Jasmonic Acid Methyl Ester Research Articles

VpUR9, a novel RING-type ubiquitin ligase gene from Vitis pseudoreticulata, is involved in powdery mildew response in transgenic V. vinifera plants

Ubiquitination plays important roles in disease resistance in plants. We report the identification and functional characterization of the RING-type ubiquitin ligase gene VpUR9 from Chinese wild Vitis pseudoreticulata accession Baihe-35-1. VpUR9, encodes 164 amino acids and possesses a RING conserved motif. It is homologously cloned from the cDNA library of the high powdery mildew (Erysiphe necator [Schw.] Burr) resistant V. pseudoreticulata accession Baihe-35-1 inoculated with E. necator. The gene is induced in response to powdery mildew and salicylic acid. VpUR9 fused with FLAG-tag controlled by 35S promoter was transformed into 15 regenerated V. vinifera L. cv. Red Globe lines via Agrobacterium tumefaciens-mediated transformation. Twelve of these lines were confirmed by Western blot of FLAG-tag. As a result, the powdery mildew-resistance of Red Globe transformed with VpUR9 was repressed. Furthermore, the expression of some disease-resistant related genes (NPR1, PR1, PR10 and PAL) of the transgenic Red Globe declined compared with wild type grapes when inoculated with powdery mildew or salicylic acid. When treated with jasmonic acid methyl ester, its PR1 gene expression decreased, while the expressions of NPR1, PR10 and PAL all increased, contrasting with the wild type grape.

Stable Internal Reference Genes for Normalizing Real-Time Quantitative PCR in Baphicacanthus cusia under Hormonal Stimuli and UV Irradiation, and in Different Plant Organs.

Baphicacanthus cusia (Nees) Bremek, the plant source for many kinds of drugs in traditional Chinese medicine, is widely distributed in South China, especially in Fujian. Recent studies about B. cusia mainly focus on its chemical composition and pharmacological effects, but further analysis of the plant's gene functions and expression is required to better understand the synthesis of its effective compounds. Real-time quantitative polymerase chain reaction (RT-qPCR) is a powerful method for gene expression analysis. It is necessary to select a suitable reference gene for expression normalization to ensure the accuracy of RT-qPCR results. Ten candidate reference genes were selected from the transcriptome datasets of B. cusia in this study, and the expression stability was assessed across 60 samples representing different tissues and organs under various conditions, including ultraviolet (UV) irradiation, hormonal stimuli (jasmonic acid methyl ester and abscisic acid), and in different plant organs. By employing different algorithms, such as geNorm, NormFinder, and BestKeeper, which are complementary approaches based on different statistical procedures, 18S rRNA was found to be the most stable gene under UV irradiation and hormonal stimuli, whereas ubiquitin-conjugating enzyme E2 was the best suitable gene for different plant organs. This novel study aimed to screen for suitable reference genes and corresponding primer pairs specifically designed for gene expression studies in B. cusia, in particular for RT-qPCR analyses.

Methyl jasmonate-dependent senescence of cotyledons in Ipomoea nil

Jasmonic acid methyl ester (JAMe) has been recently shown to play a crucial role in many physiological processes. In this paper, we focused on cotyledon senescence in Ipomoea nil and revealed that JAMe and darkness are the main factors stimulating the process examined. What is more, we showed that mefenamic acid (a jasmonate biosynthesis inhibitor) reverses the stimulatory effect of darkness on senescence. In plants growing under dark conditions, stimulation of JASMONIC ACID CARBOXYL METHYLTRANSFERASE (InJMT) expression and, consequently, an increase in JAMe content, have been observed. In turn, the level of jasmonic acid (JA) gradually decreased. Moreover, dark-grown seedlings demonstrated a lower PSII functional activity and a reduced chlorophyll content and autofluorescence. All of these data suggest that JAMe is a signal molecule controlling the senescence of cotyledons in I. nil.

Genome-Wide Identification and Analysis of the MYB Transcription Factor Superfamily in Solanum lycopersicum.

MYB proteins constitute one of the largest transcription factor families in the plant kingdom, members of which perform a variety of functions in plant biological processes. However, there are only very limited reports on the characterization of MYB transcription factors in tomato (Solanum lycopersicum). In our study, a total of 127 MYB genes have been identified in the tomato genome. A complete overview of these MYB genes is presented, including the phylogeny, gene structures, protein motifs, chromosome locations and expression patterns. The 127 SlMYB proteins could be classified into 18 subgroups based on domain similarity and phylogenetic topology. Phylogenetic analysis of SlMYBs along with MYBs from Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa) indicated 14 subfamilies. Conserved motifs outside the MYB domain may reflect their functional conservation. The identified tomato MYB genes were distributed on 12 chromosomes at various densities but mainly in chromosomes 6 and 10 (12.6% and 11.8%, respectively). Genome-wide segmental and tandem duplications were also found, which may contribute to the expansion of SlMYB genes. RNA-sequencing and microarray data revealed tissue-specific and stress-responsive expression patterns of SlMYB genes. The expression profiles of SlMYB genes in response to salicylic acid (SA) and jasmonic acid methyl ester (MeJA) were also investigated by real-time PCR. Moreover, ethylene-responsive element-binding factor-associated amphiphilic repression (EAR) motifs were found in 24 SlMYB proteins. Collectively, our comprehensive analysis of SlMYB genes will facilitate future functional studies of the tomato MYB gene family and probably other Solanaceae plants.

The Novel Gene VpPR4-1 from Vitis pseudoreticulata Increases Powdery Mildew Resistance in Transgenic Vitis vinifera L.

Pathogenesis-related proteins (PRs) can lead to increased resistance of the whole plant to pathogen attack. Here, we isolate and characterize a PR-4 protein (VpPR4-1) from a wild Chinese grape Vitis pseudoreticulata which shows greatly elevated transcription following powdery mildew infection. Its expression profiles under a number of abiotic stresses were also investigated. Powdery mildew, salicylic acid, and jasmonic acid methyl ester significantly increased the VpPR4-1 induction while NaCl and heat treatments just slightly induced VpPR4-1 expression. Abscisic acid and cold treatment slightly affected the expression level of VpPR4-1. The VpPR4-1 gene was overexpressed in 30 regenerated V. vinifera cv. Red Globe via Agrobacterium tumefaciens-mediated transformation and verified by the Western blot. The 26 transgenic grapevines exhibited higher expression levels of PR-4 protein content than wild-type vines and six of them were inoculated with powdery mildew which showed that the growth of powdery mildew was repressed. The powdery mildew-resistance of Red Globe transformed with VpPR4-1 was enhanced inoculated with powdery mildew. Moreover, other powdery mildew resistant genes were associated with feedback regulation since VpPR4-1 is in abundance. This study demonstrates that PR-4 protein in grapes plays a vital role in defense against powdery mildew invasion.

Drought-responsive WRKY transcription factor genes TaWRKY1 and TaWRKY33 from wheat confer drought and/or heat resistance in Arabidopsis.

BackgroundDrought stress is one of the major causes of crop loss. WRKY transcription factors, as one of the largest transcription factor families, play important roles in regulation of many plant processes, including drought stress response. However, far less information is available on drought-responsive WRKY genes in wheat (Triticum aestivum L.), one of the three staple food crops.ResultsForty eight putative drought-induced WRKY genes were identified from a comparison between de novo transcriptome sequencing data of wheat without or with drought treatment. TaWRKY1 and TaWRKY33 from WRKY Groups III and II, respectively, were selected for further investigation. Subcellular localization assays revealed that TaWRKY1 and TaWRKY33 were localized in the nuclei in wheat mesophyll protoplasts. Various abiotic stress-related cis-acting elements were observed in the promoters of TaWRKY1 and TaWRKY33. Quantitative real-time PCR (qRT-PCR) analysis showed that TaWRKY1 was slightly up-regulated by high-temperature and abscisic acid (ABA), and down-regulated by low-temperature. TaWRKY33 was involved in high responses to high-temperature, low-temperature, ABA and jasmonic acid methylester (MeJA). Overexpression of TaWRKY1 and TaWRKY33 activated several stress-related downstream genes, increased germination rates, and promoted root growth in Arabidopsis under various stresses. TaWRKY33 transgenic Arabidopsis lines showed lower rates of water loss than TaWRKY1 transgenic Arabidopsis lines and wild type plants during dehydration. Most importantly, TaWRKY33 transgenic lines exhibited enhanced tolerance to heat stress.ConclusionsThe functional roles highlight the importance of WRKYs in stress response.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-016-0806-4) contains supplementary material, which is available to authorized users.

Salt tolerance function of the novel C2H2-type zinc finger protein TaZNF in wheat

The expression profile chip of the wheat salt-tolerant mutant RH8706-49 was investigated under salt stress in our laboratory. Results revealed a novel gene induced by salt stress with unknown functions. The gene was named as TaZNF (Triticum aestivum predicted Dof zinc finger protein) because it contains the zf-Dof superfamily and was deposited in GenBank (accession no. KF307327). Further analysis showed that TaZNF significantly improved the salt-tolerance of transgenic Arabidopsis. Various physiological indices of the transgenic plant were improved compared with those of the control after salt stress. Non-invasive micro-test (NMT) detection showed that the root tip of transgenic Arabidopsis significantly expressed Na+ excretion. TaZNF is mainly localized in the nucleus and exhibited transcriptional activity. Hence, this protein was considered a transcription factor. The TaZNF upstream promoter was then cloned and was found to contain three salts, one jasmonic acid methyl ester (MeJA), and several ABA-responsive elements. The GUS staining and quantitative results of different tissues in the full-length promoter in the transgenic plants showed that the promoter was not tissue specific. The promoter activity in the root, leaf, and flower was enhanced after induction by salt stress. Moreover, GUS staining and quantitative measurement of GUS activity showed that the promoter sequence contained the positive regulatory element of salt and MeJA after their respective elements were mutated in the full-length promoter. RNA-Seq result showed that 2727 genes were differentially expressed; most of these genes were involved in the metabolic pathway and biosynthesis of secondary metabolite pathway.

Eugenol confers resistance to Tomato yellow leaf curl virus (TYLCV) by regulating the expression of SlPer1 in tomato plants

Tomato yellow leaf curl virus (TYLCV) is one of the most devastating plant diseases, and poses a significant agricultural concern because of the lack of an efficient control method. Eugenol is a plant-derived natural compound that has been widely used as a food additive and in medicine. In the present study, we demonstrated the potential of eugenol to enhance the resistance of tomato plants to TYLCV. The anti-TYLCV efficiency of eugenol was significantly higher than that of moroxydine hydrochloride (MH), a widely used commercial antiviral agent. Eugenol application stimulated the production of endogenous nitric oxide (NO) and salicylic acid (SA) in tomato plants. The full-length cDNA of SlPer1, which has been suggested to be a host R gene specific to TYLCV, was isolated from tomato plants. A sequence analysis suggested that SlPer1 might be a nucleobase-ascorbate transporter (NAT) belonging to the permease family. The transcript levels of SlPer1 increased markedly in response to treatment with eugenol or TYLCV inoculation. The results of this study also showed that SlPer1 expression was strongly induced by SA, MeJA (jasmonic acid methyl ester), and NO. Thus, we propose that the increased transcription of SlPer1 contributed to the high anti-TYLCV efficiency of eugenol, which might involve in the generation of endogenous SA and NO. Such findings provide the basis for the development of eugenol as an environmental-friendly agricultural antiviral agent.

Auxin increases the InJMT expression and the level of JAMe – inhibitor of flower induction in Ipomoea nil

Interactions among jasmonates and auxin in the photoperiodic flower induction of a short-day plant <em>Ipomoea nil</em> were examined. Therefore, we measured changes in jasmonic acid (JA) and jasmonic acid methyl ester (JAMe) levels in the cotyledons of <em>I. nil</em> during the inductive night, as well as the effects of indole-3-acetic acid (IAA) on their content. We noticed an interesting result, that IAA applied on the cotyledons of <em>I. nil</em> is an effective stimulator of JAMe production in seedlings cultivated under inductive night conditions. IAA treatment also significantly increased the transcriptional activity of <em>InJMT</em> (<em>JASMONIC ACID CARBOXYL METHYLTRANSFERASE</em>), while did not affect the expression of JA biosynthesis genes (lipoxyganease, allene oxide synthase, 12-oxophytodienoate reductase). These data, as well as the results of our previous research, suggest that exogenous IAA participates in <em>I. nil</em> flower induction process by stimulating <em>InJMT</em> expression and, as a consequence of that, enhancing the level of JAMe, a flowering inhibitor.

Differences in gene expression between natural and artificially induced leaf senescence in barley

Senescence is the last step of leaf development in the life span of an annual plant. Senescence can be induced prematurely by treating leaf tissues with jasmonic acid methyl ester (methyl jasmonate, MeJA). During both senescence programmes, drastic changes occur at the biochemical, cellular and ultra-structural levels that were compared here for primary leaves of barley (Hordeum vulgare L.). Our findings indicate that both types of senescence are similar with respect to the morphological changes including the loss of chlorophyll, disintegration of thylakoids, and formation of plastoglobules. However, the time elapsed for reaching senescence completion was different and ranged from 7 to 8 days for artificially senescing, MeJA-treated plants to 7–8 weeks for naturally senescing plants. Pulse-labelling studies along with RNA and protein gel blot analyses showed differential changes in the expression of both plastid and nuclear genes coding for photosynthetic proteins. Several unique messenger products accumulated in naturally and artificially senescing, MeJA-treated leaves. Detailed expression and crosslinking studies revealed that pheophorbide a oxygenase (PAO), a previously implicated key enzyme of chlorophyll breakdown, is most likely not rate-limiting for chlorophyll destruction under both senescence conditions. Metabolite profiling identified differential changes in the composition of carotenoid derivatives and prenyl-lipids to occur in naturally senescing and artificially senescing plants that underscored the differences between both senescence programmes.

Bacteria-triggered systemic immunity in barley is associated with WRKY and ETHYLENE RESPONSIVE FACTORs but not with salicylic acid.

Leaf-to-leaf systemic immune signaling known as systemic acquired resistance is poorly understood in monocotyledonous plants. Here, we characterize systemic immunity in barley (Hordeum vulgare) triggered after primary leaf infection with either Pseudomonas syringae pathovar japonica (Psj) or Xanthomonas translucens pathovar cerealis (Xtc). Both pathogens induced resistance in systemic, uninfected leaves against a subsequent challenge infection with Xtc. In contrast to systemic acquired resistance in Arabidopsis (Arabidopsis thaliana), systemic immunity in barley was not associated with NONEXPRESSOR OF PATHOGENESIS-RELATED GENES1 or the local or systemic accumulation of salicylic acid. Instead, we documented a moderate local but not systemic induction of abscisic acid after infection of leaves with Psj. In contrast to salicylic acid or its functional analog benzothiadiazole, local applications of the jasmonic acid methyl ester or abscisic acid triggered systemic immunity to Xtc. RNA sequencing analysis of local and systemic transcript accumulation revealed unique gene expression changes in response to both Psj and Xtc and a clear separation of local from systemic responses. The systemic response appeared relatively modest, and quantitative reverse transcription-polymerase chain reaction associated systemic immunity with the local and systemic induction of two WRKY and two ETHYLENE RESPONSIVE FACTOR (ERF)-like transcription factors. Systemic immunity against Xtc was further associated with transcriptional changes after a secondary/systemic Xtc challenge infection; these changes were dependent on the primary treatment. Taken together, bacteria-induced systemic immunity in barley may be mediated in part by WRKY and ERF-like transcription factors, possibly facilitating transcriptional reprogramming to potentiate immunity.

Jasmonic acid negatively regulates resistance to Tobacco mosaic virus in tobacco

Nicotiana tabacum (tobacco) cultivars possessing the N resistance gene to Tobacco mosaic virus (TMV) induce a hypersensitive response, which is accompanied by the production of phytohormones such as salicylic acid (SA) and jasmonic acid (JA), to enclose the invaded virus at the initial site of infection, which inhibits viral multiplication and spread. SA functions as a positive regulator of TMV resistance. However, the role of JA in TMV resistance has not been fully elucidated. Exogenously applied methyl jasmonate, a methyl ester of JA, reduced local resistance to TMV and permitted systemic viral movement. Furthermore, in contrast to a previous finding, we demonstrated that silencing of CORONATINE-INSENSITIVE 1 (COI1), a JA receptor, reduced viral accumulation in a tobacco cultivar possessing the N gene, as did that of allene oxide synthase, a JA biosynthetic enzyme. The reduction in viral accumulation in COI1-silenced tobacco plants was correlated with an increase in SA, and lowering SA levels by introducing an SA hydroxylase gene attenuated this reduction. Viral susceptibility did not change in a COI1-silenced tobacco cultivar lacking the N gene. These results suggest that JA signaling is not directly responsible for susceptibility to TMV, but is indirectly responsible for viral resistance through the partial inhibition of SA-mediated resistance conferred by the N gene, and that a balance between endogenous JA and SA levels is important for determining the degree of resistance.

Manipulation of methyl jasmonate esterase activity renders tomato more susceptible to Sclerotinia sclerotiorum.

Jasmonic acid methyl ester has been discussed as a stress signal in plants. To investigate the relevance of reversible methylation of jasmonic acid, stress responses of transgenic tomato lines with altered expression and activity of methyl jasmonate esterase were analysed. No consistent changes in levels of methyl jasmonate, 12-oxo-phytodienoic acid, jasmonic acid, jasmonic acid isoleucine and expression of the jasmonate-responsive genes AOC and PINII between control line and RNAi as well as overexpressing lines were detectable under basal and wound-induced conditions. In contrast, reduction as well as enhancement of methyl jasmonate esterase activity resulted in increased susceptibility to the fungal pathogen Sclerotinia sclerotiorum despite higher levels of the hormonal active jasmonic acid isoleucine conjugate. Results suggest that methyl jasmonate esterase has a function in vivo in plant defence, which appears not to be related to its in vitro capacity to hydrolyse methyl jasmonate.

Expression of the 1-SST and 1-FFT genes and consequent fructan accumulation in Agave tequilana and A. inaequidens is differentially induced by diverse (a)biotic-stress related elicitors

The expression of genes coding for sucrose:sucrose 1-fructosyltransferase (1-SST; EC 2.4.1.99) and fructan:fructan 1-fructosyltransferase (1-FFT; EC 2.4.1.100), both fructan biosynthesizing enzymes, characterization by TLC and HPAEC-PAD, as well as the quantification of the fructo-oligosaccharides (FOS) accumulating in response to the exogenous application of sucrose, kinetin (cytokinin) or other plant hormones associated with (a)biotic stress responses were determined in two Agave species grown in vitro, domesticated Agave tequilana var. azul and wild A. inaequidens. It was found that elicitors such as salicylic acid (SA), and jasmonic acid methyl ester (MeJA) had the strongest effect on fructo-oligosaccharide (FOS) accumulation. The exogenous application of 1mM SA induced a 36-fold accumulation of FOS of various degrees of polymerization (DP) in stems of A. tequilana. Other treatments, such as 50mM abscisic acid (ABA), 8% Sucrose (Suc), and 1.0mgL−1 kinetin (KIN) also led to a significant accumulation of low and high DP FOS in this species. Conversely, treatment with 200μM MeJA, which was toxic to A. tequilana, induced an 85-fold accumulation of FOS in the stems of A. inaequidens. Significant FOS accumulation in this species also occurred in response to treatments with 1mM SA, 8% Suc, and 10% polyethylene glycol (PEG). Maximum yields of 13.6 and 8.9mg FOS per g FW were obtained in stems of A. tequilana and A. inaequidens, respectively. FOS accumulation in the above treatments was tightly associated with increased expression levels of either the 1-FFT or the 1-SST gene in tissues of both Agave species.

A special pair of phytohormones controls excitability, slow closure, and external stomach formation in the Venus flytrap

Venus flytrap's leaves can catch an insect in a fraction of a second. Since the time of Charles Darwin, scientists have struggled to understand the sensory biology and biomechanics of this plant, Dionaea muscipula. Here we show that insect-capture of Dionaea traps is modulated by the phytohormone abscisic acid (ABA) and jasmonates. Water-stressed Dionaea, as well as those exposed to the drought-stress hormone ABA, are less sensitive to mechanical stimulation. In contrast, application of 12-oxo-phytodienoic acid (OPDA), a precursor of the phytohormone jasmonic acid (JA), the methyl ester of JA (Me-JA), and coronatine (COR), the molecular mimic of the isoleucine conjugate of JA (JA-Ile), triggers secretion of digestive enzymes without any preceding mechanical stimulus. Such secretion is accompanied by slow trap closure. Under physiological conditions, insect-capture is associated with Ca(2+) signaling and a rise in OPDA, Apparently, jasmonates bypass hapto-electric processes associated with trap closure. However, ABA does not affect OPDA-dependent gland activity. Therefore, signals for trap movement and secretion seem to involve separate pathways. Jasmonates are systemically active because application to a single trap induces secretion and slow closure not only in the given trap but also in all others. Furthermore, formerly touch-insensitive trap sectors are converted into mechanosensitive ones. These findings demonstrate that prey-catching Dionaea combines plant-specific signaling pathways, involving OPDA and ABA with a rapidly acting trigger, which uses ion channels, action potentials, and Ca(2+) signals.

Effect of jasmonic acid–methyl ester on the composition of carbohydrates and germination of yellow lupine (Lupinus luteus L.) seeds

Mature seeds of yellow lupine contained sucrose, raffinose family oligosaccharides (RFOs), and galactosyl cyclitols as major soluble carbohydrates. The study showed that RFOs dominated in lupine seeds (16% DW). The disappearance of both types of alpha-d-galactosides in germinating lupine seeds was strongly inhibited by the presence of jasmonic acid-methyl ester (JA-Me) at a concentration of 10(-3)M in the incubation medium. JA-Me inhibited the activity of alpha-D-galactosidase (fraction I) during seed germination. Anatomical studies of lupine roots have shown certain cell structure differences between control and JA-Me-treated seedlings. The cross-sections of plant roots treated with JA-Me showed a characteristic folding of the cell walls in all root tissues, starting from the rhyzodermis, cortex and vascular cylinder. In water-treated (control) plants, the cell walls were rounded with no folding.

The Magnesium-Chelatase H Subunit Binds Abscisic Acid and Functions in Abscisic Acid Signaling: New Evidence in Arabidopsis

Using a newly developed abscisic acid (ABA)-affinity chromatography technique, we showed that the magnesium-chelatase H subunit ABAR/CHLH (for putative abscisic acid receptor/chelatase H subunit) specifically binds ABA through the C-terminal half but not the N-terminal half. A set of potential agonists/antagonists to ABA, including 2-trans,4-trans-ABA, gibberellin, cytokinin-like regulator 6-benzylaminopurine, auxin indole-3-acetic acid, auxin-like substance naphthalene acetic acid, and jasmonic acid methyl ester, did not bind ABAR/CHLH. A C-terminal C370 truncated ABAR with 369 amino acid residues (631-999) was shown to bind ABA, which may be a core of the ABA-binding domain in the C-terminal half. Consistently, expression of the ABAR/CHLH C-terminal half truncated proteins fused with green fluorescent protein (GFP) in wild-type plants conferred ABA hypersensitivity in all major ABA responses, including seed germination, postgermination growth, and stomatal movement, and the expression of the same truncated proteins fused with GFP in an ABA-insensitive cch mutant of the ABAR/CHLH gene restored the ABA sensitivity of the mutant in all of the ABA responses. However, the effect of expression of the ABAR N-terminal half fused with GFP in the wild-type plants was limited to seedling growth, and the restoring effect of the ABA sensitivity of the cch mutant was limited to seed germination. In addition, we identified two new mutant alleles of ABAR/CHLH from the mutant pool in the Arabidopsis Biological Resource Center via Arabidopsis (Arabidopsis thaliana) Targeting-Induced Local Lesions in Genomes. The abar-2 mutant has a point mutation resulting in the N-terminal Leu-348-->Phe, and the abar-3 mutant has a point mutation resulting in the N-terminal Ser-183-->Phe. The two mutants show altered ABA-related phenotypes in seed germination and postgermination growth but not in stomatal movement. These findings support the idea that ABAR/CHLH is an ABA receptor and reveal that the C-terminal half of ABAR/CHLH plays a central role in ABA signaling, which is consistent with its ABA-binding ability, but the N-terminal half is also functionally required, likely through a regulatory action on the C-terminal half.

An advanced method for the determination of carboxyl methyl esterase activity using gas chromatography–chemical ionization–mass spectrometry

We developed a quantitative method for the determination of methyl esterase activity, analyzing substrate specificity against three major signal molecules, jasmonic acid methyl ester (MeJA), salicylic acid methyl ester (MeSA), and indole-3-acetic acid methyl ester (MeIAA). We used a silylation reagent for chemical derivatization and used gas chromatography (GC)–mass spectroscopy in analyses, for high precision. To test this method, an Arabidopsis esterase gene, AtME8, was expressed in Escherichia coli, and then the kinetic parameters of the recombinant enzyme were determined for three substrates. Finally, this method was also applied to the direct quantification of phytohormones in petals from lilies and roses.

Hydroxylated jasmonates are commonly occurring metabolites of jasmonic acid and contribute to a partial switch‐off in jasmonate signaling

In potato 12-hydroxyjasmonic acid (12-OH-JA) is a tuber-inducing compound. Here, it is demonstrated that 12-OH-JA, as well as its sulfated and glucosylated derivatives, are constituents of various organs of many plant species. All accumulate differentially and usually to much higher concentrations than jasmonic acid (JA). In wounded tomato leaves, 12-OH-JA and its sulfated, as well as glucosylated, derivative accumulate after JA, and their diminished accumulation in wounded leaves of the JA-deficient mutants spr2 and acx1 and also a JA-deficient 35S::AOCantisense line suggest their JA-dependent formation. To elucidate how signaling properties of JA/JAME (jasmonic acid methyl ester) are affected by hydroxylation and sulfation, germination and root growth were recorded in the presence of the different jasmonates, indicating that 12-OH-JA and 12-hydroxyjasmonic acid sulfate (12-HSO(4)-JA) were not bioactive. Expression analyses for 29 genes showed that expression of wound-inducible genes such as those coding for PROTEINASE INHIBITOR2, POLYPHENOL OXIDASE, THREONINE DEAMINASE or ARGINASE was induced by JAME and less induced or even down-regulated by 12-OH-JA and 12-HSO(4)-JA. Almost all genes coding for enzymes in JA biosynthesis were up-regulated by JAME but down-regulated by 12-OH-JA and 12-HSO(4)-JA. The data suggest that wound-induced metabolic conversion of JA/JAME into 12-OH-JA alters expression pattern of genes including a switch off in JA signaling for a subset of genes.

Molecular cloning and differential expression of an aldehyde dehydrogenase gene in rice leaves in response to infection by blast fungus

Aldehyde dehydrogenase (ALDH) superfamily is a group of enzymes metabolizing endogenous and exogenous aldehydes. Using differential display RT-PCR and cDNA library screening, a full-length aldehyde dehydrogenase cDNA (ALDH7B7) was isolated from rice leaves infected by incompatible race of blast fungus Magnaporthe grisea. The deduced amino acid sequence consists of 509 amino acid residues and shares 74∼81% identity with those of ALDH7Bs from other plants. ALDH7B7 expression was induced by blast fungus infection, ultraviolet, mechanical wound in rice leaves and was not detected in untreated rice organs. This gene has also been found to be inducible after exogenous phytohormones application, such as salicylic acid, methyl ester of jasmonic acid and abscisic acid. The function of ALDH7B7 in the interaction process between blast fungus and rice is discussed.