利用dsRNA干涉方法研究水稻WRKY转录因子的抗病性  

Study on Pathogen Resistance of OsWRKY Transcription Factor by dsRNA Interference Method

作　　者：邬亚文[1] 谢科[1] 李靖[1] 于永红[2] 胡国成[2] 傅亚萍[2] 斯华敏[2] 窦彩虹[1] 郭泽建[3] 孙宗修[2] 

机构地区：[1]浙江大学农业与生物技术学院,浙江杭州310029 [2]中国水稻研究所水稻生物学国家重点实验室,浙江杭州310006 [3]中国农业大学植物病理系,北京100094

出　　处：《中国水稻科学》2005年第6期489-494,共6页Chinese Journal of Rice Science

基　　金：国家自然科学基金资助项目(30471122;30370139);浙江省自然科学基金资助项目(Y304210);'水稻生物学国家重点实验室创新引导项目。

摘　　要：为了研究植物WRKY基因编码的转录因子在水稻上的功能,构建了包含WRKY保守结构域的dsRNA发夹结构干涉载体,用农杆菌介导法转野生型水稻中花11,得到9个有干涉表型的株系。PCR和Southern结果表明dsRNA已整合入中花11的基因组中,且多数为单拷贝。结合TDNA插入的WRKY突变体植株T456,研究了其中的3个干涉苗和T456对稻瘟病和白叶枯病的抗性,发现3个干涉苗对这两种水稻主要病害的抗性均明显强于T456和中花11,且T456比中花11略抗稻瘟病和白叶枯病,表明dsRNA干涉是成功的,它可能干涉或抑制了某些OsWRKY家族中负向调控抗病基因的成员。To study the functions of the WRKY superfamily of transcription factors in rice, a hairpin dsRNA interference vector involving in a sequence of conserved WRKY domain, constituted by about 60 amino acid residues, were constructed. The vector was transformed into the embryos of wild parent Zhonghua 11 by Agrobacterium mediated co-cultivation method. A total of nine transgenic lines with interference phenotypes were identified from 65 independent transgenic rice plants. PCR and Southern blotting analysis indicated that the foreign DNA had been integrated into rice genome of Zhonghua 11, and most of them were single copy. The three selected interference lines showed more resistant to rice blast （Magnaporthe grisea） and bacterial blight （Xanthomonas oryzae pv. oryzae） than WRKY T DNA insertion mutant T456 and Zhonghua 11, and T456 was slightly more resistant to these two diseases than Zhanghua 11, suggesting that dsRNA interference was successful and it might have suppressed some OsWRKY transcription factors which were highly associated with regulating pathogen genes.

关 键 词：水稻 T-DNA插入 转录因子 RNA干涉 稻瘟病 白叶枯病 

分 类 号：S511[农业科学—作物学]