灰葡萄孢非经典效应蛋白BcRicin调控植物免疫机制研究

灰霉病是蔬菜、水果上常见的重要病害之一，能造成严重的经济损失。病原菌通过分泌效应蛋白来调控寄主免疫反应以促进侵染。目前效应蛋白的研究主要以经典外泌效应蛋白为主，而对非经典外泌效应蛋白很少涉及，其功能和调控机制尚不清楚。申请人前期在灰葡萄孢菌外泌组中鉴定到一个小分子非经典外泌蛋白BcRicin，作为质外体效应蛋白能引起本氏烟过敏性细胞死亡，并诱发烟草免疫反应。本项目拟通过病毒诱导的基因沉默技术，在沉默烟草膜受体蛋白植株中瞬时表达BcRicin，分析其在本氏烟体内的互作蛋白；在番茄中寻找其同源蛋白，利用CRISPR/Cas9技术敲除番茄受体蛋白，分析其在番茄生长发育及调控植物免疫的功能。该研究不仅有助于我们深入了解死体营养型病原菌非经典效应蛋白的功能，还将丰富非经典效应蛋白与受体蛋白互作新机制，对完善病原菌与植物互作模式具有重要的科学意义。

Botrytis cinerea can infect various plant species, causing enormous financial impact. But so far, little is known about the roles of non-classical effectors produced by broad host-range necrotrophic phytopathogens during infection. Our previous study has identified a small, non-classical secret protein BcRicin in the fungus, which lack N-terminal signal peptide. BcRicin could induce significant plant cell death and transient expression of BcRicin in tobacco could improved plant resistance to the pathogens. Based on these preliminary results, we will further explore biological functions of BcRicin through gene knockout and infection-related morphogenesis; identify the receptor protein in host plant through virus-induced gene silencing (VIGS) and yeast two-hybrid (Y2H); reveal the role of the receptor protein in tomato through CRISPR/Cas9 and preliminary clarify the interaction between BcRicin and the receptor protein. Expected results from this project will be helpful for revealing the role of non-classical effector in virulence of broad host-range necrotrophic phytopathogens like B. cinerea, providing scientific basis for novel drug development against B. cinerea.