柑橘绿霉病在贮藏保鲜过程中危害严重，且该菌具有持续的致病性和抗药性等问题，这就极大地制约了柑橘贮藏保鲜的发展。本课题组在前期利用根癌农杆菌介导的转化（ATMT）构建了柑橘绿霉菌的T-DNA插入突变体库，经in vivo果实接种实验筛选出一株丧失致病能力的绿霉菌突变体P66-1976，该突变体为研究绿霉菌致病机理提供了很好的切入点。本项目拟通过致病力分析比较柑橘绿霉菌株及突变菌株在乙烯、柠檬烯等挥发性物质的释放量、柠檬酸等酸性物质的产生以及抗病蛋白的表达差异；应用TAIL-PCR等技术分离绿霉菌的致病基因，以酵母单双杂交分离其上下游的调控基因， RNAi及超量表达研究该基因功能，进而揭示这个基因在柑橘果实致病中的作用，顺着上下游基因的相互作用，找到更多致病相关的基因并揭示基因与环境、基因与基因网络调控的机理，为最终解析柑橘绿霉菌的致病机制奠定基础。

It can be dangerous for the infection of green mold during storage and keep-freshing in citrus, and it exited many problems such as persistent pathogenicity and fungicide- resistence. So it greatly restricted the development of storage and keep-freshing in citrus. T-DNA insertional mutants pool from P. digitatum was constructed through Agrobacterium tumefaciens mediated transformation (ATMT) based on preliminary work in our lab. One strain with loss of pathogenic ability in T-DNA insertional mutant P66-1976 from green mold in citrus was screened by in vivo experiment, and the mutant P66-1976 was a perfect experimental material to study the pathogenic mechanism of citrus green mold. Difference of green mold and its mutant P66-1976 was studied in the release of volatile substances (Ethylene, limonene etc), citric acid production, and antiviral protein expression. Target gene of mutant P66-1976 was cloned by TAIL-PCR, and we carried out target gene function analysis by yeast double hybrid, RNAi and Over-expression. And the role of the gene was revealed in the pathogenic mechanism of citrus green mold. Along with interaction of the upstream and downstream of the gene, the more pathogenic gene were found, and the mechanism of gene regulation networks were revealed between gene and gene, gene and environment. At last, it would lay the foundation for analyzing the the pathogenic mechanism of green mold in citrus.