马铃薯PYL5基因对非生物胁迫的响应分析及其启动子的活性鉴定OA北大核心CSTPCD
Response analysis of potato PYL5 gene to abiotic stress and activity identification of its promoter
[目的]脱落酸(ABA)作为一种"应激激素"在植物生长发育和响应干旱、盐等非生物胁迫过程中发挥重要作用,PYR/PYL/RCARs(以下简称"PYL")作为ABA受体在多种植物中也被广泛研究.基于对马铃薯StPYL5基因生物信息学及表达模式分析,并通过对其启动子活性鉴定,为进一步揭示马铃薯StPYL5功能及抗逆育种提供依据.[方法]根据转录组数据克隆得到StPYL5基因,通过DNAMAN、MEGA等软件分析了StPYL5的分子特征;通过qPCR检测了StPYL5基因的组织特异性及其对非生物胁迫的响应;利用PlantCARE网站对StPYL5基因启动子进行了分析,并通过瞬时转化烟草对其活性进行了鉴定.[结果]StPYL5基因全长534 bp,共编码177个氨基酸,蛋白质分子量20.19 ku,理论等电点(p I)为5.97.系统进化分析显示,StPYL5与SpPYL9-like亲缘关系较近.组织特异性分析结果显示,青薯9号(QS9)中StPYL5在根和叶中表达量较高,其次分别是茎和花,在块茎中表达量较低.不同胁迫下StPYL5表达量分析表明,青薯9号(QS9)中StPYL5在干旱、低温、盐和ABA胁迫下的表达量先升高后降低,且StPYL5的表达受MeJA和SA的诱导.此外,笔者成功克隆得到2 000 bp的StPYL5基因启动子.在烟草中的瞬时转化后的组织化学染色结果表明,StPYL5基因启动子具有成功启动下游GUS报告基因表达的启动子活性.[结论]全面分析了StPYL5基因的分子特征及其在多种非生物胁迫下的表达谱,并成功克隆到具有活性的pStPYL5启动子,该结果为深入研究StPYL5基因的功能以及马铃薯抗逆育种提供依据.
[Objective]Abscisic acid(ABA),as a stress hormone,plays an important role in plant growth,development,and response to abiotic stresses such as drought and salt.PYR/PYL/RCARs(referred to as"PYL"hereafter)as ABA receptors in various plants,are widely studied.Based on the bioinformatics and expression pattern analysis of potato StPYL5 gene and the identification of its promoter activity,this study provides a basis for further revealing the function of potato StPYL5 and resistance breeding.[Method]In this study,StPYL5 gene was cloned based on transcriptome data.The molecular characteristics of StPYL5 was analyzed using software such as DNAMAN and MEGA.The tissue specificity of the StPYL5 gene and its response to abiotic stress were examined by qPCR.The StPYL5 gene promoter was analyzed using the PlantCARE website,and its activity was identified through transient transformation in tobacco.[Result]The results showed that the full-length of the StPYL5 gene was 534 bp,encoding 177 amino acids.The protein had a molecular weight of 20.19 ku,and the theoretical isoelectric point(pI)was 5.97.Phylogenetic analysis revealed a close relationship between StPYL5 and SpPYL9-like.Tissue specificity analysis showed that the expression pattern of StPYL5 in potato cultivar Qingshu 9(QS9)was the highest in leaves and roots,followed by stems,flowers,and tubers.Expression analysis under different stresses showed that the expression level of StPYL5 in QS9 increased initially and then decreased under drought,low temperature,salt,and ABA stress.The expression of StPYL5 was induced by MeJA and SA.In addition,a 2 000 bp StPYL5 gene promoter was successfully cloned.Tissue staining after transient transformation in tobacco demonstrated that the StPYL5 gene promoter had promoter activity to drive downstream GUS reporter gene expression.[Conclusion]This study comprehensively analyzed the molecular characteristics of the StPYL5 gene and its expression profile under various abiotic stresses.The successful cloning of the active pStPYL5 promoter provides a foundation for further investigating the function of the StPYL5 gene and potato breeding for stress tolerance.
张春利;白江平;解潇飞;张莹;张锋;孙超;毕真真;刘玉汇;刘震;姚攀锋
甘肃农业大学 农学院,甘肃 兰州 730070||甘肃农业大学 省部共建干旱生境作物学国家重点实验室,甘肃 兰州 730070甘肃农业大学 省部共建干旱生境作物学国家重点实验室,甘肃 兰州 730070
农业科学
马铃薯ABA信号通路StPYL5基因克隆非生物胁迫表达分析
Solanum tuberosum L.ABA signaling pathwayStPYL5gene cloningabiotic stressexpression analysis
《江西农业大学学报》 2024 (002)
302-313 / 12
国家马铃薯产业技术体系(CARS-09-P10)和甘肃省科技计划项目(23JRRA1414) Project supported by National Potato Industry Technology System(CARS-09-P10)and Science and Technology Program of Gansu Province(23JRRA1414)
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