VPD modifies CO_(2) fertilization effect on tomato plants via abscisic acid and jasmonic acid signaling pathways

Atmospheric CO_(2)concentration is elevated globally,which has“CO_(2)fertilization effects”and potentially improves plant photosynthesis,yield,and productivity.Despite the beneficial effect of CO_(2)fertilization being modulated by vapor pressure deficit(VPD),the underlying mechanism is highly uncertain.In the present study,the potential roles of hormones in determining CO_(2)fertilization effects under contrasting high and low VPD conditions were investigated by integrated physiological and transcriptomic analyses.Beneficial CO_(2)fertilization effects were offset under high VPD conditions and were constrained by plant water stress and photosynthetic CO_(2)utilization.High VPD induced a large passive water driving force,which disrupted the water balance and consequently caused plant water deficit.Leaf water potential,turgor pressure,and hydraulic conductance declined under high VPD stress.The physiological evidence combined with transcriptomic analyses demonstrated that abscisic acid(ABA)and jasmonic acid(JA)potentially acted as drought-signaling molecules in response to high VPD stress.Increased foliar ABA and JA content triggered stomatal closure to prevent excessive water loss under high VPD stress,which simultaneously increased the diffusion resistance for CO_(2)uptake from atmosphere to leaf intercellular space.High VPD also significantly increased mesophyll resistance for CO_(2)transport from stomatal cavity to fixation site inside chloroplast.The chloroplast“sink”CO_(2)availability was constrained by stomatal and mesophyll resistance under high VPD stress,despite the atmospheric“source”CO_(2)concentration being elevated.Thus,ABA-and JA-mediated drought-resistant mechanisms potentially modified the beneficial effect of CO_(2)fertilization on photosynthesis,plant growth,and yield productivity.This study provides valuable information for improving the utilization efficiency of CO_(2)fertilization and a better understanding of the physiological processes.

High-temperature stress suppresses allene oxide cyclase 2 and causes male sterility in cotton by disrupting jasmonic acid signaling

Cotton(Gossypium spp.) yield is reduced by stress. In this study, high temperature(HT) suppressed the expression of the jasmonic acid(JA) biosynthesis gene allene oxide cyclase 2(GhAOC2), reducing JA content and causing male sterility in the cotton HT-sensitive line H05. Anther sterility was reversed by exogenous application of methyl jasmonate(MeJA) to early buds. To elucidate the role of GhAOC2 in JA biosynthesis and identify its putative contribution to the anther response to HT, we created gene knockout cotton plants using the CRISPR/Cas9 system. Ghaoc2 mutant lines showed male-sterile flowers with reduced JA content in the anthers at the tetrad stage(TS), tapetum degradation stage(TDS), and anther dehiscence stage(ADS). Exogenous application of MeJA to early mutant buds(containing TS or TDS anthers) rescued the sterile pollen and indehiscent anther phenotypes, while ROS signals were reduced in ADS anthers. We propose that HT downregulates the expression of GhAOC2 in anthers, reducing JA biosynthesis and causing excessive ROS accumulation in anthers, leading to male sterility. These findings suggest exogenous JA application as a strategy for increasing male fertility in cotton under HT.

Jasmonic acid-mediated stress responses share the molecular mechanism underlying male sterility induced by deficiency of ZmMs33 in maize

Plant male reproduction is a fine-tuned developmental process that is susceptible to stressful environments and influences crop grain yields.Phytohormone signaling functions in control of plant normal growth and development as well as in response to external stresses,but the interaction or crosstalk among phytohormone signaling,stress response,and male reproduction in plants remains poorly understood.Cross-species comparison among 514 stress-response transcriptomic libraries revealed that ms33-6038,a genic male sterile mutant deficient in the Zm Ms33/Zm GPAT6 gene,displayed an excessive drought stress-like transcriptional reprogramming in anthers triggered mainly by disturbed jasmonic acid(JA)homeostasis.An increased level of JA appeared in Zm Ms33-deficient anthers at both meiotic and postmeiotic stages and activated genes involved in JA biosynthesis and signaling as well as genes functioning in JA-mediated drought response.Excessive accumulation of JA elevated expression level of a gene encoding a WRKY transcription factor that activated the Zm Ms33 promoter.These findings reveal a feedback loop of Zm Ms33-JA-WRKY-Zm Ms33 in controlling male sterility and JA-mediated stress response in maize,shedding light on the crosstalk of stress response and male sterility mediated by phytohormone homeostasis and signaling.

Localized Effects of Mechanical Wounding and Exogenous Jasmonic Acid on the Induction of Secondary Laticifer Differentiation in Relation to the Distribution of Jasmonic Acid in Hevea brasiliensis

The relationship between the effect of exogenous jasmonic acid (JA) on the induction of secondary laticifer differentiation and the distribution of JA in the seedling of Hevea brasiliensis Mull. Arg. was investigated with the aid of experimental morphological and radioisotope technique. Most radioactivity of H-3-JA sustained in treated site within one hour while no radioactivity was detected in new shoot and the radioactivity in upper leaf was much less than that in the parts below the treated site, suggesting that JA was mainly transported downwards in the shoot of H brasiliensis. Mechanical wounding hindered the entrance of exogenous JA remarkably while held back the entered JA to the regions around wounded site. The effect of exogenous JA and mechanical wounding on the induction of the secondary laticifer differentiation was limited to treated site where high level of JA was expected. Mechanical wounding reduced the effect of exogenous JA on the differentiation of secondary laticifer, which could be ascribed to the hindrance of mechanical wounding to the entrance of exogenous JA. It was concluded from the combined data that a high accumulation of JA was required for inducing the secondary laticifer differentiation in H. brasiliensis.

Effect of Jasmonic Acid on Lateral Root Formation in Rice Seedling

实验材料为水稻 (OryzasativaL .)栽培品种“IR8”(国际水稻所 8号 )及其少侧根突变体MT10。将 2d水稻幼苗种子根全部浸入 0 .0 16～ 5 0 μmol/L茉莉酸 (JA)溶液处理 2d ,结果表明JA显著抑制种子根的伸长 ,其抑制程度与JA浓度成正比。不高于 2 μmol/L的JA显著促进侧根的发生 ,每cm的侧根数目随浓度的增加而增加 ,最多可增加到原来的 16 8% (“IR8”)和 2 85 % (MT10 )。 10 μmol/L的JA仍促进处理过程中和处理后生成根区段的侧根数目的增加 ,但明显抑制处理前生成根区段侧根的发生 。

Effect of Localized Scorch on the Transport and Distribution of Exogenous Jasmonic Acid in Vicia faba

Exogenous jasmonic acid (JA) has been showed to be able to induce stomatal closure in Vicia faba L. in previous investigations. The transport and distribution of 3H-JA affected by localized scorch on V. faba seedling were studied with radioisotope technique. The results showed that 3H-JA could be transported up or down at the rate of 4-5 cm·min -1 following feeding into root or shoot tip. The transport of 3H-JA in shoot reached a relative stable rate at 30 min after being fed through root. Wounding by scorch in the youngest leaf caused an increase in the transport of 3H-JA from root to shoot and enhanced the distribution of 3H-JA in the wounded leaf. However, distribution of 3H-JA in unwounded leaves increased after 5 h being fed through the youngest leaf. It was noticed that wounding improved accumulation of 3H-JA in abaxial epidermis. Consistent results were obtained: wounding prevented transport of 3H-JA out from the youngest leaf to root; These observations suggest that JA plays an important role as a defense signal and might be involved in the regulation of the stomatal movement in response to wounding stress.

Research on Expression of LeWRKY1 in Tomato Induced by Jasmonic Acid and Other Two Factors

[Objective]The aim was to explore the molecular mechanism of plant resistance to various stress response.[Method]The expression of LeWRKY1 in tomato seedlings under treatment with B.cinerea,exogenous JA and SA were explored by real time quantitative RT-PCR technology.[Result]JA induced the expression of LeWRKY1,but SA did not.LeWRKY1 expression was up-regulated under B.cinerea infection.[Conclusion]LeWRKY1 might be involved in the tomato defense response to B.cinerea through JA dependent but SA independent signal pathway.

Effects of Calcium and Jasmonic Acid on CBF Expression in Spinach

[Objective] This experiment aimed to evaluate the effects of calcium and Jasmonic acid（JA） on the expression of CBF in spinach. [Methods] The seedlings of spinach were treated with low temperature （4 ℃）, JA or A23187, then used for detecting the expression of CBF by northern blotting. [Results] The results showed that the CBF expression was regulated by low temperature and JA positively. [Conclusions] Low temperature may increase the JA content of the cell firstly, then JA induced the increase of cytosolic calcium concentration （[Ca2＋]cyt）, and the JA induced Ca2＋ transmitted the low temperature signal through CaM or CaM related proteins, regulating the CBF expression.

IP_3 Sensitive Calcium Channel Involved in the Jasmonic Acid Induced Calcium Mobilization

[Objective] This study was to investigate the role of IP3 sensitive calcium channel in the JA-induced calcium mobilization pathway.[Method] Arabidopsis thaliana leaves were labeled by Fluorescent probe Fluo-3/AM under low temperature at 4 ℃ to measure the fluorescent intensity of intracellular Ca2＋ which was pretreated with heparin on jasmonic acid（JA）-induced.[Results] When A.thaliana leaf cells were pretreated with 10,50 or 100 ng/ml heparin,intercellular free Ca2＋ fluorescence intensity was reduced in comparison with negative control.Once the heparin-pretreated A.thaliana leaf cells were stimulated with 100 μmol/L JA,intercellular Ca2＋ fluorescence intensity increased gradually and tended to be stable at a degree equivalent with that in negative control.[Conclusion] The experiment showed that the pretreatment with heparin could inhibit the increase of the intracellular Ca2＋ concentration significantly which JA-induced in leaves of Arabidopsis thaliana.

Role of the Arabidopsis thafiana NAC transcription factors ANAC019 and ANAC055 in regulating jasmonic acid-signaled defense responses

Jasmonic acid （JA） is an important phytohormone that regulates plant defense responses against herbivore attack, pathogen infection and mechanical wounding. In this report, we provided biochemical and genetic evidence to show that the Arabidopsis thaliana NAC family proteins ANAC019 and ANAC055 might function as transcription activators to regulate JA-induced expression of defense genes. The role of the two NAC genes in JA signaling was examined with the anacO19 anac055 double mutant and with transgenic plants overexpressing ANACO19 or ANAC055. The anacO19 anac055 double mutant plants showed attenuated JA-induced VEGETATIVE STORAGE PROTEIN1 （VSP1） and LIPOXYGENASE2 （LOX2） expression, whereas transgenic plants overexpressing the two NAC genes showed enhanced JA-induced VSP1 and LOX2 expression. That the JA-induced expression of the two NAC genes depends on the function of COIl and AtMYC2, together with the finding that overexpression of ANACO19 partially rescued the JA-related phenotype of the atmyc2-2 mutant, has led us to a hypothesis that the two NAC proteins act downstream of AtMYC2 to regulate JA-signaled defense responses. Further evidence to substantiate this idea comes from the observation that the response of the anacO19 anac055 double mutant to a necrotrophic fungus showed high similarity to that of the atmyc2-2 mutant.

Abscisic acid and jasmonic acid are involved in drought priming-induced tolerance to drought in wheat

Drought stress is a limiting factor for wheat production and food security.Drought priming has been shown to increase drought tolerance in wheat.However,the underlying mechanisms are unknown.In the present study,the genes encoding the biosynthesis and metabolism of abscisic acid(ABA)and jasmonic acid(JA),as well as genes involved in the ABA and JA signaling pathways were up-regulated by drought priming.Endogenous concentrations of JA and ABA increased following drought priming.The interplay between JA and ABA in plant responses to drought priming was further investigated using inhibitors of ABA and JA biosynthesis.Application of fluridone(FLU)or nordihydroguaiaretic acid(NDGA)to primed plants resulted in lower chlorophyll-fluorescence parameters and activities of superoxide dismutase and glutathione reductase,and higher cell membrane damage,compared to primed plants(PD)under drought stress.NDGA+ABA,but not FLU+JA,restored priming-induced tolerance,as indicated by a finding of no significant difference from PD under drought stress.Under drought priming,NDGA induced the suppression of ABA accumulation,while FLU did not affect JA accumulation.These results were consistent with the expression of genes involved in the biosynthesis of ABA and JA.They suggest that ABA and JA are required for priming-induced drought tolerance in wheat,with JA acting upstream of ABA.

Effect of Jasmonic Acid on Photosynthetic Pigments and Stress Markers in <i>Cajanus cajan</i>(L.) Millsp. Seedlings under Copper Stress

Jasmonates are class of plant growth regulators act as signal molecule that intercede various components in physiological and metabolic regulation, stress responses and possibly communication through signal transduction. Oxidative stress due to heavy metal exposure stimulates synthesis and activity of antioxidant metabolites and enhances antioxidant enzyme activities that could protect plant tissues. The aim of this study was to investigate the exogenous effect of JA at seed level which can transduce throughout seedling growth and regulate antioxidant activities such as superoxide dismutase (SOD;EC 1.15.1.1) and guaiacol peroxidase (POD;EC 1.11.1.7) in 12 days old seedlings of pigeon pea (Cajanus cajan (L.) Millsp.) in presence and/or absence of copper. The activity of SOD and POD increased significantly in presence of Cu2+ after seed priming with JA. JA also helps in chlorophyll and carotenoid accumulation and neutralizes the toxic effect of Cu2+ on seedlings. This is the first report of JA effect on photosynthetic pigment accumulation and H2O2 mitigating enzymes i.e. SOD and POD and it could be recommended that seed priming with JA help in ameliorating toxic effect of Cu2+.

JA-mediated MYC2/LOX/AOS feedback loop regulates osmotic stress response in tea plant

Osmotic stress caused by low-temperature,drought and salinity was a prevalent abiotic stress in plant that severely inhibited plant development and agricultural yield,particularly in tea plant.Jasmonic acid(JA)is an important phytohormone involving in plant stress.However,underlying molecular mechanisms of JA modulated osmotic stress response remains unclear.In this study,high concentration of mannitol induced JA accumulation and increase of peroxidase activity in tea plant.Integrated transcriptome mined a JA signaling master,MYC2 transcription factor is shown as a hub regulator that induced by mannitol,expression of which positively correlated with JA biosynthetic genes(LOX and AOS)and peroxidase genes(PER).CsMYC2 was determined as a nuclei-localized transcription activator,furthermore,ProteinDNA interaction analysis indicated that CsMYC2 was positive regulator that activated the transcription of CsLOX7,CsAOS2,CsPER1 and CsPER3via bound with their promoters,respectively.Suppression of CsMYC2 expression resulted in a reduced JA content and peroxidase activity and osmotic stress tolerance of tea plant.Overexpression of CsMYC2 in Arabidopsis improved JA content,peroxidase activity and plants tolerance against mannitol stress.Together,we proposed a positive feedback loop mediated by CsMYC2,CsLOX7 and CsAOS2 which constituted to increase the tolerance of osmotic stress through fine-tuning the accumulation of JA levels and increase of POD activity in tea plant.

The laccase gene Gh Lac1 modulates fiber initiation and elongation by coordinating jasmonic acid and flavonoid metabolism

Cotton fibers are single cells originating in the epidermis of cotton ovules,and serve as the largest natural fiber source for the textile industry.In theory,all epidermal cells have the potential to develop into fibers,but only 15%–25%of epidermis cells develop into commercially viable lint fibers.We previously showed that Gh Lac1 participates in cotton defense against biotic stress.Here we report that Gh Lac1 also has a role in cotton fiber development.Gh Lac1 RNAi lines in cotton showed increased differentiation of fiber initials from epidermis and shortened fiber length,resulting in unchanged lint percentage.Suppression of Gh Lac1 expression led to constitutively hyperaccumulated jasmonic acid(JA)and flavonoids in ovules and fiber cells.In vitro ovule culture experiments confirmed the distinct roles of JA and flavonoids in fiber initiation and elongation,and showed that fiber development is spatially regulated by these chemicals:the increased fiber initiation in Gh Lac1 RNAi lines is caused by hyperaccumulated JA and rutin content during the fiber initiation stage and shortened fiber length is caused by constitutively increased JA and naringenin content during the fiber elongation stage.

番茄SlJAZ7的抗病功能及与SlTGA7的互作

[目的]细菌性斑点病是导致番茄Solanum lycopersicum减产的主要因素之一,丁香假单胞菌(Pseudomonas syringae pv. tomato DC3000,Pst DC3000)是细菌性斑点病的致病因子之一。瞬时沉默番茄JAZ7基因导致其对Pst DC3000的敏感性增加,然而,验证JAZ7基因抗细菌性斑点病的直接证据及其作用机理的报道较少。本研究从番茄叶片中克隆得到SlJAZ7基因,创制稳定遗传的过表达SlJAZ7的转基因番茄,分析其抗病功能和机理,研究其在转录水平和蛋白水平上与抗病性密切相关的SlTGA7的关系,为有效防治细菌性斑点病提供理论基础。[方法]通过野生型和转基因番茄接种Pst DC3000的表型差异分析,鉴定SlJAZ7基因的抗病功能。使用RT-qPCR分析SlJAZ7和SlTGA7基因在Pst DC3000、茉莉酸甲酯(Methyl jasmonate,MeJA)和水杨酸(Salicylic acid,SA)处理下的表达模式和组织特异性。利用烟草瞬时表达的方法研究SIJAZ7和SlTGA7蛋白的亚细胞定位。利用酵母双杂交(Y2H)试验、双分子荧光互补(BiFC)试验和蛋白下拉(pull down)试验研究SlJAZ7蛋白与SlTGA7蛋白的互作关系,验证SlJAZ7的抗病功能和可能的抗病机理。[结果]过表达SlJAZ7基因的转基因番茄叶片在Pst DC3000处理时受到的过氧化损伤较野生型更少,转基因株系中SlTGA7表达量升高,SlJAZ7基因在营养器官中高表达,且受到Pst DC3000诱导,响应MeJA、SA处理,SlTGA7基因在同样的处理下呈相反的变化趋势。SlJAZ7和SlTGA7蛋白均定位于细胞核。Y2H、BiFC和pull down试验同时证明SlJAZ7蛋白和SlTGA7蛋白存在互作关系。[结论]过表达SlJAZ7基因有利于减少活性氧积累,提高番茄抗病性,同时SlJAZ7在转录水平正调控SlTGA7基因表达。SlJAZ7与SlTGA7存在互作,推测SIJAZ7基因可能通过提高Pst DC3000病原菌侵染时SITGA7基因的表达量,启动SITGA7下游抗病基因的表达来提高转基因番茄的抗病性,也可能通过与SITGA7蛋白结合,影响其调控MYC等转录因子的活性。这为进一步研究SlJAZ7的作用机制奠定了基础。

Effects of Jasmonic Acid on the Expression of Capsaicin Biosynthesis Related Genes in Chili Pepper(Capsicum annuum)

To explore the regulatory effects of jasmonic acid （JA） on the expression of the genes involved in capsaicin biosynthesis pathway, the mRNA expression of the genes was determined by fluorescence-based quantitative PCR 0, 4, 12, 24 and 48 h after chili peppers （Capsicum annuum var. conoides） were treated with 100 μmol/L JA, and the content of capsaicin was measured by high performance liquid chromatography （HPLC） 0, 1, 3, 6, 10 and 15 d after JA treatment. The results showed that JA upregulated the mRNA levels of pal, C4h, Comt, 4Cl, Hct, Paint, Bcat, FatA and pun1 in chili pepper, thus promoting the synthesis of capsaicin to different extents.

Genome-wide association study reveals that JASMONATE ZIM-DOMAIN 5 regulates seed germination in rice

Seed germination is a complex trait regulated by multiple genes in rice.However,the regulators of rice seed germination have yet to be sufficiently determined.Here,a quantitative trait locus(QTL)for rice seed germination was identified in a genome-wide association study.The candidate gene JASMONATE ZIM-DOMAIN 5(OsJAZ5)of the QTL was verified that positively regulates seed germination.OsJAZ5 regulation of seed germination involves an OsABI3-mediated abscisic acid pathway.Overexpression of OsJAZ5 facilitated seed germination.The application of OsJAZ5 might be useful for increasing seed germination for rice direct seeding.

Overexpression of auxin/indole-3-acetic acid gene MdIAA24 enhances Glomerella leaf spot resistance in apple(Malus domestica)

Auxin is throughout the entire life process of plants and is involved in the crosstalk with other hormones,yet its role in apple disease resistance remains unclear.In this study,we investigated the function of auxin/indole-3-acetic acid(IAA)gene Md IAA24 overexpression in enhancing apple resistance to Glomerella leaf spot(GLS)caused by Colletotrichum fructicola(Cf).Analysis revealed that,upon Cf infection,35S::Md IAA24 plants exhibited enhanced superoxide dismutase(SOD)and peroxidase(POD)activity,as well as a greater amount of glutathione(reduced form)and ascorbic acid accumulation,resulting in less H_(2)O_(2)and superoxide anion(O_(2)^(-))in apple leaves.Furthermore,35S::Md IAA24 plants produced more protocatechuic acid,proanthocyanidins B1,proanthocyanidins B2 and chlorogenic acid when infected with Cf.Following Cf infection,35S::Md IAA24 plants presented lower levels of IAA and jasmonic acid(JA),but higher levels of salicylic acid(SA),along with the expression of related genes.The overexpression of Md IAA24 was observed to enhance the activity of chitinase andβ-1,3-glucanase in Cfinfected leaves.The results indicated the ability of Md IAA24 to regulate the crosstalk between IAA,JA and SA,and to improve reactive oxygen species(ROS)scavenging and defense-related enzymes activity.This jointly contributed to GLS resistance in apple.

R2R3-type Lo MYB21 affects jasmonate-regulated development and dehiscence of anthers in lily(Lilium oriental hybrids)

Lilies are widely cultivated for cut flowers,but their large anthers carry a considerable amount of colored pollen that is dispersed easily.Studying the molecular mechanism of anther development and dehiscence could help solve this problem.LoMYB21,encoding a putative R2R3v-myb avian myeloblastosis viral oncogene homolog(MYB)transcription factor,was identified from oriental lilies(Lilium‘Siberia’).Real-time quantitative PCR analysis showed that LoMYB21 was mainly expressed in the anther,filament and stigma and had high expression during the late stages of lily anther development.LoMYB21 had transactivation activity and was located in the nucleus through yeast one-hybrid assays and transient expression in Nicotiana benthamiana.Suppression of LoMYB21 by virus-induced gene silencing(VIGS)in Lilium‘Siberia’led to anther indehiscence and reduced the expression of genes related to Jasmonate acid(JA)biosynthesis and signal transduction.Induction of LoMYB21 in DEX::LoMYB21 transgenic Arabidopsis caused procumbent inflorescences that became infertile,accompanied by higher expression of JA biosynthetic and signaling genes.These results demonstrated that JA content and signaling were abnormal in silenced lily and transgenic LoMYB21 Arabidopsis,which affected anther development.Our study indicated that LoMYB21 could regulate lily anther dehiscence through JA biosynthesis and signaling during the late stages of anther development.

Effects of Exogenous Jasmonic Acid on Concentrations of Direct-Defense Chemicals and Expression of Related Genes in Bt (Bacillus thuringiensis) Corn (Zea mays)

Bt corn is one of the top three large-scale commercialized transgenic crops around the world. It is increasingly clear that the complementary durable approaches for pest control, which combine the endogenous defense of the crop with the introduced foreign genes, are promising alternative strategies for pest resistance management and the next generation of insect-resistant transgenic crops. In the present study, we tested the inducible effects of exogenous jasmonic acid （JA） on direct-defense chemical content, Bt protein concentration, and related gene expression in the leaves of Bt corn cultivar 34B24 and non-Bt cultivar 34B23 by chemical analysis, ELISA, and RT-PCR. The results show that the expression of LOX, PR-2a, MPI, and PR-I genes in the treated leaf （the first leaf） was promoted by exogenous JA both in 34B24 and 34B23. As compared with the control, the concentration of DIMBOA in the treated leaf was significantly increased by 63 and 18% for 34B24 and 34B23, respectively. The total phenolic acid was also increased by 24 and 12% for both 34B24 and 34B23. The Bt protein content of 34B24 in the treated leaf was increased by 13% but decreased significantly by 27% in the second leaf. The induced response of 34B24 was in a systemic way and was much stronger than that of 34B23. Those findings indicated that there is a synergistic interaction between Bt gene and internally induced chemical defense system triggered by externally applied JA in Bt corn.