油菜黑胫病抗性相关基因BnWRKY70的功能研究  

Functionality of BnWRKY70 gene of oilseed rape ( Brassica napus L. ) in relation to blackleg

作　　者：燕孟娇 石嵘 郝丽芬[1,3] 皇甫海燕[1] 宋培玲[1] 贾晓清[1] 皇甫九茹[1] 李子钦[1] 

机构地区：[1]内蒙古农牧业科学院植物保护研究所,内蒙古呼和浩特010031 [2]内蒙古大学生命科学院,内蒙古呼和浩特010021 [3]内蒙古农业大学生命科学学院,内蒙古呼和浩特010019

出　　处：《中国油料作物学报》2018年第1期127-133,共7页Chinese Journal of Oil Crop Sciences

基　　金：内蒙古科技厅计划项目(201505018);国家国际科技合作基地建设项目

摘　　要：WRKY转录因子是植物信号传导过程中的重要成员,在植物抗病过程中发挥着重要作用。为了解BnWRKY70在油菜黑胫病抗性反应中的作用机理,本研究对油菜响应黑胫病菌侵染产生的特异基因BnWRKY70进行序列分析;将克隆得到BnWRKY70基因CDS 308bp特异区段,利用烟草脆裂病毒(Tobacco rattle virus,TRV)载体构建了基于病毒诱导的油菜基因沉默体系;利用qRT-PCR验证目的基因的表达水平;最后进一步通过表型观察和病情指数统计分析评价BnWRKY70基因沉默植株在接种黑胫病菌之后油菜的抗病性。结果表明,BnWRKY70基因最长ORF为858bp,编码285个氨基酸,含有一个WRKY保守结构域,属于Ⅲ类WRKY转录因子。BnWRKY70基因沉默后的表达量显著下降,说明在油菜中成功构建了VIGS体系。表型观察发现基因沉默油菜株系叶片比未沉默株系更易感病,病斑面积更大,且病情指数显著高于对照,说明BnWRKY70基因沉默减弱了油菜的抗病性。本研究为揭示BnWRKY70在油菜病原菌分子互作机制中的作用奠定基础。WRKY transcription factors is the important member of plant signal transduction pathway,plays an important role in the process of plant resistance,in order to better understand BnWRKY70 in rapeseed blackleg disease resistance. The sequence of BnWRKY70 specific gene to response to blackleg was analyzed,and the 308 bp CDS specific section of BnWRKY70 gene cloned,using the Tobacco rattle virus carrier constructed induced gene silencing system of oilseed,then validated target gene expression quantity with qRT-PCR,finally evaluation resistance of BnWRKY70 gene silencing rape after inoculation of blackleg pathogens by phenotypic observation and statistical analysis of disease index. The results showed that BnWRKY70 gene had the longest ORF of 858 bp,encoding 285 amino acids,containing a WRKY conservative domain structure,belonging to the class Ⅲ WRKY transcription factors,and BnWRKY70 gene expression was significantly decreased after silence by successful VIGS system constructed in oilseed rape,the leaf of gene silencing oilseed rape is more susceptible than target gene silence oilseed rape,the disease spot area is larger,and the disease index was significantly higher than that of control. It all showed that the silence of BnWRKY70 genes decreased the oilseed rape resistance,to further reveal pathogen molecular interactions mechanism of oilseed rape.

关 键 词：油菜 黑胫病 病毒诱导的基因沉默(VIGS) BnWRKY70基因 

分 类 号：S565.4[农业科学—作物学]