葡萄白粉病是一种危害最严重的世界性真菌病害。中国野生华东葡萄白河-35-1具有广谱持久的白粉病抗性。本项目前期发现白河-35-1VpCSN5蛋白可特异识别白粉菌EnNAFU1效应蛋白CSEP118，诱导过敏性细胞坏死，但不清楚是VpCSN5独立发挥功能还是通过白河-35-1特有的R蛋白激活免疫反应；在此基础上，拟进一步解析VpCSN5与CSEP118互作机制，分析CSEP118对VpCSN5蛋白生化特征和防御相关分子功能的影响，鉴定白河-35-1特异识别VpCSN5-CSEP118复合体的R蛋白，通过超量表达、RNAi干扰和CRISPR/cas9编辑VpCSN5和候选R基因，分析其抗病功能。项目的完成不仅有利于加深理解葡萄与白粉菌互作机制，而且可以为定向改良葡萄或其他果树白粉病抗性提供优良的基因资源，最终为实现葡萄或其他果树作物抗白粉病精准分子育种提供理论依据与技术支撑。

Powdery mildew is the most important fungal disease of cultivated grapevines worldwide. The wild Chinese grapevine Vitis pseudoreticulata accession Baihe-35-1 is well known for its broad-spectrum and durable resistance to powdery mildew, but the genetic basis still waits to be elicited. Recently, we found that an effector protein CSEP118 from En NAFU1could induce hypersensitive-like cell death when transiently expressed in leaves of Baihe-35-1 whereas no effects had on the leaves of Vitis vinifera cv. Thompson seedless. A yeast-two-hybrid screening proved that CSEP118 could interact with VpCSN5 of Baihe-35-1, and a further bimolecular fluorescence complementation assay also support their interaction, indicating that VpCSN5 might be essential for the immune response for Baihe-35-1. Thus, it is of significance to elucidate whether VpCSN5 directly induces the cell death or triggers innate immune response via interacting with R proteins. In this proposal, we will further investigate the structural basis of the interaction between VpCSN5 and CSEP118, and define the effects that CSEP118 has on the biochemical characterizations and defense-associated molecular roles of VpCSN5. Next, we plan to screen and indentify the potential R proteins which could specifically recognize VpCSN5-CSEP118 complex in Baihe-35-1. Then, we intend to obtain transgenic grapevines in which VpCSN5 and R genes were over-expressed, silenced, and mutated with ectopic overexpression, RNA interference, and CRISPR/Cas9 targeted gene editing technologies, respectively. Finally, to better define the roles of VpCSN5 and R genes in Baihe-35-1 response to powdery mildew, we will examine the cellular and molecular responses of these transgenic grapevines post En NAFU1 inoculation. The obtained results will not only provide useful information for further understanding the molecular mechanism of the interaction between grapevine and Erysiphe necator, but also provide new ways or ideas to identify R genes in wild Vitis spp. In practice view, it can provide better gene resource of resistance to powdery mildew for improvement European grape or other fruit trees.