灰葡萄孢是重要的植物病原真菌，分生孢子在其病害循环中发挥关键作用，抑制分生孢子萌发（包括分生孢子膨大、萌发、芽管延伸及芽管转化为菌丝体的过程）是防治灰霉病的有效途径之一。申请者前期将核盘菌双分病毒SsPV1转染至灰葡萄孢KY-1后，发现携带SsPV1的菌株KY-1V分生孢子萌发异常。本项目拟在此基础上，通过KY-1和KY-1V生物学性状比较，解析SsPV1灰葡萄孢的分生孢子结构、芽管形态、ROS积累和核分裂等孢子萌发相关性状的影响；明确SsPV1在分生孢子、芽管中的亚细胞定位及孢子携带SsPV1频率；对KY-1和KY-1V分生孢子萌发前后转录组、蛋白质组和代谢组数据关联分析，筛选差异表达基因；明确与SsPV1互作的寄主蛋白，鉴定新的孢子萌发相关基因，并构建分生孢子萌发相关信号途径的调控网络模型。研究期望阐明SsPV1抑制灰葡萄孢分生孢子萌发机制，为灰霉病绿色高效防治提供科学依据和理论指导。

Botrytis cinerea is an important plant pathogen fungus and its conidia play important roles in disease cycle. Inhibition of conidia germination (this stage includes spores inflation, germination, germ tube elongation, and conversion into mycelium) is one of effective control strategies for control gray mold disease. Previous research the partitivirus SsPV1 isolated from Sclerotinia sclerotiorum was successfully induced into B. cinerea strain KY-1. It is interesting that conidia germination of the SsPV1-infected strain KY-1V is inhibited and fails to conversion from germ tube to mycelium. In the present project, serial experiments were designed to decipher the mechanisms of conidia germination via SsPV1-Botrytis cinerea interaction system. Firstly, the biological features of conidia and germinated germ tube, ROS accumulation and nuclear division during conidia germination will be compared between strain KY-1 and KY-1V. Secondly, subcellular localization of SsPV1 in conidia and germ tube will be confirmed via FISH or immunogold techniques, and frequency of conidia carrying SsPV1 will be calculated. Thirdly, differential expression genes will be obtained based on levels of proteome, transcriptome and metabolome. Finally, combining the S. sclerotiorum proteins directly interacted with SsPV1, the specifically expressed new genes in conidia germination will be mined based on the bioinformatics methods, and an expected model of genes interaction net for conidia germination will be constructed. Our research results expect to elucidate the molecular mechanisms on inhibition of conidia germination by partitivirus SsPV1 in B. cinerea and will supply theory evidence for SsPV1 application to control gray mold disease.