条锈菌诱导的小麦候选感病基因TaHIPP1的感病机制解析

条锈病是我国小麦生产上的重要病害，随着强致病性小种的出现和发展，小麦生产上的多数主栽品种均已“丧失”抗病性，使得我国小麦再度处于条锈病大流行的威胁中，研究小麦抗条锈菌的分子机制进而培育抗性品种是目前急需解决的重大问题。针对条锈菌无法人工培养以及缺乏分子水平对小麦抗条锈病机理的系统研究的现状，本项目在前期构建的小麦与CYR31构建的亲和互作的cDNA文库中获得的2个候选的小麦条锈病感病基因（TaHIPP），通过电子克隆获得TaHIPP全长序列，而且利用酵母过表达验证了TaHIPP蛋白响应金属离子胁迫。在此基础上，拟通过VIGS技术和CRISPR/Cas9系统来验证候选感病基因TaHIPP1的功能，并利用酵母双杂交系统寻找与其互作的下游靶标蛋白，为解析小麦与条锈菌互作中的感病机制，进而为通过沉默感病基因创制持久、广谱抗病品种奠定基础。

Wheat stripe rust is an important disease of wheat production. With the emergence and development of virulent races, most wheat cultivars have been lost disease resistance, which make wheat be in the threat of pandemic stripe rust. It is urgently needed to resolve the major issues to research the molecular mechanism of resistance to stripe rust in wheat cultivation of resistant varieties. Because stripe rust fungus couldn’t be cultured and lack of systematic study of the molecular status of wheat stripe rust resistance mechanism, the foundation of the project in the early work of the two candidates obtaining wheat stripe rust susceptible gene (TaHIPP) from the cDNA library of compatible interaction between wheat and CYR31. In addition, The full-length sequences of TaHIPP genes were cloned, and TaHIPP genes was verified to response to the stress on the metal ions by the yeast over expression. On these bases, the study researches the function of candidate disease gene TaHIPP1 by VIGS technology and CRISPR/Cas9 system, and searches its interactions downstream target proteins using the yeast two-hybrid system, which analysis the susceptible mechanism in the interaction between wheat and stripe rust, and then establish the foundation of creating lasting and broad-spectrum disease resistant varieties through susceptible gene silencing.