水稻黑条矮缩病毒非结构蛋白P7-1与水稻GAPDH互作机理

项目名称： 水稻黑条矮缩病毒非结构蛋白P7-1与水稻GAPDH互作机理

项目编号： No.31201484

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 植物保护学、园艺学与植物营养学

项目作者： 孙枫

作者单位： 江苏省农业科学院

项目金额： 23万元

中文摘要： 水稻黑条矮缩病毒(RBSDV)侵染水稻导致植株矮化、育性下降，严重威胁我国水稻生产。但对病毒蛋白与寄主因子互作导致病害症状的机理还了解甚少。前期研究表明，拟南芥过量表达RBSDV P7-1引起雄性不育，P7-1与拟南芥GAPDH(甘油醛-3磷酸脱氢酶)互作；RBSDV侵染水稻诱导GAPDH表达，酵母双杂交表明病毒P7-1也与水稻GAPDH互作。本项目拟采用Pull down、BiFC进一步验证病毒P7-1与水稻GAPDH互作；构建P7-1过表达和基因沉默转基因水稻，研究P7-1在病毒致病过程中功能；构建GAPDH过表达、基因沉默转基因水稻及T-DNA突变体，研究GAPDH在育性和抗病毒过程中的作用；通过分析GAPDH酶活、mRNA转录、蛋白表达及亚细胞定位揭示病毒P7-1调控GAPDH机理。本项目所解析的RBSDV P7-1与水稻GAPDH互作机理，将为进一步理解病毒致病机理提供理论依据。

中文关键词： 水稻黑条矮缩病毒 P7-1；甘油醛-3磷酸脱氢酶；抗性；细胞定位；

英文摘要： Rice black-streaked dwarf virus (RBSDV), a member of the genus Fijivirus in the family Reoviridae, is propagatively transmitted by the small brown planthopper (Laodelphax striatellus Fallé and causes rice black-streaked dwarf and maize rough dwarf diseases, which lead to severe yield losses of crops in China. Although several RBSDV proteins have been investigated in details, functions of the nonstructural protein P7-1 are still largely unknown. To investigate the role of the P7-1 protein in virus pathogenicity, transgenic Arabidopsis thaliana plants were generated in which the P7-1 transgene was expressed under the 35S promoter. The RBSDV P7-1-transgenic Arabidopsis were male sterility.Flowers and pollen from P7-1-transgenic plants were of normal size and shape as in wild type, and anthers developed to normal size but fail to dehisce. Yeast two hybrid assay indicated that P7-1 interacted with GAPA (glyceraldehyde-3-phosphate dehydrogenase) NAD binding domain. Rice GAPDH (Os04g38600,Os03g03720)mRNA transcripts were induced by inoculated with RBSDV, and rice GAPDH protein (Os04g38600,Os03g03720)interacted with RBSDV P7-1 in yeast two hybrid assay. In this project, RBSDV P7-1 interacted with rice GAPDH protein will be tested by yeast two hybrid, pull down and BiFC (bimolecular fluorescence complementation) assays

英文关键词： RBSDV P7-1；GAPDH；resistance；cell localization；