分泌蛋白VdASP F2参与大丽轮枝菌微菌核形成分子机理的研究

Verticillium dahliae is a vascular fungal pathogen and leads to severe economic losses in agriculture production. The microsclerotia is resting structures and primary infection source of V. dahliae, which enable long-term survival of this plant pathogen in soil, so the elicitation of molecular mechanism in microsclerotial formation is crucial to develop control measures of V. dahlia. In previous work, VdASP F2 secretory factor was identified involving in microsclerotial formation of V. dahliae. The VdASP F2 deletion mutant exhibited a significant delay in melanized microsclerotial formation under limited nutrient conditions or at lower temperature. This project plans to reveal the regulatory mechanism of VdASP F2 during microsclerotial formation. For one thing, to determine the location of VdASP F2 in cell wall; for another thing, to find proteins interacting with VdASP F2 by co-immunoprecipitation; last, to deeply dig the function, sub-cellular location and expression pattern of proteins interacting with VdASP F2. The project will contribute to find more genes related with microsclerotial formation and clarify VdASP F2 how to interact with other proteins and regulate the microsclerotial formation. The results will provide the theory base for developing possible control measures of V. dahliae.

大丽轮枝菌是重要的维管束病害，严重影响农业生产。微菌核是大丽轮枝菌的休眠结构及土壤中主要的初侵染源，它能够在土壤中长期存活。因此大丽轮枝菌微菌核形成机理的研究对开发新的大丽轮枝菌防控技术极其重要。在项目的前期研究工作中，申请人发现分泌蛋白VdASP F2与大丽轮枝菌微菌核形成紧密相关。VdASP F2基因被敲除后，大丽轮枝菌在逆境条件下产微菌核的能力明显降低。本项目拟对VdASP F2在大丽轮枝菌微菌核形成过程中的调控机制进行深入研究，首先明确VdASP F2定位于大丽轮枝菌细胞壁；然后通过CO-IP寻找VdASP F2的互作蛋白；最后对互作蛋白在微菌核形成过程中的功能，亚细胞定位及表达模式等进行深入研究。项目的研究有助于挖掘更多与微菌核形成相关的基因，阐明VdASP F2如何与其它蛋白互作并参与大丽轮枝菌微菌核形成的分子机理，期望项目的研究为设计新的大丽轮枝菌防治方案提供理论基础。

大丽轮枝菌是重要的植物病原真菌。微菌核是大丽轮枝菌的休眠结构及土壤中主要的初侵染源，它能够在土壤中长期存活。因此大丽轮枝菌微菌核形成机理的研究对开发新的大丽轮枝菌防控技术极其重要。在项目的前期研究工作中，申请人发现分泌蛋白VdASP F2与大丽轮枝菌微菌核形成紧密相关。本项目主要是对VdASP F2在大丽轮枝菌微菌核形成过程中的调控机制进行深入研究。研究内容主要包括：VdASP F2的亚细胞定位，寻找VdASP F2的互作蛋白；研究互作蛋白在微菌核形成过程中的功能。结果明确了VdASP F2定位于大丽轮枝菌细胞壁，找到并验证了与VdASP F2互作的细胞膜蛋白PNH31232，阐明了PNH31232与大丽轮枝菌微菌核形成相关。研究结果对于分析大丽轮枝逆境条件下形成微菌核的分子机理具有重要意义。