constituents in olive oil. Determination of these pigments in olive oil samples was effected from the excitation spectra of dissolutions o suitable aliquots in acetone. Recovery values from olive oil, spiked with chlorophylls a and b and pheophytins a and b, were in the ranges of 70-112, 71-111, 76-105, and 82-109%, respectively.

Topics: Chlorophyll; Chlorophyll A; Olive Oil; Pheophytins; Plant Oils; Spectrometry, Fluorescence

A complete ferredoxin (Fd) cDNA clone was isolated from potato (Solanum tuberosum L. cv Desiree) leaves. By molecular and immunoblot analysis, the gene was identified as the leaf-specific Fd isoform I. Transgenic potato plants were constructed by introducing the homologous potato fed 1 cDNA clone as an antisense construct under the control of the constitutive cauliflower mosaic virus 35S promoter. Stable antisense lines with Fd contents between 40% and 80% of the wild-type level were selected by northern- and western-blot analysis. In short-term experiments, the distribution of electrons toward their stromal acceptors was altered in the mutant plants. Cyclic electron transport, as determined by the quantum yields of photosystems I and II, was enhanced. The CO2 assimilation rate was decreased, but depending on the remaining Fd content, some lines showed photoinhibition. The leaf protein content remained largely constant, but the antisense plants had a lower total chlorophyll content per unit leaf area and an increased chlorophyll a/b ratio. In the antisense plants, the redox state of the quinone acceptor A in photosystem II (QA) was more reduced than that of the wild-type plants under all experimental conditions. Because the plants with lower Fd amounts reacted as if they were grown under a higher light intensity, the possibility that the altered chloroplast redox state affects light acclimation is discussed.

Topics: Agrobacterium tumefaciens; Amino Acid Sequence; Chlorophyll; Chlorophyll A; Cloning, Molecular; DNA, Complementary; Ferredoxins; Gene Expression Regulation, Plant; Light; Molecular Sequence Data; Photosynthesis; Plant Leaves; Plant Proteins; Plants; Plants, Genetically Modified; Reverse Transcriptase Polymerase Chain Reaction; Sequence Alignment; Sequence Homology, Amino Acid; Solanum tuberosum

Mapping of QTLs controlling chrolophyll content was conducted with the method of multiple-trait composite interval mapping, based on a recombinant inbred (RI) population consisting of 131 lines (F10) derived from a cross between two indica rice varieties, Acc8558 and H359, and on a corresponding genetic map comprising 147 RFLP and 78 AFLP markers. Six QTLs for contents of chlorophyll a and chlorophyll b were detected, respectively. Among them, five QTLs were the same between the two characters. These QTLs were mainly distributed on chromosomes 1 and 4, indicating that the two chromosomes were more important for chlorophyll content. qChlA1c and qChlB1b (these two QTLs were mapped at the same location) showed relatively large effects at all the four observation times and showed the largest effects at the sword leaf period. Another two QTLs (qChlA4a/qChlB4a and qChlA4b/qChlB4b) exhibited significant effects only at the second observation time, suggesting that they might act only at specific developmental stages.

Topics: Chlorophyll; Chlorophyll A; Chromosome Mapping; Crosses, Genetic; Oryza; Quantitative Trait Loci

Three plant xanthophylls are components of the xanthophyll cycle in which, upon exposure of leaves to high light, the enzyme violaxanthin de-epoxidase (VDE) transforms violaxanthin into zeaxanthin via the intermediate antheraxanthin. Previous work () showed that xanthophylls are bound to Lhc proteins and that substitution of violaxanthin with zeaxanthin induces conformational changes and fluorescence quenching by thermal dissipation. We have analyzed the efficiency of different Lhc proteins to exchange violaxanthin with zeaxanthin both in vivo and in vitro. Light stress of Zea mays leaves activates VDE, and the newly formed zeaxanthin is found primarily in CP26 and CP24, whereas other Lhc proteins show a lower exchange capacity. The de-epoxidation system has been reconstituted in vitro by using recombinant Lhc proteins, recombinant VDE, and monogalactosyl diacylglycerol (MGDG) to determine the intrinsic capacity for violaxanthin-to-zeaxanthin exchange of individual Lhc gene products. Again, CP26 was the most efficient in xanthophyll exchange. Biochemical and spectroscopic analysis of individual Lhc proteins after de-epoxidation in vitro showed that xanthophyll exchange occurs at the L2-binding site. Xanthophyll exchange depends on low pH, implying that access to the binding site is controlled by a conformational change via lumenal pH. These findings suggest that the xanthophyll cycle participates in a signal transduction system acting in the modulation of light harvesting versus thermal dissipation in the antenna system of higher plants.

Topics: beta Carotene; Chlorophyll; Chlorophyll A; Epoxide Hydrolases; Glycerides; Light; Light-Harvesting Protein Complexes; Oxidation-Reduction; Photosynthetic Reaction Center Complex Proteins; Plant Leaves; Plastids; Spectrophotometry; Xanthophylls; Zea mays

Topics: Algal Proteins; Animals; Chlamydomonas; Chlorophyll; Chlorophyll A; Light; Photosynthesis; Pigments, Biological; Xanthophylls

Under high-light conditions, photoprotective mechanisms minimize the damaging effects of excess light. A primary photoprotective mechanism is thermal dissipation of excess excitation energy within the light-harvesting complex of photosystem II (LHCII). Although roles for both carotenoids and specific polypeptides in thermal dissipation have been reported, neither the site nor the mechanism of this process has been defined precisely. Here, we describe the physiological and molecular characteristics of the Chlamydomonas reinhardtii npq5 mutant, a strain that exhibits little thermal dissipation. This strain is normal for state transition, high light-induced violaxanthin deepoxidation, and low light growth, but it is more sensitive to photoinhibition than the wild type. Furthermore, both pigment data and measurements of photosynthesis suggest that the photosystem II antenna in the npq5 mutant has one-third fewer light-harvesting trimers than do wild-type cells. The npq5 mutant is null for a gene designated Lhcbm1, which encodes a light-harvesting polypeptide present in the trimers of the photosystem II antennae. Based on sequence data, the Lhcbm1 gene is 1 of 10 genes that encode the major LHCII polypeptides in Chlamydomonas. Amino acid alignments demonstrate that these predicted polypeptides display a high degree of sequence identity but maintain specific differences in their N-terminal regions. Both physiological and molecular characterization of the npq5 mutant suggest that most thermal dissipation within LHCII of Chlamydomonas is dependent on the peripherally associated trimeric LHC polypeptides.

Topics: Algal Proteins; Amino Acid Sequence; Animals; beta Carotene; Carotenoids; Chlamydomonas; Chlorophyll; Chlorophyll A; Fluorescence; Light; Light-Harvesting Protein Complexes; Molecular Sequence Data; Mutation; Organisms, Genetically Modified; Oxygen; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Pigments, Biological; Sequence Alignment; Sequence Homology, Amino Acid; Temperature; Xanthophylls

The ChlH gene coding the H subunit of magnesium chelatase, an enzyme involved in chlorophyll biosynthesis, was silenced in Nicotiana benthamiana plants by infection with tobacco mosaic virus vectors (pTMV-30b) containing 67,214 or 549 nt long ChlH inserts. Silencing of the nuclear ChlH gene induced a chimeric phenotype with green and yellow/white leaves associated with alterations of chloroplast shape and ultrastructure. The symptoms became first evident around veins of young leaves, and only later in the mesophyll tissues. The efficiency of gene silencing was not dependent on the insert orientation, but was strongly correlated with the size of the ChlH insert, providing a flexible method to modulate the level of gene suppression. Silencing efficiency seemed to be strongly dependent on endogenous ChlH mRNA level of the target tissue. Silencing of the ChlH gene with the longest fragment of 549 nt also lowered the accumulation of ChlD and chlorophyll synthetase mRNAs, i.e. other genes involved in chlorophyll biosynthesis.

Topics: Carbon-Oxygen Ligases; Chlorophyll; Chlorophyll A; Chloroplasts; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Plant; Gene Silencing; Lyases; Microscopy, Electron; Nicotiana; Oxidoreductases; Oxidoreductases Acting on CH-CH Group Donors; Phenotype; Plant Leaves; Plant Proteins; Protein Subunits; RNA, Messenger; RNA, Viral; Tobacco Mosaic Virus

The effects of low growth temperature (15 degrees C) on the photosynthetic apparatus of maize were investigated in a set of 233 recombinant inbred lines by means of chlorophyll fluorescence, gas exchange measurements and analysis of photosynthetic pigments. A quantitative trait loci (QTL) analysis of five traits related to the functioning of the photosynthetic apparatus revealed a total of eight genomic regions that were significantly involved in the expression of the target traits. Four of these QTLs, located on chromosomes 1 (around 146 cM), 2 (around 138 cM), 3 (around 70 cM), and 9 (around 62 cM), were identified across several traits and the phenotypic correlation observed among those traits confirmed at the genetic level. The two QTLs on chromosomes 1 and 9 were also expressed in leaves developed at near-optimal temperature (25 degrees C) whilst the two QTLs on chromosomes 2 and 3 were specific to leaves developed at sub-optimal temperature. A QTL analysis conducted on traits related to the pigment composition of the leaves developed at 15 degrees C detected the QTL on chromosome 3 around 70 cM in 7 of the 11 traits analysed. This QTL accounted for up to 28% of the phenotypic variance of the quantum yield of electron transport at PSII in the fourth leaf after about 3 weeks at a sub-optimal temperature. The results presented here suggest that key gene(s) involved in the development of functional chloroplasts of maize at low temperature should be located on chromosome 3, close to the centromere.

Topics: Adaptation, Physiological; beta Carotene; Carbon Dioxide; Chlorophyll; Chlorophyll A; Chloroplasts; Chromosome Mapping; Cold Temperature; Light-Harvesting Protein Complexes; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Pigments, Biological; Plant Leaves; Polymorphism, Restriction Fragment Length; Quantitative Trait, Heritable; Xanthophylls; Zea mays; Zeaxanthins

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Endpoint Determination; Lethal Dose 50; Magnoliopsida; Pesticides; Risk Assessment

Chlorophyllide a oxygenase (CAO) catalyzes two-step oxygenation reactions and converts chlorophyllide a to chlorophyllide b. When CAO was introduced into the Synechocystis sp. PCC6803 genome, chlorophyll b was synthesized and incorporated into P700-chlorophyll a-protein complexes. Curve analysis of photosystem I particles showed that Ca687 was decreased with a concomitant increase in Cb652 suggesting that chlorophyll b was incorporated into Ca687-binding sites. When the level of chlorophyll b was high, Ca704, which is known as red chlorophyll and photosystem I trimers were decreased. Formation of photosystem I trimers is discussed in relation to red chlorophyll and chlorophyll b accumulation.

Topics: Binding Sites; Chlorophyll; Chlorophyll A; Chlorophyllides; Cyanobacteria; Genes, Bacterial; Mutation; Oxygenases; Photosynthetic Reaction Center Complex Proteins; Protein Structure, Quaternary; Spectrophotometry

The aim of this work was to examine the role of cytosolic phosphoglucomutase (cPGM; EC 5.4.2.2) in photosynthetic carbon partitioning. We have previously described the generation and characterisation of the tuber metabolism of transgenic potato ( Solanum tuberosum cv. Desiree) lines expressing the StcPGM gene in the antisense orientation under the control of the 35S promoter. Here we extend the characterisation of leaf metabolism within these lines, examining properties of gas exchange, carbon partitioning, and the effect of the genetic manipulation on a wide range of metabolites including metabolites of the sucrose-starch transition, glycolysis, the Krebs cycle and amino acid metabolism. The data acquired in the present study surprisingly reveal that the photosynthetic sucrose synthetic capacity of the leaves is largely unaltered but that these plants display a reduced rate of photosynthesis, a dramatic reduction in nucleotide levels, and a general decline of biosynthesis. We conclude that these lines exhibit only moderate changes in sucrose synthesis but more complex changes on a range of diverse metabolic pathways.

Topics: Amino Acids; Biological Transport; Carbohydrate Metabolism; Carbon; Carbon Dioxide; Carbon Radioisotopes; Carboxylic Acids; Carotenoids; Chlorophyll; Chlorophyll A; Cytosol; Enzymes; Nucleotides; Phosphoglucomutase; Photosynthesis; Pigments, Biological; Plant Leaves; Plants, Genetically Modified; Solanum tuberosum; Sucrose

Absorption, fluorescence, and fluorescence excitation spectra have been measured from CP26, CP29, and monomeric and trimeric LHCIIb light-harvesting complexes isolated from Photosystem II subchloroplast particles from spinach. The complexes were purified using a combination of isoelectric focusing and sucrose gradient ultracentrifugation. The chlorophyll (Chl) and xanthophyll pigment compositions were measured using high-performance liquid chromatography (HPLC). Using the pigment compositions from the HPLC analysis as a starting point, the absorption spectral profiles of the complexes have been reconstructed from the individual absorption spectra obtained for each of the pigments. Also, the fluorescence excitation spectra of the complexes have been deconvoluted. The data reveal the energy transfer efficiencies between Chl b and Chl a and between specific xanthophylls and Chl a in the complexes. The spectral analyses reveal the underlying features of the highly congested spectral profiles associated with the complexes and are expected to be beneficial to researchers employing spectroscopic methods to investigate the mechanisms of energy transfer between the pigments bound in these complexes.

Topics: Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Chromatography, High Pressure Liquid; Fluorescence Resonance Energy Transfer; Light-Harvesting Protein Complexes; Macromolecular Substances; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plant Proteins; Spectrometry, Fluorescence; Spectrophotometry; Spinacia oleracea; Xanthophylls

Low methanol concentrations (about 0.5% v/v) induce biomass production in cultures of the unicellular green alga Scenedesmus obliquus by more than 300%, compared to controls without this solvent. This effect on the microalgal growth was found to be dependent on the solvent concentration, the packed cell volume (PCV), light intensity and light quality. It could be shown that methanol addition leads to a decrease in size of the light harvesting complex (LHC) on the basis of chlorophylls and proteins, and thus to changes in structure and functioning of the photosynthetic apparatus. These alterations lead to enhanced photosynthesis and respiration rates. The action of methanol on the photosynthetic apparatus is thus comparable to the effect of enhanced CO(2) concentrations. These findings support the previously proposed pathway for methanol metabolization with CO(2) as the final product. We conclude that the subsequent assimilation of the increased CO(2) amounts by the Calvin-Benson cycle is a possible explanation for the methanol-mediated increase in biomass production in terms of PCV. The methanol effect is observed only in the light and in the presence of a functioning photosynthetic apparatus. Preliminary action spectra suggest that the primary photoreceptor is a chlorophyll-protein complex with two absorption maxima at 680 and 430 nm, which may possibly be attributed to the reaction center of photosystem II (PSII).

Topics: Biomass; Cell Size; Chlorophyll; Chlorophyll A; Chlorophyta; Light; Light-Harvesting Protein Complexes; Methanol; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Time Factors

Acclimation to irradiance was measured in terms of light-saturated photosynthetic carbon assimilation rates (P(max)), Rubisco, and pigment content in mature field-grown rice (Oryza sativa) plants in tropical conditions. Measurements were made at different positions within the canopy alongside irradiance and daylight spectra. These data were compared with a second experiment in which acclimation to irradiance was assessed in uppermost leaves within whole-plant shading regimes (10% low light [LL], 40% medium light [ML], and 100% high light [HL] of full natural sunlight). Two varieties, japonica (tropical; new plant type [NPT]) and indica (IR72) were compared. Values for Rubisco amount, chlorophyll a/b, and P(max) all declined from the top to the base of the canopy. In the artificial shading experiment, acclimation of P(max) (measured at 350 microL L(-1) CO(2)) occurred between LL and ML for IR72 with no difference observed between ML and HL. The Rubisco amount increased between ML and HL in IR72. A different pattern was seen for NPT with higher P(max) (measured at 350 microL L(-1) CO(2)) at LL than IR72 and some acclimation of this parameter between ML and HL. Rubisco levels were higher in NPT than IR72 contrasting with P(max). Comparison of data from both experiments suggests a leaf aging effect between the uppermost two leaf positions, which was not a result of irradiance acclimation. Results are discussed in terms of: (a) acclimation of photosynthesis and radiation use efficiency at high irradiance in rice, and (b) factors controlling photosynthetic rates of leaves within the canopy.

Topics: Acclimatization; Chlorophyll; Chlorophyll A; Crops, Agricultural; Light; Oryza; Photosynthesis; Pigments, Biological; Plant Leaves; Ribulose-Bisphosphate Carboxylase

The photosynthetic rate, the leaf characteristics related to photosynthesis, such as the chlorophyll content, chlorophyll a/b ratio and density of the stomata, the leaf area and the dry weight in seedlings of Japanese flowering cherry grown under normal gravity and simulated microgravity conditions were examined. No significant differences were found in the photosynthetic rates between the two conditions. Moreover, leaf characteristics such as the chlorophyll content, chlorophyll a/b ratio and density of the stomata in the seedlings grown under the simulated microgravity condition were not affected. However, the photosynthetic product of the whole seedling under the simulated microgravity condition increased compared with the control due to its leaf area increase. The results suggest that dynamic gravitational stimulus controls the partitioning of the products of photosynthesis.

Topics: Chlorophyll; Chlorophyll A; Gravitation; Photosynthesis; Plant Leaves; Prunus; Rotation; Seedlings; Weightlessness Simulation

The trimeric main light-harvesting complex (LHC-II) is the only antenna complex of higher plants of which a high-resolution 3D structure has been obtained (Kühlbrandt, W., Wang, D., and Fujiyoshi, Y. (1994) Nature 367, 614-621) and which can be refolded in vitro from its components. Four different recombinant forms of LHC-II, each with a specific chlorophyll (Chl) binding site removed by site-directed mutagenesis, were refolded from heterologously overexpressed apoprotein, purified pigments, and lipid. Absorption spectra of mutant LHC-II were measured in the temperature range from 4 to 300 K and compared to likewise refolded wild-type complex and to native LHC-II isolated from pea chloroplasts. Chls at different binding sites have characteristic, well-defined absorption sub-bands. Mixed occupation of binding sites with Chls a and b is not observed. Temperature-dependent changes of the mutant absorption spectra reveal a consistent shift of the major difference bands but an irregular behavior of minor bands. A model of the spectral substructure of LHC-II is proposed which accounts for the different absorption properties of the 12 individual Chls in the complex, thus establishing a first consistent correlation between the 3D structure of LHC-II and its spectral properties. The spectral substructure is valid for recombinant and native LHC-II, indicating that both have the same spatial arrangement of Chls and that the refolded complex is fully functional.

Topics: Binding Sites; Carotenoids; Chlorophyll; Chlorophyll A; Freezing; Light-Harvesting Protein Complexes; Mutagenesis, Site-Directed; Photosynthetic Reaction Center Complex Proteins; Pisum sativum; Protein Folding; Spectrometry, Fluorescence; Spectrophotometry, Ultraviolet; Spinacia oleracea

To elucidate the mechanism of an irradiance-dependent adjustment in the chlorophyll (Chl) antenna size of Dunaliella salina, we investigated the regulation of expression of the Chl a oxygenase (CAO) and light-harvesting complex b (Lhcb) genes as a function of Chl availability in the photosynthetic apparatus. After a high-light to low-light shift of the cultures, levels of both CAO and Lhcb transcripts were rapidly induced by about 6-fold and reached a high steady-state level within 1.5 h of the shift. This was accompanied by repair of photodamaged photosystem II (PSII) reaction centers, accumulation of Chl a and Chl b (4:1 ratio), photosystem I (PSI), light-harvesting complex, and by enlargement of the Chl antenna size of both photosystems. In gabaculine-treated cells, induction of CAO and Lhcb transcripts was not affected despite substantial inhibition in de novo Chl biosynthesis. However, cells were able to synthesize and accumulate some Chl a and Chl b (1:1 ratio), resulting in a marked lowering of the Chl a to Chl b ratio in the presence of this inhibitor. Assembly incorporation of light-harvesting complex and a corresponding Chl antenna size increase, mostly for the existing photosystems, was noted in the presence of gabaculine. Repair of photodamaged PSII was not affected by gabaculine. However, assembly accumulation of new PSI was limited under such conditions. These results suggest a coordinate regulation of CAO and Lhcb gene transcription by irradiance, independent of Chl availability. The results are discussed in terms of different signal transduction pathways for the regulation of the photosynthetic apparatus organization by irradiance.

Topics: Apoproteins; Cells, Cultured; Chlorophyll; Chlorophyll A; Chlorophyta; Cyclohexanecarboxylic Acids; Light; Light-Harvesting Protein Complexes; Oxygenases; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Signal Transduction; Thylakoids

The effect of enhanced UV-B radiation on buckwheat (Fagopyrum esculentum Moench. variety 'Darja'), an important high elevation crop, was studied in order to estimate its vulnerability in changing UV-B environment. Plants were grown in outdoor experiments from July to October under reduced and ambient UV-B levels, and an UV-B level simulating 17% ozone depletion in Ljubljana. During the development the following parameters were monitored: light saturated photosynthetic activity, transpiration, potential and effective photochemical efficiencies of photosystem II, the contents of photosynthetic pigments and methanol soluble UV-B absorbing compounds. At the end of the experiment, growth rate and production of seeds were estimated. In the following growth season the seeds collected from plants exposed to different UV-B treatments were tested for germination capacity. Total UV-B absorbing compounds during plant development were increased by UV-B radiation, photosynthetic pigments (chlorophyll a and b and carotenoids) decreased. Photosynthetic rate was lowered in an early stage of development. UV-B treatment resulted in the increase in the transpiration rate and consequently the decrease in water use efficiency (WUE). The disturbances in water economy and in photosynthesis affected the reproduction potential negatively; the production of seeds in plants cultivated under ambient and enhanced UV-B was 57 and 39% of the production of specimens treated with reduced UV-B, respectively. The germination of seeds collected from treated plants revealed on average about 95% success, independently of the treatment, but the time needed for germination was the shortest for seeds developed under enhanced UV-B level treatment. Enhanced UV-B radiation affected water relations and production of buckwheat, but not the potential of seeds for germination.

Topics: Chlorophyll; Chlorophyll A; Crops, Agricultural; Fagopyrum; Photosynthesis; Plant Transpiration; Seeds; Ultraviolet Rays

Trichloroacetic acid (TCA) has been detected in rain, snow, and river samples throughout the world. It may enter into natural water systems via herbicide use, as a by-product of water disinfection, from emissions of spent bleach liquor of kraft pulp mills, and as a natural fungal product. This compound is phytotoxic and likely to accumulate in aquatic environments. A study to assess the fate of TCA in semi-natural aquatic environments and the toxicity of TCA to rooted aquatic macrophytes was conducted. The experiment involved exposing three replicate 12000 l aquatic microcosms at the University of Guelph Microcosm Facility to 0.05, 0.5, 3, and 10 mg/l of TCA for 35 days in a one-way analysis of variance design. Each microcosm was stocked with 14 individual 5 cm apical shoots of Myriophyllum spicatum and M. sibiricum. The plants were sampled at regular intervals and assessed for the somatic endpoints of plant length, root growth, number of nodes and wet and dry mass and the biochemical endpoints of chlorophyll-a and chlorophyll-b, carotenoid content, and citric acid levels. TCA half-lives in the microcosms ranged from 190 to 296 h depending on the initial concentration of TCA. Myriophyllum spp. results indicate that while there were some statistically significant differences from controls, there were no biologically significant effects of TCA for any of the endpoints examined. These data suggest that TCA does not pose a significant risk to these macrophytes up to 10 mg/l, which typically exceeds environmentally relevant concentrations by several orders of magnitude.

Topics: Biomass; Carotenoids; Chlorophyll; Chlorophyll A; Citric Acid; Ecosystem; Environmental Monitoring; Half-Life; Magnoliopsida; Plant Roots; Trichloroacetic Acid; Water Pollutants, Chemical

CP29 (the lhcb4 gene product), a minor photosystem II antenna complex, binds six chlorophyll (Chl) a, two Chl b, and two to three xanthophyll molecules. The Chl a/b Q(y) absorption band substructure of CP29 (purified from spinach) was investigated by nonlinear polarization spectroscopy in the frequency domain (NLPF) at room temperature. A set of NLPF spectra was obtained at 11 probe wavelengths. Seven probe wavelengths were located in the Q(y) spectral region (between 630 and 690 nm) and four in the Soret band (between 450 and 485 nm). Evaluation of the experimental data within the framework of global analysis leads to the following conclusions: (i) The dominant Chl a absorption (with a maximum at 674 nm) splits into (at least) three subbands (centered at 660, 670, and 681.5 nm). (ii) In the Chl b region two subbands can be identified with maxima located at 640 and 646 nm. (iii) The lowest energy Q(y) transition (peaking at 681.5 nm) is assigned to a Chl a which only weakly interacts with other Chl aor b molecules by incoherent Förster-type excitation energy transfer. (iv) Pronounced excitonic interaction exists between certain Chl a and Chl b molecules, which most likely form a Chl a/b heterodimer. The subbands centered at 640 and 670 nm constitute a strongly coupled Chl a/b pair. The findings of the study indicate that the currently favored view of spectral heterogeneity in CP29 being due essentially to pigment-protein interactions has to be revised.

Topics: Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plant Proteins; Protein Structure, Tertiary; Spectrophotometry, Atomic; Spinacia oleracea

To assess effects of the environmental stress on polyphenol compounds (polyphenols) in plants, the polyphenol contents were investigated in the seedlings of safflower (Carthamus tinctrius L.) and cucumber (Cucumis sativus L.) grown under three types of growth conditions: control; light stress, irradiated with strong light in the visible wavelength range; and light/water stress, irradiated with strong visible light with a limited water supply. The total polyphenol contents and the amounts of the major polyphenols, especially luteolin 7-O-glucoside in safflower cotyledons, and luteolin 7-O-glucoside and luteolin in safflower foliage leaves, increased in response to both stresses. The polyphenol increasing effect of light/water stress was clearly observed in safflower compared to cucumber, suggesting that plants that are resistant to these stresses can accumulate substantial amounts of polyphenols compared to the plants which respond weakly to the stresses.

Topics: Asteraceae; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Cucumis sativus; Flavonoids; Glucosides; Light; Luteolin; Phenols; Plant Leaves; Polymers; Seeds; Ultraviolet Rays; Water

Stepwise two-photon excitation of chlorophyll a and b in the higher plant main light-harvesting complex (LHC II) and the minor complex CP29 (as well as in organic solution) with 100-fs pulses in the Q(y) region results in a weak blue fluorescence. The dependence of the spectral shape of the blue fluorescence on excitation wavelength offers a new approach to elucidate the long-standing problem of the origin of spectral "chlorophyll forms" in pigment-protein complexes, in particular the characterization of chlorophyll a/b-heterodimers. As a first result we present evidence for the existence of strong chlorophyll a/b-interactions (excitonically coupled transitions at 650 and 680 nm) in LHC II at ambient temperature. In comparison with LHC II, the experiments with CP29 provide further evidence that the lowest energy chlorophyll a transition (at approximately 680 nm) is not excitonically coupled to chlorophyll b.

Topics: Bacterial Proteins; Chlorophyll; Chlorophyll A; Dimerization; Light; Light-Harvesting Protein Complexes; Photons; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Spectrometry, Fluorescence; Temperature

Trichloroacetic acid (TCA) and trifluoroacetic acid (TFA) have been detected together in environmental water samples throughout the world. TCA may enter into aquatic systems via rainout as the degradation product of chlorinated solvents, herbicide use, as a by-product of water disinfection and from emissions of spent bleach liquor of kraft pulp mills. Sources of TFA include degradation of hydrofluorocarbons (HFCs) refrigerants and pesticides. These substances are phytotoxic and widely distributed in aquatic environments. A study to assess the risk of a binary mixture of TCA and TFA to macrophytes in aquatic microcosms was conducted as part of a larger study on haloacetic acids. M. spicatum and M. sibiricum were exposed to 0.1, 1, 3 and 10 mg/l of both TCA and TFA (neutralized with sodium hydroxide) in replicate (n = 3) 12000 l aquatic microcosms for 49 days in an one-way analysis of variance design. Each microcosm was stocked with 14 individual apical shoots per species. The plants were sampled at regular intervals and assessed for the somatic endpoints of plant length, root growth, number of nodes and wet and dry mass and the biochemical endpoints of chlorophyll-a, chlorophyll-b, carotenoid content and citric acid levels. Results indicate that there were statistically significant effects of the TCA/TFA mixture on certain pigment concentrations immediately after the start of exposure (2-7 days), but the plants showed no signs of stress thereafter. These data suggest that TCA/TFA mixtures at environmentally relevant concentrations do not pose a significant risk to these aquatic macrophytes.

Topics: Biomass; Caustics; Chlorophyll; Chlorophyll A; Disinfectants; Herbicides; Plant Development; Plants; Rain; Solvents; Trichloroacetic Acid; Trifluoroacetic Acid; Water Pollutants, Chemical

Aging has received considerable attention in biomedicine, but little is known about the regulatory mechanisms responsible for the aging not associated with senescence in plants. This study provides new insights into the relationship between oxidative stress and plant aging, and points out chloroplasts as one of the target organelles of age-associated oxidative stress in plants. We simultaneously analyzed lipid oxidation, photosynthesis, chlorophyll content, de-epoxidation state of the xanthophyll cycle, and levels of chloroplastic antioxidant defenses such as beta-carotene and alpha-tocopherol in leaves of the same age in 1-, 3- and 7-year-old Cistus clusii Dunal plants growing under Mediterranean field conditions. Enhanced formation of malondialdehyde in leaves (2.7-fold) and chloroplasts (2.8-fold), decreased photosynthetic activity (25%), and lower chlorophyll (ca. 20%) and chloroplastic antioxidant defense levels (ca. 25%-85%) were observed in 7-year-old plants, when compared with 1- and 3-year-old plants. The differences observed, which were associated with plant aging, were only noticeable in mature nonsenescing plants (7-year-old plants). No differences were observed between pre-reproductive (1-year-old plants) and young plants (3-year-old plants). This study shows that from a certain age, oxidative stress increases progressively in chloroplasts as plants age, whereas photosynthesis is reduced. The results indicate that the oxidative stress associated with the aging in plants accumulates progressively in chloroplasts, and that the contribution of oxidative stress to aging increases as plants age.

Topics: alpha-Tocopherol; Antioxidants; beta Carotene; Chlorophyll; Chlorophyll A; Chloroplasts; Cistus; Lipid Peroxidation; Oxidative Stress; Photosynthesis; Pigments, Biological; Plant Leaves; Reactive Oxygen Species; Time Factors; Xanthophylls

Transgenic plants of Arabidopsis thaliana (L.) Heynh. (ecotype Columbia) expressing the antisense AtMECT gene, encoding 2- C-methyl- D-erythritol 4-phosphate cytidylyltransferase, were generated to elucidate the physiological role of the nonmevalonate pathway for production of ent-kaurene, the latter being the plastidic precursor of gibberellins. In transformed plants pigmentation and accumulation of ent-kaurene were reduced compared to wild-type plants. Fosmidomycin, an inhibitor of 1-deoxy- D-xylulose 5-phosphate reductoisomerase (DXR), caused a similar depletion of these compounds in transgenic plants. These observations suggest that both AtMECT and DXR are important in the synthesis of isopentenyl diphosphate and dimethylallyl diphosphate and that ent-kaurene is mainly produced through the nonmevalonate pathway in the plastid.

Topics: Aldose-Ketose Isomerases; Arabidopsis; Carotenoids; Chlorophyll; Chlorophyll A; Chloroplasts; Cloning, Molecular; Diterpenes; Diterpenes, Kaurane; DNA, Antisense; Fosfomycin; Hemiterpenes; Mevalonic Acid; Molecular Sequence Data; Multienzyme Complexes; Nucleotidyltransferases; Organophosphorus Compounds; Oxidoreductases; Phenotype; Pigments, Biological; Plants, Genetically Modified

The chlorophyll a/b-xanthophyll-protein CP26 complex belongs to the Lhc protein family. It binds nine chlorophylls and two xanthophylls per 26.6 kDa polypeptide. Determination of the characteristics of each binding site is needed for the understanding of functional organization of individual proteins belonging to the photosystem II supramolecular complex. The biochemical and spectroscopic features of native CP26 are presented here together with identification of pigment binding and energy transitions in different sites. The analysis has been performed via a new approach using recombinant CP26 complexes in which the chromophore content has been experimentally modified. Data were interpreted on the basis of homology with CP29 and LHCII complexes, for which detailed knowledge is available from mutation analysis. We propose that one additional Chl b is present in CP26 as compared to CP29 and that it is located in site B2. We also found that in CP26 three chlorophyll binding sites are selective for Chl a, one of them being essential for the folding of the pigment-protein complex. Two xanthophyll binding sites were identified, one of which (L1) is essential for protein folding and specifically binds lutein. The second site (L2) has lower selectivity and can bind any of the xanthophyll species present in thylakoids.

Topics: Binding Sites; Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Circular Dichroism; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plant Proteins; Protein Denaturation; Protein Folding; Recombinant Proteins; Spectrometry, Fluorescence; Spectrophotometry; Thylakoids; Xanthophylls; Zea mays

The Willstätter allomerization reaction of chlorophylls (Chl) has posed a difficult problem in Chl and photosynthesis research over the past 90 years. Here, we present strong additional evidence, based on (18)O-labeling and mass spectrometry, for the previously published free-radical allomerization (FRA) mechanism (Hynninen, Z. Naturforsch. 1981, 36b, 1010-1016). This mechanism is also complemented now by describing two alternative pathways for the formation of 13(2)(S/R)-hydroxy-Chl a. The results from the (18,18)O(2)-experiments suggest that the predominant route for the formation of the 13(2)(S/R)-hydroxy-Chl a under essentially anhydrous conditions (anhydrous Chl and thoroughly dried methanol) is the homolytic cleavage of the C-13(2)-hydroperoxide intermediate. However, if Chl dihydrate and undried methanol are used in the reaction mixture, the direct route from the Chl C-13(2) radical to 13(2)(S/R)-hydroxy-Chl a can be predicted to become significant. The results from the (18,18)O(2)-allomerization experiments described in this paper also verified that the 13(2)(S/R)-methoxy-lactone derivatives and the 15-glyoxylic acid derivative of Chl a incorporated each a single (18)O-atom, whereas 13(2)(R/S)-methoxy-Chl a remained unlabeled. Consequently, these allomers are formed via the pathways previously suggested in the original FRA mechanism. The possible factors contributing to the control of the allomerization reactions are considered. Finally, the relationship between the allomerization reactions of Chl a and those of Chl b and BChl a is briefly discussed.

Topics: Bacteriochlorophyll A; Carbon Isotopes; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Dimerization; Energy Transfer; Kinetics; Mass Spectrometry; Molecular Structure; Oxygen Isotopes; Photosynthesis; Phytolacca dodecandra; Stereoisomerism; Structure-Activity Relationship

Guard cells of the orchid genus, Paphiopedilum have been reported to lack developed chloroplasts and detectable chlorophyll a autofluorescence. Paphiopedilum stomata lack a photosynthesis-dependent opening response but have a blue light-specific opening. The present study found that low fluence rate green and red light elicited stomatal opening in Paphiopedilum and this opening was reversed by far red light, indicating the presence of a phytochrome-mediated opening response. Phytochrome-dependent, red light-stimulated opening was largest under low fluence rates and decreased to near zero as fluence rate increased. A recently discovered green light reversibility of blue light-specific stomatal opening was used to probe the properties of the blue light response in Paphiopedilum stomata. Blue light-stimulated opening was completely reversed by green light in the presence of far red light. Red light enhanced the blue light response of Paphiopedilum guard cells when given as a pretreatment or together with blue light. Analysis of guard cell pigments showed that guard cells have small amounts of chlorophyll a and b, zeaxanthin, violaxanthin, antheraxanthin and lutein. Zeaxanthin content increased in response to blue light or ascorbate and declined in the dark or under illumination in the presence of dithiothreitol, indicating the presence of an active xanthophyll cycle. Thus Paphiopedilum stomata possess both a blue light-mediated opening response with characteristics similar to species with normal chloroplast development and a novel phytochrome-mediated opening response.

Topics: Adaptation, Physiological; Ascorbic Acid; beta Carotene; Chlorophyll; Chlorophyll A; Dithiothreitol; Light; Lutein; Orchidaceae; Photosynthesis; Phytochrome; Pigments, Biological; Plant Epidermis; Xanthophylls; Zeaxanthins

Light-harvesting complexes (LHC) of higher plants are composed of at least 10 different proteins. Despite their pronounced amino acid sequence homology, the LHC of photosystem II show differences in pigment binding that are interpreted in terms of partly different functions. By contrast, there is only scarce knowledge about the pigment composition of LHC of photosystem I, and consequently no concept of potentially different functions of the various LHCI exists. For better insight into this issue, we isolated native LHCI-730 and LHCI-680. Pigment analyses revealed that LHCI-730 binds more chlorophyll and violaxanthin than LHCI-680. For the first time all LHCI complexes are now available in their recombinant form; their analysis allowed further dissection of pigment binding by individual LHCI proteins and analysis of pigment requirements for LHCI formation. By these different approaches a correlation between the requirement of a single chlorophyll species for LHC formation and the chlorophyll a/b ratio of LHCs could be detected, and indications regarding occupation of carotenoid-binding sites were obtained. Additionally the reconstitution approach allowed assignment of spectral features observed in native LHCI-680 to its components Lhca2 and Lhca3. It is suggested that excitation energy migrates from chlorophyll(s) fluorescing at 680 (Lhca3) via those fluorescing at 686/702 nm (Lhca2) or 720 nm (Lhca3) to the photosystem I core chlorophylls.

Topics: beta Carotene; Binding Sites; Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Pigments, Biological; Plant Leaves; Solanum lycopersicum; Spectrometry, Fluorescence; Xanthophylls

Toxins are secondary metabolites produced by several plant pathogenic microorganisms. These toxins play a major role in the development of disease symptoms. Coronatine, the non specific toxin, was extracted and purified from the culture medium of Pseudomonas syringae pv. glycinea. In this study, the effect of coronatine on the development of the chlorotic lesion on the Phaseolus vulgaris L. leaves, indicates that coronatine induced chlorosis on treated leaves as well as or untreated leaves on the same plant. An other effect of this toxin is the reduction of amount of both chlorophyll a and b. These results, provide that the development of chlorotic lesion is a primarily mode of action of coronatine.

Topics: Amino Acids; Chlorophyll; Chlorophyll A; Indenes; Phaseolus; Plant Leaves; Pseudomonas

This study aimed to examine the electron transport rates in the thylakoids, isolated from leaves of pea plants grown under clinorotation and in vertical control, to measure the chlorophyll a/b (Chl a/b) ratio in such thylakoids and in photosystem I (PSI) particles isolated from them, to elucidate if there are any differences in changes of PS II activity in thylakoids and Chl a/b ratio in PS I particles under phosphorylation of polypeptides of thylakoid pigment-protein complexes.

Topics: Chlorophyll; Chlorophyll A; Electron Transport; Light-Harvesting Protein Complexes; Phosphorylation; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Pisum sativum; Plant Leaves; Rotation; Thylakoids; Weightlessness Simulation

The steady state absorption and fluorescence spectroscopic properties of the xanthophylls, violaxanthin, zeaxanthin, and lutein, and the efficiencies of singlet energy transfer from the individual xanthophylls to chlorophyll have been investigated in recombinant CP26 protein overexpressed in Escherichia coli and then refolded in vitro with purified pigments. Also, the effect of the different xanthophylls on the extents of static and dynamic quenching of chlorophyll fluorescence has been investigated. Absorption, fluorescence, and fluorescence excitation demonstrate that the efficiency of light harvesting from the xanthophylls to chlorophyll a is relatively high and insensitive to the particular xanthophyll that is present. A small effect of the different xanthophylls is observed on the extent of quenching of Chl fluorescence. The data provide the precise wavelengths of the absorption and fluorescence features of the bound pigments in the highly congested spectral profiles from these light-harvesting complexes. This information is important in assessing the mechanisms by which higher plants dissipate excess energy in light-harvesting proteins.

Topics: beta Carotene; Chlorophyll; Chlorophyll A; Energy Transfer; Escherichia coli; Light-Harvesting Protein Complexes; Lutein; Photochemistry; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Pigments, Biological; Recombinant Proteins; Spectrometry, Fluorescence; Spinacia oleracea; Xanthophylls; Zeaxanthins

Chlorophyll and glycoalkaloids are synthesized in different parts of the potato plant including leaves, tubers, and sprouts. Although light stimulates the biosynthesis of both constituents, the question of whether the two biosynthetic pathways are under the same genetic control has not been resolved. This study investigated the dynamics of incorporation of labeled [2-(14)C]-DL-mavalonate into chlorophyll a, chlorophyll b, and the glycoalkaloids alpha-chaconine and alpha-solanine in potato sprouts after 7 and 14 days of storage in the light and in the dark. No chlorophyll synthesis occurred in the dark. Fractionation of the "glycoalkaloid" extract followed by high-performance liquid chromatography produced four peaks. The fractions were collected and analyzed for radioactivity. About 80% of the radioactivity resided in fraction 1, the composition of which is unknown. Two of the fractions, with 1-14% of the original label, were alpha-chaconine and alpha-solanine. The radioactivity derived from mevalonate largely resides in unidentified compound(s) eluting as a single peak on the HPLC column before the peaks associated with the glycoalkaloids. The specific radioactivity of alpha-chaconine and alpha-solanine increased approximately 2-fold in going from 7 to 14 days of exposure in the light and in the dark. These and additional observations point to the near identity of the dynamics of biosynthesis of the two glycoalkaloids. These data also implicate a non-mevalonate pathway for the synthesis of both chlorophylls and the glycoalkaloids and are consistent with independent genetic control of the concurrent formation of the two classes of compounds during greening of potatoes.

Topics: Carbon Radioisotopes; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Mevalonic Acid; Solanine; Solanum tuberosum

Evidence is presented, by means of both fluorescence and action spectroscopy, that a small, spectroscopically heterogeneous population of both Chl a and Chl b molecules is present in isolated spinach thylakoids and is active in photoinhibition. The broadness of the action spectrum suggests that degraded or incompletely assembled pigment-protein complexes may be involved.

Topics: Chlorophyll; Chlorophyll A; Light; Light-Harvesting Protein Complexes; Mass Spectrometry; Photosynthetic Reaction Center Complex Proteins; Plant Leaves; Spectrometry, Fluorescence; Spinacia oleracea; Thylakoids

Resonance Raman spectra of the native Lhcb4 antenna protein are compared with those of a recombinant protein prepared by in vitro refolding of its polypeptide, over-expressed in Escherichia coli, with added pigments [Giuffra et al. (1996) Eur. J. Biochem. 238, 112-120]. The results indicate that the native pigment conformation is reproduced almost perfectly in the reconstituted protein, with only small differences which are attributed to a slight shift in the Soret absorption peak of two or more chlorophylls. This procedure therefore represents a model system for the investigation of site-directed mutant LHC proteins, which are otherwise very difficult to obtain.

Topics: Carotenoids; Carrier Proteins; Chlorophyll; Chlorophyll A; Mutagenesis, Site-Directed; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plant Proteins; Recombinant Proteins; Spectrum Analysis, Raman; Zea mays

Topics: Bacteria; Chlorophyll; Chlorophyll A; Chloroplasts; Photosynthesis; Plants; Proteins; Thylakoids

Empetrum nigrum L. (crowberry) is one of the plants surviving near the Cu-Ni smelters in Finland and Russia. According to field observations, the fine roots of E. nigrum are situated below 40 cm depth and the root biomass is reduced in the polluted sites. This could cause a reduced root uptake of macronutrients and trace elements in the field and, therefore, the possible element uptake by aboveground parts of E. nigrum was studied in a greenhouse. Six different treatment solutions containing various heavy metal and macronutrient concentrations were applied to the stems and leaves of E. nigrum and the chemical composition and ecophysiological parameters were measured. Heavy metal concentrations in the leaves and stem bark, and Cu concentrations in the stems, increased with increasing metal concentrations in the spraying solutions. The bark and leaves had higher heavy metal concentrations than the stems of comparable age classes. The macronutrient and Mn concentrations in E. nigrum did not change significantly with increasing element concentrations in the spraying solution. Neither the stem water potential nor the leaf chlorophyll concentrations showed any clear response to element applications. Therefore, the element uptake by aboveground parts of E. nigrum was not confirmed by this study. However, there was a tendency to a decrease in CO2 exchange rate and increase in foliar abscisic acid content in plants treated with the highest element concentrations.

Topics: Abscisic Acid; Carbon Dioxide; Chlorophyll; Chlorophyll A; Copper; Finland; Metallurgy; Nickel; Photosynthesis; Plant Leaves; Plant Physiological Phenomena; Plant Stems; Plants; Soil; Statistics, Nonparametric; Water

The major light-harvesting chlorophyll a/b complex (LHCIIb) of photosystem II in higher plants can be reconstituted with pigments in lipid-detergent micelles. The pigment-protein complexes formed are functional in that they perform efficient internal energy transfer from chlorophyll b to chlorophyll a. LHCIIb formation in vitro, can be monitored by the appearance of energy transfer from chlorophyll b to chlorophyll a in time-resolved fluorescence measurements. LHCIIb is found to form in two apparent kinetic steps with time constants of about 30 and 200 seconds. Here we report on the dependence of the LHCIIb formation kinetics on the composition of the pigment mixture used in the reconstitution. Both kinetic steps slow down when the concentration of either chlorophylls or carotenoids is reduced. This suggests that the slower 200 seconds formation of functional LHCIIb still includes binding of both chlorophylls and carotenoids. LHCIIb formation is accelerated when the chlorophylls in the reconstitution mixture consist predominantly of chlorophyll a although the complexes formed are thermally less stable than those reconstituted with a chlorophyll a:b ratio < or = 1. This indicates that although chlorophyll a binding is more dominant in the observed rate of LHCIIb formation, the occupation of (some) chlorophyll binding sites with chlorophyll b is essential for complex stability. The accelerating effect of various carotenoids (lutein, zeaxanthin, violaxanthin, neoxanthin) on LHCIIb formation correlates with their affinity to two lutein-specific binding sites. We conclude that the occupation of these two carotenoid binding sites but not of the third (neoxanthin-specific) binding site is an essential step in the assembly of LHCIIb in vitro.

Topics: Binding Sites; Carotenoids; Chlorophyll; Chlorophyll A; Fluorometry; Kinetics; Light-Harvesting Protein Complexes; Lutein; Micelles; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Pisum sativum; Plant Leaves; Protein Binding; Protein Folding; Thermodynamics

Chlorophyll (Chl) molecules attached to plant light-harvesting complexes (LHC) differ in their spectral behavior. While most Chl a and Chl b molecules give rise to absorption bands between 645 nm and 670 nm, some special Chls absorb at wavelengths longer than 700 nm. Among the Chl a/b-antennae of higher plants these are found exclusively in LHC I. In order to assign this special spectral property to one chlorophyll species we reconstituted LHC of both photosystem I (Lhca4) and photosystem II (Lhcb1) with carotenoids and only Chl a or Chl b and analyzed the effect on pigment binding, absorption and fluorescence properties. In both LHCs the Chl-binding sites of the omitted Chl species were occupied by the other species resulting in a constant total number of Chls in these complexes. 77-K spectroscopic measurements demonstrated that omission of Chl b in refolded Lhca4 resulted in a loss of long-wavelength absorption and 730-nm fluorescence emission. In Lhcb1 with only Chl b long-wavelength emission was preserved. These results clearly demonstrate the involvement of Chl b in establishing long-wavelength properties.

Topics: Absorption; Binding Sites; Centrifugation, Density Gradient; Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Pigments, Biological; Plant Leaves; Protein Binding; Solanum lycopersicum; Spectrometry, Fluorescence; Spinacia oleracea; Temperature

The light-harvesting efficiency of a photosystem is thought to be largely dependent on its photosynthetic antenna size. It has been suggested that antenna size is controlled by the biosynthesis of chlorophyll b. To verify this hypothesis, we overexpressed the enzyme for chlorophyll b biosynthesis, chlorophyllide a oxygenase (CAO), in Arabidopsis thaliana by transforming the plant with cDNA for CAO under the control of the 35S cauliflower mosaic virus promoter. In the early de-etiolation phase, when the intrinsic CAO expression is very low, the chlorophyll a: b ratio was drastically decreased from 28 to 7.3, indicating that enhancement of chlorophyll b biosynthesis had been successfully achieved. We made the following observations in full-green rosette leaves of transgenic plants. (1) The chlorophyll a : b ratio was reduced from 2.85 to 2.65. (2) The ratio of the peripheral light-harvesting complexes (LHCII) to the core antenna complex (CPa) resolved with the green-gel system increased by 20%. (3) The ratio of the light-harvesting complex II apoproteins (LHCP) to 47-kDa chlorophyll a protein (CP47), which was estimated by the results of immunoblotting, increased by 40%. These results indicated that the antenna size increased by at least 10-20% in transgenic plants, suggesting that chlorophyll b biosynthesis controls antenna size. To the best of our knowledge, this is the first report on enlargement of the antenna size by genetic manipulations.

Topics: Apoproteins; Arabidopsis; Chlorophyll; Chlorophyll A; Light; Light-Harvesting Protein Complexes; Oxygenases; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plant Leaves; Plant Proteins; Plants, Genetically Modified; Recombinant Proteins; Thylakoids

Photoinduced electron-transfer processes in the systems of chlorophylls (Chl) (chlorophyll-a [Chl-a] and chlorophyll-b) and fullerenes (C60/C70) in both polar and non-polar solvents have been investigated with nanosecond laser photolysis technique, observing the transient spectra in the visible/near-IR regions. By the excitation of Chl in benzonitrile (BN) it has been proved that electron transfer takes place from the triplet excited states of Chl to the ground states of C60/C70. By the excitation of C70 in BN electron transfer takes place from the ground states of Chl to the triplet excited state of C70. In both Chl the rate constants and quantum yields for the electron-transfer processes are as high as those of zinc porphyrins and zinc phthalocyanines, indicating that the long alkyl chains of Chl play no role in retarding the electron transfer. The rate constant for the electron-mediating process from the radical anion of C70 to octylviologen dication yielding the octylviologen radical cation was evaluated. The back electron-transfer process from the viologen radical cation to the radical cation of Chl-a takes place in a longer time-scale, indicating that a photosensitized electron-transfer/electron-mediating cycle is achieved.

Topics: Carbon; Chlorophyll; Chlorophyll A; Electron Transport; Fullerenes; Lasers; Photochemistry; Photolysis; Photosensitizing Agents

Directional reflectance (R) spectra from 380 to 780 nm for nadir illuminated leaves of four different plants (croton, Codiaeum variegatum; spotted eleagnus, Eleagnus pungens Maculata; Japanese pittosporum, Pittosporum tobira and Benjamin fig, Ficus benjamina Starlight) were acquired at a viewing angle of 30 degrees from the nadir direction. Chlorophyll-a and -b content of leaves covered a range of 1-60 and 0.5-21 microg/cm(2), respectively. In contrast with previous results from hemispherical reflectance measurements, directional reflectance data does not correlate well with chlorophyll concentration. This is mainly due to the external reflectance (R(E)) at the leaf epidermis, caused by the mismatch of the refractive index at the air-epidermis and epidermis-inner layer boundary. The external reflectance can be identified with the blue flat reflectance between 380 and 480 nm. The inner reflectance (R(I)), obtained by subtracting the external reflectance from the measured spectra, was found to be linearly related to the logarithm of the chlorophyll content. Good fitting of the log (Chl) versus R(I)(lambda) curves were obtained for R(I) in the green band (around 550 nm) and close to the inflection point in the red edge (around 700 nm). The coefficient of determination, r(2), of curve fitting improved (up to 0.97) when the normalised inner reflectance NR(I)(lambda)=R(I)(lambda)/R(I)(lambda(0)), with lambda(0)>or=750 nm, was used instead of the absolute reflectance. The best indices for Chl, Chl-a and Chl-b determination were R(I)(542)/R(I)(750), R(I)(706)/R(I)(750) and R(I)(556)/R(I)(750), respectively. However, since the content of Chl-a relative to Chl-b was almost constant for the plants investigated, the two last indices must be further validated on leaves with a high variability in the Chl-a:Chl-b ratio. The error in the determination of chlorophyll content was found to be of the order of 10%. This value was lower than those obtained by applying the vegetation indices previously suggested. Therefore, the normalised inner reflectance in the green and in the red edge represents a more suitable index for the chlorophyll determination than those up to now used.

Topics: Chlorophyll; Chlorophyll A; Croton; Ficus; Magnoliopsida; Mass Spectrometry; Plant Leaves

The Lhcb gene family in green plants encodes several light-harvesting Chl a/b-binding (LHC) proteins that collect and transfer light energy to the reaction centers of PSII. We comprehensively characterized the Lhcb gene family in the unicellular green alga, Chlamydomonas reinhardtii, using the expressed sequence tag (EST) databases. A total of 699 among over 15,000 ESTs related to the Lhcb genes were assigned to eight, including four new, genes that we isolated and sequenced here. A sequence comparison revealed that six of the Lhcb genes from C. reinhardtii correspond to the major LHC (LHCII) proteins from higher plants, and that the other two genes (Lhcb4 and Lhcb5) correspond to the minor LHC proteins (CP29 and CP26). No ESTs corresponding to another minor LHC protein (CP24) were found. The six LHCII proteins in C. reinhardtii cannot be assigned to any of the three types proposed for higher plants (Lhcb1-Lhcb3), but were classified as follows: Type I is encoded by LhcII-1.1, LhcII-1.2 and LhcII-1.3, and Types II, III and IV are encoded by LhcII-2, LhcII-3 and LhcII-4, respectively. These findings suggest that the ancestral LHC protein diverged into LHCII, CP29 and CP26 before, and that LHCII diverged into multiple types after the phylogenetic separation of green algae and higher plants.

Topics: Algal Proteins; Amino Acid Sequence; Animals; Biological Evolution; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Chlorophyll Binding Proteins; Cloning, Molecular; DNA, Complementary; Expressed Sequence Tags; Light-Harvesting Protein Complexes; Molecular Sequence Data; Multigene Family; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Phylogeny; Plant Proteins; RNA, Messenger; Sequence Homology, Amino Acid; Solanum lycopersicum

Chlorophyll and its derivatives are examples of plant compounds (purified and/or extracted) which appear to protect DNA from damage caused by chemical or physical agents, although some studies have identified clastogenic activity of these compounds. This study was carried out to assess the genotoxic activity of chlorophyll-a (Chl-a), -b (Chl-b) and chlorophyllin (Chl) and their antigenotoxic activity against the DNA damage induced by methyl methanesulphonate (MMS) under conditions of simultaneous, pre-, post-treatment, and simultaneous treatment after pre-incubation of the chemical with MMS. The micronucleus (MN) test was used in binucleated cells (induced by cytochalasin-B) of a mammalian cell line (V79). The three concentrations of Chl-a, Chl-b or Chl (0.1375, 0.275, 0.55microM) were not genotoxic and the genotoxic action of MMS (400microM) decreased (74-117%) under all treatment conditions. The results showed that there was no significant difference among the treatment types, the concentration or the nature of chlorophyll used. The data obtained suggest that Chl-a, Chl-b and Chl when associated with the DNA damaging agent, MMS, may protect the DNA by desgenotoxic action and/or by bio-antigenotoxic mechanisms, with the similar efficiency.

Topics: Animals; Antimutagenic Agents; Cell Line; Chlorophyll; Chlorophyll A; Chlorophyllides; Cricetinae; DNA Damage; Methyl Methanesulfonate; Micronucleus Tests; Mutagens

Photochemical characteristics and lipid composition of thylakoid membranes from 12 day-old pea leaves that were exposed to slow clino-rotation were examined and compared with a vertical control. Proton permeability of thylakoid membranes was estimated from light-induced proton uptake (delta H+) and post-illumination proton efflux in chloroplast suspensions. The delta pH magnitude was calculated from the level of light-induced quenching of 9-aminoacridine fluorescence. Proton permeability of thylakoid membranes increased during exposure to clino-rotation. When subsequently transferred to darkness, proton efflux increased almost 2-fold in clinorotated leaves. The results were compared with data on pigment and polar lipid composition of photosynthetic membranes in clino-rotated and control plants. It was concluded that both the increase of proton permeability and the decrease of polar lipid content in chloroplasts were induced by clino-rotation.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Chloroplasts; Glycolipids; Gravitation; Hydrogen-Ion Concentration; Light; Lipid Bilayers; Lipid Metabolism; Phospholipids; Pisum sativum; Proton Pumps; Rotation; Thylakoids; Weightlessness Simulation

The capacity for photosynthetic acclimation in Arabidopsis thaliana (L.) Heynh. cv. Landsberg erecta was assessed during growth over a broad range of irradiance. Discontinuities in the response to growth irradiance were revealed for the light- and CO2-saturated rate of photosynthesis (Pmax) and the ratio of chlorophyll a to chlorophyll b (Chl a/b). Three separate phases in the response of Pmax and Chl a/b to growth light were evident, with increases at low and high irradiance ranges and a plateau at intermediate irradiance. By measuring all chlorophyll-containing components of the thylakoid membrane that contribute to Chl a/b we reveal that distinct strategies for growth at low and high irradiance underlie the discontinuous response. These strategies include, in addition to changes in the major light-harvesting complexes of photosystem II (LHCII), large shifts in the amounts of both reaction centres as well as significant changes in the levels of minor LHCII and LHCI components.

Topics: Acclimatization; Arabidopsis; Carbon Dioxide; Chlorophyll; Chlorophyll A; Light; Light-Harvesting Protein Complexes; Oxygen Consumption; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Ribulose-Bisphosphate Carboxylase

An Arabidopsis thaliana chlorophyll(ide) a oxygenase gene (cao), which is responsible for chlorophyll b synthesis from chlorophyll a, was introduced and expressed in a photosystem I-less strain of the cyanobacterium Synechocystis sp. PCC 6803. In this strain, most chlorophyll is associated with the photosystem II complex. In line with observations by Satoh et al. [Satoh, S., Ikeuchi, M., Mimuro, M. & Tanaka, A. (2001) J. Biol. Chem. 276, 4293-4297], chlorophyll b was made but accounted for less than 10% of total chlorophyll. However, when lhcb encoding light-harvesting complex (LHC)II from pea was present in the same strain (lhcb(+)/cao(+)), chlorophyll b accumulated in the cell to levels exceeding those of chlorophyll a, although LHCII did not accumulate. In the lhcb(+)/cao(+) strain, the total amount of chlorophyll, the number of chlorophylls per photosystem II center, and the oxygen-evolving activity on a per-chlorophyll basis were similar to those in the photosystem I-less strain. Furthermore, the chlorophyll a/b ratio of photosystem II core particles (retaining CP47 and CP43) and of whole cells of the lhcb(+)/cao(+) strain was essentially identical, and PS II activity could be obtained efficiently by chlorophyll b excitation. These data indicate that chlorophyll b functionally substitutes for chlorophyll a in photosystem II. Therefore, the availability of chlorophylls, rather than their binding specificity, may determine which chlorophyll is incorporated at many positions of photosystem II. We propose that the transient presence of a LHCII/chlorophyll(ide) a oxygenase complex in the lhcb(+)/cao(+) strain leads to a high abundance of available chlorophyll b that is subsequently incorporated into photosystem II complexes. The apparent LHCII requirement for high chlorophyll(ide) a oxygenase activity may be instrumental to limit the occurrence of chlorophyll b in plants to LHC proteins.

Topics: Arabidopsis; Chlorophyll; Chlorophyll A; Cyanobacteria; Genes, Plant; Light-Harvesting Protein Complexes; Oxygenases; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Pigments, Biological

Light-harvesting complexes (Lhc) catalyse sunlight harvesting for photosynthesis as well as other essential functions, including photoprotection by quenching of harmful chlorophyll triplet states and prevention of photoinhibition by dissipation of excitation energy in excess. In addition, folding of Lhc proteins depends on the availability of both xanthophylls and carotenoids, thus preventing the potential formation of harmful chlorophyll-protein complexes lacking photoprotectors. We have used the mutation analysis in order to study the association of the different functions to three protein domains, each composed of a xanthophyll molecule and of neighbour chlorophylls a and b, within the major antenna complex of photosystem II, i.e. LHCII. We have found that the xanthophyll to chlorophyll energy transfer is a shared property of the whole pigment-protein complex, and occurs with similar efficiency in each of the three structural domains. Photoprotection by quenching of chlorophyll triplets is catalysed mainly by lutein bound to site L1, and occurs via energy transfer from chlorophylls A1 and B1. This domain is essential for pigment-induced protein folding. The domains L2 and N1 weakly influence either the protein stability or the photoprotection; however, replacement of xanthophyll species bound to these structural domains modulates the fluorescence quantum yield of LHCII, and suggests that non-radiative dissipation of excess energy can be regulated through allosteric modification of the protein structure by exchanging xanthophylls in these sites.

Topics: Binding Sites; Chlorophyll; Chlorophyll A; Energy Transfer; Fluorescence; Light; Light-Harvesting Protein Complexes; Models, Molecular; Mutation; Photochemistry; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Protein Folding; Protein Structure, Tertiary; Structure-Activity Relationship; Thermodynamics; Xanthophylls; Zea mays

Antioxidant activity of green tea extract or tea-derived polyphenols has been extensively studied. However, antioxidant activity in the non-polyphenolic fraction of green tea has been poorly analyzed. In the present study, we analyzed the antioxidant activity of the non-polyphenolic fraction of the residual green tea (Camellia sinensis) after hot water extraction using the aluminum chloride method. The non-polyphenolic fraction of residual green tea caused a significant suppression against hydroperoxide generation from oxidized linoleic acid in a dose-dependent manner. When the concentrate of the non-polyphenolic fraction was applied to a silica gel TLC plate and developed, six color spots were observed, which were considered to be chlorophylls a and b, pheophytins a and b, carotenoids, such as beta-carotene and lutein according to their specific colors, Rf values of silica gel TLC and spectrophotometric properties. Among these pigments, pheophytins a and b showed relatively abundant amounts, and the second major group of the pigment was chlorophylls a and b, and carotenoids such as beta-carotene and lutein indicated lower concentrations. Although all these pigments exhibited significant antioxidant activities, the ranks of suppressive activity against hydroperoxide generation were chlorophyll a > lutein > pheophytin a > chlorophyll b > beta-carotene > pheophytin b. These results suggest that the non-polyphenolic fraction of residual green tea has a potent suppressive activity against hydroperoxide generation from oxidized linoleic acid, which is derived from the antioxidant activities of chlorophylls a and b, pheophytins a and b, beta-carotene and lutein. This finding also implies that the combined intake of polyphenols in water-soluble fraction and antioxidative pigments in the non-polyphenolic fraction of green tea will be more efficient to prevent life style-related chronic diseases.

Topics: Acetone; Antioxidants; beta Carotene; Camellia sinensis; Chlorophyll; Chlorophyll A; Dose-Response Relationship, Drug; Hot Temperature; Lipid Peroxides; Lutein; Pheophytins; Pigments, Biological; Tea; Water

Effect of different concentrations of CdSO4 (10(-4) M, 5 x 10(-4) M and 10(-3) M) on leaf area, stomatal frequency, stomatal index and pigment content in 6 days old mungbean seedlings was studied under laboratory conditions. Except at 10(-4) M CdSO4, leaf area decreased with an increase in the concentration of CdSO4. Stomatal frequency on the leaf surface increased at 5 x 10(-4) M and 10(-3) M CdSO4 in comparison to untreated control. However, reverse was the case for stomatal index. Chlorophyll-a, chlorophyll-b, total chlorophyll, and chlorophyll a/b ratio were increased at 10(-4) M CdSO4, and decreased thereafter with an increase in the concentration of Cd2+. Possible reasons for the stimulatory effect at low concentrations and inhibitory effect with higher concentrations are discussed.

Topics: Cadmium; Chlorophyll; Chlorophyll A; Dose-Response Relationship, Drug; Fabaceae; Plant Leaves

The major light-harvesting complex of photosystem II can be reconstituted in vitro from its bacterially expressed apoprotein with chlorophylls a and b and neoxanthin, violaxanthin, lutein, or zeaxanthin as the only xanthophyll. Reconstitution of these one-carotenoid complexes requires low-stringency conditions during complex formation and isolation. Neoxanthin complexes (containing 30-50% of the all-trans isomer) disintegrate during electrophoresis, exhibit a largely reduced resistance against proteolytic attack; in addition, energy transfer from Chl b to Chl a is easily disrupted at elevated temperature. Complexes reconstituted in the presence of either zeaxanthin or lutein contain nearly two xanthophylls per 12 chlorophylls and are more resistant against trypsin. Lutein-LHCIIb also exhibits an intermediate maintenance of energy transfer at higher temperature. Violaxanthin complexes approach a xanthophyll/12 chlorophyll ratio of 3, similar to the ratio in recombinant LHCIIb containing all xanthophylls. On the other hand, violaxanthin-LHCIIb exhibits a low thermal stability like neoxanthin complexes, but an intermediate accessibility towards trypsin, similar to lutein-LHCIIb and zeaxanthin-LHCIIb. Binary competition experiments were performed with two xanthophylls at varying ratios in the reconstitution. Analysis of the xanthophyll contents in the reconstitution products yielded information about relative carotenoid affinities of three assumed binding sites. In lutein/neoxanthin competition experiments, two binding sites showed a strong preference (> 200-fold) for lutein, whereas the third binding site had a higher affinity (25-fold) to neoxanthin. Competition between lutein and violaxanthin gave a similar result, although the specificities were lower: two binding sites have a 36-fold preference for lutein and one has a fivefold preference for violaxanthin. The lowest selectivity was between lutein and zeaxanthin: two binding sites had a fivefold higher affinity for lutein and one has a threefold higher affinity to zeaxanthin.

Topics: Apoproteins; beta Carotene; Binding Sites; Binding, Competitive; Carotenoids; Chlorophyll; Chlorophyll A; Electrophoresis, Polyacrylamide Gel; Energy Transfer; Light-Harvesting Protein Complexes; Lutein; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Pigments, Biological; Plant Proteins; Plants; Protein Precursors; Substrate Specificity; Trypsin; Xanthophylls; Zeaxanthins

In order to obtain information on the organization of the pigment molecules in chlorophyll (Chl) a/b/c-containing organisms, we have carried out circular dichroism (CD), linear dichroism (LD) and absorption spectroscopic measurements on intact cells, isolated thylakoids and purified light-harvesting complexes (LHCs) of the prasinophycean alga Mantoniella squamata. The CD spectra of the intact cells and isolated thylakoids were predominated by the excitonic bands of the Chl a/b/c LHC. However, some anomalous bands indicated the existence of chiral macrodomains, which could be correlated with the multilayered membrane system in the intact cells. In the red, the thylakoid membranes and the LHC exhibited a well-discernible CD band originating from Chl c, but otherwise the CD spectra were similar to that of non-aggregated LHC II, the main Chl a/b LHC in higher plants. In the Soret region, however, an unusually intense (+) 441 nm band was observed, which was accompanied by negative bands between 465 and 510 nm. It is proposed that these bands originate from intense excitonic interactions between Chl a and carotenoid molecules. LD measurements revealed that the Q(Y) dipoles of Chl a in Mantoniella thylakoids are preferentially oriented in the plane of the membrane, with orientation angles tilting out more at shorter than at longer wavelengths (9 degrees at 677 nm, 20 degrees at 670 nm and 26 degrees at 662 nm); the Q(Y) dipole of Chl c was found to be oriented at 29 degrees with respect to the membrane plane. These data and the LD spectrum of the LHC, apart from the presence of Chl c, suggest an orientation pattern of dipoles similar to those of higher plant thylakoids and LHC II. However, the tendency of the Q(Y) dipoles of Chl b to lie preferentially in the plane of the membrane (23 degrees at 653 nm and 30 degrees at 646 nm) is markedly different from the orientation pattern in higher plant membranes and LHC II. Hence, our CD and LD data show that the molecular organization of the Chl a/b/c LHC, despite evident similarities, differs significantly from that of LHC II.

Topics: Bacterial Proteins; Chlorophyll; Chlorophyll A; Circular Dichroism; Eukaryota; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Pigments, Biological; Spectrum Analysis; Thylakoids

Plastids of Cuscuta reflexa Roxb., C. subinclusa D. et H., C. gronovii Willd. and C. campestris Yunck. possess thylakoids and contain both chlorophyll a and b in a ratio similar to that of stem tissue of the systematically closely related but 'normal' green Ipomoea tricolor. In contrast, plastids of C. odorata R. et P. and C. grandiflora H.B.K. do not contain any chlorophyll or possess thylakoids. Light-driven electron transport, as measured by oxygen evolution and indicated by analysis of chlorophyll fluorescence, was present in all chlorophyll-containing species. The photosystem II efficiency was low and ranged from 0.511 to 0.687. The plastid rbcL gene could not be detected in C. odorata, but was present in all other tested species. Neither rbcL transcripts nor the large subunit of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) could be detected in C. odorata and C. grandiflora. Low amounts of the large subunit of Rubisco were detected immunologically in all other Cuscuta species. Apparently, the genus Cuscuta comprises species with different degrees of plastid functionality, ranging from intact chloroplasts, via plastids with impaired protein production and gene expression to plastids with reduced plastome gene content.

Topics: Blotting, Northern; Blotting, Southern; Blotting, Western; Carotenoids; Chlorophyll; Chlorophyll A; DNA, Plant; Light-Harvesting Protein Complexes; Magnoliopsida; Oxygen; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plant Proteins; Plastids; Ribulose-Bisphosphate Carboxylase; RNA, Plant; Species Specificity

PSI-K is a subunit of photosystem I. The function of PSI-K was characterized in Arabidopsis plants transformed with a psaK cDNA in antisense orientation, and several lines without detectable PSI-K protein were identified. Plants without PSI-K have a 19% higher chlorophyll a/b ratio and 19% more P700 than wild-type plants. Thus, plants without PSI-K compensate by making more photosystem I. The photosystem I electron transport in vitro is unaffected in the absence of PSI-K. Light response curves for oxygen evolution indicated that the photosynthetic machinery of PSI-K-deficient plants have less capacity to utilize light energy. Plants without PSI-K have less state 1-state 2 transition. Thus, the redistribution of absorbed excitation energy between the two photosystems is reduced. Low temperature fluorescence emission spectra revealed a 2-nm blue shift in the long wavelength emission in plants lacking PSI-K. Furthermore, thylakoids and isolated PSI without PSI-K had 20-30% less Lhca2 and 30-40% less Lhca3, whereas Lhca1 and Lhca4 were unaffected. During electrophoresis under mildly denaturing conditions, all four Lhca subunits were partially dissociated from photosystem I lacking PSI-K. The observed effects demonstrate that PSI-K has a role in organizing the peripheral light-harvesting complexes on the core antenna of photosystem I.

Topics: Arabidopsis; Chlorophyll; Chlorophyll A; DNA, Antisense; Electron Transport; Kinetics; Light; Light-Harvesting Protein Complexes; Oxygen; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Plant Leaves; Plant Proteins; Thylakoids

A novel type of degradation of photosystem I peripheral antenna complexes has been observed in rice leaves under darkness in the present study. Photosynthesis, chlorophyll content, the chlorophyll a/b ratio, and relative amounts of ribulose-1,5-bisphosphate carboxylase/oxygenase decrease during dark treatment. The levels of photosystem II reaction-center complex and cytochrome f on the basis of units of chlorophyll also decline rapidly under darkness. In contrast, the levels of photosystem I reaction-center complex remain stable under darkness for six days. Low-temperature fluorescence emission spectra ascribed to photosystem I antennae clearly show a blue shift. A similar shift is also observed in the photosystem I complexes resolved with dodecyl maltoside-polyacrylamide gel electrophoresis. Moreover, polypeptide analysis of the thylakoids and photosystem I complexes isolated from the green gels shows that some polypeptides originating from photosystem I peripheral antenna complexes disappear during the dark treatment. A curve-fitting method also displays remarkable changes in the chlorophyll components between the light and dark treatments. It is likely that these results indicate the disconnection/disassembly of the photosystem I antenna as well as the photosystem II complexes induced by dark treatment. Moreover, these findings also imply the existence of different degradation mechanisms for the photosystem I and II complexes.

Topics: Chlorophyll; Chlorophyll A; Darkness; Electron Transport; Kinetics; Light; Light-Harvesting Protein Complexes; Oryza; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Ribulose-Bisphosphate Carboxylase; Time Factors

Three forms of light-harvesting chlorophyll a/b-protein complexes of photosystem II (LHC II) were isolated from the thylakoid membranes of Dunaliella salina grown under different irradiance conditions. Cells grown under a low intensity light condition (80 micromol quanta m(-2) s(-1)) contained one form of LHC II, LHC-L. Two other forms of LHC II, LHC-H1 and LHC-H2, were separated from the cells grown under a high intensity light condition (1,500 micromol quanta m(-2) s(-1)). LHC-L and LHC-H1 showed an apparent particle size of 310 kDa and contained four polypeptides of 31, 30, 29 and 28 kDa. LHC-H2, with a particle size of 110 kDa, consisted of 30 and 28 kDa polypeptides. LHC-L contained 7.5 molecules of Chl a, 3.2 of Chl b and 2.1 of lutein per polypeptide, analogous to the content in higher plants. LHC-H1, with 5.6 molecules of Chl a, 2.5 of Chl b and 1.8 of lutein per polypeptide was similar to that in the green alga Bryopsis maxima. LHC-L and LHC-H1 maintained high efficiency energy transfer from Chl b and lutein to Chl a molecules. LHC-H2 showed a high Chl a/b ratio of 7.5 and contained 3.4 molecules of Chl a, 0.5 of Chl b and 1.4 of lutein per polypeptide. Chl b and lutein could not completely transfer the excitation energy to Chl a in LHC-H2.

Topics: Chlorophyll; Chlorophyll A; Chlorophyta; Chromatography, Gel; Electrophoresis, Polyacrylamide Gel; Light; Lighting; Molecular Weight; Photosynthetic Reaction Center Complex Proteins; Spectrometry, Fluorescence; Spectrophotometry

NADP-dependent malic enzyme (NADP-ME) is a major decarboxylating enzyme in NADP-ME-type C4 species such as maize and Flaveria. In this study, chloroplastic NADP-ME was transferred to rice (Oryza sativa L.) using a chimeric gene composed of maize NADP-ME cDNA under the control of rice light-harvesting chlorophyll-a/b-binding protein (Cab) promoter. There was a 20- to 70-fold increase in the NADP-ME activity in leaves of transgenic rice compared to that in wild-type rice plants. Immunocytochemical studies by electron microscopy showed that maize NADP-ME was mostly localized in chloroplasts in transgenic rice plants, and that the chloroplasts were agranal without thylakoid stacking. Chlorophyll content and photosystem II activity were inversely correlated with the level of NADP-ME activity. These results suggest that aberrant chloroplasts in transgenic plants may be caused by excessive NADP-ME activity. Based on these results and the known fact that only bundle sheath cells of NADP-ME species, among all three C4 subgroups, have agranal chloroplasts, we postulate that a high level of chloroplastic NADP-ME activity could strongly affect the development of chloroplasts.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Light-Harvesting Protein Complexes; Malate Dehydrogenase; Oryza; Phenotype; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plant Leaves; Plants, Genetically Modified; Promoter Regions, Genetic; Recombinant Fusion Proteins; Reverse Transcriptase Polymerase Chain Reaction; Zea mays

The assembly, organization and function of the photosynthetic apparatus was investigated in the wild type and a chlorophyll (Chl) b-less mutant of the unicellular green alga Chlamydomonas reinhardtii, generated via DNA insertional mutagenesis. Comparative analyses were undertaken with cells grown photoheterotrophically (acetate), photomixotrophically (acetate and HCO3-) or photoautotrophically (HCO3-). It is shown that lack of Chl b diminished the photosystem-II (PSII) functional Chl antenna size from 320 Chl (a and b) to about 95 Chl a molecules. However, the functional Chl antenna size of PSI remained fairly constant at about 290 Chl molecules, independent of the presence of Chl b. Western blot and kinetic analyses suggested the presence of inner subunits of the Chl a-b light-harvesting complex of PSII (LHCII) and the entire complement of the Chl a-b light-harvesting complex of PSI (LHCI) in the mutant. It is concluded that Chl a can replace Chl b in the inner subunits of the LHCII and in the entire complement of the LHCI. Growth of cells on acetate as the sole carbon source imposes limitations in the photon-use efficiency and capacity of photosynthesis. These are manifested as a lower quantum yield and lower light-saturated rate of photosynthesis, and as lower variable to maximal (Fv/Fmax) chlorophyll fluorescence yield ratios. This adverse effect probably originates because acetate shifts the oxidation-reduction state of the plastoquinone pool, and also because it causes a decrease in the amount and/or activity of Rubisco in the chloroplast. Such limitations are fully alleviated upon inclusion of an inorganic carbon source (e.g. bicarbonate) in the cell growth medium. Further, the work provides evidence to show that transformation of green algae can be used as a tool by which to generate mutants exhibiting a permanently truncated Chl antenna size and a higher (per Chl) photosynthetic productivity of the cells.

Topics: Animals; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Intracellular Membranes; Light-Harvesting Protein Complexes; Mutagenesis, Insertional; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Thylakoids

The formal description of Prochlorococcus marinus Chisholm et al. 1992, 299 was based on the non-axenic nomenclatural type, strain CCMP 1375T. The purification and properties of the axenic strain PCC 9511, derived from the same primary culture (SARG) as the type species, are reported here. Prochlorococcus PCC 9511 differs from the latter in possessing horseshoe-shaped thylakoids, exhibiting a low chlorophyll b2 content and lacking phycoerythrin, but shares these phenotypic properties with Prochlorococcus strain CCMP 1378. This relationship was confirmed by 16S rRNA sequence analyses, which clearly demonstrated that the axenic isolate is not co-identic with the nomenclatural type. Strain PCC 9511 has a low mean DNA base composition (32 mol% G+C) and harbours the smallest genome of all known oxyphotobacteria (genome complexity 1.3 GDa = 2 Mbp). Urea and ammonia are the preferred sources of nitrogen for growth, whereas nitrate is not utilized. Several different organic phosphorus compounds efficiently replace phosphate in the culture medium, indicative of ecto-phosphohydrolase activity. In order to distinguish strain PCC 9511 from the nomenclatural type, a new subspecies is proposed, Prochlorococcus marinus Chisholm et al. 1992 subsp. pastoris subsp. nov.

Topics: Bacterial Typing Techniques; Base Composition; Carotenoids; Chlorophyll; Chlorophyll A; Culture Media; Cyanobacteria; DNA, Bacterial; DNA, Ribosomal; Molecular Sequence Data; Phycoerythrin; Phylogeny; Pigments, Biological; RNA, Ribosomal, 16S; Seawater; Sequence Analysis, DNA; Spectrophotometry

Analysis of reflectance can be used to estimate foliar concentrations of photosynthetic pigments, thus providing information on the physiological status of green plants. We compared several methods of reflectance analysis for the capacity to differentiate among effects of fertilization treatments across different irradiances on seedlings of Engelmann spruce (Picea engelmanii Parry ex Engelm.). Seedlings were grown in two light treatments (0 and 60% shade) and three nitrogen (N) treatments (10, 25 and 100 mg N l-1) for one growing season, after which foliar reflectance of the needles was measured. Five indices were tested: R550 (% reflectance at 550 nm); red edge position; the ratio R698:R760; the structure independent pigment index (SIPI); and the photochemical reflectance index (PRI). Both the light and nutrient treatments significantly affected foliar chlorophyll a and b and carotenoid concentrations. Among the indices tested, R550, red edge position and R698:R760 ratio were related to chlorophyll concentration, and were significantly affected by both light and N treatments. Both SIPI and PRI were related to chlorophyll and carotenoid concentrations. Among these relationships, PRI was affected by both treatments, whereas SIPI was sensitive to N treatment but not to light treatment. All five indices were weakly but significantly correlated with growth as measured by dry weight.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Light; Nitrogen; Plant Leaves; Trees

The spectral characteristics of chlorophyll fluorescence and absorption during linear heating of barley leaves within the range 25-75 degreesC (fluorescence temperature curve, FTC) were studied. Leaves with various content of light harvesting complexes (green, Chl b-less chlorina f2 and intermittent light grown) revealing different types of FTC were used. Differential absorption, emission and excitation spectra documented four characteristic phases of the FTC. The initial two FTC phases (a rise in the 46-49 degreesC region and a subsequent decrease to about 55 degreesC) mostly reflected changes in the fluorescence quantum yield peaking at about 685 nm. A steep second fluorescence rise at 55-61 degreesC was found to originate from a short-wavelength Chl a spectral form (emission maximum at 675 nm) causing a gradual blue shift of the emission spectra. In this temperature range, a clear correspondence of the blue shift in the emission and absorption spectra was found. We suggest that the second fluorescence rise in FTC reflects a weakening of the Chl a-protein interaction in the thylakoid membrane.

Topics: Chlorophyll; Chlorophyll A; Fluorescence; Heating; Hordeum; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Plant Leaves; Spectrometry, Fluorescence

Photosynthesis, the conversion of light energy into chemical energy, is a critical biological process, whereby plants synthesize carbohydrates from light, carbon dioxide (CO2) and water. The influence of gravity on this biological process, however, is not well understood. Thus, centrifugation was used to alter the gravity environment of Euglena gracilis grown on nutritive agar plates illuminated with red and blue light emitting diodes. The results showed that hypergravity (up to 10xg) had an inhibitory effect on photosynthetic CO2 fixation. Chlorophyll accumulation per cell was essentially unaffected by treatment; however, Chl a/Chl b ratios decreased in hypergravity when compared to 1xg controls. Photosynthesis in Euglena appears to have limited tolerance for even moderate changes in gravitational acceleration.

Topics: Animals; Carbon Dioxide; Centrifugation; Chlorophyll; Chlorophyll A; Euglena gracilis; Hypergravity; Photosynthesis; Pigments, Biological

The morphological differences, chlorophyll contents, fluorescence and diurnal course of leaf gas exchange between terrestrial banyan and amphibious banyan were compared with pot culture. The amphibious banyan possesses well developed aerial and hydro-adventitious roots, and wider leaf with inclination of evolution toward mesophytic traits. The chlorophyll content of terrestrial banyan was higher than that of amphibious banyan. The diurnal course of leaf gas exchange indicated that net photosynthetic rate of terrestrial banyan was slightly higher than that of amphibious banyan grown in water, but much higher than that grown in soil. The amphibious banyan grown in water had the highest transpiration rate, the terrestrial banyan had a lower one, and the amphibious banyan grown in soil had the lowest. Linear regression analysis showed a positive correlation between net photosynthetic rate and stomatal conductance, implying that the stomatal conductance was dominant factor controlling the gas exchange. In this study, the term of intrinsic water use efficiency (net photosynthetic rate/stomatal conductance ratio, Intrinsic WUE) was applied to describe the photosynthesis and water properties, and the result showed that it was a more suitable measure compared to the usual WUE(net photosynthetic rate/transpiration rate). Among the three banyan plants examined, the amphibious banyan had the highest intrinsic WUE.

Topics: Adaptation, Physiological; Carbon Dioxide; Carotenoids; Chlorophyll; Chlorophyll A; Circadian Rhythm; Ficus; Fluorescence; Oxygen; Photosynthesis

Trimeric (bT) and monomeric (bM) light-harvesting complex II (LHCII) with a chlorophyll a/b ratio of 0.03 were reconstituted from the apoprotein overexpressed in Escherichia coli. Chlorophyll/xanthophyll and chlorophyll/protein ratios of bT complexes and 'native' LHCII are rather similar, namely, 0.28 vs 0. 27 and 10.5 +/- 1.5 vs 12, respectively, indicating the replacement of most chlorophyll a molecules with chlorophyll b, leaving one chlorophyll a per trimeric complex. The LD spectrum of the bT complexes strongly suggests that the chlorophyll b molecules adopt orientations similar to those of the chlorophylls a that they replace. The circular dichroism (CD) spectra of bM and bT complexes indicate structural arrangements resembling those of 'native' LHCII. Thermolysin digestion patterns demonstrate that bT complexes are folded and organized like 'native' trimeric LHCII. Surprisingly, in the bT complexes at 77 K, half of the excitations that are created on either chlorophyll b or xanthophyll are transferred to chlorophyll a. No or very limited triplet transfer from chlorophyll b to xanthophyll appears to take place. However, the efficiency of triplet transfer from chlorophyll a to xanthophyll is close to 100%, even higher than in 'native' LHCII at 77 K. It is concluded from the triplet-minus-singlet and CD results that the single chlorophyll a molecule that on the average is present in each bT complex binds preferably next to a xanthophyll molecule at the interface between the monomers.

Topics: Absorption; Chlorophyll; Chlorophyll A; Circular Dichroism; Energy Transfer; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Pigmentation; Protein Folding; Spectrometry, Fluorescence; Spectrophotometry; Structure-Activity Relationship

This study reveals by in vivo deuterium labeling that in higher plants chlorophyll (Chl) b is converted to Chl a before degradation. For this purpose, de-greening of excised green primary leaves of barley (Hordeum vulgare) was induced by permanent darkness in the presence of heavy water (80 atom % (2)H). The resulting Chl a catabolite in the plant extract was subjected to chemical degradation by chromic acid. 3-(2-Hydroxyethyl)-4-methyl-maleimide, the key fragment that originates from the Chl catabolite, was isolated. High resolution (1)H-, (2)H-NMR and mass spectroscopy unequivocally demonstrates that a fraction of this maleimide fragment consists of a mono-deuterated methyl group. These results suggest that Chl b is converted into Chl a before degradation. Quantification proves that the initial ratio of Chl a:Chl b in the green plant is preserved to about 60-70% in the catabolite composition isolated from yellowing leaves. The incorporation of only one deuterium atom indicates the involvement of two distinguishable redox enzymes during the conversion.

Topics: Chlorophyll; Chlorophyll A; Darkness; Deuterium Oxide; Hordeum; Kinetics; Magnetic Resonance Spectroscopy; Models, Molecular; Molecular Structure; Plant Leaves; Spectrometry, Mass, Secondary Ion

Four amino acids in the major light-harvesting chlorophyll (Chl) a/b complex (LHCII) that are thought to coordinate Chl molecules have been exchanged with amino acids that presumably cannot bind Chl. Amino acids H68, Q131, Q197, and H212 are positioned in helixes B, C, A, and D, respectively, and, according to the LHCII crystal structure [Kühlbrandt, W., et al. (1994) Nature 367, 614-621], coordinate the Chl molecules named a(5), b(6), a(3), and b(3). Moreover, a double mutant was analyzed carrying exchanges at positions E65 and H68, presumably affecting Chls a(4) and a(5). All mutant proteins could be reconstituted in vitro with pigments, although the thermal stability of the resulting mutant versions of recombinant LHCII varied significantly. All complexes reconstituted with the mutant proteins contained fewer chlorophyll molecules per two lutein molecules than complexes reconstituted with the wild-type protein. However, the chlorophyll-binding amino acids could not be unambiguously assigned to binding either chlorophyll a or b, as in most cases more than one chlorophyll molecule was lost due to the mutation. The changes in Chl stoichiometries suggest that in LHCII some chlorophyll positions can be filled with either Chl a or b. Only some of the point mutations in LHCII affected the ability of the apoprotein to assemble into trimeric LHCII upon insertion into isolated thylakoid membranes. Among these were exchanges of H68 with either F or L, suggesting that the stability of the LHCII trimer significantly depends on this amino acid or the Chl molecule named a(5) that is attached to it and is located close to the center of the trimeric complex. The ion pair bridge between E65 and R185 in LHCII does not appear to be essential for the proper folding of the protein.

Topics: Amino Acid Sequence; Amino Acid Substitution; Amino Acids; Binding Sites; Carrier Proteins; Chlorophyll; Chlorophyll A; Chloroplasts; Light-Harvesting Protein Complexes; Macromolecular Substances; Molecular Sequence Data; Mutagenesis, Site-Directed; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Pisum sativum; Plant Proteins

Degradation of chlorophyll in broccoli juice occurred at temperatures exceeding 60 degrees C. Chemical analysis revealed that degradation of chlorophyll a and b to pheophytin a and b, respectively, followed first-order kinetics and that chlorophyll a was more heat sensitive than chlorophyll b. Temperature dependencies of chlorophyll a and b degradation rate constants could be described by Arrhenius equations with activation energies (E(a)) of 71.04 +/- 4.89 and 67.11 +/- 6.82 kJ/mol, respectively. Objective greenness measurements, using the -a value as the physical property, together with a fractional conversion kinetic analysis, indicated that green color degradation followed a two-step process. Kinetic parameters for the first degradation step were in accordance with the kinetic parameters for pheophytinization of the total chlorophyll content, as determined by chemical analysis (E(a) approximately 69 kJ/mol). The second degradation step, that is, the subsequent decomposition of pheophytins, was characterized by an activation energy of 105.49 +/- 4.74 kJ/mol.

Topics: Beverages; Brassica; Chlorophyll; Chlorophyll A; Color; Hot Temperature; Kinetics

Changes in photosynthesis of wheat (pigment content and chlorophyll fluorescence) have been measured to study effect of gaseous sulfur dioxide on wheat cenosis. Several biochemical and biophysical techniques have been estimated to evaluate their ability for early detection of destructive effect of SO2. A mathematical model of toxic gas absorption in a closed system with plants has been proposed.

Topics: Absorption; Air Pollutants; Chlorophyll; Chlorophyll A; Dose-Response Relationship, Drug; Ecological Systems, Closed; Fluorescence; Light-Harvesting Protein Complexes; Models, Biological; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Sulfur Dioxide; Triticum

During colonization of space by humans, the bioregenerative life-support systems on board the space ships will require the plants with a highly efficient photosynthesis, a process producing food and O2 and removing CO2-Therefore, in recent years the scientists increasingly focus the their attention to study on photosynthetic apparatus of plants grown in space. Although the available data are quite scanty and, at times, controversial, it is Considered that the space grown plants differ from around control plants by growth and development, many structural and functional indices and metabolism. Data exist showing changes in the chlorophyll (Chl) content, structure and number of chloroplasts in the cell, swelling of thylakoids and decrease in the number and size of starch grains in the chloroplasts (for reviews, see Halstead and Dutcher, 1987; Kordyum, 1997). The decrease of shoot fresh weight and reduction of CO2-saturated photosynthetic rate at saturating light intensities in space grown wheat plants in comparison with ground control have been reported by Tripathy et al. (1996). The thylakoids isolated from space grown plants displayed lower rates of electron transport through photosystems I and II (PSI and PSII) and in a whole chain. This study aimed to examine the electron transport rates through PSI and PSII in the isolated thylakoids, to elucidate if there are any differences in accumulation of thylakoid membranes between space grown Brassica rapa plants and ground control plants (based on Chl quantity) and to measure the Chl a/b ratio in isolated thylakoids. These studies were part of the Collaborative US/Ukrainian program during the STS-87 mission (1997).

Topics: Brassica; Chlorophyll; Chlorophyll A; Electron Transport; Light-Harvesting Protein Complexes; Oxygen; Photosynthetic Reaction Center Complex Proteins; Pigments, Biological; Plant Leaves; Space Flight; Thylakoids; Weightlessness

The influence of aggregation on triplet formation in the light-harvesting pigment-protein complex of photosystem II of green plants (LHCII) has been studied with time-resolved laser flash photolysis. The aggregation state of LHCII has been varied by changing the detergent concentration. The triplet yield increases upon disaggregation and follows the same dependence on the detergent concentration as the fluorescence yield. The rate constant of intersystem crossing is not altered by disaggregation, and variations of the triplet yield appear to be due to aggregation-dependent quenching of singlet excited states. The efficiency of triplet transfer in LHCII aggregates from chlorophyll (Chl) to carotenoid (Car) is 92 +/- 7% at room temperature and 82 +/- 6% at 5 K, and does not change upon disaggregation. The Chl's that do not transfer their triplets to Car's seem to be bound to LHCII and are capable of transfering/accepting their singlet excitations to/from other Chl's. Two spectral contributions of Car triplets are observed: at 525 and 506 nm. Disaggregation of macroaggregates to small aggregates reduces by 10% the relative contribution of Car triplets absorbing at 525 nm. This effect most likely originates from a decreased efficiency of intertrimer Chl-to-Car triplet transfer. At the critical micelle concentration, at which small aggregates are disassembled into trimers, the interactions between Chl and Car are changed. At room temperature, this effect is much more pronounced than at 5 K.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Cold Temperature; Light; Light-Harvesting Protein Complexes; Photolysis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Protein Conformation; Spectrophotometry

The energy transfer process in the minor light-harvesting antenna complex CP29 of green plants was probed in multicolor transient absorption experiments at 77 K using selective subpicosecond excitation pulses at 640 and 650 nm. Energy flow from each of the chlorophyll (Chl) b molecules of the complex could thus be studied separately. The analysis of our data showed that the "blue" Chl b (absorption around 640 nm) transfers excitation to a "red" Chl a with a time constant of 350 +/- 100 fs, while the 'red' Chl b (absorption at 650 nm) transfers on a picosecond time scale (2.2 +/- 0.5 ps) toward a "blue" Chl a. Furthermore, both fast (280 +/- 50 fs) and slow (10-13 ps) equilibration processes among the Chl a molecules were observed, with rates and associated spectra very similar to those of the major antenna complex, LHC-II. Based on the protein sequence homology between CP29 and LHC-II, a basic modelling of the observed kinetics was performed using the LHC-II structure and the Förster theory of energy transfer. Thus, an assignment for the spectral properties and orientation of the two Chl's b, as well as for their closest Chl a neighbors, is put forward, and a comparison is made with the previous assignments and models for LHC-II and CP29.

Topics: Chlorophyll; Chlorophyll A; Energy Transfer; Lasers; Light; Light-Harvesting Protein Complexes; Models, Chemical; Models, Molecular; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Spectrophotometry; Spinacia oleracea; Time Factors

We studied the role of added thylakoid lipids in the light-induced reversible structural changes in isolated macroaggregates of the main light-harvesting chlorophyll a/b complex of photosystem II (LHCII). Loosely stacked lamellar macroaggregates were earlier shown to undergo light-induced reversible structural changes and changes in the photophysical pathways, which resembled those in thylakoid membranes exposed to excess light [Barzda, V., et al. (1996) Biochemistry 35, 8981-8985]. This structural flexibility of LHCII depends critically on the lipid content of the preparations [Simidjiev, I., et al. (1997) Anal. Biochem. 250, 169-175]. It is now reported that lamellar aggregates of LHCII are capable of incorporating substantial amounts of different thylakoid lipids. The long-range order of the chromophores is retained, while the ultrastructure of the lipid-protein macroaggregates can be modified significantly. Addition of thylakoid lipids to the preparations significantly enhances the ability of the LHCII macroaggregates to undergo light-induced structural changes. The lipid environment of the LHCII complexes therefore plays a significant role in determining the structural flexibility of the macroaggregates. As concerns the mechanism of these changes, it is proposed that the absorption of light and the dissipation of its energy in the macrodomains induces thermal fluctuations which bring about changes in the shape or in the stacking interactions of the membranes, this in turn affecting the long-range order of the embedded chromophores. In thylakoids, a similar mechanism is likely to explain the light-induced structural changes which are largely independent of the photochemical activity of the membranes.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Circular Dichroism; Diglycerides; Galactolipids; Glycolipids; Light-Harvesting Protein Complexes; Lipid Bilayers; Macromolecular Substances; Microscopy, Electron; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Pisum sativum

The chlorophyll (Chl) a/b proteins of the photosynthetic prokaryotes appear to have evolved by gene duplication and divergence of the core Chl a antenna family, which also includes CP43 and CP47 and the iron-stress induced Chl a-binding IsiA proteins. We show here that Prochlorothrix hollandica has a cluster of three pcb (prochlorophyte chlorophyll b) genes which are co-transcribed. The major antenna polypeptides of 32 and 38 kDa are encoded by pcbA and pcbC respectively. The pcbC gene is significantly divergent from the other two and may have originated by a gene duplication independent of the one that led to isiA and the other prochlorophyte pcb genes. The distant relatedness of the three prochlorophyte genera implies that not only the ability to make Chl b and use it for light-harvesting arose independently in the three lineages, but also that the pcb genes may have arisen as the result of independent gene duplications in each lineage.

Topics: Amino Acid Sequence; Chlorophyll; Chlorophyll A; DNA, Bacterial; Evolution, Molecular; Genes, Bacterial; Models, Genetic; Molecular Sequence Data; Multigene Family; Phylogeny; Prochlorothrix; Sequence Homology, Amino Acid; Transcription, Genetic

The light-harvesting chlorophyll a/b protein CP24, a minor subunit of the photosystem II antenna system, is a major violaxanthin-binding protein involved in the regulation of excited state concentration of chlorophyll a. This subunit is poorly characterized due to the difficulty in isolation and instability during purification procedures. We have used an alternative approach in order to gain information on the properties of this protein; the Lhcb6 cDNA has been overexpressed in bacteria in order to obtain the CP24 apoprotein, which was then reconstituted in vitro with xanthophylls, chlorophyll a, and chlorophyll b, yielding a pigment-protein complex with properties essentially identical to the native protein extracted from maize thylakoids. Although all carotenoids were supplied during refolding, the recombinant holoprotein exhibited high selectivity in xanthophyll binding by coordinating violaxanthin and lutein but not neoxanthin or beta-carotene. Each monomer bound a total of 10 chlorophyll a plus chlorophyll b and two xanthophyll molecules. Moreover, the protein could be refolded in the presence of different chlorophyll a to chlorophyll b ratios for yielding a family of recombinant proteins with different chlorophyll a/b ratios but still binding the same total number of porphyrins. A peculiar feature of CP24 was its refolding capability in the absence of lutein, contrary to the case of other homologous proteins, thus showing higher plasticity in xanthophyll binding. These characteristics of CP24 are discussed with respect to its role in binding zeaxanthin in high light stress conditions. The spectroscopic analysis of a recombinant CP24 complex binding eight chlorophyll b molecules and a single chlorophyll a molecule by Gaussian deconvolution allowed the identification of four subbands peaking at wavelengths of 638, 645, 653, and 659 nm, which have an increased amplitude with respect to the native complex and therefore identify the chlorophyll b absorption in the antenna protein environment. Gaussian subbands at wavelengths 666, 673, 679, and 686 nm are depleted in the high chlorophyll b complex, thus suggesting they derive from chlorophyll a.

Topics: Amino Acid Sequence; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Cloning, Molecular; Escherichia coli; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Pigments, Biological; Protein Binding; Recombinant Proteins; Spectrometry, Fluorescence; Zea mays

CP26 is the most recently described antenna protein in higher plants which has been reported to be involved in xanthophyll-dependent regulation of the light-harvesting function but is largely unknown due to the difficulties of purification. In this study we have overexpressed in Escherichia coli the Lhcb5 gene product and reconstituted CP26 in vitro by refolding the recombinant protein in the presence of chlorophyll a, chlorophyll b and xanthophylls. The resulting pigment-protein complex is stable enough to be isolated by partially denaturing gel electrophoresis. Reconstitution and isolation conditions for CP26 are similar to those used for other chlorophyll a/b complexes like the major light-harvesting complex of photosystem II (LHCII) and CP29; however, CP26 differs with regard to its lower specificity in carotenoid binding. Most significantly, rather stable recombinant CP26 can be reconstituted containing violaxanthin as the only carotenoid. This enhanced plasticity with respect to carotenoid binding is consistent with CP26 being the major binding protein of violaxanthin involved in the xanthophyll cycle. The availability of recombinant CP26 opens the way to a better characterisation of this pigment-protein complex with regard to its biochemistry and its physiological functions.

Topics: Apoproteins; Carotenoids; Chlorophyll; Chlorophyll A; Cloning, Molecular; Escherichia coli; Lutein; Photosynthetic Reaction Center Complex Proteins; Recombinant Proteins

Potent antigenotoxic and anti-tumor promoting activities of chlorophyll a from green tea (camellia sinensis) have been shown using in vitro cell culture experiments (Okai Y. et al. (1996) Mutation Res., 370, 11-17). In the present study, the authors analyzed in vivo effects of chlorophyll a and b from green tea on tumor promotion in mouse skin in the following ways. 1. When chlorophyll a and b from green tea were applied before each treatment by a tumor promoter, 12-O-tetradecanoyl-phorbol-13-acetate (TPA) on BALB/c mouse skin initiated by 7, 12-dimethylbenz [a] an-thracene (DMBA), they caused significant suppression in a dose-dependent manner against BALB/c mouse skin tumorigenesis. 2. Chlorophyll a and b showed significant suppressive effects against TPA-induced inflammatory reaction such as edema formation in BALB/c mouse ear skin in a dose-dependent fashion. These results suggest that chlorophyll a and b possess potent suppressive activities against tumor promotion in mouse skin.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinogens; Chlorophyll; Chlorophyll A; Edema; Mice; Mice, Inbred BALB C; Pigments, Biological; Skin Neoplasms; Tea; Tetradecanoylphorbol Acetate

Chlorophyll b is an ubiquitous accessory pigment in land plants, green algae, and prochlorophytes. Its biosynthesis plays a key role in the adaptation to various light environments. We isolated six chlorophyll b-less mutants by insertional mutagenesis by using the nitrate reductase or argininosuccinate lyase genes as tags and examined the rearrangement of mutant genomes. We found that an overlapping region of a nuclear genome was deleted in all mutants and that this encodes a protein whose sequence is similar to those of methyl monooxygenases. This coding sequence also contains putative binding domains for a [2Fe-2S] Rieske center and for a mononuclear iron. The results demonstrate that a chlorophyll a oxygenase is involved in chlorophyll b formation. The reaction mechanism of chlorophyll b formation is discussed.

Topics: Amino Acid Sequence; Chlorophyll; Chlorophyll A; Genetic Complementation Test; Molecular Sequence Data; Mutagenesis, Insertional; Oxygenases; Phenotype; Restriction Mapping

The reduction of chlorophyllide b and its analogue zinc pheophorbide b in etioplasts of barley (Hordeum vulgare L.) was investigated in detail. In intact etioplasts, the reduction proceeds to chlorophyllide a and zinc pheophorbide a or, if incubated together with phytyldiphosphate, to chlorophyll a and zinc pheophytin a, respectively. In lysed etioplasts supplied with NADPH, the reduction stops at the intermediate step of 7(1)-OH-chlorophyll(ide) and Zn-7(1)-OH-pheophorbide or Zn-7(1)-OH-pheophytin. However, the final reduction is achieved when reduced ferredoxin is added to the lysed etioplasts, suggesting that ferredoxin is the natural cofactor for reduction of chlorophyll b to chlorophyll a. The reduction to chlorophyll a requires ATP in intact etioplasts but not in lysed etioplasts when reduced ferredoxin is supplied. The role of ATP and the significance of two cofactors for the two steps of reduction are discussed.

Topics: Adenosine Triphosphate; Alcohol Oxidoreductases; Chlorophyll; Chlorophyll A; Ferredoxin-NADP Reductase; Ferredoxins; Hordeum; Intracellular Membranes; Oxidation-Reduction; Pheophytins; Plastids; Subcellular Fractions; Zinc

Dried seeds of maize inbred lines were carried by recoverable satellite to space conditions at an altitude of 175-253 km from sea level for 15 d. The effects of space flight on ultrastructure and photosynthetic pigment contents of maize leaves were studied. Various changes in structure were observed. Chloroplast envelope membrane notch, swelling of thylakoids, less granum stacks and stroma thylakoid membranes, and changes of chloroplast contour were often seen. The contents of chlorophyll a, chlorophyll b, and chlorophyll a+b and carotenoids tended to be lower, but Ca/Cb ratio tended to be higher in the leaves from space flight maize in comparison with ground control.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Microscopy, Electron; Pigments, Biological; Plant Leaves; Seeds; Space Flight; Thylakoids; Weightlessness; Zea mays

A water-soluble Chl a/b-protein (CP673) was isolated and purified from Brussels sprouts (Brassica oleracea L. var. gemmifera DC). The protein had a molecular mass of 78 kDa and an isoelectric point of 4.7, consisted of three or four subunits of 22 kDa and was extremely heat-stable. Although CP673 contained about one Chl a per protein, the blue and red absorption bands of Chl a that consisted of three or four Chl a forms with different absorption maxima suggested that there are several different modes or sites of binding for Chl a. Chl a/b ratio of larger than 10 also indicated that Chl b is present only in a small fraction of CP673. The heterogeneity of CP673 in terms of composition and binding of Chl suggests that Chl is not an intrinsic component of the Chl-protein. Homology search showed that the N-terminal amino acid sequence of CP673 is highly homologous with that of a 22 kDa protein that accumulates in water-stressed leaves of two Brassicaceae plants, rapeseed and radish, but not with those of the light-harvesting Chl a/b-proteins of photosynthesis. A possible function of the water-soluble Chl-protein was discussed.

Topics: Amino Acid Sequence; Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Molecular Sequence Data; Photosynthetic Reaction Center Complex Proteins; Plant Proteins; Protease Inhibitors; Sequence Homology, Amino Acid; Vegetables

Detergent effects on the pre-crystallization of light-harvesting chlorophyll a/b-protein complex (LHCII) were investigated through the fluorescence depolarization method. Stable LHCII crystals were formed in the media containing Triton X-100 (TX) or n-nonyl-beta-D-glucopyranoside (NG) and the crystallization efficients were dependent on their concentrations. The second virial coefficient of crystallizing system, estimated by the fluorescence depolarization analysis of LHCII, showed the most harmonious value under the condition yielding the most efficient crystallization of LHCII to suggest that some specific molecular interaction leading to the crystal growth would be induced according to the concentration of TX or NG.

Topics: Chlorophyll; Chlorophyll A; Crystallization; Detergents; Fluorescence Polarization; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Spinacia oleracea

Light-harvesting chlorophyll a/b-binding protein, LHCP, or its precursor, pLHCP, cannot be stably inserted into barley etioplast membranes in vitro. However, when these etioplast membranes are supplemented with the chlorophyll analogs Zn-pheophytin a/b, synthesized in situ from Zn-pheophorbide a/b and digeranyl pyrophosphate, pLHCP is inserted into a protease-resistant state. This proves that chlorophyll is the only component lacking in etioplast membranes that is necessary for stable LHCP insertion. Synthesis of Zn-pheophytin b alone promotes insertion of LHCP in vitro into a protease-resistant state, whereas synthesis of Zn-pheophytin a alone does not. Insertion of pLHCP into etioplast membranes can also be stimulated by adding chlorophyll a and chlorophyll b to the membranes, albeit at a significantly lower efficiency as compared with Zn-pheophytin a/b synthesized in situ. When pLHCP is inserted into chlorophyll- or Zn-pheophytin-supplemented etioplast membranes and then assayed with protease, only the protease digestion product indicative of the monomeric major light-harvesting chlorophyll a/b complex (LHCII) is found but not the one indicating trimeric complexes. In this respect, chlorophyll- or Zn-pheophytin-supplemented etioplast membranes resemble thylakoid membranes at an early greening stage: pLHCP inserted into plastid membranes from greening barley is assembled into trimeric LHCII only after more than 1 h of greening.

Topics: Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Pheophytins; Photosynthetic Reaction Center Complex Proteins; Plants; Zinc

The minor photosystem II antenna complex CP29(Lhcb-4) has been reconstituted in vitro with the Lhcb-4 apoprotein, overexpressed in Escherichia coli, and the native pigments. Modulation of the pigment composition during reconstitution yields binding products with markedly different chlorophyll a/b binding ratios even though the total number of bound chlorophylls (a plus b) remains constant at eight. A thermodynamic analysis of steady state absorption and fluorescence spectra demonstrates that all chlorophylls are energetically coupled, while the kinetics of chlorophyll photooxidation indicate that triplet chlorophyll-carotenoid coupling is also conserved during pigment binding in vitro. The influence of the chlorophyll a/b binding ratio on the absorption spectra measured at 72 and 300 K is analyzed for the Qy absorption region. Increased chlorophyll b binding leads to large increases in absorption in the 640-660 nm region, while absorption in the 675-690 nm interval decreases markedly. These changes are analyzed in terms of a Gaussian decomposition description in which the eight subbands display a temperature-dependent broadening in agreement with the weak electron-phonon coupling demonstrated for other antenna chlorophyll spectral forms. In this way, we demonstrate that increased chlorophyll b binding leads to increased absorption intensity associated with the subbands at 640, 648, 655, and 660 nm and decreased intensity for the long wavelength subbands at 678 and 684 nm. The wavelength position of all subbands is unchanged. The above data are interpreted to indicate that CP29 has eight chlorophyll binding sites, many or all of which can be occupied by either chlorophyll a or chlorophyll b according to the conditions in which pigment binding occurs. Chlorophyll b absorption is primarily associated with four subbands located at 640, 648, 655, and 660 nm. The invariance of the wavelength position of the absorption bands in recombinant products with different chlorophyll a/b binding stoichiometries is discussed in terms of the mechanism involved in the formation of spectral bands. We conclude that pigment-protein interactions dominate in the determination of spectral heterogeneity with probably only minor effects on absorption associated with pigment-pigment interactions.

Topics: Apoproteins; Binding Sites; Chlorophyll; Chlorophyll A; Cloning, Molecular; Escherichia coli; Kinetics; Light; Light-Harvesting Protein Complexes; Oxidation-Reduction; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Pigments, Biological; Recombinant Proteins; Spectrometry, Fluorescence; Spectrophotometry

Energy transfer from chlorophyll b (Chl b) to chlorophyll a (Chl a) in monomeric preparations of light-harvesting complex II (LHCII) from spinach was studied at 77 K using pump-probe experiments. Sub-picosecond excitation pulses centered at 650 nm were used to excite preferentially Chl b and difference absorption spectra were detected from 630 to 700 nm. Two distinct Chl b to Chl a transfer times, approximately 200 fs and 3 ps, were found. A clearly distinguishable energy transfer process between Chl a molecules occurred with a time constant of 18 ps. The LHCII monomer data are compared to previously obtained LHCII trimer data, and both data sets are fitted simultaneously using a global analysis fitting routine. Both sets could be described with the following time constants: 140 fs, 600 fs, 8 ps, 20 ps, and 2.9 ns. In both monomers and trimers 50% of the Chl b to Chl a transfer is ultrafast (<200 fs). However, for monomers this transfer occurs to Chl a molecules that absorb significantly more toward shorter wavelengths than for trimers. Part of the transfer from Chl b to Chl a that occurs with a time constant of 600 fs in trimers is slowed down to several picoseconds in monomers. However, it is argued that observed differences between monomers and trimers should be ascribed to the loss of some Chl a upon monomerization or a shift of the absorption maximum of one or several Chl a molecules. It is concluded that Chl b to Chl a transfer occurs only within monomeric subunits of the trimers and not between different subunits.

Topics: Chlorophyll; Chlorophyll A; Cold Temperature; Energy Transfer; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Spectrum Analysis; Spinacia oleracea

Prochlorothrix hollandica is an oxygenic photosynthetic prokaryote that differs from the cyanobacteria in having chlorophyll a/b-protein complexes instead of phycobilisomes as major light-harvesting antennae. We report the isolation and culturing of an axenic strain of P. hollandica, available from the Pasteur Culture Collection of Cyanobacteria as strain PCC 9006. The strain has a mean DNA base composition of 51.6 +/- 0.1 mol% G+C and a genomic complexity of 3.37 +/- 0.17 x 10(9) daltons (5,505 kb). A reiterated DNA sequence represents approximately 4.4% of the genome. Restriction enzyme isoschizomers with different sensitivities to base methylation were used to demonstrate that most A residues in the sequence GATC are methylated in P. hollandica DNA and that this methylation increases with culture age. Furthermore, some C residues are methylated, although the specificity of the C methylation system does not match that of well-characterized C methylases. Nucleotide analysis showed that up to approximately 3.5% of both dA and dC residues are methylated in P. hollandica DNA.

Topics: Base Pairing; Chlorophyll; Chlorophyll A; DNA Methylation; DNA, Bacterial; Phycobilisomes; Prochlorothrix; Repetitive Sequences, Nucleic Acid; Time Factors

Excess light energy absorbed by the chloroplast membranes of green plants is dissipated by nonradiative de-excitation in order to protect against photodamage. This is observed as the nonphotochemical quenching of chlorophyll fluorescence, which has been suggested to result from an alteration in the structure and function of the chlorophyll a/b light-harvesting complexes of photosystem II (LHCII) due to the combined effects of protonation and the de-epoxidation of bound violaxanthin to form zeaxanthin. In agreement with this hypothesis, it is shown that the light-harvesting chlorophyll a/b proteins purified from spinach leaves exhibit pH-stimulated quenching of chlorophyll fluorescence; this quenching shares all the key features observed for the nonphotochemical quenching of chlorophyll fluorescence in vivo. In the case of the two minor complexes, LHCIIa (CP29) and LHCIIc (CP26), quenching is much greater than in the bulk complex LHCIIb and is strongly inhibited by the reagent dicyclohexylcarbodiimide. The carotenoids violaxanthin and zeaxanthin cause strong inhibition and stimulation of quenching, respectively, in these complexes. The results of this study are consistent with the suggestion that the minor light-harvesting complexes play a crucial role in photoprotective energy dissipation in the photosynthetic membrane of green plants. Moreover, for the first time, a system using isolated LHCIIa and LHCIIc for the study of the regulation of light harvesting is described.

Topics: beta Carotene; Carotenoids; Carrier Proteins; Chlorophyll; Chlorophyll A; Fluorescence; Light-Harvesting Protein Complexes; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plants; Xanthophylls

Chlorophyll b is synthesized from chlorophyll a by the oxidation of the methyl group on the ring B of the tetrapyrrole ring to the formyl group. Previously, we reported that chlorophyllide b could be converted to chlorophyll a in isolated cucumber etioplasts indicating the conversion of chlorophyll b to chlorophyll a. To identify the intermediate molecule, we used barley etioplasts instead of cucumber. Chlorophyll a and an additional pigment were found after incubation of chlorophyllide b with isolated barley etioplasts. The pigment has the same retention time and absorption spectrum as 7-hydroxymethyl chlorophyll, which has the hydroxymethyl group on ring B instead of the formyl group of chlorophyll b. Authentic 7-hydroxymethyl chlorophyll was prepared by reduction of chlorophyll b by NaBH4. Chlorophyll a accumulated during the incubation of 7-hydroxymethyl chlorophyllide with etioplasts. These findings indicate that chlorophyll b is converted to chlorophyll a via 7-hydroxymethyl chlorophyll. Chlorophyll b and 7-hydroxymethyl chlorophyll accumulated within a short period of incubation of chlorophyllide b with etioplasts. However, chlorophyll a accumulated with a concomitant decrease of chlorophyll b and 7-hydroxymethyl chlorophyll. These observations also suggest that chlorophyll b is converted to 7-hydroxymethyl chlorophyll and then to chlorophyll a. Both steps required ATP.

Topics: Carbon Radioisotopes; Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Hordeum; Kinetics; Pigments, Biological; Radioisotope Dilution Technique; Spectrophotometry; Time Factors

The minor light-harvesting chlorophyll-a/b-binding protein CP29 (Lhcb4), overexpressed in Escherichia coli, has been reconstituted in vitro with pigments. The recombinant pigment-protein complexes show biochemical and spectral properties identical to the native CP29 purified from maize thylakoids. The xanthophyll lutein is the only carotenoid necessary for reconstitution, a finding consistent with the structural role of two lutein molecules/polypeptide suggested by the crystallographic data for the homologous protein light-harvesting chlorophyll-a/b-binding protein of photosystem II (LHCII). The CP29 protein scaffold can accommodate different chromophores. This conclusion was deduced by the observation that the pigment composition of the reconstituted protein depends on the pigments present in the reconstitution mixture. Thus, in addition to a recombinant CP29 identical to the native one, two additional forms of the complex could be obtained by increasing chlorophyll b content. This finding is typical of CP29 because the major LHCII complex shows an absolute selectivity for chromophore binding [Plumley, F. G. & Schmidt, G. W. (1987) Proc. Natl Acad. Sci. USA 84, 146-150; Paulsen, H., Rümler, U. & Rüdiger, W. (1990) Planta (Heidelb.) 181, 204-211], and it is consistent with the higher stability of CP29 during greening and in chlorophyll b mutants compared with LHCII.

Topics: Apoproteins; Base Sequence; Chlorophyll; Chlorophyll A; Circular Dichroism; Escherichia coli; Light-Harvesting Protein Complexes; Molecular Sequence Data; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Pigments, Biological; Protein Binding; Recombinant Proteins; Spectrometry, Fluorescence; Zea mays

Photosystem II, the photosynthetic water-oxidizing complex, can be isolated from both plants and cyanobacteria. A variety of methods have been developed for purification of this enzyme, which can be isolated in several functional and structural forms. Knowledge of the pigment content of photosystem II preparations is important for precise spectroscopic, biochemical, and functional analysis. We have determined pigment stoichiometries in oxygen-evolving photosystem II preparations from plants and cyanobacteria. We have employed a solvent system for the isocratic elution of a reverse phase HPLC column in which we have determined the extinction coefficients of the relevant pigments. Pigments were extracted from four photosystem II preparations. These preparations included spinach photosystem II membranes [Berthold, D. A., Babcock, G. T., & Yocum, C. F. (1981) FEBS Lett. 134, 231-234], spinach photosystem II reaction center complexes [Ghanotakis, D. F., & Yocum, C. F. (1986) FEBS Lett. 197, 244-248], spinach photosystem II complexes [MacDonald, G. M., & Barry, B. A. (1992) Biochemistry 31, 9848-9856], and photosystem II particles isolated from the cyanobacterium, Synechocystis sp. PCC 6803 [Noren, G. H., Boerner, R. J., & Barry, B. A. (1991) Biochemistry 30, 3943-3950]. Pigment stoichiometries were determined using two different methods of data analysis and were based on the assumption that there are two pheophytin a molecules per photosystem II reaction center. The pigment stoichiometries obtained were comparable for the two methods of data analysis and agreed with previous biophysical and biochemical characterizations of the preparations. The average pigment stoichiometries (chlorophyll:plastoquinone-9 per 2 pheophytin a) determined using the two data analysis methods were as follows: photosystem II membranes, 274:3.2; photosystem II reaction center complexes, 78:2.5; Synechocystis PS II particles, 55:2.4; photosystem II complexes, 121:2.0.

Topics: Chlorophyll; Chlorophyll A; Chromatography, High Pressure Liquid; Cyanobacteria; Light-Harvesting Protein Complexes; Pheophytins; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plastoquinone; Spectrophotometry; Spinacia oleracea

Prochlorococcus marinus CCMP 1375, a ubiquitous and ecologically important marine prochlorophyte, was bound to possess functional genes coding for the alpha and beta subunits of a phycobiliprotein. The latter is similar to phycoerythrins (PE) from marine Synechococcus cyanobacteria and bind a phycourobilin-like pigment as the major chromophore. However, differences in the sequences of the alpha and beta chains compared with known PE subunits and the presence of a single bilin attachment site on the alpha subunit designate it as a novel PE type, which we propose naming PE-III. P. marinus is the sole prokaryotic organisms known so far that contains chlorophylls a and b as well as phycobilins. These data strongly suggest that the common ancestor of prochlorophytes and the Synechococcus cyanobacteria contained phycobilins. Flow cytometric data from the tropical Pacific Ocean provide evidence that deep populations of Prochlorococcus possess low amounts of a PE-like pigment, which could serve either in light harvesting or nitrogen storage or both.

Topics: Amino Acid Sequence; Bacterial Proteins; Chlorophyll; Chlorophyll A; Cyanobacteria; Genes, Bacterial; Molecular Sequence Data; Phycoerythrin; Phylogeny; Sequence Alignment; Sequence Homology, Amino Acid

Analyses of a series of allelic chlorina mutants of wheat (Triticum aestivum L.), which have partial blocks in chlorophyll (Chl) synthesis and, therefore, a limited Chl supply, reinforce the principle that Chl is required for the stable accumulation of Chl-binding proteins and that only reaction centers accumulate when the supply of Chl is severely limited. Depending on the rate of Chl accumulation (determined by the severity of the mutation) and on the rate of turnover of Chl and its precursors (determined by the environment in which the plant is grown), the mutants each reach an equilibrium of Chl synthesis and degradation. Together these mutants generate a spectrum of phenotypes. Under the harshest conditions (high illumination), plants with moderate blocks in Chl synthesis have membranes with very little Chl and Chl-proteins and membrane stacks resembling the thylakoids of the lethal xantha mutants of barely grown at low to medium light intensities (which have more severe blocks). In contrast, when grown under low-light conditions the same plants with moderate blocks have thylakoids resembling those of the wild type. The wide range of phenotypes of Chl b-deficient mutants has historically produced more confusion than enlightenment, but incomparable growth conditions can now explain the discrepancies reported in the literature.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Dose-Response Relationship, Radiation; Intracellular Membranes; Light; Light-Harvesting Protein Complexes; Mutation; Phenotype; Photosynthetic Reaction Center Complex Proteins; Protoporphyrins; Solanum lycopersicum; Species Specificity; Triticum

Monospecific polyclonal antibodies have been raised against synthetic peptides derived from the primary sequences from different plant light-harvesting Chl a/b-binding (LHC) proteins. Together with other monospecific antibodies, these were used to quantify the levels of the 10 different LHC proteins in wild-type and chlorina f2 barley (Hordeum vulgare L.), grown under normal and intermittent light (ImL). Chlorina f2, grown under normal light, lacked Lhcb1 (type I LHC II) and Lhcb6 (CP24) and had reduced amounts of Lhcb2, Lhcb3 (types II and III LHC II), and Lhcb4 (CP 29). Chlorina f2 grown under ImL lacked all LHC proteins, whereas wild-type ImL plants contained Lhcb5 (CP 26) and a small amount of Lhcb2. The chlorina f2 ImL thylakoids were organized in large parallel arrays, but wild-type ImL thylakoids had appressed regions, indicating a possible role for Lhcb5 in grana stacking. Chlorina f2 grown under ImL contained considerable amounts of violaxanthin (2-3/reaction center), representing a pool of phototransformable xanthophyll cycle pigments not associated with LHC proteins. Chlorina f2 and the plants grown under ImL also contained early light-induced proteins (ELIPs) as monitored by western blotting. The levels of both ELIPs and xanthophyll cycle pigments increased during a 1 h of high light treatment, without accumulation of LHC proteins. These data are consistent with the hypothesis that ELIPs are pigment-binding proteins, and we suggest that ELIPs bind photoconvertible xanthophylls and replace "normal" LHC proteins under conditions of light stress.

Topics: Amino Acid Sequence; Antibodies; Chlorophyll; Chlorophyll A; Hordeum; Intracellular Membranes; Light; Light-Harvesting Protein Complexes; Microscopy, Electron; Molecular Sequence Data; Photosynthetic Reaction Center Complex Proteins; Pigments, Biological

The mutant, C-2A'-34, lacks the beta, epsilon-carotenoids, alpha-carotene, lutein and loroxanthin. When grown under heterotrophic or mixotropic conditions this strain develops significantly higher levels of beta-carotene and violaxanthin than does the original developmental mutant of Scenedesmus, C-2A'. The decrease in chlorophyll a and chlorophyll b observed in C-2A'-34 is accompanied by the near absence of the LHC. The light intensity dependence of greening of this strain is comparable to that of C-2A'; the loss of the beta,epsilon-carotenoids and modification of the pool of beta,beta-carotenoids neither prevent the proximal pigment-protein complexes of photosystems I and II from developing nor cause any short term photosensitivity. The increase in the beta,beta-carotenoids in C-2A'-34 apparently compensates for the loss of the beta,epsilon-carotenoids required in formation of the proximal and distal antennae systems but not in the LHCs.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Chlorophyta; Intramolecular Lyases; Isomerases; Light; Light-Harvesting Protein Complexes; Mutation; Photosynthetic Reaction Center Complex Proteins

Resonance Raman spectroscopy was performed on photosystem II light-harvesting complex (LHCII) in trimeric and oligomeric states with various excitation conditions. From these studies, it can be concluded that the structure of LHCII is altered during the trimer/oligomer transition. These structural changes affect the conformation of a population of carotenoid within the complex, which becomes twisted in the oligomeric form. Moreover, the interactions assumed by a chlorophyll a and a chlorophyll b are modified during the oligomerization process. This induces the formation of an H-bond to a formyl group of a chlorophyll b molecule and to a keto group of a chlorophyll a molecule. The extent to which these H-bonds to chlorophyll relate to the formation of the quencher cannot yet be precisely established. However, the structural changes they evidence may play a role in the control of the energy flux by LHCII complexes.

Topics: Carotenoids; Chlorophyll; Chlorophyll A; Hydrogen Bonding; In Vitro Techniques; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Spectrum Analysis, Raman; Spinacia oleracea

We describe a procedure to separate stacked and unstacked membranes from the prochlorophyte Prochlorothrix hollandica that is based on methods used for the separation of grana and stroma thylakoids from chloroplasts. Stacked membranes were isolated from Triton X-100-treated whole thylakoid preparations, unstacked membranes from French press disrupted cells. Membrane fractions were isolated by differential centrifugation. The stacked membranes were enriched in photosystem (PS) II and a chlorophyll a/b-binding antenna complex, whereas the unstacked membranes contained PS I and the ATP synthase. No evidence for a PS I-associated chlorophyll a/b antenna system was obtained. The PS II-associated chlorophyll a/b antenna complex is composed of several apoproteins in the molecular mass range from 32 to 38 kDa. The 38-kDa protein of this complex becomes phosphorylated on its stromal surface by a light-activated kinase, but, unlike the light-harvesting II complex of chloroplasts, it does not migrate from grana to stroma membrane regions. Overall, the thylakoid membranes of Prochlorothrix exhibit a remarkably similar organization to those of chloroplasts, especially in terms of the non-random distribution of protein complexes between grana and stroma thylakoid membranes. However, in contrast to the rapidly reversible phosphorylation-dependent state 1-state 2 response of light-harvesting complex associated with PS II in chloroplasts, the slowly reversible phosphorylation in Prochlorothrix leads only to a functional uncoupling from PS II but not to its redistribution between stacked and unstacked membrane regions.

Topics: Chlorophyll; Chlorophyll A; Chlorophyta; Chloroplasts; Cyanobacteria; Membrane Proteins; Phosphorylation; Plant Proteins

The phosphatidylglycerol containing the unusual delta 3-trans hexadecenoic fatty acid is specifically found in photosynthetic membranes of eukaryotic organisms. Its involvement in the biogenesis and the structure of the light-harvesting chlorophyll a/b-protein complex has been evidenced by in vivo targeting this lipid to photosynthetic membranes of Chlamydomonas reinhardtii mutants lacking this lipid. In the mf1 and mf2 mutants, this deficiency results in (a) the absence of the oligomeric light-harvesting complex of photosystem 2; (b) an extensive destacking of thylakoid membranes; (c) a very low 77-K fluorescence emission in the photosystem-2 region. We show in this paper that these deficiencies result from modifications in the pigment and polypeptide compositions of the photosystem-2 light-harvesting complex; it contains less chlorophyll b and some of its constitutive polypeptides are absent or reduced in amount, while immunologically related polypeptides of lower molecular mass accumulate. The direct involvement of the lack of trans-C16: 1-phosphatidylglycerol in these deficiencies is evidenced by the partial restoration of normal characteristics of the light-harvesting complex (pigment and polypeptide composition, oligomerization) after liposome-mediated, in vivo incorporation of this lipid into the photosynthetic membranes of the mf2 mutant. Trans-C16:1-phosphatidylglycerol, therefore, is involved in the biogenesis of the photosystem-2 light-harvesting chlorophyll a/b-protein complex through a mechanism that may prevent degradation processes. Its contribution to the structural conformation of neosynthesized monomers and to their organization into stable oligomeric form is discussed.

Topics: Animals; Centrifugation, Density Gradient; Chlamydomonas reinhardtii; Chlorophyll; Chlorophyll A; Chloroplasts; Electrophoresis, Polyacrylamide Gel; Isoelectric Focusing; Light-Harvesting Protein Complexes; Liposomes; Membrane Lipids; Mutation; Palmitic Acids; Peptides; Phosphatidylglycerols; Photosynthetic Reaction Center Complex Proteins; Structure-Activity Relationship

Temperature dependence in electronic energy transfer steps within light-harvesting antenna trimers from photosystem II was investigated by studying Chl a pump-probe anisotropy decays at several wavelengths from 675 to 682 nm. The anisotropy lifetime is markedly sensitive to temperature at the longest wavelengths (680-682 nm), increasing by factors of 5 to 6 as the trimers are cooled from room temperature to 13 K. The temperature dependence is muted at 677 and 675 nm. This behavior is modeled using simulations of temperature-broadened Chl a absorption and fluorescence spectra in spectral overlap calculations of Förster energy transfer rates. In this model, the 680 nm anisotropy decays are dominated by uphill energy transfers from 680 nm Chl a pigments at the red edge of the LHC-II spectrum; the 675 nm anisotropy decays reflect a statistical average of uphill and downhill energy transfers from 676-nm pigments. The measured temperature dependence is consistent with essentially uncorrelated inhomogeneous broadening of donor and acceptor Chl a pigments.

Topics: Anisotropy; Biophysical Phenomena; Biophysics; Chlorophyll; Chlorophyll A; Cold Temperature; Energy Metabolism; Energy Transfer; Light; Light-Harvesting Protein Complexes; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Protein Conformation; Vegetables

Chlorophyll a accumulated when chlorophyllide b was incubated with isolated cucumber etioplasts in the dark. When [14C]chlorophyllide b was used as the substrate for chlorophyll synthesis, [14C]chlorophyll a was formed, showing that chlorophyll a was synthesized from the exogenously added chlorophyllide b, not by light-independent reduction of endogenous protochlorophyllide. The reaction studied showed an optimal pH of 7.5 and required both the soluble and membrane fractions of etioplasts together with ATP. Time course experiments showed that chlorophyll a began to accumulate later than chlorophyll b. Chlorophyllide b esterified with geranylgeraniol accumulated first, but there was little accumulation of chlorophyll a with an unhydrogenated prenyl side chain. No chlorophyllide a was detected during incubation. These observations indicate that esterified chlorophyll b was converted to chlorophyll a. This conversion would play an important role in the use of chlorophyll b for the synthesis of chlorophyll a in the reconstruction of photosystems.

Topics: Chlorophyll; Chlorophyll A; Chlorophyllides; Organelles; Vegetables

The mechanism of formation of the formyl group of chlorophyll b has long been obscure but, in this paper, the origin of the 7-formyl-group oxygen of chlorophyll b in higher plants was determined by greening etiolated maize leaves, excised from dark-grown plants, by illumination under white light in the presence of either H2(18)O or 18O2 and examining the newly synthesized chlorophylls by mass spectroscopy. To minimize the possible loss of 18O label from the 7-formyl substituent by reversible formation of chlorophyll b-7(1)-gem-diol (hydrate) with unlabelled water in the cell, the formyl group was reduced to a hydroxymethyl group during extraction with methanol containing NaBH4: chlorophyll a remained unchanged during this rapid reductive extraction process. Mass spectra of chlorophyll a and [7-hydroxymethyl]-chlorophyll b extracted from leaves greened in the presence of either H2(18)O or 18O2 revealed that 18O was incorporated only from molecular oxygen but into both chlorophylls: the mass spectra were consistent with molecular oxygen providing an oxygen atom not only for incorporation into the 7-formyl group of chlorophyll b but also for the well-documented incorporation into the 13(1)-oxo group of both chlorophylls a and b [see Walker, C. J., Mansfield, K. E., Smith, K. M. & Castelfranco, P. A. (1989) Biochem. J. 257, 599-602]. The incorporation of isotope led to as much as 77% enrichment of the 13(1)-oxo group of chlorophyll a: assuming identical incorporation into the 13(1) oxygen of chlorophyll b, then enrichment of the 7-formyl oxygen was as much as 93%. Isotope dilution by re-incorporation of photosynthetically produced oxygen from unlabelled water was negligible as shown by a greening experiment in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. The high enrichment using 18O2, and the absence of labelling by H2(18)O, unequivocally demonstrates that molecular oxygen is the sole precursor of the 7-formyl oxygen of chlorophyll b in higher plants and strongly suggests a single pathway for the formation of the chlorophyll b formyl group involving the participation of an oxygenase-type enzyme.

Topics: Chlorophyll; Chlorophyll A; Light; Molecular Structure; Oxidation-Reduction; Oxygen; Oxygen Isotopes; Oxyquinoline; Spectrometry, Mass, Fast Atom Bombardment; Spectrophotometry; Zea mays

The excitation energy transfer between chlorophyll b (Chl b) and chlorophyll a (Chl a) in the isolated trimeric chlorophyll-a/b-binding protein complex of spinach photosystem 2 (LHC II) has been studied by femtosecond spectroscopy. In the main absorption band of Chl b the ground state recovery consists of two components of 0.5 ps and 2.0 ps, respectively. Also in the Chl a absorption band, at 665 nm, the ground state recovery is essentially bi-exponential. In this case is, however, the fastest relaxation lifetime is a 2.0 ps component followed by a slower component with a lifetime in the order of 10-20 ps. In the Chl b absorption band a more or less constant anisotropy of r = 0.2 was observed during the 3 ps the system was monitored. In the Chl a absorption band there was, however, a relaxation of the anisotropy from r = 0.3 to a quasi steady state level of r = 0.18 in about 1 ps. Since the 0.5 ps component is only seen upon selective excitation of Chl b we assign this component to the energy transfer between Chl b and Chl a. The other components most likely represents redistribution processes of energy among spectrally different forms of Chl a. The energy transfer process between Chl b and Chl a can well be explained by the Förster mechanism which also gives a calculated distance of 13 A between interacting chromophores. The organisation of chlorophylls in LHC II is discussed in view of the recent crystal structure data (1991) Nature 350, 130].

Topics: Chlorophyll; Chlorophyll A; Energy Transfer; Fluorescence Polarization; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Plants; Spectrometry, Fluorescence

Chlorophyllide b was converted to chlorophyll a when incubated with cucumber etioplasts in the presence of phytol, ATP, and MgCl2. Chlorophyll a was identified by retention time on HPLC, absorption, fluorescence, and excitation spectra. It is suggested that the interconversion of chlorophyll a and b plays a significant role in the establishment of the required chlorophyll a/b ratio during the adaptation of leaves to high and low light conditions.

Topics: Chlorophyll; Chlorophyll A; Chlorophyllides; Chromatography, High Pressure Liquid; Organelles; Spectrometry, Fluorescence; Spectrophotometry; Vegetables

The conformational behaviour of the light-harvesting chlorophyll a/b-binding protein (LHCP), the apoprotein of the major light-harvesting complex (LHCII) of photosystem II in plants, has been studied. According to the circular dichroism in the ultraviolet range measured with isolated LHCII, the protein in the complex adopts a folded structure with a high content of alpha helix (about 60%), whereas the non-pigmented, solubilized protein has a less ordered structure (about 20% alpha helix). LHCP-pigment complexes that have been reconstituted from the overexpressed protein and isolated pigments in the presence of detergents display a protein CD signal similar to that of authentic LHCII, indicating that LHCP folds into the native structure during the reconstitution procedure. Renaturation of LHCP in these experiments is dependent on the presence of pigments and the formation of stable LHCP-pigment complexes. Pigment-induced engagement of LHCP in a compact structure has also been shown by two additional experimental approaches. (a) Upon complex formation, LHCP or its precursor (pLHCP) becomes resistant to trypsin digestion with the exception of an N-terminal segment of the protein; the same protection of LHCP is known to occur in intact thylakoids. (b) Pigment binding renders a cysteine residue within the N-proximal hydrophobic domain of the protein as well as a newly introduced cysteine four amino acid positions from the C terminus inaccessible to modification with a sulfhydryl-specific label whereas the N terminus stays susceptible to specific labelling. These observations support the notion that only the N terminus protrudes from a compact protein-pigment structure in LHCII. The fact that the major part of LHCP is trypsin-resistant in pigmented complexes reconstituted in the absence of a membrane or even lipids justifies caution in using protection against trypsin as a criterion for the integration of LHCP into the thylakoid membrane.

Topics: Apoproteins; Base Sequence; Chlorophyll; Chlorophyll A; DNA, Single-Stranded; Endopeptidases; Fabaceae; Hydrolysis; Intracellular Membranes; Light-Harvesting Protein Complexes; Lutein; Molecular Sequence Data; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plants, Medicinal; Protein Binding; Protein Folding

Using mass spectroscopy, we demonstrate as much as 93% enrichment of the 7-formyl group oxygen of chlorophyll b when dark-grown, etiolated maize leaves are greened under white light in the presence of 18O2. This suggests that a mono-oxygenase is involved in the oxidation of its methyl group precursor. The concomitant enrichment of about 75% of the 13(1)-oxygen confirms the well-documented finding that this oxo group, in both chlorophyll a and b, also arises from O2. High 18O enrichment into the 7-formyl oxygen relative to the substrate 18O2 was achieved by optimization of the greening conditions in combination with a reductive extraction procedure. It indicates not only a single pathway for Chl b formyl group formation, but also unequivocally demonstrates that molecular oxygen is the sole precursor of the 7-formyl oxygen.

Topics: Chlorophyll; Chlorophyll A; Mass Spectrometry; Mixed Function Oxygenases; Molecular Structure; Oxygen; Zea mays

In order to identify segments of light-harvesting chlorophyll a/b-binding protein (LHCP) that are important for pigment binding, we have tested various LHCP mutants regarding their ability to form stable pigment-protein complexes in an in vitro reconstitution assay. Deletion of 10 C-terminal amino acids in the LHCP precursor, pLHCP, did not significantly affect pigment binding, whereas deletion of one additional amino acid, a tryptophan, completely abolished the formation of stable pigment-protein complexes. This tryptophan, however, can be exchanged with other amino acids in full-length pLHCP without noticeably altering the stability or spectroscopic properties of pigment complexes made with these mutants. Thus, the tryptophan residue is not likely to be involved in a highly specific interaction stabilizing the complex. A double mutant of LHCP lacking 66 N-terminal and 6 C-terminal amino acids still forms pigmented complexes that are virtually identical to those formed with the full-length protein concerning their pigment composition and spectroscopic properties. We conclude that about 30% of the polypeptide chain in LHCP is not involved in pigment binding.

Topics: Amino Acid Sequence; Base Sequence; Chlorophyll; Chlorophyll A; Cloning, Molecular; Escherichia coli; Molecular Sequence Data; Mutagenesis, Site-Directed; Oligodeoxyribonucleotides; Photosynthetic Reaction Center Complex Proteins; Plants; Restriction Mapping

The structure and supramolecular assembly of the soybean photosystem 1 (PS 1) chlorophyll a/b-binding antenna (LHC 1) was examined. We identified the subunit composition of LHC 1 in soybean and followed the accumulation of individual subunits during light-induced assembly. We observed four LHC 1 subunits, at 23, 22, 21 and 20.5 kDa, obtained partial sequence information by amino-terminal sequence analysis, and classified the 20.5, 22, and 21 kDa subunits as being encoded by type I, II, and IV chlorophyll a/b binding protein genes, respectively. Antisera against LHC 1 subunits were used to follow the accumulation of individual subunits during the light-initiated transition from etioplast to chloroplast. Several points are noteworthy. First, monospecific antibody against the 22 kDa subunit decorated a 25 kDa peptide in etiolated tissue, which declined during maturation. This decline correlated with the light-induced appearance of mature 22 kDa peptide, suggesting a precursor/product relationship. Second, the same antibody identified a 22 kDa protein in mature corn, but not a larger band in etiolated corn, suggesting that LHC 1 accumulation is regulated differently between species before the onset of chlorophyll biosynthesis. Third, the mature 22 kDa subunit appeared somewhat later than the other LHC 1 peptides during greening, implying that this subunit is less intimately associated with the PS1 core than are the subunits appearing earlier in development.

Topics: Antibodies; Apoproteins; Carrier Proteins; Chlorophyll; Chlorophyll A; Electrophoresis, Polyacrylamide Gel; Glycine max; Immune Sera; Intracellular Membranes; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Pigments, Biological; Zea mays

Thylakoid membranes were isolated from pea seedlings grown under intermittent light (2-min light/118-min dark cycles). These preparations differed from controls (thylakoids from plants grown under 16-h light/8-h dark cycles) in the following respects: 15 times smaller chlorophyll/protein ratio, 10 times greater chlorophyll a/b ratio, absence of light-harvesting chlorophyll a/b binding proteins, and 2-3-fold greater ratio of photosystem II over photosystem I. In addition we found the following: (1) Electrogenic electron transfer around cytochrome b6/f under flashing light was greatly enhanced, probably as a consequence of the greater photosystem II/photosystem I ratio. (2) The rate of proton uptake from the medium at the acceptor side of photosystem II was enhanced, probably by unshielding of the quinone binding domain. (3) The N,N'-dicyclohexylcarbodiimide sensitivity of the proton-pumping activity of photosystem II was absent, which was consistent with the attribution of a N,N'-dicyclohexylcarbodiimide-induced protonic short circuit to chlorophyll a/b binding proteins. (4) The sensitivity of oxygen evolution under continuous light to variations of pH or the concentration of Ca2+ was altered. Chlorophyll a/b binding proteins serve as light-harvesting antennas. We found in addition that they modulated the activity of water oxidation and, in particular, the proteolytic reactions around photosystem II.

Topics: Cell Fractionation; Chlorophyll; Chlorophyll A; Fabaceae; Hydrogen-Ion Concentration; Intracellular Membranes; Kinetics; Light; Light-Harvesting Protein Complexes; Membrane Potentials; Organelles; Oxygen; Photosynthesis; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Plants, Medicinal; Spectrophotometry

The cab and psb A RNA transcript levels have been determined in Pisum sativum leaves exposed to supplementary ultraviolet-B radiation. The nuclear-encoded cab transcripts are reduced to low levels after only 4 h of UV-B treatment and are undetectable after 3 days exposure. In contrast, the chloroplast-encoded psb A transcript levels, although reduced, are present for at least 3 days. After short periods of UV-B exposure (4 h or 8 h), followed by recovery under control conditions, cab RNA transcript levels had not recovered after 1 day, but were re-established to ca. 60% of control levels after 2 more days. Increased irradiance during exposure to UV-B reduced the effect upon cab transcripts, although the decrease was still substantial. These results indicate rapid changes in the cellular regulation of gene expression in response to supplementary UV-B and suggest increased UV-B radiation may have profound consequences for future productivity of sensitive crop species.

Topics: Blotting, Northern; Carrier Proteins; Chlorophyll; Chlorophyll A; Fabaceae; Genes, Plant; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Plants, Medicinal; RNA; Transcription, Genetic; Ultraviolet Rays

Topics: Amino Acid Sequence; Base Sequence; Chlorophyll; Chlorophyll A; DNA; Fabaceae; Light-Harvesting Protein Complexes; Molecular Sequence Data; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plants, Medicinal

Topics: Amino Acid Sequence; Base Sequence; Chlorophyll; Chlorophyll A; Consensus Sequence; DNA; Fabaceae; Light-Harvesting Protein Complexes; Molecular Sequence Data; Photosynthetic Reaction Center Complex Proteins; Plants, Medicinal

cDNAs encoding three different LHC I polypeptides (Type I, Type II and Type III) from the gymnosperm Scots pine (Pinus sylvestris L.) were isolated and sequences. Comparisons of the deduced amino acid sequences with the corresponding tomato sequences showed that all three proteins were highly conserved although less so than the LHC II proteins. The similarities between mature Scots pine and tomato Types I, II and III LHC I proteins were 80%, 87% and 85%, respectively. Two of the five His residues that are found in AXXXH sequences, which have been identified as putative chlorophyll ligands in the Type I and Type II proteins, were not conserved. The same two regions of high homology between the different LHC proteins, which have been identified in tomato, were also found in the Scots pine proteins. Within the conserved regions, the Type I and Type II proteins had the highest similarity; however, the Type II and Type III proteins also showed a similarity in the central region. The results suggest that all flowering plants (gymnosperms and angiosperms) probably have the same set of LHC polypeptides. A new nomenclature for the genes encoding LHC polypeptides (formerly cab genes) is proposed. The names lha and lhb are suggested for genes encoding LHC I and LHC II proteins, respectively, analogous to the nomenclature for the genes encoding other photosynthetic proteins.

Topics: Amino Acid Sequence; Base Sequence; Biological Evolution; Chlorophyll; Chlorophyll A; Light-Harvesting Protein Complexes; Molecular Sequence Data; Photosynthetic Reaction Center Complex Proteins; Plant Proteins; Plants; Sequence Alignment; Sequence Homology, Nucleic Acid

A 30-kDa chlorophyll-a/b-binding protein was purified from photosystem II membrane fragments using Ca(2+)-chelating Sepharose 6B chromatography. The protein binds approximately four chlorophyll a molecules, one chlorophyll b molecule and carotenoids. Its 77-K fluorescence-emission spectrum exhibits a maximum at 680 +/- 1 nm. The protein has a high tendency to form a dimer in the presence of Ca2+.Ca2+ binding affects the low-temperature fluorescence-emission maximum, leading to a decrease in its intensity and a blue shift of 1 nm. Similar spectral changes were obtained in the presence of Mg2+, possibly indicating a common binding domain for both cations. We interpret these observations as cation-induced conformational changes of the protein, which were reversible upon subsequent incubation in EDTA. Evidence is presented for the involvement of carboxyl groups in the coordination sphere of the bivalent cations. The possible structural and functional role of the protein is discussed.

Topics: Blotting, Western; Calcium; Chlorophyll; Chlorophyll A; Chromatography, Gel; Electrophoresis, Polyacrylamide Gel; Hydrolysis; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plant Proteins; Plants; Spectrometry, Fluorescence

Large two-dimensional crystals of the light-harvesting chlorophyll a/b-protein complex (LHC-II) from the photosynthetic membrane of pea chloroplasts were grown by a new method from detergent solution. The structure of these crystals was examined by electron crystallography, using three different media to preserve high-resolution detail: vitrified water, glucose and tannin. The crystals diffracted electrons to at least 3.2 A resolution in all three media. R-factors between the three data sets of electron diffraction amplitudes ranged from 6.4% to 14.3%. Fourier difference maps were generated and compared to a projection map of the complex at 3.4 A resolution. No significant differences were found, proving that all three media preserved the native structure of LHC-II at high resolution. The probability of recording high-quality electron diffraction patterns with tannin was 90%. With glucose and water this probability was lower by a factor of 10 to 20, suggesting that tannin may be preferable as a preserving medium for sensitive biological specimens.

Topics: Chlorophyll; Chlorophyll A; Crystallization; Crystallography; Detergents; Fabaceae; Fourier Analysis; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Plants, Medicinal; Solutions; X-Ray Diffraction

The structure of the light-harvesting chlorophyll a/b-protein complex, a membrane protein serving as the major antenna of solar energy in plant photosynthesis, has been determined at 6 A resolution by electron crystallography. Within the complex, three membrane-spanning alpha helices and 15 chlorophyll molecules are resolved. There is an intramolecular diad relating two of the alpha helices and some of the chlorophylls. The spacing of the chlorophylls suggests energy transfer by delocalized exciton coupling and Förster mechanisms.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Crystallography; Light-Harvesting Protein Complexes; Lipid Bilayers; Models, Molecular; Molecular Structure; Photosynthetic Reaction Center Complex Proteins; Protein Conformation

Photosystem (PS) II membranes, obtained by the method of Berthold et al. (Berthold, D. A., Babcock, G. T., and Yocum, C. F. (1981) FEBS Lett. 134, 231-234), have been fractionated by a sucrose gradient ultracentrifugation method which allows the quantitative separation of the three major chlorophyll binding complexes in these membranes: the chlorophyll (chl) a binding PSII reaction center core, the major light-harvesting complex II, and the minor chl a/b proteins called CP26, CP29, and CP24. Each fraction has been analyzed for its subunit stoichiometry by quantitative sodium dodecyl sulfate-polyacrylamide gel electrophoresis methods. The results show that 12 mol of light-harvesting complex II and 1.5 mol of each of the minor chl a/b proteins are present per mol of the PSII reaction center complex in PSII membranes. These data suggest a dimeric organization of PSII, in agreement with a recent crystallographic study (Bassi, R., Ghiretti Magaldi, A., Tognon, G., Giacometti, G. M., and Miller, K. (1989) Eur. J. Cell Biol. 50, 84-93) and imply that such a dimeric complex is served by antenna chl a/b proteins whose minimal aggregation state includes three polypeptides. This was confirmed by covalent cross-linking of purified antenna complexes.

Topics: Centrifugation, Density Gradient; Chlorophyll; Chlorophyll A; Chloroplasts; Cross-Linking Reagents; Electrophoresis, Polyacrylamide Gel; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Zea mays

A cDNA library was constructed from mRNA prepared from light-treated seedlings of Scots pine (Pinus sylvestris L.) and cDNAs for the chlorophyll a/b-binding protein LHC-II were identified using a pea gene as the heterologous probe. Three cDNA clones were sequenced. The deduced amino acid sequences of two of the genes corresponded to Type I and one to Type II LHC-II proteins which were ca. 90% homologous to their angiosperm counterparts. The transit peptides of the Scots pine preLHC-II showed features common to angiosperm transit peptides. The three cDNAs had a 70 to 75% preference for G + C in the third base position. CpG and GpC profiles and degenerate codon position bias suggested that two of the corresponding genes lie within CpG islands.

Topics: Amino Acid Sequence; Base Sequence; Carrier Proteins; Chlorophyll; Chlorophyll A; Codon; DNA; Light-Harvesting Protein Complexes; Molecular Sequence Data; Photosynthetic Reaction Center Complex Proteins; Pinus sylvestris; Plants; Species Specificity

Topics: Amino Acid Sequence; Base Sequence; Carrier Proteins; Chlorophyll; Chlorophyll A; DNA; Light-Harvesting Protein Complexes; Molecular Sequence Data; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Sequence Homology, Nucleic Acid; Zea mays

We have cloned and characterized members of a gene family encoding polypeptide constituents of the fucoxanthin, chlorophyll a/c protein complex, a light-harvesting complex associated with photosystem II of diatoms and brown algae. Three cDNA clones encoding proteins associated with this complex in the diatom Phaeodactylum tricornutum have been isolated. As deduced from the nucleotide sequences, these light-harvesting proteins show homology to the chlorophyll a/b binding polypeptides of higher plants. Specifically, the N-terminal regions of the fucoxanthin, chlorophyll a/c-binding proteins are homologous to the chlorophyll a/b binding proteins in both the third membrane-spanning domain and the stroma-exposed region between membrane-spanning domains 2 and 3. Like the chlorophyll a/b-binding proteins, the mature fucoxanthin, chlorophyll a/c polypeptides have three hydrophobic alpha-helical domains which could span the membrane bilayer. The similarities between the two light-harvesting proteins might reflect the fact that both bind chlorophyll molecules and/or might be important for maintaining certain structural features of the complex. There is little similarity between the N-terminal sequences of the primary translation products of the fucoxanthin, chlorophyll a/c proteins and any transit sequences that have been characterized. Instead, the N-terminal sequences have features resembling those of signal sequences. Thus either transit peptides used in P. tricornutum show little resemblance to those of higher plants and green algae or the nuclear-encoded plastid proteins enter the organelle via a mechanism different from that used in higher plants.

Topics: Amino Acid Sequence; Base Sequence; Biological Transport; Carotenoids; Carrier Proteins; Chlorophyll; Chlorophyll A; Chloroplasts; Eukaryota; Light-Harvesting Protein Complexes; Molecular Sequence Data; Multigene Family; Phaeophyceae; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plants; Restriction Mapping; Sequence Homology, Nucleic Acid; Xanthophylls

The structure of the light-harvesting chlorophyll a/b-protein complex has been determined at 3.7 A resolution in projection by electron diffraction, electron microscopy and image analysis. Diffraction patterns and high-resolution spotscan images of two-dimensional crystals stabilized with tannin were recorded at low temperature. Phases of structure factors were obtained directly by image processing, after correction of the images for lattice distortions, defocus and beam tilt. Amplitudes were measured by electron diffraction. The projection map shows the detailed structure of the trimeric complex, suggesting the positions of two domains of potential structural and functional homology, of one membrane-spanning alpha-helix approximately perpendicular to the membrane plane and of several tightly bound lipid molecules.

Topics: Chlorophyll; Chlorophyll A; Macromolecular Substances; Pigments, Biological; Plant Proteins; X-Ray Diffraction

The chlorophyll-protein complexes of the thylakoid membrane from Prochlorothrix hollandica were identified following electrophoresis under nondenaturing conditions. Five complexes, CP1-CP5, were resolved and these green bands were analyzed by spectroscopic and immunological methods. CP1 contains the photosystem I (PSI) reaction center, as this complex quenched fluorescence at room temperature, and had a 77 K fluorescence emission peak at 717 nm. CP4 contains the major chlorophyll-a-binding proteins of the photosystem II (PSII) core, because this complex contained polypeptides which cross-reacted to antibodies raised against Chlamydomonas PSII proteins 5 and 6. Furthermore, fluorescence excitation studies at 77 K indicated that only a Chl a is bound to CP4. Complexes CP2, CP3 and CP5 contained functionally bound Chl a and b as judged by absorption spectroscopy at 20 degrees C and fluorescence excitation spectra at 77 K. CP2, CP3 and CP5 all contain polypeptides of 30-33 kDa which are immunologically distinct from the LHC-II complex of higher plant thylakoids.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Electrophoresis, Polyacrylamide Gel; Immunoassay; Light-Harvesting Protein Complexes; Peptides; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Pigments, Biological; Plant Proteins; Protein Binding; Protein Denaturation; Spectrometry, Fluorescence

Phycobiliproteins obtained by dissociation of phycobilisomes were reassociated in vitro with intact thylakoids or isolated photosystems I and II preparations obtained from cyanophytes (prokaryotes) or green algae (eukaryotes) to form bound phycobilisome complexes. Energy transfer from Fremyella diplosiphon phycobiliproteins to chlorophyll a of reaction centers I and II was measured in: complexes containing intact thylakoids of the cyanophytes F. diplosiphon or Anacystis nidulans and the eukaryotic algae Euglena gracilis and mutants of Chlamydomonas reinhardtii; complexes containing isolated photosystem II particles of A. nidulans or C. reinhardtii; and complexes containing reaction center I of F. diplosiphon or C. reinhardtii. Energy transfer from phycoerythrin to chlorophyll a of photosystem II could be demonstrated in complexes containing phycobilisomes bound to cyanophyte thylakoids or isolated photosystem II particles of A. nidulans or C. reinhardtii. Bound phycobilisomes did not transfer energy to photosystem II within green algae thylakoids containing altered forms of light-harvesting chlorophyll a/b-protein complex (LHC) II antenna, reduced amounts of LHC II, or chlorophyll b, or chlorophyll b-less mutants, nor to chlorophyll a of photosystem I of intact thylakoids or isolated reaction centers. We conclude that phycobilisomes can form a specific and functional association with photosystem II particles of both cyanophytes and eukaryotic thylakoids. This interaction appears to be hindered by the presence of LHC II antenna in the eukaryotic thylakoids.

Topics: Chlamydomonas; Chlorophyll; Chlorophyll A; Chlorophyta; Chloroplasts; Electrophoresis, Polyacrylamide Gel; Energy Transfer; Light-Harvesting Protein Complexes; Photosynthetic Reaction Center Complex Proteins; Photosystem I Protein Complex; Photosystem II Protein Complex; Phycobilisomes; Plant Proteins

Several compounds, occurring in food, were tested for antimutagenic activity towards cigarette-smoke condensate (CSC) and benzo[a]pyrene (BaP). Antimutagenicity was determined in the Salmonella/microsome test, with tester strain TA98, in the presence of rat-liver homogenate. Dose-response curves did show reduction of CSC- and BaP-induced mutagenicity by ellagic acid, riboflavin and chlorophyllin. Chlorophyll a and chlorophyll b, although less distinct, also inhibited CSC- and BaP-induced mutagenicity. Ascorbic acid, beta-carotene, tocopherol acetate, chlorogenic acid and butyl hydroxyanisole did not have any influence on the mutagenicity of CSC and BaP. The similarity in results for cigarette-smoke condensate and for BaP indicates that a general mechanism may be involved in the inhibition of CSC- and BaP-induced mutagenicity.

Topics: Animals; Anisoles; Ascorbic Acid; Benzo(a)pyrene; Benzopyrans; beta Carotene; Butylated Hydroxyanisole; Carotenoids; Chlorogenic Acid; Chlorophyll; Chlorophyll A; Chlorophyllides; Dose-Response Relationship, Drug; Ellagic Acid; Male; Mutagenicity Tests; Mutation; Nicotiana; Plants, Toxic; Rats; Riboflavin; Salmonella typhimurium; Smoke; Vitamin E; Vitamins

A chlorophyll deficient mutant of clover has been examined by SDS-PAGE, spectrophotometric and electron microscopic methods. By a comparison of the absorption and first derivative spectra of acetone extracts from the mutant and normal biotypes, we observed a deficiency in chlorophyll b for the mutant biotype. The calculated chlorophyll a/b ratios, using the method of Arnon (Plant Physiol. 24, 1-15, 1949), approached infinity for the mutant whereas it ranged from 3.0-3.3 in the wild-type. The low temperature (77 degrees K) fluorescence emission bands in the 685-695 nm region could not be differentiated for the biotypes; however, the long wavelength emission band (near 740 nm in the wild-type) was shifted to shorter wavelengths (ca 720-725 nm) in the mutant indicating loss of photosystem I antenna. The SDS-PAGE profile of the mutant biotype showed a dramatic decline in the Coomassie stained polypeptides of apparent molecular weights similar to those of LHC II. Transmission electron micrographs of the mutant and normal tissue exhibited similar extents of grana-stacking, indicating that a component(s) other than the LHC II may be responsible for membrane adhesion in this mutant.

Topics: Chlorophyll; Chlorophyll A; Chloroplasts; Electrophoresis, Polyacrylamide Gel; Microscopy, Electron; Mutation; Photosynthesis; Plants; Spectrometry, Fluorescence; Spectrophotometry

Chlorophyll a and chlorophyll b have been inserted into reconstituted envelopes of Sendai virus particles. Fluorescence measurements indicated a high efficiency of energy transfer between the two chlorophyll molecules due to their close proximity in the viral envelope. Fusion of reconstituted, pigmented virus envelopes with various biological cell membranes at 37 degrees C resulted in a significant decrease in the yield of energy transfer. Reduction in the efficiency of energy transfer was temperature and time dependent, and was also dependent upon the ratio between the reconstituted Sendai virus envelopes (donor) and recipient cells (acceptor). No reduction in the efficiency of energy transfer was observed when non-fusogenic, reconstituted viral envelopes were incubated with cell membranes.

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Membrane; Chlorophyll; Chlorophyll A; Energy Transfer; Erythrocyte Membrane; Humans; Membrane Fusion; Mice; Parainfluenza Virus 1, Human; Rats; Spectrometry, Fluorescence; Synaptic Membranes

On the edge of Don Juan Pond in the Wright Valley of Antarctica lies a mat of mineral and detritus cemented by organic matter. In spite of a CaCl2 concentration of about 33% (w/v), the mat contains Oscillatoria and other cyanobacteria, unicellular forms, colonial forms rich in carotenoids, and diatoms. Bacteria are rare; fungal filaments are not. Oscillatoria showed motility but only at temperatures <10 degrees C. Acetone extracts of the mat and nearby muds yielded visible spectra similar to those of laboratory grown O. sancta, with 50- to 70-fold molar ratio of chlorophyll a to b. Although rare, tardigrades were also found. The algal mat had enzymatic activities characteristic of peroxidase, catalase, dehydrogenase, and amylase. Cellulose, chitin, protein, lipid and ATP were present. Previously, algae in the Wright Valley have been described in melt water, not in the brine itself. Wright Valley has been used as a near sterile Martian model. It obviously contains an array of hardy terrestrial organisms.

Topics: Adenosine Triphosphate; Animals; Antarctic Regions; Calcium Chloride; Cellulose; Chitin; Chlorophyll; Chlorophyll A; Cyanobacteria; Desert Climate; Environmental Microbiology; Geologic Sediments; Invertebrates; Lipids; Mars; Proteins

The increase in the ratio [chlorophyll a]/[chlorophyll b] which occurs in expanding bean leaves in the dark, is a result of chlorophyll a formation with a concomitant loss in chlorophyll b. The analytical methods for assay of these pigments were examined closely and found to be adequate for this purpose when proper safeguards are taken.

Topics: Chlorophyll; Chlorophyll A; Darkness; Plant Leaves; Plants

Topics: Chemistry Techniques, Analytical; Chlorophyll; Chlorophyll A; Pheophytins; Research; Water

Topics: Bacteria; Bacteriochlorophylls; Biochemical Phenomena; Biochemistry; Chlorobium; Chlorophyll; Chlorophyll A; Hydrogen; Magnetic Resonance Spectroscopy; Research; Rhodospirillum

This paper shows that the "second Emerson effect"1 exists not only in photosynthesis, but also in the quinone reduction (Hill reaction), in Chlorella pyrenoidosa and Anacystis nidulans. The peaks at 650 mmu, 600 mmu, 560 mmu, 520 mmu, and 480 mmu, observed in the action spectrum of this effect in the Hill reaction in Chorella, are attributable to chlorophyll b; the occurrence of an additional peak at 670 mmu, 620 mmu, and of two (or three) peaks in the blueviolet region suggests that (at least) one form of chlorophyll a contributes to it. In analogy to suggestions made previously in the interpretation of the Emerson effect in photosynthesis, these results are taken as indicating that excitation by light preferentially absorbed by one (or two) forms of chlorophyll a (Chl a 690 + 700), needs support by simultaneous absorption of light in another form of chlorophyll a (Chl a 670)-directly or via energy transfer from chlorophyll b-in order to produce the Hill reaction with its full quantum yield. In Anacystis, the participation of phycocyanin in the Emerson effect in the Hill reaction is revealed by the occurrence, in the action spectrum of this effect, of peaks at about 560 mmu, 610 mmu, and 640 mmu; a peak at 670 mmu, due to Chl a 670, also is present.

Topics: Chlorophyll; Chlorophyll A; Chlorophyta; Energy Transfer; Eukaryota; Light; Photosynthesis; Phycocyanin; Quinones

Action spectra of the Emerson effect in Chlorella and Navicula reveal peaks at 670 mut, in addition to those at 650 mu (Chlorella) and 630 mmicro (Navicula) attributable to chlorophylls b and c. Thus, excitation of chlorophyll a form "chlorophyll a 670" can supplement, in these algae, the excitation of the form "chlorophyll a 680-700." The effect of the auxiliary pigments in these algae may be mediated by energy transfer to "chlorophyll a 670."

Topics: Chlorophyll; Chlorophyll A; Energy Transfer; Plants