大丽轮枝菌微菌核发育的分子机理研究

项目名称： 大丽轮枝菌微菌核发育的分子机理研究

项目编号： No.31201462

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 植物保护学、园艺学与植物营养学

项目作者： 林玲

作者单位： 江苏省农业科学院

项目金额： 24万元

中文摘要： 大丽轮枝菌是引起多种重要作物黄萎病的主要土传病原真菌，微菌核是其在土壤中的主要存活结构和初侵染源，也是防治的直接靶标。但微菌核形成发育的机制尚不清楚。本项目在农杆菌介导的双向启动子诱捕载体插入的大丽轮枝菌菌核型、中间型和菌丝型菌株的3个突变体库的基础上，筛选出单拷贝插入的微菌核发育受影响的突变菌株。结合大丽轮枝菌基因组数据库，从这些突变菌株中克隆参与调控微菌核形成发育的基因并进行回补验证。并且通过核酸序列和氨基酸序列的比较分析进行基因功能预测，通过对突变菌株和野生菌株在不同培养条件下进行相关基因实时表达特性分析和表达产物的细胞定位进行表达特性研究，以及通过致病性鉴定分析微菌核发育和致病性的关系，从而阐明微菌核发育相关基因的作用机制。本研究对明确黄萎病流行规律和制定防治措施具重要意义，为开发新型、低毒、环境友好的杀菌剂提供靶标。同时，也将有助于揭示大丽轮枝菌微菌核发育的群体遗传变异规律。

中文关键词： 大丽轮枝菌；T-DNA 插入突变体库；微菌核发育；基因功能；

英文摘要： Verticillium dahliae is the primary causal agent of Verticillium wilt and is able to infect many important crops. Because the microsclerotia are the most important structures of the pathogen for survival and causing initial infections, the microsclerotia are regarded as one of the direct targets of control of this disease. But the mechanism of microsclerotial formation and development is unknown. A promoter trap vector carrying GFP gene was used to optimize the Agrobacterium tumefaciens-mediated transformation (ATMT) of V. dahliae. The T-DNA insertion mutant libraries of a sclerotium type, a hyphal type and a intermediate type strain were constructed. Based on these results, in this project we will screen more single copy T-DNA integration mutants those microsclerotial phenotype changed. Combined with the genome sequence database of V. dahliae, microsclerotial development related genes will be cloned and complemented. Then the molecular mechanisms of microsclerotial development related genes will be clarified through the gene function prediction via the nucleic acid sequences analysis and amino acid sequences analysis, studying on the gene expression characteristics when the mutants and wild strains will be cultured in different culture conditions via assay of gene expression level and cellular localization of

英文关键词： Verticillium dahliae；T-DNA insertion mutant libraries；microsclerotial development；gene function；