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.

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.

The Role of Jasmonates as Antibulbing Substances in the Bulb Formation of Onion

Onion plants form spherical bulbs under long-day conditions.Substances regulating bulb formation remain unknown.In the course of chemical studies on the bulb formation,α-linolenic acid was isolated from onion extracts as an antibulbing substance,the amount of which was synchronized with the bulb formation.Since allene oxide synthase inhibitor canceled the antibulbing activity ofα-linolenic acid,it was disclosed that jasmonic acid concerns this regulation.Structure-activity-relationship study revealed that its(3R,7S)stereochemistry is necessary for showing its antibulbing activity.It is concluded that(3R,7S)-jasmonate derived fromα-linolenic acid actually participates in the regulation of bulb formation.

GhWRKY33 negatively regulates jasmonate-mediated plant defense to Verticillium dahliae

Verticillium wilt,caused by Verticillium dahliae,seriously restricts the yield and quality improvement of cotton.Previous studies have revealed the involvement of WRKY members in plant defense against V.dahliae,but the underlying mechanisms involved need to be further elucidated.Here,we demonstrated that Gossypium hirsutum WRKY DNA-binding protein 33(GhWRKY33) functions as a negative regulator in plant defense against V.dahliae.GhWRKY33 expression is induced rapidly by V.dahliae and methyl jasmonate,and overexpression of GhWRKY33 reduces plant tolerance to V.dahliae in Arabidopsis.Quantitative RT-PCR analysis revealed that expression of several JA-associated genes was significantly repressed in GhWRKY33 overexpressing transgenic plants.Yeast one-hybrid analysis revealed that GhWRKY33 may repress the transcription of both AtERF1 and GhERF2 through its binding to their promoters.Protein-protein interaction analysis suggested that GhWRKY33 interacts with G.hirsutum JASMONATE ZIM-domain protein 3(GhJAZ3).Similarly,overexpression of GhJAZ3 also decreases plant tolerance to V.dahliae.Furthermore,GhJAZ3 acts synergistically with GhWRKY33 to suppress both AtERF1 and GhERF2 expression.Our results imply that GhWRKY33 may negatively regulate plant tolerance to V.dahliae via the JA-mediated signaling pathway.

打顶和激素对上部烟叶烟碱的影响及源库关系分析

为明确打顶和激素调控烤烟烟碱合成与积累的生理机制,以烤烟品种K326为材料,研究了不同打顶措施与激素互作处理对烤烟根系、上部叶片性状,上部叶烟碱、总氮含量,根系氮代谢关键酶活性及烟碱合成与运输基因表达的影响,并分析烟碱积累的源库关系。结果表明,打顶显著提高上部烟叶烟碱含量与积累量,分别是不打顶处理的1.41倍、1.85倍;不打顶时喷施茉莉酸甲酯(MeJA)显著增加烟碱含量,增幅为14.60%,喷施N-1-氨甲酰苯甲酸萘酯(NPA)显著降低烟碱含量,降幅为10.01%;打顶后喷施水杨苷异羟肟酸(SHAM)、萘乙酸(NAA)显著降低烟碱含量,降幅分别为23.07%、16.92%。茉莉酸(JA)对根系发育的影响有限,主要通过直接调控烟碱合成与转运基因的表达促进烟碱合成,而生长素(IAA)主要通过调控根系发育间接影响烟碱合成与积累。通径分析显示,根系性状中根尖数对烟碱积累的直接作用最大,根表面积对烟碱积累的总作用最大,是影响烟碱合成的主要“源”指标;叶片性状中叶面积对烟碱积累的直接作用最大,叶干质量对烟碱积累的总作用最大,是影响烟碱积累的主要“库”指标。综合分析,打顶后根系IAA含量提高,促进根尖数的增加,即增加了烟碱合成“源”数量,同时增强硝酸还原酶(NR)、谷氨酰胺合成酶(GS)活性,促进氮代谢与氮吸收;打顶后根系JA含量增加,直接增强烟碱合成与转运基因PMT、QPT、A622和NUP1的表达,提高了烟碱合成强度;此外,打顶促进叶面积、叶干质量等“库”容的增大,提高了烟碱积累效率。

外源茉莉酸甲酯对BNS366雄性育性的影响

为探究外源茉莉酸甲酯(methyl jasmonate,MeJA)对小麦温敏雄性不育系BNS366雄性育性的影响,以BNS366及其近等基因系郑麦366为试验材料,在正常秋季播种(2020年10月12日)和晚播(2020年12月2日)条件下,分别于2021年3月22日和3月31日开始至开花期前,设置0(清水,对照)、50、100、200、300、400和500μmol·L^(-1) MeJA(水溶液,喷施)7个处理,比较分析了不同处理间花粉可育率和自交结实率的差异。结果表明:在正常秋季播种条件下,郑麦366花粉可育率、国内法自交结实率和国际法自交结实率分别为86.31%、70.36%和112.22%,在晚播条件下分别为82.53%、92.53%和166.18%,均正常可育;在正常秋季播种条件下,200μmol·L^(-1) MeJA处理的国际法自交结实率为70.15%,比对照显著降低了42.07个百分点;在晚播条件下,200、300和500μmol·L^(-1) MeJA处理的国内法自交结实率分别为74.71%、74.01%和73.45%,比对照显著降低了17.82、18.52和19.08个百分点;在两个播期下,郑麦366其他浓度MeJA处理的上述3个指标与对照差异均不显著。在正常秋季播种条件下,BNS366花粉可育率为零,达到全不育水平,国内法自交结实率和国际法自交结实率均为0.24%,100μmol·L^(-1) MeJA处理的花粉可育率为88.25%,达到正常可育水平,国内法自交结实率和国际法自交结实率分别为56.41%和94.01%,能正常结实,与对照差异达到显著水平;在晚播条件下,BNS366的上述3个指标分别为51.72%、41.23%和93.08%,正常可育,100μmol·L^(-1) MeJA处理的国际法自交结实率为39.72%,比对照显著降低了53.36个百分点;在两个播期下,BNS366其他浓度MeJA处理的上述3个指标与对照差异均不显著。由此说明,在2020-2021年,外源MeJA对郑麦366和BNS366可育植株的雄性育性可能具有降低效应,对BNS366不育植株的雄性不育性具有较强恢复效应。

A change of phenolic acids content in poplar leaves induced by methyl sali-cylate and methyl jasmonate

The contents of seven different phenolic acids such as gallic acid, catechinic acid, pyrocatechol, caffeic acid, coumaric acid, ferulic acid and benzoic acid in the poplar leaves （Populus Simonii×Populus Pyramibalis c.v and Populus deltoids） suffocated by Methyl jasmonate （MeJA） and Methyl salicylate （MeSA） were monitored for analyzing their functions in interplant communications by using high-pressure liquid chromatography （HPLC）.The results showed that the contents of phenolic acids had obviously difference in leaves exposed to either MeSA or MeJA.When P.deltoides leaves exposed to MeJA or MeSA, the level of gallic acid, coumaric acid, caffeic acid, ferulic acid and benzoic acid was increased, gallic acid in leaves treated with MeJA comes to a peak at 24 h while to a peak at 12-d having leaves treated with MeSA.When P.Simonii ×P.Pyramibalis c.v leaves were exposed to MeJA or MeSA, the level of gallic acid, pyrocatechol and ferulic acid was increased; The catechinic acid and benzoic acid had a little drop; The caffeic acid and coumaric acid were undetected in both suffocated and control leaves.This changed pattern indicated that MeJA and MeSA can act as airborne signals to induce defense response of plants.

The Relationship of Methyl Jasmonate Enhanced Powdery Mildew Resistance in Wheat and the Expressions of 9 Disease Resistance Related Genes

[Objective] This study was carried out to determine the induction effect of jasmonic acid（JA）on powdery mildew resistance in wheat,the activation effect on the expressions of plant disease resistance related genes,and to investigate the relationship between the induced resistance and the gene expression patterns.[Method] Three powdery mildew susceptible cultivars of ＂Chinese Spring＂,＂Pumai 9＂ and ＂Zhoumai 18＂ typically representing different phenotypes in the field were employed.The powdery mildew was assessed by detached leaf assay,and real time quantitative RT-PCR was used to determine the expression patterns of 9 disease resistance related genes of PR1（PR1.1）,PR2（β,1-3 glucanase）,PR3（chitinase）,PR4（wheatwin1）,PR5（thaumatin-like protein）,PR9（TaPERO,peroxidase）,PR10,TaGLP2a（germin-like）and Ta-JA2（jasmonate-induced protein）in leaf of the three cultivars.[Result] MeJA application enhanced the powdery mildew resistances of ＂Chinese Spring＂,＂Pumai 9＂ and ＂Zhoumai 18＂.The induced powdery mildew resistance could be detected from 12 h to 96 h after MeJA treatment,and the peak value was at 24 h.Though there were differences between the three cultivars,MeJA significantly effect on the expressions of the 8 disease resistance related genes except TaGLP2a,and the peak values were at 12 h,24 h or 48 h after treatments.The strongest activation of MeJA was on PR9 and PR1 that their expressions could reach more than 100 times of the untreated samples.MeJA strongly activated PR2、PR4、PR5、PR3、PR10 and Ta-JA2,their expression could reach 10 to 70 times,and there was almost no activation effect on TaGLP2a.The induced powdery mildew resistance positively correlated with the induced expressions of the 8 disease related genes.[Conclusion] The induced powdery mildew resistance positively correlated with the induced expressions of the disease related genes.Jasmonate signalling plays a role in defence against Blumeria graminis f.sp.tritici.and future manipulation of this pathway may improve powdery mildew resistance in wheat.

Increased Endogenous Methyl Jasmonate Altered Leaf and Root Development in Transgenic Soybean Plants

Methyl jasmonate （MeJA） is a plant-signaling molecule that regulates plant morphogenesis and expression of plant defense genes. To determine the role of the endogenous MeJA levels in the development of plants, transgenic soybean [Glycine max （L.） Merrill] plants harboring NTR1 gene encoding for jasmonic acid carboxyl methyltransferase （JMT） were produced. The activation of NTR1 gene expression resulted in the production of MeJA. Overexpression of the NTR1 cDNA under the regulation of cauliflower mosaic virus （CaMV） 35S promoter in the transgenic soybean plants was confirmed using Northern blot analysis. The significant differences in leaf and root growth patterns were observed between the transgenic plants and the wild-type plants. The leaves of the transgenic plants were slightly elongated in length but dramatically narrowed in width compared with the nontrans-formed wild-type plants. In addition, elongation of primary root was inhibited in the overexpressed transgenic soybean plantlets, whereas the development of lateral root was stimulated relative to the nontransformed plants. The leaves of the transgenic plants showed 2-2.5-fold higher levels of MeJA than the control plants. These results indicated that the increased endogenous levels of MeJA is involved in regulation of morphogenesis in soybean plants.

Transcription factor OsSPL10 interacts with OsJAmyb to regulate blast resistance in rice

Transcription factors(TFs)play essential roles in transcriptional reprogramming during activation of plant immune responses to pathogens.OsSPL10(SQUAMOSA promoter binding protein-like10)is an important TF regulating trichome development and salt tolerance in rice.Here we report that knockout of OsSPL10 reduces whereas its overexpression enhances rice resistance to blast disease.OsSPL10 positively regulates chitin-induced immune responses including reactive oxygen species(ROS)burst and callose deposition.We show that OsSPL10 physically associates with OsJAmyb,an important TF involved in jasmonic acid(JA)signaling,and positively regulates its protein stability.We then prove that OsJAmyb positively regulates resistance to blast.Our results reveal a molecular module consisting of OsSPL10 and OsJAmyb that positively regulates blast resistance.

Expression Patterns of a Vernalization-related Genes Responding to Jasmonate

利用RNA原位杂交技术分析了春化相关基因ver2 0 3F在冬小麦 (TriticumaestivumL .cv .JingdongNo .1 )胚芽组织内的表达模式。结果表明 ,ver2 0 3F基因的转录本在春化处理的冬小麦胚芽幼叶中有明显的积累 ,而在胚芽鞘以及生长点处却未见杂交信号。茉莉酸诱导冬小麦胚芽后的基因表达模式与春化处理后的模式相似。实验结果暗示感受春化作用的信号部位可能是胚芽的幼叶 ,细胞对春化的应答与茉莉酸介导的信号转导途径有关。

阳春砂茉莉酸生物合成酶基因鉴定与时空表达分析

为探究阳春砂(Amomum villosum)花器中茉莉酸(JA)生物合成酶基因表达与雌雄蕊生长的关系,本研究利用cDNA末端快速扩增(RACE)技术克隆阳春砂JA生物合成酶基因的全长序列,并对各基因进行生物信息学分析、表达模式测定及各基因表达与阳春砂雌雄蕊生长的相关性分析,以探究阳春砂花器中JA生物合成酶基因表达与雌雄蕊生长的关系。结果表明,阳春砂中JA生物合成酶基因AvLOX、AvAOS、AvAOC、AvOPR3序列全长分别为2733、1798、930、1400 bp,分别包含2577、1485、744、1179 bp的开放阅读框(ORF),其编码蛋白分别含有858、494、247、392个氨基酸残基。4个JA生物合成酶基因的表达随阳春砂雄蕊的发育进程逐渐增加,但在雌蕊中的表达则呈现多样性。在雌蕊中,AvLOX、AvAOS基因在开花后1天表达最高,AvAOC在标记后第9天表达最高,AvOPR3基因在标记后第6天表达最高。除AvAOS基因在小花在标记后第3天以及AvOPR3基因在小花标记后第6天外,4个JA生物合成酶基因在雄蕊中的表达在各发育阶段显著高于同时期在雌蕊中的表达。与对照组相比,在喷施茉莉酸甲酯(MeJA)后,4个JA生物合成酶基因在雄蕊中的表达模式,以及AvLOX和AvOPR3基因在雌蕊中的表达模式没有改变,但AvAOS和AvAOC基因在雌蕊中的表达模式有变化。在对照组和MeJA处理组中,4个JA生物合成酶基因的表达与阳春砂小花花丝和花柱的长度有极显著的相关性。本研究结果为进一步探究JA与阳春砂花器官生长发育的关系奠定了基础。

茉莉酸在L-谷氨酸诱导的番茄果实抗性中的作用

为解析植物激素茉莉酸在化学激发子L-谷氨酸诱导抗性机制中的作用,以番茄果实为对象,经0.1 mg/mL L-谷氨酸和0.1 mmol/L茉莉酸甲酯处理后接种链格孢菌孢子悬浮液,观察果实的发病率。采用实时荧光定量PCR方法从转录水平上分析L-谷氨酸对果实茉莉酸合成路径和信号路径上关键基因表达的影响。结果显示:番茄果实经L-谷氨酸处理后其发病率较对照组下降了35%,而在茉莉酸甲酯处理组中,L-谷氨酸与对照组发病率无显著差异;茉莉酸路径上编码脂氧合酶、丙二烯氧化物合酶、丙二烯氧化物环化酶、冠菌素不敏感蛋白1、转录因子MYC2和蛋白酶抑制剂PI-II的基因不同程度地被L-谷氨酸下调表达,同时茉莉酸信号路径的负调控因子JAZ1则被诱导上调表达。综上,在番茄果实与链格孢菌互作中,L-谷氨酸可能通过抑制果实的茉莉酸路径来调控对病原菌的防御反应,这解析了L-谷氨酸作为激发子响应病原菌侵染的抗性机制,也为其商业化应用提供理论依据。

基于转录组分析茉莉酸甲酯诱导猕猴桃果实抗采后软腐病作用机制

【目的】茉莉酸甲酯(MeJA)作为生物信号分子广泛存在于植物体内,可激发果实抵御生物或非生物因子胁迫,在贮藏保鲜中发挥重要作用。转录组技术已广泛应用于探索采后病害防治的相关调控机制,它能够在分子水平上深入阐明病原菌与植物之间互作的机制,有利于鉴定和挖掘胁迫响应基因。前期研究表明,MeJA能够通过调控防御系统诱导猕猴桃果实抗采后软腐病,并分析了苯丙烷途径、膜脂代谢以及能量代谢与MeJA诱导猕猴桃果实抗软腐病的关系,从生理和分子水平初步阐述了MeJA介导的猕猴桃果实抗病途径。基于茉莉酸甲酯(MeJA)诱导猕猴桃果实抵御软腐病菌(Botryosphaeria dothidea)的转录组数据,分析猕猴桃果实响应病原侵染过程中抗病相关基因的表达,为研究MeJA介导的猕猴桃抗软腐病的分子机制提供理论参考。【方法】以‘红阳’猕猴桃为试验材料,选择接种B.dothidea组和MeJA处理并接种组,将0,1,5 d的猕猴桃样本进行转录组测序。以log2FC≥2,Qvalue≤0.05和FPKM>2为标准筛选差异表达基因,进行实时荧光定量PCR验证,并进行GO功能注释和KEGG通路富集分析。【结果】转录组测序共获得约114.97 GB数据,各样本Clean data均达到6.39 GB,Q30碱基百分比在90.41%以上。对各个时期差异表达基因数量统计发现,猕猴桃果实DEGs随着B.dothidea侵染时间延长逐渐增加;而响应MeJA诱导的DEGs主要基中在预处理时,接种病原菌后DEGs数量逐渐减少,表明外源MeJA在早期诱导了大量基因参与果实抗病性。未处理接种组和MeJA处理接种组在0,1,5 d共同差异表达基因分别为164个和314个。GO富集结果表明,差异表达基因主要与生物过程中的细胞过程、代谢过程、对刺激的反应和生物调节,细胞组分中的细胞解剖实体和胞内,以及分子功能中的结合、催化活性和转录调节活性相关。KEGG通路富集结果表明,猕猴桃果实经MeJA处理后接种B.dothidea,其植物-病原互作途径、α-亚麻酸代谢途径、谷胱甘肽代谢途径、苯并恶嗪类生物合成途径、抗坏血酸和醛酸代谢途径特有富集;苯丙烷生物合成途径、芪类化合物、二芳基庚烷和姜醇生物合成途径、类黄酮生物合成途径和MAPK信号通路在两个处理组中共同富集。共筛选出了31个同时响应MeJA和B.dothidea的差异表达基因,推测这些基因与MeJA介导的抗病过程有关。【结论】对外源MeJA处理的猕猴桃果实在接种B.dothidea后的不同时期(0,1,5 d)进行RNASeq分析,结果显示MeJA诱导猕猴桃果实抵御软腐病过程中,大量的生理代谢途径的基因发生变化,有助于增强果实信号分子对防御系统的启动,提高抗菌物质的合成和抗氧化能力,进而提高了猕猴桃果实的抗病性,延长其贮藏时间。

外源茉莉酸甲酯诱导大麦叶斑病抗性研究

【目的】探讨不同浓度外源茉莉酸甲酯(MeJA)诱导大麦抗叶斑病效应差异及其分子机制,为应用MeJA防治大麦叶斑病提供理论依据。【方法】以‘蒙啤麦3号’大麦品种幼苗为材料,设置不接菌(无菌水处理叶片)、接菌(无菌水处理叶片接种麦根腐平脐蠕孢菌)和接菌+MeJA(0.5,1.0,1.5,2.0,2.5mmol/LMeJA喷施叶片后接菌)3组处理,于三叶期调查叶斑病发病情况,并据此筛选最适MeJA浓度,然后测定不接菌、接菌及接菌+MeJA(最适浓度)下不同处理时间叶片的抗氧化酶、抗病相关酶活性、丙二醛含量、渗透调节物质含量以及相关基因表达水平。【结果】(1)叶面喷施外源MeJA提高了大麦对叶斑病的抗性,1.5mmol/LMeJA处理叶片的病情指数较对照显著降低19.03%,诱导抗性效果最佳;(2)与单独接菌处理相比,1.5mmol/LMeJA处理大麦叶片超氧化物歧化酶、过氧化物酶、过氧化氢酶、几丁质酶和β-1,3-葡聚糖酶活性均显著提高,而其丙二醛、脯氨酸、可溶性糖和可溶性蛋白含量显著降低,同时受MeJA调控转录因子及编码抗病相关酶的基因表达量显著上调。【结论】外源喷施1.5mmol/LMeJA通过调节抗病相关酶活性和渗透调节物质含量,以及调控抗病相关酶基因及茉莉酸信号途径关键转录因子基因表达,进而提高大麦植株的叶斑病抗性。

基于全长转录组测序的掌叶大黄bZIP基因家族鉴定分析

bZIP转录因子在调控植物生长发育、次生代谢产物合成以及胁迫响应中发挥着重要作用。本研究基于全长转录组测序分析,系统筛选掌叶大黄(Rheum palmatum)bZIP基因家族,分析其编码蛋白的理化性质、保守基序及系统进化等特性,利用RNA-seq进行基因组织表达分析和茉莉酸甲酯(MeJA)处理基因表达分析。结果表明,掌叶大黄bZIP基因家族有63个成员(RpbZIP1~RpbZIP63);根据其与拟南芥(Arabidopsis thaliana)bZIP系统发育关系被分为A~I和K、S等11个亚族;RpbZIPs基因主要编码不稳定的亲水蛋白,均定位于细胞核;RpbZIPs蛋白二级结构主要为α螺旋和无规卷曲。基于RNA-seq数据,筛选出掌叶大黄根及根茎中高表达且响应MeJA的RbZIP16、RpbZIP17、RpbZIP61等6个基因,qPCR验证了其中4个与转录组测序表达结果一致。本研究系统分析掌叶大黄bZIP基因家族,为后续进一步研究基因功能及大黄次生代谢转录调控机制提供了科学依据。

茉莉酸甲酯对猕猴桃果实抗葡萄座腔菌过程中能量代谢和膜脂代谢的影响

【背景】由葡萄座腔菌(Botryosphaeria dothidea)引起的软腐病是猕猴桃果实贮藏期的重要病害,对猕猴桃产业造成了严重的经济损失。茉莉酸甲酯(MeJA)是一类广泛存在于植物体内的生物信号分子,前期研究发现MeJA能够有效诱导猕猴桃果实抵御B.dothidea的侵染。【目的】分析MeJA对果实能量代谢和膜脂代谢的影响,深入解析MeJA介导的猕猴桃果实抗病机理。【方法】以‘红阳’猕猴桃(Actinidia chinensis cv.Hongyang)为试验材料,分3组进行试验:接种组(Inoculation),果实不经过MeJA处理,但接种B.dothidea;MeJA+接种组(MeJA+inoculation),果实经过0.1 mmol·L^(-1)MeJA熏蒸处理24 h并接种B.dothidea;对照组(Control),不作任何处理,即未经MeJA处理且不接种B.dothidea。所有果实于培养箱((20±1)℃、相对湿度90%—95%)中贮藏8 d。利用2,7-二氯二氢荧光素乙酰乙酸(H2DCFDA)染色分析果实活性氧(ROS)积累情况,测定丙二醛(MDA)含量和相对电导率,分析磷脂酶D(PLD)、脂肪酶(LPS)、脂氧合酶(LOX)、琥珀酸脱氢酶(SDH)、苹果酸脱氢酶(MDH)、H^(+)-ATP酶(H+-ATPase)、Ca^(2+)-ATP酶(Ca^(2+)-ATPase)和细胞色素C氧化酶(CCO)活性及其相应基因的表达,利用高效液相色谱仪(HPLC)检测三磷酸腺苷(ATP)、二磷酸腺苷(ADP)、单磷酸腺苷(AMP)、草酰乙酸和延胡索酸的含量,并对接种组和MeJA+接种组猕猴桃果实膜脂代谢和能量代谢指标进行相关性分析。【结果】与接种组相比,MeJA处理抑制了接种B.dothidea果实中活性氧(ROS)的过量累积,减缓了膜脂过氧化程度,并有效降低了PLD、LPS和LOX活性及AcLOXs、AcPLD和AcLPS的表达,但提高了SDH、MDH、H^(+)-ATPase、Ca^(2+)-ATPase和CCO的活性及AcSDH、AcMDH、AcH^(+)-ATPase、AcCa^(2+)-ATPase和AcCCO转录水平,促进了ATP、ADP、草酰乙酸和延胡索酸的产生,延缓了果实能荷的下降,保障了果实抵御病原菌所需的能量。相关性分析表明,猕猴桃果实膜脂代谢与ROS的积累呈正相关,而与能荷呈负相关。【结论】MeJA诱导猕猴桃果实对B.dothidea的抗性与其调控果实能量水平和减缓ROS参与的膜脂代谢进程有关。

苗期喷施茉莉酸甲酯对香稻品种象牙香占香气的影响

茉莉酸甲酯能够提高植物的抗逆境胁迫能力,但在对香稻香气物质2-乙酰基-1-吡咯啉(2-AP)的影响研究还鲜有报道。以香稻品种象牙香占为试验材料,设置茉莉酸甲酯3种喷施浓度(0.25、0.50和1.00 mmol/L),研究苗期喷施对香稻幼苗2-AP合成与累积、香气前体物质含量、香气相关酶活性、香气相关基因表达量调控、抗性酶活性变化和秧苗素质的影响。结果表明,与对照(CK)相比,喷施适当的茉莉酸甲酯浓度(0.50~1.00 mmol/L),香稻幼苗2-AP含量增加,香气合成前体物质1-吡咯啉、丙酮醛和1-吡咯啉-5-羧酸(P5C)含量提高,二胺氧化酶2基因(DAO4)、鸟氨酸转氨酶基因(OAT)和脯氨酸脱氢酶基因(ProDH)表达量提升。茉莉酸甲酯处理还提高了香稻秧苗素质,增加了抗性酶活性,提高了抗氧化能力。适度的茉莉酸甲酯能提高香稻2-AP含量和秧苗素质,归因于相关酶活性和基因表达量的提高。

茉莉酸甲酯类生长调节剂对‘蛇龙珠’葡萄果实品质及脂氧合酶代谢途径的影响

以‘蛇龙珠’葡萄果实为试材,通过测定其基本理化指标,筛选茉莉酸甲酯(methyljasmonate,MeJA)类物质最适浓度,同时结合顶空固相微萃取-气相色谱-质谱联用技术及酶联免疫吸附试验分析MeJA类物质响应的挥发性关键代谢物及关键酶活性,探究采前MeJA类物质处理对‘蛇龙珠’葡萄果实品质及脂氧合酶(lipoxygenase,LOX)代谢途径中绿叶挥发物的影响。结果表明,与对照组相比,0.1 mmol/L MeJA和5 mmol/L二氢茉莉酸甲酯处理可显著提高‘蛇龙珠’葡萄果实百粒质量、总糖质量浓度、可溶性固形物质量分数、pH值、总酚及总花色苷含量,在采收期时分别提高了10.24%、27.83%、28.24%、4.80%、77.63%、85.29%和9.15%、24.01%、24.41%、8.30%、69.21%、71.57%,果实可滴定酸质量浓度分别降低40.0%及30.24%,果实品质得到显著提高和改善。此浓度处理后,‘蛇龙珠’葡萄果实C6类化合物含量明显提高,更加突出其果实香气的典型性。同时,LOX代谢途径中各个关键酶活性与代谢产物之间具有显著相关性,说明较适浓度的MeJA类物质处理对‘蛇龙珠’葡萄果实中绿叶挥发物的积累具有积极作用。

茉莉酸调控马铃薯离体块茎发育的主要代谢物变化

【目的】茉莉酸(jasmonic acid,JA)是马铃薯块茎发育过程中重要调控激素之一,研究JA调控块茎发育机理将为块茎产量及品质性状形成提供重要理论基础。【方法】采用马铃薯离体匍匐茎作为供试材料,外源添加JA(0、0.5、5和50μmol·L^(-1))处理,分析JA诱导离体块茎表型形态、组织显微结构、碳水化合物积累及蛋白质组变化。【结果】随JA处理浓度升高,单个匍匐茎形成块茎数目、块茎直径及干鲜重、环髓区细胞面积、淀粉及可溶性糖含量在0.5和5μmol·L^(-1)JA处理后逐渐增加(P<0.05),而50μmol·L^(-1)JA处理后块茎直径及干鲜重、淀粉含量显著减少(P<0.05);脂氧合酶活性随JA浓度升高逐渐降低(P<0.05)。采用双向凝胶电泳(2-DE)和MALDI-TOF/TOF-MS质谱技术鉴定到JA调控块茎发育密切相关的35个差异表达蛋白质(P<0.05且差异表达倍数≥2.5倍),主要涉及生物能量与代谢(28.6%)、细胞防御(28.6%)、蛋白质生物合成与贮藏(11.4%)、信号转导(8.6%)、转录(8.6%)、未知功能(8.6%)和混杂(5.6%)。通过聚类分析将这些蛋白质差异表达模式聚为三类:第一类包括17个蛋白质,在0.5μmol·L^(-1)JA处理后下调表达,而在5μmol·L^(-1)JA处理后上调表达,主要参与生物能量与代谢、蛋白质生物合成与贮藏、信号转导、转录;第二类包括10个蛋白质,随JA浓度升高上调表达,主要参与细胞防御、生物能量与代谢、转录;第三类包括8个蛋白质,JA处理后均下调表达,主要参与生物能量与代谢、细胞防御、蛋白质生物合成与贮藏。【结论】低浓度JA(0.5和5μmol·L^(-1))主要通过诱导块茎环髓区细胞膨大、细胞内蔗糖及多糖积累、细胞防御能力来促进块茎形态建成,而高浓度JA(50μmol·L^(-1))则表现为抑制作用。