jasmonic-acid and zeatin-riboside

Fermentation broth and biomass from three strains of Botryodiplodia theobromae were characterized by high performance liquid chromatography-electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) method, in order to quantify different phytohormones and to identify amino acid conjugates of jasmonic acid (JA) present in fermentation broths. A liquid-liquid extraction with ethyl acetate was used as sample preparation. The separation was carried out on a C18 reversed-phase HPLC column followed by analysis via ESI-MS/MS. The multiple reaction monitoring mode was used for quantitative measurement. For the first time, indole-3-acetic acid, indole-3-propionic acid, indole-3-butyric acid and JA were identified and quantified in the ethyl acetate extracts from the biomass, after the separation of mycelium from supernatant. The fermentation broths showed significantly higher levels of JA in relation to the other phytohormones. This is the first report of the presence of gibberellic acid, abscisic acid, salicylic acid and the cytokinins zeatin, and zeatin riboside in fermentation broths of Botryodiplodia sp. The presence of JA-serine and JA-threonine conjugates in fermentation broth was confirmed using HPLC-ESI tandem mass spectrometry in negative ionization mode, while the occurrence of JA-glycine and JA-isoleucine conjugates was evidenced with the same technique but with positive ionization. The results demonstrated that the used HPLC-ESI-MS/MS method was effective for analysing phytohormones in fermentation samples.

Topics: Abscisic Acid; Ascomycota; Chromatography, High Pressure Liquid; Cyclopentanes; Fermentation; Gibberellins; Indoleacetic Acids; Indoles; Isopentenyladenosine; Oxylipins; Plant Growth Regulators; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Zeatin

Organogenesis in peach (Prunus persica L. Batsch) and peach rootstocks (P. persica × Prunus dulcis) has been achieved and the action of the regeneration medium on 7 phytohormones, zeatin (Z), zeatin riboside (ZR), indole-3-acetic acid (IAA), abscisic acid (ABA), ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), salicylic acid (SA), and jasmonic acid (JA), has been studied using High performance liquid chromatography - mass spectrometry (HPLC-MS/MS). Three scion peach cultivars, 'UFO-3', 'Flariba' and 'Alice Bigi', and the peach × almond rootstocks 'Garnem' and 'GF677' were cultured in two different media, Murashige and Skoog supplemented with plant growth regulators (PGRs) (regeneration medium) and without PGRs (control medium), in order to study the effects of the media and/or genotypes in the endogenous hormones content and their role in organogenesis. The highest regeneration rate was obtained with the peach × almond rootstocks and showed a lower content of Z, IAA, ABA, ACC and JA. Only Z, ZR and IAA were affected by the action of the culture media. This study shows which hormones are external PGRs-dependent and what is the weight of the genotype and hormones in peach organogenesis that provide an avenue to manipulate in vitro organogenesis in peach.

Topics: Abscisic Acid; Amino Acids, Cyclic; Cyclopentanes; Cytokinins; Indoleacetic Acids; Isopentenyladenosine; Organogenesis; Oxylipins; Plant Growth Regulators; Prunus; Salicylic Acid; Zeatin

A method for the simultaneous determination of indole-3-acetic acid, N6-(2-isopentenyl) adenosine, N6-(2-isopentenyl) adenine, trans-zeatin riboside, zeatin, strigolactone, abscisic acid, salicylic acid, gibberellin A3 and jasmonic acid in five different parts of main branch, lateral branch, primary leaf, secondary leaf and stem of Sargasum fusiforme (Hary.) Seichell was established by high performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-TQMS). The samples were extracted with methanol/water/formic acid (15 : 4 :1, v/v/v) (containing 0.5% 2, 6-di-tert-butyl-4-methylphenol, BHT) after vacuum freeze-drying. The separation was performed on a Hypersil Gold C18 column by using methanol and water as mobile phases with gradient elution. The analytes were detected by tandem mass spectrometry in selected reaction monitoring (SRM) mode. The electrospray ionization (ESI) source was used for the quantitative analysis in the positive mode or negative mode. Under the optimized conditions, the correlation coefficients (r) of the ten phytohormones were from 0. 9989 to 1. 0000 in the linear ranges. The detection limits of the ten phytohormones were 0. 001 2-4. 651 2 μ/L. The average recoveries were 72. 24% -91. 31% with the relative standard deviations not more than 6. 59%. In the five parts of fresh Sargasum fusiforme (Hary.) Seichell samples, the contents of the ten phytohormones were from not detected to 4 041. 431 ng/g. This method has good sensitivity, precision, recovery, and can be used to simultaneously determine the phytohormones.

Topics: Abscisic Acid; Chromatography, High Pressure Liquid; Cyclopentanes; Gibberellins; Indoleacetic Acids; Isopentenyladenosine; Oxylipins; Plant Growth Regulators; Reproducibility of Results; Sargassum; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Pinus radiata D. Don is one of the most abundant species in the north of Spain. Knowledge of drought response mechanisms is essential to guarantee plantation survival under reduced water supply as predicted in the future. Tolerance mechanisms are being studied in breeding programs, because information on such mechanisms can be used for genotype selection. In this paper, we analyze the changes of leaf water potential, hydraulic conductance (K(leaf)), stomatal conductance and phytohormones under drought in P. radiata breeds (O1, O2, O3, O4, O5 and O6) from different climatology areas, hypothesizing that they could show variable drought tolerance. As a primary signal, drought decreased cytokinin (zeatin and zeatin riboside-Z + ZR) levels in needles parallel to K(leaf) and gas exchange. When Z + ZR decreased by 65%, indole-3-acetic acid (IAA) and abscisic acid (ABA) accumulation started as a second signal and increments were higher for IAA than for ABA. When plants decreased by 80%, Z + ZR and K(leaf) doubled their ABA and IAA levels, the photosystem II yield decreased and the electrolyte leakage increased. At the end of the drought period, less tolerant breeds increased IAA over 10-fold compared with controls. External damage also induced jasmonic acid accumulation in all breeds except in O5 (P. radiata var. radiata × var. cedrosensis), which accumulated salicylic acid as a defense mechanism. After rewatering, only the most tolerant plants recovered their K(leaf,) perhaps due to an IAA decrease and 1-aminocyclopropane-1-carboxylic acid maintenance. From all phytohormones, IAA was the most representative 'water deficit signal' in P. radiata.

Topics: Abscisic Acid; Adaptation, Physiological; Amino Acids, Cyclic; Breeding; Climate; Cyclopentanes; Droughts; Electrolytes; Genotype; Indoleacetic Acids; Isopentenyladenosine; Oxylipins; Photosynthesis; Photosystem II Protein Complex; Pinus; Plant Growth Regulators; Plant Leaves; Plant Stomata; Salicylic Acid; Signal Transduction; Spain; Stress, Physiological; Water; Zeatin

Through analysis of variation and function of 5 main endogenous hormones in the formation of microtuber of Dioscorea opposite in vitro to explore the physiological and biochemical mechanism of microtuber development.. When microtubers were induced on MS + 6-BA 1.5 mg x L(-1) + NAA 1.5 mg x L(-1) + sucrose 5% medium, the endogenous hormones were isolated during different formation stages of microtubers, then purified and detected with enzyme-linked immunosorbent assays (ELISA).. The results showed that GA3 slightly decreased in initial period, rose suddenly 20 days later, and than decreased. IAA showed a dropping tendency in the total course, ABA and ZR increased in a long period, dropped at last. JA continuously rose and never dropped, GA3 and ABA and the ratio of GA3 and JA varied obviously.. IAA, ABA, JA , ZR and GA3 play an important role in controlling formation of microtubers in D. opposite in vitro.

Topics: Abscisic Acid; Cyclopentanes; Dioscorea; Gibberellins; Indoleacetic Acids; Isopentenyladenosine; Oxylipins; Plant Growth Regulators; Plant Tubers

Stinging nettle (Urtica dioica) is a herbaceous, dioecious perennial that is widely distributed around the world, reproduces both sexually and asexually, and is characterized by rapid growth. This work was aimed at evaluating the effects of plant maturity, shoot reproduction and sex on the growth of leaves and shoots.. Growth rates of apical shoots, together with foliar levels of phytohormones (cytokinins, auxins, absicisic acid, jasmonic acid and salicylic acid) and other indicators of leaf physiology (water contents, photosynthetic pigments, alpha-tocopherol and F(v)/F(m) ratios) were measured in juvenile and mature plants, with a distinction made between reproductive and non-reproductive shoots in both males and females. Vegetative growth rates were not only evaluated in field-grown plants, but also in cuttings obtained from these plants. All measurements were performed during an active vegetative growth phase in autumn, a few months after mature plants reproduced during spring and summer.. Vegetative growth rates in mature plants were drastically reduced compared with juvenile ones (48 % and 78 % for number of leaves and leaf biomass produced per day, respectively), which was associated with a loss of photosynthetic pigments (up to 24 % and 48 % for chlorophylls and carotenoids, respectively) and increases of alpha-tocopherol (up to 2.7-fold), while endogenous levels of phytohormones did not differ between mature and juvenile plants. Reductions in vegetative growth were particularly evident in reproductive shoots of mature plants, and occurred similarly in both males and females.. It is concluded that (a) plant maturity reduces vegetative growth in U. dioica, (b) effects of plant maturity are evident both in reproductive and non-reproductive shoots, but particularly in the former, and (c) these changes occur similarly in both male and female plants.

Topics: Abscisic Acid; alpha-Tocopherol; Chlorophyll; Cyclopentanes; Cytokinins; Indoleacetic Acids; Isopentenyladenosine; Oxylipins; Plant Leaves; Plant Shoots; Reproduction; Salicylic Acid; Urtica dioica; Zeatin

To study the change of endogenous hormone (ABA, IAA, JA, GA3, ZR) in the leaves, petioles, tubers of Pinellia ternate around sprout tumble. It also provided some valuable information to prevent sprout tumble and increase production.. Tubers of P. ternata were cultured firstly at (23 +/- 1) degree C for certain days, and then they were coerced under (30 +/- 1 ) degree C stress in the same artificial climate boxes. The endogenous hormones in leaves, petioles and tubers during different stages of high temperature stress were determined with Enzyme-linked Immunosorbent Assays (ELISA).. After under high temperature stress, ABA content in leaves, petioles and tubers increased obviously. Similarly, JA content rose all in the leaves, petioles and tubers. But in the same conditions IAA content declined significantly in the leaves and petioles. In the tubers, IAA content also decreased, but not quickly. With the extension of high temperature coercion, the leaves, petioles, tubers, ZR content were gradually falling off. In the leaves of GA3 content rose markedly at the third day, fell down at the sixth day, but remained higher than before treatment. With the extension of the processing time, GA3 content fell off in the petioles and tubers.. ABA, JA, ZT and GA3 played an important role in controlling sprout tumble of P. ternata.

Topics: Abscisic Acid; Cyclopentanes; Enzyme-Linked Immunosorbent Assay; Gibberellins; Hot Temperature; Indoleacetic Acids; Isopentenyladenosine; Oxylipins; Pinellia; Plant Growth Regulators; Plant Leaves; Plant Tubers; Plants, Medicinal