chlorophyll-a and 3-3-dimethylallyl-pyrophosphate

Role of terpenes and isoprenoids has been pivotal in the survival and evolution of higher plants in various ecoregions. These products find application in the pharmaceutical, flavor fragrance, and biofuel industries. Fitness of plants in a wide range of environmental conditions entailed (i) evolution of secondary metabolic pathways enabling utilization of photosynthate for the synthesis of a variety of biomolecules, thereby facilitating diverse eco-interactive functions, and (ii) evolution of structural features for the sequestration of such compounds away from the mainstream primary metabolism to prevent autotoxicity. This review summarizes features and applications of terpene and isoprenoid compounds, comprising the largest class of secondary metabolites. Many of these terpene and isoprenoid biomolecules happen to be high-value bioproducts. They are essential components of all living organisms that are chemically highly variant. They are constituents of primary (quinones, chlorophylls, carotenoids, steroids) as well as secondary metabolism compounds with roles in signal transduction, reproduction, communication, climatic acclimation, defense mechanisms and more. They comprise single to several hundreds of repetitive five-carbon units of isopentenyl diphosphate (IPP) and its isomer dimethylallyl diphosphate (DMAPP). In plants, there are two pathways that lead to the synthesis of terpene and isoprenoid precursors, the cytosolic mevalonic acid (MVA) pathway and the plastidic methylerythritol phosphate (MEP) pathway. The diversity of terpenoids can be attributed to differential enzyme and substrate specificities and to secondary modifications acquired by terpene synthases. The biological role of secondary metabolites has been recognized as pivotal in the survival and evolution of higher plants. Terpenes and isoprenoids find application in pharmaceutical, nutraceutical, synthetic chemistry, flavor fragrance, and possibly biofuel industries.

Topics: Carotenoids; Chlorophyll; Hemiterpenes; Organophosphorus Compounds; Quinones; Terpenes

Both OsIPPI1 and OsIPPI2 enzymes are found in the endoplasmic reticulum, providing novel important insights into the role of this compartment in the synthesis of MVA pathway isoprenoids. Isoprenoids are synthesized from the precursor's isopentenyl diphosphate (IPP) and dimethylallyl diphosphosphate (DMAPP), which are interconverted by the enzyme isopentenyl diphosphate isomerase (IPPI). Many plants express multiple isoforms of IPPI, the only enzyme shared by the mevalonate (MVA) and non-mevalonate (MEP) pathways, but little is known about their specific roles. Rice (Oryza sativa) has two IPPI isoforms (OsIPPI1 and OsIPPI2). We, therefore, carried out a comprehensive comparison of IPPI gene expression, protein localization, and isoprenoid biosynthesis in this species. We found that OsIPPI1 mRNA was more abundant than OsIPPI2 mRNA in all tissues, and its expression in de-etiolated leaves mirrored the accumulation of phytosterols, suggesting a key role in the synthesis of MVA pathway isoprenoids. We investigated the subcellular localization of both isoforms by constitutively expressing them as fusions with synthetic green fluorescent protein. Both proteins localized to the endoplasmic reticulum (ER) as well as peroxisomes and mitochondria, whereas only OsIPPI2 was detected in plastids, due to an N-terminal transit peptide which is not present in OsIPPI1. Despite the plastidial location of OsIPPI2, the expression of OsIPPI2 mRNA did not mirror the accumulation of chlorophylls or carotenoids, indicating that OsIPPI2 may be a redundant component of the MEP pathway. The detection of both OsIPPI isoforms in the ER indicates that DMAPP can be synthesized de novo in this compartment. Our work shows that the ER plays an as yet unknown role in the synthesis of MVA-derived isoprenoids, with important implications for the metabolic engineering of isoprenoid biosynthesis in higher plants.

Topics: Carbon-Carbon Double Bond Isomerases; Carotenoids; Chlorophyll; Endoplasmic Reticulum; Gene Expression Regulation, Plant; Hemiterpenes; Mevalonic Acid; Mitochondria; Organophosphorus Compounds; Oryza; Peroxisomes; Plant Leaves; Plants, Genetically Modified; Plastids; Terpenes

When inoculated onto maize leaves at the onset of senescence, the hemibiotroph Colletotrichum graminicola causes green islands that are surrounded by senescing tissue. Taking advantage of green islands as indicators of sites of the establishment of successful infection and of advanced high-performance liquid chromatography tandem mass spectrometry methodology, we analyzed changes in the patterns and levels of cytokinins (CK) at high spatial and analytical resolution. Twenty individual CK were detected in green islands. Levels of cis-zeatin-9-riboside and cis-zeatin-9-riboside-5'-monophosphate increased drastically, whereas that of the most prominent CK, cis-zeatin-O-glucoside, decreased. The fungus likely performed these conversions because corresponding activities were also detected in in vitro cultures amended with CK. We found no evidence that C. graminicola is able to synthesize CK entirely de novo in minimal medium but, after adding dimethylallyl diphosphate, a precursor of CK biosynthesis occurring in plants, a series of trans-zeatin isoforms (i.e., trans-zeatin-9-riboside-5'-monophosphate, trans-zeatin-9-riboside, and trans-zeatin) was formed. After applying CK onto uninfected leaves, transcripts of marker genes for senescence, photosynthesis, and assimilate distribution were measured by quantitative reverse-transcribed polymerase chain reaction; furthermore, pulse-amplitude modulation chlorophyll fluorometry and single-photon avalanche diode analyses were conducted. These experiments suggested that modulation of CK metabolism at the infection site affects host physiology.

Topics: Chlorophyll; Chromatography, High Pressure Liquid; Colletotrichum; Culture Media; Cytokinins; Gene Expression Regulation, Plant; Hemiterpenes; Host-Pathogen Interactions; Isopentenyladenosine; Organophosphorus Compounds; Photosynthesis; Plant Diseases; Plant Leaves; Purine Nucleotides; Tandem Mass Spectrometry; Zea mays

The physiological role of isoprene emission in plants is a matter of much debate. One of the most widely propagated hypotheses suggests a function of isoprene in the protection of leaf physiological processes against thermal and oxidative stress. To test this hypothesis, we developed transgenic Grey poplar (Populusxcanescens) plants in which gene expression of isoprene synthase (ISPS) was either silenced by RNA interference (RNAi) or upregulated by over-expression of the ISPS gene. Despite increased ISPS mRNA levels, we did not observe consistent increases in isoprene emission in the over-expressing lines, indicating post-transcriptional control of ISPS by co-suppression. In the RNAi lines, levels of isoprene emission were effectively suppressed to virtually zero. Transgenic plants were subjected to temperature stress with three transient heat phases of 38-40 degrees C, each followed by phases of recovery at 30 degrees C. Parallel measurements of gas exchange, chlorophyll fluorescence and isoprene emission provided new insights into the physiological link between isoprene and enhanced temperature tolerance. Transgenic non-isoprene-emitting poplars showed reduced rates of net assimilation and photosynthetic electron transport during heat stress, but not in the absence of stress. The decrease in the efficiency of photochemistry was inversely correlated with the increase in heat dissipation of absorbed light energy, measured as NPQ (non-photochemical quenching). Isoprene-repressed poplars also displayed an increased formation of the xanthophyll cycle pigment zeaxanthin in the absence of stress, which can cause increased NPQ or may indicate an increased requirement for antioxidants. In conclusion, using a molecular genetic approach, we show that down-regulation of isoprene emission affects thermotolerance of photosynthesis and induces increased energy dissipation by NPQ pathways.

Topics: Acclimatization; Alkyl and Aryl Transferases; Butadienes; Carbon Dioxide; Chlorophyll; Electron Transport; Hemiterpenes; Hot Temperature; Organophosphorus Compounds; Pentanes; Photosynthesis; Pigments, Biological; Plants, Genetically Modified; Populus; RNA Interference

In cyanobacteria many compounds, including chlorophylls, carotenoids, and hopanoids, are synthesized from the isoprenoid precursors isopentenyl diphosphate (IPP) and dimethylallyl diphosphate. Isoprenoid biosynthesis in extracts of the cyanobacterium Synechocystis strain PCC 6803 grown under photosynthetic conditions, stimulated by pentose phosphate cycle substrates, does not appear to require methylerythritol phosphate pathway intermediates. The sll1556 gene, distantly related to type 2 IPP isomerase genes, was disrupted by insertion of a Kanr cassette. The mutant was fully viable under photosynthetic conditions although impaired in the utilization of pentose phosphate cycle substrates. Compared to the parental strain the Deltasll1556 mutant (i) is deficient in isoprenoid biosynthesis in vitro with substrates including glyceraldehyde-3-phosphate, fructose-6-phosphate, and glucose-6-phosphate; (ii) has smaller cells (diameter ca. 13% less); (iii) has fewer thylakoids (ca. 30% less); and (iv) has a more extensive fibrous outer wall layer. Isoprenoid biosynthesis is restored with pentose phosphate cycle substrates plus the recombinant Sll1556 protein in the Deltasll1556 supernatant fraction. IPP isomerase activity could not be demonstrated for the purified Sll1556 protein under our in vitro conditions. The reduction of thylakoid area and the effect on outer wall layer components are consistent with an impairment of isoprenoid biosynthesis in the mutant, possibly via hopanoid biosynthesis. Our findings are consistent with an alternate metabolic shunt for biosynthesis of isoprenoids.

Topics: Bacterial Proteins; Carotenoids; Cell Wall; Chlorophyll; Cyanobacteria; Fructosephosphates; Gene Silencing; Genes, Bacterial; Glucose-6-Phosphate; Glyceraldehyde 3-Phosphate; Hemiterpenes; Mutagenesis, Insertional; Organophosphorus Compounds; Pentose Phosphate Pathway; Terpenes; Thylakoids