水稻LysM结构域包含基因OsEMSA1的克隆及过表达株系构建  

Cloning and Overexpression Strain Construction of a LysM Domain-Containing Gene OsEMSA1 in Rice

作　　者：朱骞[1,2] 张小玲[1,2] Sadia Nadir Henry Akrofi Doku 陈丽娟[1,2] 李东宣[1,2] 

机构地区：[1]云南农业大学稻作研究所,昆明650201 [2]云南农业大学省部共建云南生物资源保护与利用国家重点实验室,昆明650201 [3]Department of Chemistry,University of Science and Technology,Barmu,KPK,Pakistan,28100 [4]CSIR-Crops Research Institute,P.O.Box 3785,Kumasi,Ghana,West Africa

出　　处：《分子植物育种》2018年第1期22-30,共9页Molecular Plant Breeding

基　　金：“十三五”国家重点研发计划项目(2016YFD0101101-5);NSFC-云南联合基金重点项目(U11-36604);国家自然科学基金(31560115);省部共建云南生物资源保护与利用国家重点实验室开放课题基金;云南省博士研究生学术新人奖(A3007962)共同资助

摘　　要：LysM结构域包含蛋白(lysM domain containing protein)为植物中公认的病原菌信号受体蛋白。本研究在水稻雌性不育基因FST遗传调控网中筛选获得一个关键靶基因,暂命名为OsEMSA1,该基因编码包含一个LysM结构域的未知功能蛋白质。蛋白质序列分析表明,OsEMSA1蛋白N端包含一个信号肽序列,具备跨膜结构,LysM结构域位于蛋白C端,为胞外结构。启动子顺式作用元件分析表明,光响应元件、激素应答元件、生长调节元件在OsEMSA1启动子区有很高的分布。电子表达谱分析表明,OsEMSA1基因在野生型水稻日本晴多组织中均有不同程度的表达,而根中和开花前的胚囊中表达量相对较高,可能参与调控水稻根和雌配子发育,同时逆境胁迫、激素信号以及病菌侵害也能不同程度的诱导OsEMSA1基因的表达。基因共表达分析显示,OsEMSA1基因与激素信号传导响应、逆境胁迫应答以及抵御真菌病害的基因存在互作。本研究成功构建了由OsEMSA1基因自身启动子驱动的过表达转基因水稻株系,为进一步分析OsEMSA1基因功能奠定了实验基础,并为发掘LysM结构域包含蛋白的潜在功能提供了一定的理论依据。LysM domain-c ontaining protein is generally recognized as a pathogen signal receptor protein in plants.In this study,a key target gene was screened from the genetic regulatory network of female sterile gene FST in rice,which was temporarily named as OsEMSA1.The gene encoded a function-unknown protein that contained a LysM domain.Protein sequence analysis showed that the N-terminus of OsEMSA1 protein contained a signal peptide sequence with a transmembrane structure.The LysM domain was located at the C-terminus of OsEMSA1 protein,which was an extracellular structure.The cis-acting element analysis for promoter indicated that the lightresponsive elements,hormone responsive elements and growth regulatory elements were highly distributed in the OsEMSA1 promoter region.The electronic expression profile analysis suggested that OsEMSA1 was expressed with various degrees in different tissues of Nipponbare,and the expression levels of OsEMSA1 in root as well as in pre-flowering embryosac were higher than those in others,which implied that OsEMSA1 might be involved in the regulation of root and female gamete development.Moreover,adversity stress,hormone signaling and pathogen infections could induce the expression of OsEMSA1 in different degrees.Gene co-expression analysis revealed that OsEMSA1 interacted with some other genes which were involved in hormone signaling,abiotic stress response and pathogen defense.In this study,an overexpression transgenic rice strain driven by OsEMSA1 promoter was successfully constructed,which established an experimental basis for further research of analysis of OsEMSA1 function and provided a theoretical basis for exploring the potential function of LysM domain-containing protein in rice.

关 键 词：水稻 基因克隆 LysM结构域 

分 类 号：Q943.2[生物学—植物学] S511[农业科学—作物学]