Application Of Methyl Jasmonate Research Articles

Herbivory resistance in dwarf shrubs combines with simulated warming to shift phenology and decrease reproduction

Abstract Research focus on the phenology of plants has accompanied current trends in climate warming, because the two are inextricably linked. Warmer temperatures have led to advanced plant phenology in a range of systems, although some responses are species specific. However, other stressors, such as herbivory, can delay or advance plant phenology, and few studies have addressed the combined impact of these drivers. We experimentally warmed plots with open top chambers (OTCs) and simulated herbivory resistance to insect outbreaks with methyl jasmonate (MeJA), along an elevational climatic gradient. We then recorded vegetative and reproductive phenology in two functionally important dwarf shrubs, the deciduous Vaccinium myrtillus (bilberry) and the evergreen V. vitis‐idaea (lingonberry), to test the responses to the combined treatments. Based on earlier findings, we expected larger phenological responses at higher elevations and for bilberry. We found that in the year after MeJA application, both bilberry and lingonberry delayed vegetative and reproductive phenology, although responses were stronger in bilberry. Warming with OTCs weakly advanced phenology in both species, but combined effects were not consistent. We further examined the consequences of this altered phenology on reproductive output with multigroup piecewise structural equation modelling and found that herbivory resistance driven changes to phenology had a strong mediating effect on berry numbers, particularly under warmed conditions and mainly in the year after MeJA application. Synthesis. These results demonstrate the need to consider combined pressures in understanding the impacts of global change on plants and highlight that multiple drivers may have unidentified synergetic effects on species phenology and reproduction.

SmJAZ1/8 inhibits the stimulation of SmbHLH59, which limits the accumulation of salvianolic acids and tanshinones in Salvia miltiorrhiza

Salvia miltiorrhiza is a model medicinal plant that is typically used to treat cardiovascular and cerebrovascular diseases. The primary active medicinal ingredients of S. miltiorrhiza are salvianolic acids and tanshinones. Jasmonate (JA) is a vital phytohormone that regulates secondary metabolism. The exogenous application of methyl jasmonate (MeJA) can promote the accumulation of active ingredients in S. miltiorrhiza. Here, we identified a MeJA-responsive SmbHLH59 gene that encodes for a bHLH IIIe family transcription factor. The overexpression of SmbHLH59 in S. miltiorrhiza increased the contents of salvianolic acids and tanshinones, while the opposite effect was observed when SmbHLH59 was knocked out via CRISPR. Meanwhile, SmbHLH59 was observed to activate the expressions of SmPAL1, SmC4H1, SmHPPR1, SmCPS1, and SmKSL1 genes by binding to the E/G-box elements of their promoters. Further investigations demonstrated that SmJAZ1 and SmJAZ8 interacted with SmbHLH59 to inhibit its activation of these five genes. In summary, a JA signaling pathway component (SmbHLH59) was identified that strongly enhanced the accumulation of salvianolic acids and tanshinones through the direct activation of multiple enzyme genes in their biosynthetic pathways. Consequently, this study enriches our knowledge toward further elucidating the molecular mechanisms behind the regulation of JA in the secondary metabolism of S. miltiorrhiza.

Foliar Methyl Jasmonate Application Activates Antioxidant Mechanisms to Counteract Water Deficits and Aluminum Stress in Vaccinium corymbosum L.

Due to climate change, water deficits (WDs) and aluminum (Al) toxicity are increasing, affecting plants, especially crops such as blueberries (Vaccinium corymbosum L.). The application of methyl jasmonate (MeJA) could mitigate these effects. This work aimed to evaluate the effective MeJA dose to overcome oxidative stress provoked by combined WD+Al stress in blueberries. Plants of Al-sensitive (Star) and Al-resistant (Legacy) cultivars were exposed to control (Al at 65 mg/Kg, 80% field capacity), WD+Al (50% field capacity; Al at 1665 mg/Kg), and WD+Al treatment with different foliar MeJA doses (10, 50, and 100 μM) during 7 and 21 days. Data revealed that plants exposed to WD+Al and treated with 50 µM MeJA reduced Al up to 3.2-fold in roots and 2.7-fold in leaves and improved water potential (Ψw) up to 2.5-fold. The sensitive cultivar decreased the relative growth rate under WD+Al, increasing by 1.9-fold with 50 µM MeJA. Under WD+Al stress, all MeJA doses mitigated the decrease in relative water content in Al-resistant cultivars, restoring values like control plants. In the sensitive cultivar, 50 µM MeJA increased photosynthesis (1.5-fold) and stomatal conductance (1.4-fold), without changes in transpiration. Lipid peroxidation decreased (1.2-fold) and increased antioxidant activity (1.8-fold), total phenols (1.6-fold), and superoxide dismutase activity (3.3-fold) under WD+Al and 50 µM-MeJA. It was concluded that the most effective dose to alleviate the WD+Al stress was 50 µM MeJA due to the activation of antioxidants in blueberry plants. Therefore, the MeJA application could be a potential strategy for enhancing the resilience of V. corynbosum exposed to WD+Al stress.

Effects of foliar applications of methyl jasmonate alone or with urea on anthocyanins content during grape ripening

Anthocyanins, the main phenolic compounds responsible for the color of red grapes and wines, exhibit antioxidant and antimicrobial properties and offer various health benefits for humans. Currently, climate change can affect grape quality by causing a decoupling between the technological and phenolic maturities of the grapes. Foliar application of methyl jasmonate (MeJ) and urea (Ur) can be a tool to mitigate the global warming effects in the vineyard. The aim of this work was to study i) the development of anthocyanin content in ‘Tempranillo’ grapes along the ripening process, and ii) the effect of foliar application of MeJ and MeJ+Ur on these phenolic compounds over two vintages (2019 and 2020). Overall, in the first vintage, anthocyanins peaked in their concentrations in the pre-harvest or harvest moments; whereas in the second vintage, the highest contents of these phenolic compounds were reached at harvest time, after that, a maintenance or decreased of their concentrations was observed. In both vintages, non-acylated anthocyanins was the most abundant family of anthocyanins. Foliar application of MeJ+Ur was more effective in enhancing anthocyanin biosynthesis than MeJ treatment, which also increased anthocyanin content compared to control grapes. Therefore, MeJ and Ur foliar treatment could present a synergy in anthocyanin biosynthesis by the plant. Moreover, the impact of these applications varied across vintages, and also the content of anthocyanins in grapes, probably due to different climatological conditions. Consequently, the two foliar treatments, MeJ and MeJ+Ur, could be a suitable tool to increase anthocyanins biosynthesis in grapes, and thus improve grape quality.

Foliar Application of Methyl Jasmonate and Chitosan Improve Growth, Yield, and Quality of Guar (Cyamopsis tetragonoloba L.) Under Water-Deficit Stress.

Guar (Cyamopsis tetragonoloba L.), a summer legume, is becoming increasingly important as an industrial crop due to its high gum and viscosity content. This study investigated the effects of methyl jasmonate (MeJA), chitosan (CH), and their combination on the growth, yield, and quality of guar under irrigation regimes. A greenhouse experiment was conducted using a factorial design to evaluate the effect of foliar spraying with MeJA (5, 25, and 50 µM), CH (100, 150, and 200 mg/L), their combination (25 µM MeJA + 150 mg/L CH), and control on two commercial guar varieties (RGC-986 and BR-2017) under different irrigation regimes (100%, 70%, and 40% field capacity). The results showed that the exogenous application of MeJA and CH, individually and in combination, significantly enhanced various morphological traits and yield components in guar, including plant height, pod characteristics, seed yield, and root development. Additionally, the combination treatments improved seed quality parameters, such as gum percentage and viscosity content. Leaf analysis revealed increased levels of total phenolic content, total flavonoid, and anthocyanin contents. The BR-2017 variety showed superior performance in most morphological and qualitative traits, demonstrating greater resistance to irrigation regimes. It maintained yield and quality characteristics under water-deficit conditions, particularly when treated with 25 µM MeJA and 150 mg/L CH. The highest gum percentage (33.67%) and viscosity (4768.5 cP) were observed in the RGC-986 variety, along with enhanced levels of secondary metabolites. This study provides new insights into how MeJA, CH, and their combination can improve the yield and quality of guar under water deficit stress conditions. The results suggest that the use of these elicitors, especially in combination, represents an innovative strategy for improving guar production and quality, with potential variety-specific responses to water-deficit stress.

Positive association between foliar silicon and extrafloral nectar in Vicia faba with application of methyl jasmonate

Plants have evolved direct and indirect defences against herbivores, which may come at a cost to other plant functions. Many plants can uptake and deposit large amounts of silicon within plant tissue, creating structures which can reduce herbivore performance. Silicon uptake can increase plant defenses against herbivores, but it has also been shown to trade-off with defensive phenolic compounds due to interference with jasmonic acid (JA) signaling. Additionally, plants can recruit predacious insects with extrafloral nectar (EFN), a sugar secretion not involved in pollination. It is currently unclear whether silicon uptake reduces other putative defences associated with the JA pathway, like EFN production. We used faba bean (Vicia faba) to identify potential trade-offs between silicon accumulation, phenolic content, and EFN production. We grew four genotypes of faba bean that varied in tannin content in control soil, or soil supplemented with silicon. After five weeks of growth, we exposed plants to either a buffer or methyl jasmonate (MeJA) solution to simulate an herbivory response. We measured EFN production at 24 and 48 hours after treatment, and harvested leaves to quantify silicon and phenolic content. We found silicon supplementation, but not MeJA treatment, increased foliar silicon concentration. Silicon supplementation did not affect foliar phenolic content or EFN sugar content. Silicon concentration (ppm) and MeJA treatment did not decrease foliar phenolic content or EFN sugar content. However, we found an interaction between silicon concentration (ppm) and MeJA treatment with EFN sugar content: across MeJA-treated plants, we detected a positive association between foliar silicon concentration and the amount of sugar (mg) in EFN. This study is the first to show MeJA can interact with leaf silicon concentration to alter EFN response, with the potential for cascading effects on other trophic levels.

Optimization of Cannabinoid Production in Hemp Through Methyl Jasmonate Application in a Vertical Farming System

Optimization of Cannabinoid Production in Hemp Through Methyl Jasmonate Application in a Vertical Farming System

In vitro application of Methyl jasmonate to induce phytochemical production and antioxidant activity in Dracocephalum polychaetum Bornm. cells

In vitro application of Methyl jasmonate to induce phytochemical production and antioxidant activity in Dracocephalum polychaetum Bornm. cells

Methyl jasmonate mitigates Fusarium graminearum infection in wheat by inhibiting deoxynivalenol synthesis.

Methyl jasmonate (MeJA), a plant growth regulator, coordinates a diverse array of physiological responses, including the inhibition of seed germination, modulation of secondary metabolite biosynthesis, and activation of defence responses. The external application of MeJA has been demonstrated to effectively diminish the severity of fungal diseases. Here, we unveil a novel mechanism through which exogenous MeJA alleviates Fusarium head blight (FHB) by inhibiting the synthesis of deoxynivalenol (DON) in Fusarium graminearum, rather than by enhancing the wheat resistance response. MeJA treatment reduced the infection by wild-type F. graminearum in wheat coleoptiles, but exhibited no significant influence on that of the DON-deficient mutant strain (∆Tri5). The production of DON in F. graminearum was significantly inhibited both in vitro and in planta. MeJA affected the expression of genes related to DON biosynthesis, without influencing the formation of toxisomes as observed under microscopic analysis. Exogenous MeJA demonstrated a limited impact on the early genes of plant jasmonic acid signalling pathway, in contrast to the wild-type pathogen strain, which induced the upregulation of these genes. The expression levels of defence marker genes induced by MeJA were notably lower compared to those induced by the pathogen. This study elucidates the molecular mechanisms of MeJA in modulating the wheat-F. graminearum interaction, providing new insights into the development of environmentally friendly strategies against fungi.

Transcriptomic Analysis of the Combined Effects of Methyl Jasmonate and Wounding on Flavonoid and Anthraquinone Biosynthesis in Senna tora.

Jasmonates, including jasmonic acid (JA) and its derivatives such as methyl jasmonate (MeJA) or jasmonly isoleucine (JA-Ile), regulate plant responses to various biotic and abiotic stresses. In this study, we applied exogenous MeJA onto Senna tora leaves subjected to wounding and conducted a transcriptome deep sequencing analysis at 1 (T1), 3 (T3), 6 (T6), and 24 (T24) h after MeJA induction, along with the pretreatment control at 0 h (T0). Out of 18,883 mapped genes, we identified 10,048 differentially expressed genes (DEGs) between the T0 time point and at least one of the four treatment times. We detected the most DEGs at T3, followed by T6, T1, and T24. We observed the upregulation of genes related to JA biosynthesis upon exogenous MeJA application. Similarly, transcript levels of genes related to flavonoid biosynthesis increased after MeJA application and tended to reach their maximum at T6. In agreement, the flavonols kaempferol and quercetin reached their highest accumulation at T24, whereas the levels of the anthraquinones aloe-emodin, emodin, and citreorosein remained constant until T24. This study highlights an increase in flavonoid biosynthesis following both MeJA application and mechanical wounding, whereas no significant influence is observed on anthraquinone biosynthesis. These results provide insights into the distinct regulatory pathways of flavonoid and anthraquinone biosynthesis in response to MeJA and mechanical wounding.

Methyl Jasmonate (MeJA) Promotes the Self-Pollen Tube Growth of Camellia oleifera by Regulating Lignin Biosynthesis

Self-incompatibility (SI) poses a significant reproductive barrier, severely impacting the yield, quality, and economic value of Camellia oleifera. In this study, methyl jasmonate (MeJA) was employed as an exogenous stimulus to alleviate SI in C. oleifera. The research findings revealed that an exogenous dose of 1000 μmol·L−1 MeJA enhanced the germination and tube growth of C. oleifera self-pollen and greatly improved ovule penetration (18.75%) and fertilization (15.81%), ultimately increasing fruit setting (18.67%). It was discovered by transcriptome analysis that the key genes (CAD, C4H) involved in the lignin production process exhibited elevated expression levels in self-pistils treated with MeJA. Further analysis showed that the lignin concentration in the MeJA-treated pistils was 31.70% higher compared with the control group. As verified by pollen germination assays in vitro, lignin in the appropriate concentration range could promote pollen tube growth. Gene expression network analysis indicated that transcription factor bHLH may be pivotal in regulating lignin biosynthesis in response to MeJA, which in turn affects pollen tubes. Further transient knockdown of bHLH (Co_33962) confirmed its important role in C. oleifera pollen tube growth. In summary, the application of MeJA resulted in the stimulation of self-pollen tube elongation and enhanced fruit setting in C. oleifera, which could be associated with the differential change in genes related to lignin synthesis and the increased lignin content.

Methyl salicylate induces endogenous jasmonic acid and salicylic acid in 'Nam Dok Mai' mango to maintain postharvest ripening and quality

Salicylic acid (SA) and jasmonic acid (JA) are indispensable phytohormones whose interaction influences the ripening process in plants. Methyl salicylate (MeSA) and methyl jasmonate (MeJA) have been utilized to elevate the endogenous levels of SA and JA in horticultural products after harvest. However, their ability to preserve mango is uncertain. Individually and combined effects of exogenous MeSA and MeJA on mango ripening quality were investigated. 'Nam Dok Mai' mangoes were fumigated with MeSA, MeJA, and MeSA plus MeJA (MeSAJA) prior to storage for 6 d at 25 °C and 80–85% relative humidity (RH). Fruit ripening attributes, respiration rate, ethylene (ET) production, total phenolics (TP), and malondialdehyde (MDA) were assessed. Endogenous SA and JA levels were measured, as were the activities of lipoxygenase (LOX) and phenylalanine ammonia-lyase (PAL), and the expression of related genes MiPAL, MiICS, and MiLOX. Individual application of MeSA or MeJA preserved the mango quality by reducing ET production, respiration rate, and MDA levels while raising TP shortly after treatment. The ripening quality mirrored the induced SA and JA levels and correlated with the high expression of biosynthetic-related genes (MiPAL, MiICS, and MiLOX). Individual treatments stimulated SA and JA biosynthesis, demonstrating that these phytohormones are functionally connected and interdependent. When combined, MeSAJA caused a tradeoff response and distinct phenotypic outcomes compared to the individual treatments. As a result, MeSA fumigation is a practical method for preserving mango quality after harvest.

Integrated volatile metabolome and transcriptome analyses provide insights into the warm aroma formation elicited by methyl jasmonate in carrot root.

Carrot is a highly significant vegetable cultivated worldwide and possesses a unique aroma with abundant edible and medicinal values. However, it remains largely unknown whether jasmonic acid could regulate aroma formation in carrot. Here, an integrated analysis of the volatile metabolome and transcriptome of carrot roots exposed to different concentrations of methyl jasmonate (MeJA) was performed. The results revealed 1,227 volatile organic compounds and 972 differential accumulated metabolites, with terpenes representing the largest portion. MeJA treatment evidently increased the relative odor activity values as well as the accumulation of most volatile compounds. In addition, 4,787 differentially expressed genes were identified and subjected to function enrichment analysis, indicating a role of terpene biosynthesis and metabolism in response to MeJA application. A network consisting of 4,680 transcription factor-structural pairs that showed highly significant positive correlations was constructed, which may be utilized as genetic targets for examining terpene accumulation and aroma formation elicited by methyl jasmonate. The results from the present work substantially improved our understanding of MeJA-mediated aroma formation in carrot.

Synergistic suppression of grey mold on cut rose flower by combined application of methyl jasmonate and lipopeptides from Bacillus altitudinis

Roses are the most popular cut flowers that are widely used in various occasions and celebrations, yet can also be easily infected by grey mold caused by Botrytis cinerea; a notorious pathogen in the cut flower industry. A wide range of methods, including fungicides, have been used to control B. cinerea. Environmentally benign methods are increasingly sought after to minimize disease incidence. In this study, we evaluated 10 strains of Bacillus across eight species for their action against B. cinerea. Strains TM22A (Bacillus altitudinis) and S2 (Bacillus subtilis) were identified to be effective in disease suppression. Lipopeptides (LPs) were isolated from two strains, and their activity for inhibiting the growth of B. cinerea was tested. Additionally, methyl jasmonate (MeJA), a natural growth regulator, was evaluated for disease suppression using a in planta assay. Subsequently, LPs from TM22A and MeJA, alone or in combination, were evaluated for disease occurrence in cut rose flowers. Results showed that either LPs or MeJA at concentrations of 250 μmol/L significantly reduced lesion size and disease incidence. The combined application of LPs from TM22A with MeJA further reduced the disease occurrence in cut rose flowers. Furthermore, the activities of POD, PPO, PAL, CAT, and SOD enzymes were much higher in flowers treated with combined application of LPs from TM22A with MeJA. The enhanced suppression of B. cinerea is attributed to the antifungal effects of LPs and MeJA and activation of antioxidant enzymes. This study suggests that the combined application of LPs from TM22A with MeJA could be an environmentally friendly approach for control of grey mold disease in cut roses.

Methyl jasmonate induced tolerance effect of Pinus koraiensis to Bursaphelenchus xylophilus.

Methyl jasmonate (MeJA) can affect the balance of hormones and regulate the disease resistance of plants. Exploring the application and mechanism of MeJA in inducing the tolerance of Pinus koraiensis to pine wood nematode (PWN) infection is of great significance for developing new strategies for pine wilt disease control. Different concentrations (0.1, 1, 5 and 10 mm) of MeJA treatment groups showed differences in relative tolerance index and relative anti-nematode index of P. koraiensis seedlings to PWN infection. The treatment of 5 mm MeJA solution induced the best tolerance effect, followed by the 1 mm MeJA solution. Transcriptome analysis indicated that many plant defense-related genes upregulated after treatment with 1, 5 and 10 mm MeJA solutions. Among them, genes such as jasmonate ZIM domain-containing protein, phenylalanine ammonia-lyase and peroxidase also continuously upregulated after PWN infection. Metabolome analysis indicated that jasmonic acid (JA) was significantly increased at 7 days postinoculation with PWN, and after treatment with both 1 and 5 mm MeJA solutions. Integrated analysis of transcriptome and metabolome indicated that differences in JA accumulation might lead to ubiquitin-mediated proteolysis, and expression changes in trans-caffeic acid and trans-cinnamic acid-related genes, leading to the abundance differences of these two metabolisms and the formation of multiple lignin and glucosides. MeJA treatment could activate the expression of defense-related genes that correlated with JA, regulate the abundance of defense-related secondary metabolites, and improve the tolerance of P. koraiensis seedlings to PWN infection. © 2024 Society of Chemical Industry.

Integrative metabolome and transcriptome analysis characterized methyl jasmonate-elicited flavonoid metabolites of Blumea balsamifera.

As a commonly used medicinal plant, the flavonoid metabolites of Blumea balsamifera and their association with genes are still elusive. In this study, the total flavonoid content (TFC), flavonoid metabolites and biosynthetic gene expression patterns of B. balsamifera after application of exogenous methyl jasmonate (MeJA) were scrutinized. The different concentrations of exogenous MeJA increased the TFC of B. balsamifera leaves after 48 h of exposure, and there was a positive correlation between TFC and the elicitor concentration. A total of 48 flavonoid metabolites, falling into 10 structural classes, were identified, among which flavones and flavanones were predominant. After screening candidate genes by transcriptome mining, the comprehensive analysis of gene expression level and TFC suggested that FLS and MYB may be key genes that regulate the TFC in B. balsamifera leaves under exogenous MeJA treatment. This study lays a foundation for elucidating flavonoids of B. balsamifera, and navigates the breeding of flavonoid-rich B. balsamifera varieties.

Promoter of COR2-like gene is a stress inducible regulatory region in banana

A promoter is a crucial component in driving the expression of a transgene of interest for biotechnological applications in crop improvement and thus characterization of varied regulatory regions is essential. Here, we identified the promoter of COR2-like (codeinone reductase-like) from banana and characterized its tissue specific and stress inducible nature. MusaCOR2-like of banana is closely related to COR2 and CHR (chalcone reductase) sequences from different plant species and contains signature sequences including a catalytic tetrad typical of proteins with aldo–keto reductase activity. Transcript level of MusaCOR2-like was strongly induced in response to drought, salinity and exposure of signaling molecules such as abscisic acid, methyl-jasmonate and salicylic acid. Induction of MusaCOR2-like under stress strongly correlated with the presence of multiple cis-elements associated with stress responses in the PMusaCOR2-like sequence isolated from Musa cultivar Rasthali. Transgenic tobacco lines harbouring PMusaCOR2-like-GUS displayed visible GUS expression in vascular tissue of leaves and stem while its expression was undetectable in roots under control conditions. Exposure to drought, salinity and cold strongly induced GUS expression from PMusaCOR2-like-GUS in transgenic tobacco shoots in a window period of 3H to 12H. Applications of salicylic acid, methyl-jasmonate, abscisic acid and ethephon also activate GUS in transgenic shoots at different period, with salicylic acid and abscisic acid being the stronger stimulants of PMusaCOR2-like. Using PMusaCOR2-like-GUS fusion and expression profiling, the current study sheds insights into a complex regulation of COR2-like, one of the least studied genes of secondary metabolite pathway in plants.

Preharvest methyl jasmonate application delays cell wall degradation and upregulates phenolic metabolism and antioxidant activities in cold stored raspberries

The effects of preharvest methyl jasmonate (MeJA) spray application on the physicochemical quality, metabolism of phenolics, and cell wall components in raspberries were investigated during a 10-day cold storage period. MeJA spray reduced firmness loss, decay incidence, and weight loss, while maintained higher levels of soluble solids content, ascorbic acid, anthocyanins and flavonoids in raspberries. Furthermore, MeJA application resulted in increased total pectin and protopectin levels, as well as lowered water-soluble pectin, and activities of pectin methyl esterase, polygalacturonase and cellulase enzymes. Additionally, MeJA treatment upregulated the phenylpropanoid pathway, leading to higher endogenous phenolics and activities of phenylalanine-ammonia lyase and shikimate dehydrogenase. In conclusion, preharvest MeJA spray application could be adopted to enhance the storage potential of cold-stored raspberries for 10 days by maintaining higher firmness, assuring better physicochemical quality, and increasing phenolic metabolism, while reducing cell wall hydrolysis.

PpBBX32 and PpZAT5 modulate temperature-dependent and tissue-specific anthocyanin accumulation in peach fruit.

Anthocyanins are important compounds for fruit quality and nutrition. The R2R3 MYB transcription factor PpMYB10.1 is known to be critical for regulating anthocyanin accumulation in peach. However, regulatory factors upstream of PpMYB10.1 which control temperature-dependent, cultivar-contrasted and tissue-specific anthocyanin accumulation remain to be determined. In this study, differential anthocyanin accumulation in the outer flesh near the peel (OF) of peach [Prunus persica (L.) Batsch] was observed between cultivars 'Zhonghuashoutao' and 'Dongxuemi', as well as among different storage temperatures and different fruit tissues of 'Zhonghuashoutao'. By cross-comparisons of RNA-Seq data of samples with differential anthocyanin accumulation, transcription factor genes PpBBX32 and PpZAT5 were identified. These were functionally characterized as two positive regulators for anthocyanin accumulation via transient expression and genetic transformation. Various interaction assays revealed that both PpBBX32 and PpZAT5 can directly activate the PpMYB10.1 promoter and meanwhile interact at protein level as a PpZAT5-PpBBX32-PpMYB10.1 complex. Furthermore, the results of in silico analysis and exogenous application of methyl jasmonate (MeJA) indicated that MeJA favored anthocyanin accumulation, while it was also found that anthocyanin accumulation as well as PpBBX32 and PpZAT5 expression correlated significantly with endogenous JA and JA-Ile in different fruit tissues. In summary, PpBBX32 and PpZAT5 are upstream activators of PpMYB10.1, allowing JAs to take part in temperature-dependent and tissue-specific anthocyanin accumulation by modulating their expression. This work enriches the knowledge of the transcriptional regulatory mechanisms for differential anthocyanin accumulation under internal and external factors.

CaWRKY22b Plays a Positive Role in the Regulation of Pepper Resistance to Ralstonia solanacearum in a Manner Associated with Jasmonic Acid Signaling.

As important transcription factors, WRKYs play a vital role in the defense response of plants against the invasion of multiple pathogens. Though some WRKY members have been reported to participate in pepper immunity in response to Ralstonia solanacearum infection, the functions of the majority of WRKY members are still unknown. Herein, CaWRKY22b was cloned from the pepper genome and its function against R. solanacearum was analyzed. The transcript abundance of CaWRKY22b was significantly increased in response to the infection of R. solanacearum and the application of exogenous methyl jasmonate (MeJA). Subcellular localization assay in the leaves of Nicotiana benthamiana showed that CaWRKY22b protein was targeted to the nuclei. Agrobacterium-mediated transient expression in pepper leaves indicated that CaWRKY22b overexpression triggered intensive hypersensitive response-like cell death, H2O2 accumulation, and the up-regulation of defense- and JA-responsive genes, including CaHIR1, CaPO2, CaBPR1, and CaDEF1. Virus-induced gene silencing assay revealed that knock-down of CaWRKY22b attenuated pepper's resistance against R. solanacearum and the up-regulation of the tested defense- and jasmonic acid (JA)-responsive genes. We further assessed the role of CaWRKY22b in modulating the expression of JA-responsive CaDEF1, and the result demonstrated that CaWRKY22b trans-activated CaDEF1 expression by directly binding to its upstream promoter. Collectively, our results suggest that CaWRKY22b positively regulated pepper immunity against R. solanacearum in a manner associated with JA signaling, probably by modulating the expression of JA-responsive CaDEF1.