果实采后灰霉菌的胞外致病因子鉴定及其功能分析

灰霉菌（Botrytis cinerea）是果蔬采后第一大病原真菌，每年造成巨大经济损失。灰霉菌具有侵染方式多样、抗逆性强、易产生抗药性等特点，导致防治非常困难。鉴定灰霉菌的致病因子并解析其与果实的互作机制将为灰霉病的防治提供理论依据。灰霉菌在侵染寄主的过程中要分泌大量的胞外蛋白，这些蛋白在病原菌与寄主的互作过程中发挥重要作用。但是，大多数胞外蛋白的功能至今仍不清楚。本研究首先将通过烟草-病原菌互作体系在灰霉菌的胞外蛋白中筛选具有致病功能的蛋白，利用基因敲除等方法在灰霉菌中验证这些蛋白的功能。同时，通过酵母文库筛选、蛋白质免疫沉淀等方法分析致病蛋白在果实中的靶标蛋白；在番茄中构建靶标蛋白的基因编辑突变株，分析靶标蛋白在果实抗病反应中的功能。本研究将为在分子水平深入理解灰霉菌与果实的互作机制提供新的证据。

Botrytis cinerea is the most important postharvest pathogenic fungus of fruits and vegetables, causing great economic losses every year. B. cinerea is difficult to control because it has a variety of modes of attack, strong stress resistance and easy to develop drug resistant. Identification of pathogenic factors of B. cinerea and analysis of its interaction mechanism with host will provide theoretical basis for controlling grey mold. B. cinerea can secrete a large amount of extracellular proteins in the process of infecting and these proteins play an important role in the interaction with the host. At present, the function of most extracellular proteins in the interaction with host cell is largely unknown. In this study, we will identify new pathogenic extracellular protein of B. cinerea through tobacco-pathogen interaction system, and then verify the function of the pathogenic proteins in B. cinerea by gene knockout and other methods. At the same time, the target protein of pathogenic protein in fruit will be analyzed by the methods of yeast library screening, protein immunoprecipitation; we will construct the gene editing mutants of the target proteins in tomato and analyze the function of the target proteins in resistance response of fruit. This study will provide new evidence for further understanding the interaction mechanism between B. cinerea and fruit at molecular level.