The plant endophytic fungus Cyanodermella asteris produces the phytohormone jasmonic acid.

The plant immune system uses several second messengers to encode information generated by the pathogen-associated molecular patterns (PAMPs) and deliver the information downstream of plant pattern recognition receptors (PRRs) to proteins which decode/interpret signals and initiate defense gene expression. Plant hormones play important role in intercellular and systemic signaling systems in plant immunity. Jasmonic acid (JA) system is a key component in the complex plant hormone signaling systems. The production of jasmonic acid (JA) in plants is a tightly regulated process. In healthy unstressed plants, JA content is very low and PAMP cues enhance the biosynthesis and accumulation of JA in the PAMP-treated plant tissues. PAMP elicitor signals activate G-proteins and the activated G-proteins further switch on calcium ion channels. Ca2+ influx and subsequent Ca2+ wave (calcium signature) may activate NADPH oxidase and H2O2 production. The Ca2+ influx-mediated NO plays important role in JA biosynthesis. Co-expression of MAPK genes with the genes involved in the JA biosynthesis pathway suggests that MAPK cascades may also be involved in JA biosynthesis. COI1 is as a key player in the jasmonate perception and signal transduction pathway. JA receptor is a three-molecule co-receptor complex, consisting of COI1, JAZ, and inositol pentakisphosphate, all of which are indispensable for high-affinity hormone binding. The bioactive jasmonate-isoleucine (JA-Ile) promotes physical interaction between JAZ1 and COI1. The JA receptor JAZ proteins have been identified as suppressors of jasmonate signaling. NINJA is an adaptor protein that interacts with the ZIM domain of most JAZs. NINJA contains an EAR (for ERF-associated amphiphilic repression) motif that recruits the corepressor TOPLESS (TPL). JAZs are a scaffold on which the NINJA–TPL corepressor complex is assembled. Repression of JA response genes involves binding of JAZ to ‑NINJA, which contains an EAR motif that recruits the corepressor TPL, which may silence gene expression. In the absence of JA signal, JAZ proteins actively repress the transcription factor MYC2, which binds to cis-acting elements of JA response genes. In response to cues that up-regulate JA-Ile synthesis, the hormone triggers the specific binding of JAZ proteins to COI1, leading to poly-ubiquitination. Subsequent degradation of JAZ by the 26S proteasome relieves repression of MYC2 and probably other transcription factors, permitting the expression of jasmonate-responsive genes. MEDIATOR25 subunit of the Mediator complex is a positive regulator of jasmonate-responsive gene expression in Arabidopsis. MED25 functions in association with transcriptional regulators of the JA pathway. Histone deacetylase may regulate JA-mediated signaling systems. JA and ET pathways appear to function cooperatively in modulating plant immune responses. Ethylene signaling may render JA signaling insensitive to subsequent suppression by SA. JA may inhibit SA signaling and SA may suppress the biosynthesis of JA. SA suppresses JA signaling system by targeting GCC-box motifs in JA-responsive promoters. JA signaling may be integrated into early ABA signaling and may affect ABA receptor complexes to regulate downstream common signal components. MicroRNA-directed RNA silencing system triggers JA signaling by specifically activating biosynthesis of JA. JA-Ile may be the mobile signaling cue involved in the induced systemic resistance (ISR). JA-Ile may be synthesized de novo and transported into systemic tissues to trigger ISR. JA-mediated priming plays an important role in the ISR.

Although many important discoveries have been made regarding the jasmonate signaling pathway, how jasmonate biosynthesis is initiated is still a major unanswered question in the field. Previous evidences suggest that jasmonate biosynthesis is limited by the availability of fatty acid precursor, such as ⍺-linolenic acid (⍺-LA). This indicates that the lipase responsible for releasing α-LA in the chloroplast, where early steps of jasmonate biosynthesis take place, is the key initial step in the jasmonate biosynthetic pathway. Nicotiana benthamiana glycerol lipase A1 (NbGLA1) is homologous to N. attenuata GLA1 (NaGLA1) which has been reported to be a major lipase in leaves for jasmonate biosynthesis. NbGLA1 was studied for its potential usefulness in a species that is more common in laboratories. Virus-induced gene silencing of both NbGLA1 and NbGLA2, another homolog, resulted in more than 80% reduction in jasmonic acid (JA) biosynthesis in wounded leaves. Overexpression of NbGLA1 utilizing an inducible vector system failed to increase JA, indicating that transcriptional induction of NbGLA1 is insufficient to trigger JA biosynthesis. However, co-treatment with wounding in addition to NbGLA1 induction increased JA accumulation several fold higher than the gene expression or wounding alone, indicating an enhancement of the enzyme activity by wounding. Domain-deletion of a 126-bp C-terminal region hypothesized to have regulatory roles increased NbGLA1-induced JA level. Together, the data show NbGLA1 to be a major lipase for wound-induced JA biosynthesis in N. benthamiana leaves and demonstrate the use of inducible promoter-driven construct of NbGLA1 in conjunction with its transient expression in N. benthamiana as a useful system to study its protein function.

Jasmonic acid (JA) biosynthesis and signaling are activated in Arabidopsis cultivated in phosphate (Pi) deprived conditions. This activation occurs mainly in photosynthetic tissues and is less important in roots. In leaves, the enhanced biosynthesis of JA coincides with membrane glycerolipid remodeling triggered by the lack of Pi. We addressed the possible role of JA on the dynamics and magnitude of glycerolipid remodeling in response to Pi deprivation and resupply. Based on combined analyses of gene expression, JA biosynthesis and glycerolipid remodeling in wild-type Arabidopsis and in the coi1-16 mutant, JA signaling seems important in the determination of the basal levels of phosphatidylcholine, phosphatidic acid (PA), monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol. JA impact on MGDG steady state level and fluctuations seem contradictory. In the coi1-16 mutant, the steady state level of MGDG is higher, possibly due to a higher level of PA in the mutant, activating MGD1, and to an increased expression of MGD3. These results support a possible impact of JA in limiting the overall content of this lipid. Concerning lipid variations, upon Pi deprivation, JA seems rather associated with a specific MGDG increase. Following Pi resupply, whereas the expression of glycerolipid remodeling genes returns to basal level, JA biosynthesis and signaling genes are still upregulated, likely due to a JA-induced positive feedback remaining active. Distinct impacts on enzymes synthesizing MGDG, that is, downregulating MGD3, possibly activating MGD1 expression and limiting the activation of MGD1 via PA, might allow JA playing a role in a sophisticated fine tuning of galactolipid variations.

GA suppresses the conversion from α-linolenic acid to JA through the GhGAI1-GhMYC3-GhLOX3 module to promote cotton fiber elongation.

Arabidopsis thaliana contains a large number of genes that encode carboxylic acid-activating enzymes, including nine long-chain fatty acyl-CoA synthetases, four 4-coumarate:CoA ligases (4CL), and 25 4CL-like proteins of unknown biochemical function. Because of their high structural and sequence similarity with bona fide 4CLs and their highly hydrophobic putative substrate-binding pockets, the 4CL-like proteins At4g05160 and At5g63380 were selected for detailed analysis. Following heterologous expression, the purified proteins were subjected to a large scale screen to identify their preferred in vitro substrates. This study uncovered a significant activity of At4g05160 with medium-chain fatty acids, medium-chain fatty acids carrying a phenyl substitution, long-chain fatty acids, as well as the jasmonic acid precursors 12-oxo-phytodienoic acid and 3-oxo-2-(2'-pentenyl)-cyclopentane-1-hexanoic acid. The closest homolog of At4g05160, namely At5g63380, showed high activity with long-chain fatty acids and 12-oxo-phytodienoic acid, the latter representing the most efficiently converted substrate. By using fluorescent-tagged variants, we demonstrated that both 4CL-like proteins are targeted to leaf peroxisomes. Collectively, these data demonstrate that At4g05160 and At5g63380 have the capacity to contribute to jasmonic acid biosynthesis by initiating the beta-oxidative chain shortening of its precursors.

PLIP lipases can initiate jasmonic acid (JA) biosynthesis. However, little is known about the transcriptional regulation of this process. In this study, an ERF transcription factor (CsESE3) was found to be co-expressed with all necessary genes for JA biosynthesis and several key genes for wax biosynthesis in transcriptomes of ‘Newhall’ navel orange. CsESE3 shows partial sequence similarity to the well-known wax regulator SHINEs (SHNs), but lacks a complete MM protein domain. Ectopic overexpression of CsESE3 in tomato (OE) resulted in reduction of fruit surface brightness and dwarf phenotype compared to the wild type. The OE tomato lines also showed significant increases in the content of wax and JA and the expression of key genes related to their biosynthesis. Overexpression of CsESE3 in citrus callus and fruit enhanced the JA content and the expression of JA biosynthetic genes. Furthermore, CsESE3 could bind to and activate the promoters of two phospholipases from the PLIP gene family to initiate JA biosynthesis. Overall, this study indicated that CsESE3 could mediate JA biosynthesis by activating PLIP genes and positively modulate wax biosynthesis. The findings provide important insights into the coordinated control of two defense strategies of plants represented by wax and JA biosynthesis.

Jasmonic acid has properties of a plant hormone, including the induction of specific genes associated with plant defense. We previously described jar1-1, an Arabidopsis jasmonate response mutant that exhibits reduced sensitivity to methyl jasmonate. We have further characterized this mutant and two new alleles; jar1-2 from a gamma irradiated population, and jar1-4 from a T-DNA mutant population. Seedling root growth in jar1-1 was equally insensitive to methyl jasmonate and jasmonic acid, indicating that the defect was not in the conversion of methyl jasmonate to the acid. None of the jar1 mutants showed an altered sensitivity to auxin, cytokinin, or the ethylene precursor 1-aminocyclopropane-1-carboxylic acid, indicating that the lesion does not affect the general uptake or transport of hormones. A soil fungus, Pythium irregulare, was found to blight jar1-1. Cultures of this organism caused the symptoms in all three jar1 mutants but not in wild type, indicating that increased susceptibility was due to the lesion in the JAR1 locus. A fatty acid desaturase triple mutant that is defective in the biosynthesis of jasmonic acid (J. Browse, Washington State University) was also susceptible, confirming that jasmonate is involved in resistance. The jar1-1 locus was mapped to the lower end of chromosome 2, about 11.4 cM from as1 and 1.6 cM from cer8. These results establish that jasmonate signaling plays an important role in resistance to soil micro-organisms in plants.

Foliar stomatal movements are critical for regulating plant water loss and gas exchange. Elevated carbon dioxide (CO2 ) levels are known to induce stomatal closure. However, the current knowledge on CO2 signal transduction in stomatal guard cells is limited. Here we report metabolomic responses of Brassica napus guard cells to elevated CO2 using three hyphenated metabolomics platforms: gas chromatography-mass spectrometry (MS); liquid chromatography (LC)-multiple reaction monitoring-MS; and ultra-high-performance LC-quadrupole time-of-flight-MS. A total of 358 metabolites from guard cells were quantified in a time-course response to elevated CO2 level. Most metabolites increased under elevated CO2 , showing the most significant differences at 10 min. In addition, reactive oxygen species production increased and stomatal aperture decreased with time. Major alterations in flavonoid, organic acid, sugar, fatty acid, phenylpropanoid and amino acid metabolic pathways indicated changes in both primary and specialized metabolic pathways in guard cells. Most interestingly, the jasmonic acid (JA) biosynthesis pathway was significantly altered in the course of elevated CO2 treatment. Together with results obtained from JA biosynthesis and signaling mutants as well as CO2 signaling mutants, we discovered that CO2 -induced stomatal closure is mediated by JA signaling.

Fungal elicitor prepared from the cell walls of Aspergillum niger induces multiple responses of Hypericum perforatum cells, including nitric oxide (NO) generation, jasmonic acid (JA) biosynthesis, and hypericin production. To determine the role of NO and JA in elicitor-induced hypericin production, we study the effects of NO scavenger 2- to 4-carboxyphenyl-4,4, 5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPITO), nitric oxide synthase inhibitor S,S'-1,3-phenylene-bis(1,2-ethanediyl)-bis-isothiourea, and inhibitors of the octadecanoid pathway on elicitor-induced NO generation, JA biosynthesis, and hypericin production. Pretreatment of the cells with cPITO and JA biosynthesis inhibitors suppresses not only the elicitor-induced NO generation and JA accumulation but also the elicitor-induced hypericin production, which suggests that both NO and JA are involved in elicitor-induced hypericin biosynthesis. S,S'-1,3-phenylene-bis(1,2-ethanediyl)-bis-isothiourea and cPITO inhibit both elicitor-induced NO generation and JA biosynthesis, while JA biosynthesis inhibitors do not affect the elicitor-induced NO generation, indicating that JA acts downstream of NO generation and that its biosynthesis is regulated by NO. External application of NO via its donor sodium nitroprusside induces hypericin production in the absence of fungal elicitor. Sodium-nitroprusside-induced hypericin production is blocked by JA biosynthesis inhibitors, showing that JA biosynthesis is essential for NO-induced hypericin production. The results demonstrate a causal relationship between elicitor-induced NO generation, JA biosynthesis, and hypericin production in H. perforatum cells and indicate a sequence of signaling events from NO to hypericin production, within which NO mediates the elicitor-induced hypericin biosynthesis at least partially via a JA-dependent signaling pathway.

Theobroxide inhibits stem elongation in Pharbitis nil by regulating jasmonic acid and gibberellin biosynthesis

Dipsacus asperoides is a perennial herb, the roots of which are abundant in asperosaponin VI, which has important medicinal value. However, the molecular mechanism underlying the biosynthesis of asperosaponin VI in D. asperoides remains unclear. In present study, a comprehensive investigation of asperosaponin VI biosynthesis was conducted at the levels of metabolite and transcript during root development. The content of asperosaponin VI was significantly accumulated in two-leaf stage roots, and the spatial distribution of asperosaponin VI was localized in the xylem. The concentration of asperosaponin VI gradually increased in the root with the development process. Transcriptome analysis revealed 3916 unique differentially expressed genes (DEGs) including 146 transcription factors (TFs) during root development in D. asperoides. In addition, α-linolenic acid metabolism, jasmonic acid (JA) biosynthesis, JA signal transduction, sesquiterpenoid and triterpenoid biosynthesis, and terpenoid backbone biosynthesis were prominently enriched. Furthermore, the concentration of JA gradually increased, and genes involved in α-linolenic acid metabolism, JA biosynthesis, and triterpenoid biosynthesis were up-regulated during root development. Moreover, the concentration of asperosaponin VI was increased following methyl jasmonate (MeJA) treatment by activating the expression of genes in the triterpenoid biosynthesis pathway, including acetyl-CoA acetyltransferase (DaAACT), 3-hydroxy-3-methylglutaryl coenzyme A synthase (DaHMGCS), 3-hydroxy-3-methylglutaryl coenzyme-A reductase (DaHMGCR). We speculate that JA biosynthesis and signaling regulates the expression of triterpenoid biosynthetic genes and facilitate the biosynthesis of asperosaponin VI. The results suggest a regulatory network wherein triterpenoids, JA, and TFs co-modulate the biosynthesis of asperosaponin VI in D. asperoides.

Jasmonates and their precursors, the octadecanoids, are signals in stress-induced alteration of gene expression. Several mRNAs coding for enzymes of jasmonic acid (JA) biosynthesis are up-regulated upon JA treatment or endogenous increase of the JA level. Here we investigated the positive feedback of endogenous JA on JA formation, as well as its beta-oxidation steps. JA-responsive gene expression was recorded in terms of proteinase inhibitor2 (pin2) mRNA accumulation. JA formed upon treatment of tomato (Lycopersicon esculentum cv. Moneymaker) leaves with JA derivatives carrying different lengths of the carboxylic acid side chain was quantified by gas chromatography-mass spectrometry (GC-MS). The data revealed that beta-oxidation of the side chain occurs up to a butyric acid moiety. The amount of JA formed from side-chain modified JA derivatives correlated with pin2-mRNA accumulation. JA derivatives with a carboxylic side chain of 3, 5 or 7 carbon atoms were unable to form JA and to express on pin2, whereas even-numbered derivatives were active. After treatment of tomato leaves with (10-(2)H)-(-)-12-oxophytoenoic acid, (4-(2)H)-(-)-JA and its methyl ester were formed and could be quantified separately from the endogenously nonlabeled JA pool by GC-MS analysis via isotopic discrimination. The level of 8 nmol per g fresh weight JA and its methyl ester originated exclusively from labeled 12-oxophytoenic acid. This and further data indicate that endogenous synthesis of the JA precursor 12-oxophytodienoic acid, as well as of JA and its methyl ester, are not induced in tomato leaves, suggesting that positive feedback in JA biosynthesis does not function in vivo.

Recent updates in JA biosynthesis, signaling pathways and the crosstalk between JA and others phytohormones in relation with plant responses to different stresses. In plants, the roles of phytohormone jasmonic acid (JA), amino acid conjugate (e.g., JA-Ile) and their derivative emerged in last decades as crucial signaling compounds implicated in stress defense and development in plants. JA has raised a great interest, and the number of researches on JA has increased rapidly highlighting the importance of this phytohormone in plant life. First, JA was considered as a stress hormone implicated in plant response to biotic stress (pathogens and herbivores) which confers resistance to biotrophic and hemibiotrophic pathogens contrarily to salicylic acid (SA) which is implicated in plant response to necrotrophic pathogens. JA is also implicated in plant responses to abiotic stress (such as soil salinity, wounding and UV). Moreover, some researchers have recently revealed that JA controls several physiological processes like root growth, growth of reproductive organs and, finally, plant senescence. JA is also involved in the biosynthesis of various metabolites (e.g., phytoalexins and terpenoids). In plants, JA signaling pathways are well studied in few plants essentially Arabidopsis thaliana, Nicotiana benthamiana, and Oryza sativa L. confirming the crucial role of this hormone in plants. In this review, we highlight the last foundlings about JA biosynthesis, JA signaling pathways and its implication in plant maturation and response to environmental constraints.

Jasmonic acid (JA) and Salicylic acid (SA) are the essential plant hormones responsible for the plant’s proper growth and development. These signaling molecules have a significant role in plants along with the regulation of defense mechanisms locally and systemically under various biotic and abiotic stresses. Among abiotic stresses, ultraviolet-B (UV-B) radiation coming to the Earth’s surface due to depletion of the stratospheric ozone layer is of serious concern to all living organisms. UV-B is an important factor, negatively influencing the growth and yield of plants on this Earth, ultimately posing a threat to food security. Therefore, understanding the signaling behavior of JA and SA under UV-B stress will be definitely beneficial for the maintenance of agricultural productivity worldwide. Plant responses related to morphological, biochemical, physiological, growth, and yield have been extensively studied under UV-B stress, although studies conducted with UV-B exposure and its impact on plant’s endogenous JA and SA contents are limited. On the other hand, some studies have also explored the regulatory impact of exogenously supplied JA and SA to the plants. More accumulation of endogenous JA and SA contents has been observed under elevated UV-B exposure in plants. JA and SA play synergistic as well as antagonistic roles during the regulation of defense responses under various stresses. An inverse relationship between JA and SA are well established under UV-B stress in pea, soybean, and mungbean cultivars. Increased JA content provided better plant resistance while increased SA level imposed higher oxidative stress to plants when exposed to elevated UV-B. Oxidative stress caused by the higher accumulation of SA is well correlated with its inhibitory impact on catalase and ascorbate peroxidase activity leading to more generation of Reactive oxygen species (ROS) under UV-B exposure. JA has an inhibitory effect on the accumulation of SA by the regulation of NAC transcription factors like ANAC019/055/072 where MYC2 binds to the promoter regions of these NAC transcription factors, which further inhibits ISOCHORISMATE SYNTHASE1 (ICS1) expression, which is responsible for initiating the expression of BSMT1 (BENZOIC ACID/SA CARBOXYL MEHYLTRANSFERASE 1) during SA biosynthesis. Therefore, the present chapter will focus on the effect of UV-B stress in plants with special emphasis on JA and SA signaling, their antagonistic and synergistic behavior in plant defense, and ROS interaction.

CORONATINE INSENSITIVE 1 (COI1) is a well-known key player in processes downstream of jasmonic acid (JA) biosynthesis: silencing COI1 in Nicotiana attenuata (ir-coi1) makes plants insensitive to JA, prevents the up-regulation of JA-mediated defenses and decreases the plant's resistance to herbivores and pathogens. In agreement with previous studies, we observed that regulation of several JA biosynthesis genes elicited by Manduca sexta oral secretions (OS) is COI1 dependent. In response to wounding and application of OS ir-coi1 plants accumulate 75% less JA compared with wild-type plants (WT), resembling JA levels found in plants silenced in the key enzyme in JA biosynthesis LIPOXYGENASE 3 (as-lox). However, while OS-elicited as-lox plants also accumulated lower levels of the JA-conjugate JA-isoleucine (JA-Ile) than did WT plants, JA-Ile accumulation in ir-coi1 was higher, prolonged and peaked with a delay of 30 min. In vivo substrate feeding experiments of N. attenuata demonstrate that the increased and prolonged JA-Ile accumulation pattern in ir-coi1 is not the result of altered substrate availability, i.e. of JA and/or Ile, but is due to an approximately 6-fold decrease in JA-Ile turnover. These results provide the first evidence for a second, novel regulatory feedback function of COI1 in enhancing JA-Ile turnover. Hence, in addition to its control over JA biosynthesis, COI1 might fine-tune the dynamics of the jasmonate response after induction by herbivore elicitors.