Jasmonic Acid-isoleucine Conjugate Research Articles

The efficacy of Azotobacter chroococcum in altering maize plant-defense responses to armyworm at elevated CO2 concentration

Elevated atmospheric carbon dioxide (eCO2) concentrations can alter the carbon:nitrogen ratio and palatability of host plants for herbivorous insects, but rhizobacteria likely mitigate the alteration and influence physiological adaptation of insects. In this study, we conducted transcriptomic analysis of maize (Zea mays) response to Azotobacter chroococcum (AC) inoculation under eCO2 conditions in contrast to ambient CO2 (aCO2), and studied the effects of plant-defense change of maize under eCO2 on the oriental armyworm, Mythimna separata. Results showed that there were 16, 14, 16 and 135 differentially expressed genes that were associated with plant-defense response in maize leaves between aCO2-CK and aCO2-AC, eCO2-CK and eCO2-AC, aCO2-CK and eCO2-CK, aCO2-AC and eCO2-AC, respectively. Moreover, A. chroococcum inoculation and eCO2 influenced plant hormone signal transduction of maize. Interestingly, A. chroococcum inoculation significantly decreased the contents of JA (jasmonic acid) and JA-Ile (isoleucine conjugate of JA) in leaves, but eCO2 markedly increased contents of JA-Ile, JA and SA (salicylic acid). Compared to aCO2, eCO2 significantly decreased activity of protective enzyme (catalase), and increased activities of digestive (lipase and protease), protective (peroxidase) and detoxifying enzymes (carboxylesterase, Mixed-functional oxidase and glutathione s-transferase), prolonged developmental time, and decreased survival rate and body weight of larvae (P < 0.05). A. chroococcum inoculation significantly increased the activity of protective enzyme (catalase), and decreased the activities of detoxifying enzymes (carboxylesterase, glutathione s-transferase and mixed-functional oxidase), thus increased the growth rate and body weight of larvae in comparison with no-inoculation of A. chroococcum (P < 0.05). The indices of M. separata were significantly correlated with the foliar contents of JA, JA-Ile and SA (|r| = 0.44–0.85, P < 0.05), indicating that A. chroococcum inoculation altered the physiological adaptation of M. separata under eCO2 by disturbing defense substances in maize. Our results in understanding effects of A. chroococcum inoculation on maize resistance to herbivorous insects will be valuable for agricultural pest control in the future at eCO2 conditions.

Plant grafting relieves asymmetry of jasmonic acid response induced by wounding between scion and rootstock in tomato hypocotyl.

Plant grafting is a sequential wound healing process. However, whether wounding induces a different jasmonic acid (JA) response within half a day (12 h) after grafting or non-grafting remains unclear. Using the tomato hypocotyl grafting method, we show that grafting alleviates the asymmetrical accumulation of JA and jasmonic acid isoleucine conjugate (JA-Ile) in scion and rootstock caused by wounding, and from 2 h after tomato micrografting, grafting obviously restored the level of JA-Ile in the scion and rootstock. Meanwhile, five JA-related genes, SlLOX11, SlAOS, SlCOI1, SlLAPA and SlJA2L, are detected and show significant changes in transcriptional expression patterns within 12 h of grafting, from asymmetrical to symmetrical, when the expression of 30 JA- and defense-related genes were analyzed. The results indicated that grafting alleviates the asymmetrical JA and defense response between scion and rootstock of the tomato hypocotyl within 12 h as induced by wounding. Moreover, we demonstrate that in the very early hours after grafting, JA-related genes may be involved in a molecular mechanism that changes asymmetrical expression as induced by wounding between scion and rootstock, thereby promoting wound healing and grafting success.

Jasmonoyl-l-isoleucine and allene oxide cyclase-derived jasmonates differently regulate gibberellin metabolism in herbivory-induced inhibition of plant growth

The trade-off between plant growth and resistance to herbivory is thought to be at least partly mediated by the interactions between jasmonates and gibberellins (GAs). Insect herbivory activates jasmonate biosynthesis and signaling, and plant growth is concomitantly inhibited. Whether or not the herbivory-induced jasmonates suppress the accumulation of GAs and thus reduce plant growth, and which jasmonates are functional in this process, remain unclear. In this study, we show that herbivory-induced stunted growth of Nicotiana attenuata was completely dependent on allene oxide cyclase (AOC) and coronatine insensitive1 (COI1), which encode a JA biosynthetic enzyme and the receptor, respectively, but only partially dependent on jasmonic acid-isoleucine conjugate (JA-Ile), the bioactive jasmonate. Quantification of GAs and exogenous treatments indicated that herbivory-induced growth inhibition was caused by GA4 deficiency, and that the reduction in GA4 accumulation was strongly associated with both decreased concentrations of GA biosynthetic gene transcripts and transcriptional activation of GA catabolic genes. We further show that JA-Ile only positively regulated the levels of GA catabolic genes, while the accumulation of GA biosynthetic gene transcripts was controlled by certain AOC-derived jasmonate(s) rather than by JA-Ile. This work sheds light on the mechanisms by which plants adapt to herbivory by using intricate phytohormone signaling and transcriptional regulatory networks.

Azotobacter chroococcum inoculation can improve plant growth and resistance of maize to armyworm, Mythimna separata even under reduced nitrogen fertilizer application.

Nitrogen (N) is essential to crop yield improvement and it can change crops' ability to defend against herbivores. To maximize economic yield, a higher amount of N-fertilizer is often applied than the minimum required. Azotobacter is a good alternative to reduce N fertilizer application. In this study, we studied the yield and secondary defensive chemicals of maize, as well as the response of the key maize insect pest, Mythimna separata, as fed on maize plants inoculated with Azotobacter chroococcum and cultivated at different N fertilizer rates (i.e. the control rate of nitrogen fertilizer (CR), 80%CR and 60%CR) from 2018 to 2019. A. chroococcum inoculation just positively increased yield production of maize at 80%CR. Moreover, reduced N-fertilizer application and A. chroococcum inoculation had opposite impacts on the foliar contents of jasmonic acid (JA), isoleucine conjugate of JA (JA-Ile) and DIMBOA in maize, and they both negatively decreased the pupation rate and fecundity, and positively increased the eclosion rate and approximate digestibility (AD) of M. separata (P < 0.05). Furthermore, reduced N-fertilizer application negatively prolonged larval life-span, and decreased pupal weight, relative growth rate (RGR), efficiency of conversion of ingested food (ECI) and efficiency of conversion of digested food (ECD) of M. separata even A. chroococcum inoculation had positive effects on these indexes of M. separata (P < 0.05). These results help in understanding of the effects of low-level N-fertilizer and A. chroococcum inoculation on maize production and maize resistance to insects. This will be conducive to the integrated control of agricultural pests. © 2020 Society of Chemical Industry.

Leaf-Derived Jasmonate Mediates Water Uptake from Hydrated Cotton Roots under Partial Root-Zone Irrigation.

Partial root-zone irrigation (PRI), a water-saving technique, improves water uptake in hydrated roots by inducing specific responses that are thought to be regulated by signals originating from leaves; however, this signaling is poorly understood. Using a split-root system and polyethylene glycol 6000 to simulate PRI in cotton (Gossypium hirsutum), we showed that increased root hydraulic conductance (L) and water uptake in the hydrated roots may be due to the elevated expression of cotton plasma membrane intrinsic protein (PIP) genes. Jasmonate (jasmonic acid [JA] and jasmonic acid-isoleucine conjugate [JA-Ile]) content and the expression of three JA biosynthesis genes increased in the leaves of the PRI plants compared with those of the polyethylene glycol-free control. JA/JA-Ile content also increased in the hydrated roots, although the expression of the three JA genes was unaltered, compared with the control. The JA/JA-Ile contents in leaves increased after the foliar application of exogenous JA and was followed by an increase in both JA/JA-Ile content and L in the hydrated roots, whereas the silencing of the three JA genes had the opposite effect in the leaves. Ring-barking the hydrated hypocotyls increased the JA/JA-Ile content in the leaves but decreased the JA/JA-Ile content and L in the hydrated roots. These results suggested that the increased JA/JA-Ile in the hydrated roots was mostly transported from the leaves through the phloem, thus increasing L by increasing the expression of GhPIP in the hydrated roots under PRI. We believe that leaf-derived JA/JA-Ile, as a long-distance signal, positively mediates water uptake from the hydrated roots of cotton under PRI.

Jasmonates: News on Occurrence, Biosynthesis, Metabolism and Action of an Ancient Group of Signaling Compounds.

Jasmonic acid (JA) and its related derivatives are ubiquitously occurring compounds of land plants acting in numerous stress responses and development. Recent studies on evolution of JA and other oxylipins indicated conserved biosynthesis. JA formation is initiated by oxygenation of α-linolenic acid (α-LeA, 18:3) or 16:3 fatty acid of chloroplast membranes leading to 12-oxo-phytodienoic acid (OPDA) as intermediate compound, but in Marchantia polymorpha and Physcomitrella patens, OPDA and some of its derivatives are final products active in a conserved signaling pathway. JA formation and its metabolic conversion take place in chloroplasts, peroxisomes and cytosol, respectively. Metabolites of JA are formed in 12 different pathways leading to active, inactive and partially active compounds. The isoleucine conjugate of JA (JA-Ile) is the ligand of the receptor component COI1 in vascular plants, whereas in the bryophyte M. polymorpha COI1 perceives an OPDA derivative indicating its functionally conserved activity. JA-induced gene expressions in the numerous biotic and abiotic stress responses and development are initiated in a well-studied complex regulation by homeostasis of transcription factors functioning as repressors and activators.

Tendril Coiling in Grapevine: Jasmonates and a New Role for GABA?

Grapevine (Vitis vinifera L., Vitaceae) belongs to the genus Vitis, and is characterized as a vine due to the presence of tendrils, which are located opposite to leaves. Tendrils are thigmo-responsive organs, able to carry out delicate mechanosensory responses upon touch and related stimuli. These organs are an adaptation of the plant to climb with the help of support to higher places and finally remain at a position with favorable light quality. In previous studies on Bryonia dioica (Cucurbitaceae), phytohormones of the jasmonate class were identified as the endogenous hormone signals to initiate coiling of the tendrils. Strikingly, this is still the only example for jasmonate-induced tendril coiling. In grapevine, three compounds (12-oxo-phytodienoic acid, jasmonic acid (JA), and JA isoleucine conjugate) of the jasmonate class were found at higher concentrations in non-coiled tendrils when compared with coiled ones. Upon treatment with phytohormones, we could confirm the activity of jasmonates on tendril coiling in grapevine. However, not jasmonates but a non-proteinogenic amino acid, γ-aminobutyric acid (GABA), was detected to accumulate in grapevine tendrils at significantly higher levels than in all other tissues, independent of their coiling status. For GABA we detected a significant, transient positive effect on tendril coiling. Use of a GABA synthesis blocker, 3-mercaptopropionic acid, caused reduced GABA- but not JA-induced coiling scores. No additive effect of JA plus GABA was detectable on the tendrils’ coiling score. Thus, for grapevine, our data demonstrate a strong activity of jasmonates in tendril coiling induction even without mechanical stimuli and, furthermore, the data also suggest that GABA can independently promote tendril coiling.

Modulation of Legume Defense Signaling Pathways by Native and Non-native Pea Aphid Clones.

The pea aphid (Acyrthosiphon pisum) is a complex of at least 15 genetically different host races that are native to specific legume plants, but can all develop on the universal host plant Vicia faba. Despite much research, it is still unclear why pea aphid host races (biotypes) are able to colonize their native hosts while other host races are not. All aphids penetrate the plant and salivate into plant cells when they test plant suitability. Thus plants might react differently to the various pea aphid host races. To find out whether legume species vary in their defense responses to different pea aphid host races, we measured the amounts of salicylic acid (SA), the jasmonic acid-isoleucine conjugate (JA-Ile), other jasmonate precursors and derivatives, and abscisic acid (ABA) in four different species (Medicago sativa, Trifolium pratense, Pisum sativum, V. faba) after infestation by native and non-native pea aphid clones of various host races. Additionally, we assessed the performance of the clones on the four plant species. On M. sativa and T. pratense, non-native clones that were barely able to survive or reproduce, triggered a strong SA and JA-Ile response, whereas infestation with native clones led to lower levels of both phytohormones. On P. sativum, non-native clones, which survived or reproduced to a certain extent, induced fluctuating SA and JA-Ile levels, whereas the native clone triggered only a weak SA and JA-Ile response. On the universal host V. faba all aphid clones triggered only low SA levels initially, but induced clone-specific patterns of SA and JA-Ile later on. The levels of the active JA-Ile conjugate and of the other JA-pathway metabolites measured showed in many cases similar patterns, suggesting that the reduction in JA signaling was due to an effect upstream of OPDA. ABA levels were downregulated in all aphid clone-plant combinations and were therefore probably not decisive factors for aphid-plant compatibility. Our results suggest that A. pisum clones manipulate plant-defense signaling to their own advantage, and perform better on their native hosts due to their ability to modulate the SA- and JA-defense signaling pathways.

Deletion of the SACPD-C Locus Alters the Symbiotic Relationship Between Bradyrhizobium japonicum USDA110 and Soybean, Resulting in Elicitation of Plant Defense Response and Nodulation Defects.

Legumes form symbiotic associations with soil-dwelling bacteria collectively called rhizobia. This association results in the formation of nodules, unique plant-derived organs, within which the rhizobia are housed. Rhizobia-encoded nitrogenase facilitates the conversion of atmospheric nitrogen into ammonia, which is utilized by the plants for its growth and development. Fatty acids have been shown to play an important role in root nodule symbiosis. In this study, we have investigated the role of stearoyl-acyl carrier protein desaturase isoform C (SACPD-C), a soybean enzyme that catalyzes the conversion of stearic acid into oleic acid, which is expressed in developing seeds and in nitrogen-fixing nodules. In-depth cytological investigation of nodule development in sacpd-c mutant lines M25 and MM106 revealed gross anatomical alteration in the sacpd-c mutants. Transmission electron microscopy observations revealed ultrastructural alterations in the sacpd-c mutants that are typically associated with plant defense response to pathogens. In nodules of two sacpd-c mutants, the combined jasmonic acid (JA) species (JA and the isoleucine conjugate of JA) were found to be reduced and 12-oxophytodienoic acid (OPDA) levels were significantly higher relative to wild-type lines. Salicylic acid levels were not significantly different between genotypes, which is divergent from previous studies of sacpd mutant studies on vegetative tissues. Soybean nodule phytohormone profiles were very divergent from those of roots, and root profiles were found to be almost identical between mutant and wild-type genotypes. The activities of antioxidant enzymes, ascorbate peroxidase, and superoxide dismutase were also found to be higher in nodules of sacpd-c mutants. PR-1 gene expression was extremely elevated in M25 and MM106, while the expression of nitrogenase was significantly reduced in these sacpd-c mutants, compared with the parent 'Bay'. Two-dimensional gel electrophoresis and matrix-assisted laser desorption-ionization time of flight mass spectrometry analyses confirmed sacpd-c mutants also accumulated higher amounts of pathogenesis-related proteins in the nodules. Our study establishes a major role for SACPD-C activity as essential for proper maintenance of soybean nodule morphology and physiology and indicates that OPDA signaling is likely to be involved in attenuation of nodule biotic defense responses.

The Recently Identified Isoleucine Conjugate of cis-12-Oxo-Phytodienoic Acid Is Partially Active in cis-12-Oxo-Phytodienoic Acid-Specific Gene Expression of Arabidopsis thaliana.

Oxylipins of the jasmonate family are active as signals in plant responses to biotic and abiotic stresses as well as in development. Jasmonic acid (JA), its precursor cis-12-oxo-phytodienoic acid (OPDA) and the isoleucine conjugate of JA (JA-Ile) are the most prominent members. OPDA and JA-Ile have individual signalling properties in several processes and differ in their pattern of gene expression. JA-Ile, but not OPDA, is perceived by the SCFCOI1-JAZ co-receptor complex. There are, however, numerous processes and genes specifically induced by OPDA. The recently identified OPDA-Ile suggests that OPDA specific responses might be mediated upon formation of OPDA-Ile. Here, we tested OPDA-Ile-induced gene expression in wild type and JA-deficient, JA-insensitive and JA-Ile-deficient mutant background. Tests on putative conversion of OPDA-Ile during treatments revealed only negligible conversion. Expression of two OPDA-inducible genes, GRX480 and ZAT10, by OPDA-Ile could be detected in a JA-independent manner in Arabidopsis seedlings but less in flowering plants. The data suggest a bioactivity in planta of OPDA-Ile.

Influence of medium and elicitors on the production of cocaine, amino acids and phytohormones by Erythroxylum coca calli

Erythroxylum coca (Erythroxylaceae) is the source of the tropane alkaloid cocaine. Several lines of evidence suggest that tropane alkaloid biosynthesis in E. coca differs from that in solanaceous species, but there are many gaps in our understanding of the pathways in both groups. The development of an E. coca cell culture that produces cocaine could provide a reproducible model system for discovering novel biosynthetic genes and study pathway regulation. Calli cultures were successfully established from young leaf explants on three different media: Anderson’s Rhododendron, Gamborg B5, and modified Murashige-Tucker, all supplemented with growth regulators: 2,4-D (0.6 mg L−1), indole butyric acid (0.06 mg L−1), and benzylaminopurine (0.5 mg L−1). All accumulated cocaine and cinnamoylcocaine at levels of 0.05–0.5 nmol per gram dry weight, as determined by LC–MS, several orders of magnitude below the concentration found in the intact plant. Anderson’s Rhododendron medium supported the highest level of tropane alkaloid production, as well as the highest level of the amino acids arginine, glutamate, proline and phenylalanine, all thought to be precursors of cocaine, but contained generally lower levels of hydroxycinnamate-quinate esters, such as chlorogenic acid. These differences may be ascribed to its relatively low content of nitrate or salts, or its high content of adenine. Addition of 100 μM salicylic acid or coronalon, an analog of the bioactive jasmonic acid-isoleucine conjugate, did not result in any increase in tropane alkaloid production. These E. coca calli could provide valuable material for studies on tropane alkaloid biosynthesis and regulation.

Simultaneous analysis of apolar phytohormones and 1-aminocyclopropan-1-carboxylic acid by high performance liquid chromatography/electrospray negative ion tandem mass spectrometry via 9-fluorenylmethoxycarbonyl chloride derivatization

A strategy to detect and quantify the polar ethylene precursor 1-aminocyclopropan-1-carboxylic acid (ACC) along with the more apolar phytohormones abscisic acid (ABA), indole-3-acetic acid (IAA), jasmonic acid (JA), jasmonic acid-isoleucine conjugate (JA-Ile), 12-oxo-phytodienoic acid (OPDA), trans-zeatin, and trans-zeatin 9-riboside using a single extraction is presented. Solid phase resins commonly employed for extraction of phytohormones do not allow the recovery of ACC. We circumvent this problem by attaching an apolar group to ACC via derivatization with the amino group specific reagent 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl). Derivatization in the methanolic crude extract does not modify other phytohormones. The derivatized ACC could be purified and detected together with the more apolar phytohormones using common solid phase extraction resins and reverse phase HPLC/electrospray negative ion tandem mass spectrometry. The limit of detection was in the low nanomolar range for all phytohormones, a sensitivity sufficient to accurately determine the phytohormone levels from less than 50mg (fresh weight) of Arabidopsis thaliana and Nicotiana benthamiana tissues. Comparison with previously published phytohormone levels and the reported changes in phytohormone levels after stress treatments confirmed the accuracy of the method.

Wound and insect‐induced jasmonate accumulation in carnivorous Drosera capensis: two sides of the same coin

Carnivorous sundew plants catch and digest insect prey for their own nutrition. The sundew species Drosera capensis shows a pronounced leaf bending reaction upon prey capture in order to form an 'outer stomach'. This formation is triggered by jasmonates, phytohormones typically involved in defence reactions against herbivory and wounding. Whether jasmonates still have this function in D. capensis in addition to mediating the leaf bending reaction was investigated here. Wounded, insect prey-fed and insect-derived oral secretion-treated leaves of D. capensis were analysed for jasmonates (jasmonic acid, JA; jasmonic acid-isoleucine conjugate, JA-Ile) using LC-MS/MS. Prey-induced jasmonate accumulation in D. capensis leaves was persistent, and showed high levels of JA and JA-Ile (575 and 55.7 pmol · g · FW(-1) , respectively), whereas wounding induced a transient increase of JA (maximum 500 pmol · g · FW(-1) ) and only low (3.1 pmol · g · FW(-1) ) accumulation of JA-Ile. Herbivory, mimicked with a combined treatment of wounding plus oral secretion (W+OS) obtained from Spodoptera littoralis larvae induced both JA (4000 pmol · g · FW(-1) ) and JA-Ile (25 pmol · g · FW(-1) ) accumulation, with kinetics similar to prey treatment. Only prey and W+OS, but not wounding alone or OS, induced leaf bending. The results indicate that both mechanical and chemical stimuli trigger JA and JA-Ile synthesis. Differences in kinetics and induced jasmonate levels suggest different sensing and signalling events upon injury and insect-dependent challenge. Thus, in Drosera, jasmonates are still part of the response to wounding. Jasmonates are also employed in insect-induced reactions, including responses to herbivory and carnivory.

Pathogen-Triggered Ethylene Signaling Mediates Systemic-Induced Susceptibility to Herbivory inArabidopsis

Multicellular eukaryotic organisms are attacked by numerous parasites from diverse phyla, often simultaneously or sequentially. An outstanding question in these interactions is how hosts integrate signals induced by the attack of different parasites. We used a model system comprised of the plant host Arabidopsis thaliana, the hemibiotrophic bacterial phytopathogen Pseudomonas syringae, and herbivorous larvae of the moth Trichoplusia ni (cabbage looper) to characterize mechanisms involved in systemic-induced susceptibility (SIS) to T. ni herbivory caused by prior infection by virulent P. syringae. We uncovered a complex multilayered induction mechanism for SIS to herbivory. In this mechanism, antiherbivore defenses that depend on signaling via (1) the jasmonic acid-isoleucine conjugate (JA-Ile) and (2) other octadecanoids are suppressed by microbe-associated molecular pattern-triggered salicylic acid (SA) signaling and infection-triggered ethylene signaling, respectively. SIS to herbivory is, in turn, counteracted by a combination of the bacterial JA-Ile mimic coronatine and type III virulence-associated effectors. Our results show that SIS to herbivory involves more than antagonistic signaling between SA and JA-Ile and provide insight into the unexpectedly complex mechanisms behind a seemingly simple trade-off in plant defense against multiple enemies.

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.

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.

The moss Physcomitrella patens contains cyclopentenones but no jasmonates: mutations in allene oxide cyclase lead to reduced fertility and altered sporophyte morphology

• Two cDNAs encoding allene oxide cyclases (PpAOC1, PpAOC2), key enzymes in the formation of jasmonic acid (JA) and its precursor (9S,13S)-12-oxo-phytodienoic acid (cis-(+)-OPDA), were isolated from the moss Physcomitrella patens. • Recombinant PpAOC1 and PpAOC2 show substrate specificity against the allene oxide derived from 13-hydroperoxy linolenic acid (13-HPOTE); PpAOC2 also shows substrate specificity against the allene oxide derived from 12-hydroperoxy arachidonic acid (12-HPETE). • In protonema and gametophores the occurrence of cis-(+)-OPDA, but neither JA nor the isoleucine conjugate of JA nor that of cis-(+)-OPDA was detected. • Targeted knockout mutants for PpAOC1 and for PpAOC2 were generated, while double mutants could not be obtained. The ΔPpAOC1 and ΔPpAOC2 mutants showed reduced fertility, aberrant sporophyte morphology and interrupted sporogenesis.

A Comparison of Two Nicotiana attenuata Accessions Reveals Large Differences in Signaling Induced by Oral Secretions of the Specialist Herbivore Manduca sexta

Genetic variation within and among populations provides the raw material for evolution. Although many studies describe inter- and intraspecific variation of defensive metabolites, little is known about variation among plant populations within early signaling responses elicited by herbivory or by herbivore oral secretions (OS) introduced into wounds during feeding. In this study, we compare the OS-elicited early responses as well as the antiherbivore defensive metabolites in two accessions of the wild tobacco Nicotiana attenuata and show that, compared with an accession collected from Utah, an Arizona accession has lower herbivore-elicited activity of the salicylic acid-induced protein kinase, an important mitogen-activated protein kinase involved in herbivore resistance. These differences in salicylic acid-induced protein kinase activity were associated with substantially different levels of OS-elicited jasmonic acid, jasmonic acid-isoleucine conjugate, and ethylene bursts. Gene expression level polymorphism (ELP) determines phenotypic variation among populations, and we found the two accessions to have significantly different ELPs in the genes involved in early signaling responses to herbivory. In addition, we found differences between the Utah and the Arizona accessions in the concentrations of several secondary metabolites that contribute to N. attenuata's direct and indirect defenses. This study demonstrates significant natural variation in regulatory elements that mediate plant responses to herbivore attack, highlighting the role of ELP in producing a diversity of plant defense phenotypes.

Tissue-specific oxylipin signature of tomato flowers: allene oxide cyclase is highly expressed in distinct flower organs and vascular bundles.

A crucial step in the biosynthesis of jasmonic acid (JA) is the formation of its correct stereoisomeric precursor, cis(+)12-oxophytodienoic acid (OPDA). This step is catalysed by allene oxide cyclase (AOC), which has been recently cloned from tomato. In stems, young leaves and young flowers, AOC mRNA accumulates to a low level, contrasting with a high accumulation in flower buds, flower stalks and roots. The high levels of AOC mRNA and AOC protein in distinct flower organs correlate with high AOC activity, and with elevated levels of JA, OPDA and JA isoleucine conjugate. These compounds accumulate in flowers to levels of about 20 nmol g-1 fresh weight, which is two orders of magnitude higher than in leaves. In pistils, the level of OPDA is much higher than that of JA, whereas in flower stalks, the level of JA exceeds that of OPDA. In other flower tissues, the ratios among JA, OPDA and JA isoleucine conjugate differ remarkably, suggesting a tissue-specific oxylipin signature. Immunocytochemical analysis revealed the specific occurrence of the AOC protein in ovules, the transmission tissue of the style and in vascular bundles of receptacles, flower stalks, stems, petioles and roots. Based on the tissue-specific AOC expression and formation of JA, OPDA and JA amino acid conjugates, a possible role for these compounds in flower development is discussed in terms of their effect on sink-source relationships and plant defence reactions. Furthermore, the AOC expression in vascular bundles might play a role in the systemin-mediated wound response of tomato.