稻瘟病菌TORC2复合体调控细胞膜张力及致病性的机制研究

稻瘟病菌作为丝状致病真菌中的模式生物，在生长发育和侵染机制方面具有典型的特征，全面系统的研究稻瘟病菌致病机理对稻瘟病以及其他植物真菌病害的防治具有重大的意义。细胞膜是细胞生存的基础。甾醇和鞘脂作为细胞膜的主要成分，对维持细胞膜稳态及细胞内信号的传递起到十分重要的作用，但细胞膜稳态的调控机制还不清楚。本研究小组前期发现甾醇结合蛋白MoVast1通过调节细胞膜甾醇和鞘脂的含量来影响TORC2活性和细胞膜张力，进而调控致病力。为了探究TORC2复合体是如何调控稻瘟病菌细胞膜张力以及致病力，本项目拟在TORC2调控因子MoVast1相关工作的基础上，通过细胞膜张力测定、自噬流检测以及药剂敏感性分析，利用荧光定位、免疫共沉淀、酵母双杂交和BiFC等分子生物学技术结合代谢组、磷酸化组等组学技术进一步解析TORC2复合体及上下游调控因子的功能，为基于细胞膜稳态的稻瘟病菌的防治提供理论基础和新的策略。

Magnaporthe oryzae (syn. Pyricularia oryzae), as a model organism among the filamentous pathogenic fungi, has typical characteristics in growth, development, and plant infection. Therefore, systematic studies of the pathogenic mechanisms in the rice blast fungus are important to prevent and control rice blast disease and even other plant fungal diseases. Sterols and sphingolipids, as the main components of plasma membrane which is fundamental for the survival of biological cells, play a vital role in maintaining plasma membrane homeostasis and intracellular signal transmission. However, the regulatory mechanisms of membrane homeostasis are still unclear. In our previous research, we uncovered that MoVast1 functions as a sterol binding protein regulating TORC2 activity and membrane tension by mediating the content of membrane sterols and sphingolipids, and thereby regulating pathogenicity. In order to explore how the TORC2 complex mediates the membrane tension and the infection in rice blast fungus, this project intends to further focusing on analyzing the functions of the TORC2 complex and its regulatory factors using filipin or Flipper-TR staining, detection of autophagic flux and chemical sensitivity assays, fluorescence localization, co-immunoprecipitation, yeast two hybrid and BiFC techniques combined with metabonomics and phosphoproteomics. Our results would contribute to developing new or specific strategies to control the rice blast disease based on targeting to membrane homeostasis regulatory factors.