Clathrin介导的内吞作用在类酵母共生菌进入褐飞虱卵巢上皮栓滤泡细胞中的作用机制

Yeast-like symbiont (YLS) plays important roles in the growth and development of brown planthopper, as well as in its adaptation to resistant rice varieties. Transovarial transmission is the prerequisite for YLS to function continuously in the brown planthopper population. Entering into the epithelial plug of ovary is the key step during the transovarial transmission of YLS, but its molecular mechanism is unclear. In this project, technologies, including gene database construction, bioinformatics analysis, experiment of protein-protein interactions, and protein expression and location analysis, are used to screen and verify the interactive proteins between YLS and the cytomembrane of epithelial plug follicular cell, thereby the molecular of YLS cell wall and the membrane receptor of epithelial plug follicular cell in the interaction will be clarified. Then, co-immunoprecipitation and mass spectrum are used to obtain the Clathrin-mediated endocytosis (CME) associated protein interacted with the membrane receptor mentioned above. Combined with other CME associated proteins, the locations and interactions of these CME associated proteins during the entry of YLS into epithelial plug follicular cell will be studied in depth, and then the regulatory roles of these CME associated proteins during the entry of YLS into epithelial plug follicular cell will be explicated. Finally, the functional mechanism of CME in the entry of YLS into the ovary epithelial plug follicular cell is elucidated. This project will reveal profoundly the molecular mechanism of the entry of YLS into the ovary of brown planthopper, and screen out target proteins for blocking the transovarial transmission of YLS.

类酵母共生菌（YLS）在褐飞虱生长发育和适应抗性水稻品种方面非常重要，经卵传播是YLS能持续地在褐飞虱种群中发挥作用的前提条件。进入卵巢上皮栓是YLS经卵传播的关键一步，但其分子机制尚不清楚。本项目拟通过基因数据库构建、生物信息学分析、蛋白互作实验、蛋白表达定位分析等技术对YLS与卵巢上皮栓滤泡细胞细胞膜的互作蛋白进行筛选和验证，从而明确相互作用的YLS胞壁分子和上皮栓滤泡细胞的膜上受体；通过免疫共沉淀和质谱分析获得与上述膜上受体互作的Clathrin介导的内吞作用（CME）相关蛋白，结合其它CME相关蛋白，深入研究在YLS进入上皮栓滤泡细胞中这些CME蛋白之间的定位与互作，从而明确它们在YLS进入上皮栓滤泡细胞中的调控作用，最终阐明CME在YLS进入卵巢上皮栓滤泡细胞中的作用机制。本项目的实施，将深刻揭示YLS进入褐飞虱卵巢的分子机理，并有望筛选到能阻断YLS经卵传播的作用靶标蛋白。

类酵母共生菌在褐飞虱生长发育和适应抗性水稻品种方面非常重要，经卵传播是其能持续地在褐飞虱种群中发挥作用的前提条件。进入卵巢上皮栓是类酵母共生菌经卵传播的关键一步，但其分子机制尚不清楚。本项目通过基因表达数据构建和生物信息学分析以及蛋白互作实验，发现类酵母共生菌胞壁蛋白烯醇酶是其经卵传播的重要入侵因子，网格蛋白可通过肌球蛋白调节轻链2和线粒体ATP合酶脂质结合蛋白等蛋白与类酵母共生菌烯醇酶互作从而影响其经卵传播。对卵巢上皮栓处特异高表达的蛋白酶体亚基NlPSMA1、NlPSMB7和溶质载体家族蛋白NlSLC26A10等蛋白进行时空表达模式和RNA干扰研究，结果表明它们在类酵母共生菌经卵传播中起着重要作用，可作为今后防治褐飞虱的潜在靶标。