核盘菌SsNep2响应并促进盾壳霉重寄生的分子机制研究

盾壳霉是核盘菌属真菌的专一性重寄生真菌，寄生其菌核和菌丝，对由核盘菌引起的作物菌核病有显著的防治效果。乙烯/坏死诱导蛋白（NLPs）是一类广泛存在于卵菌、细菌和真菌中的分泌蛋白质。申请者前期研究时发现核盘菌基因SsNep2在接触盾壳霉后表达显著上调（log2Fold = 4.91）；有意思的是，沉默该基因却显著增强了核盘菌抵抗盾壳霉寄生的能力。申请者推测SsNep2在盾壳霉寄生核盘菌过程中扮演了“感病基因”的功能，并有潜在的应用价值。本项目拟采用基因敲除、互补等技术解析SsNep2在盾壳霉-核盘菌互作中的作用；通过蛋白亚细胞定位、酵母双杂交等明确SsNep2促进盾壳霉寄生的分子机制；在盾壳霉中超量表达SsNep2，明确它是否可以提高盾壳霉寄生核盘菌的能力。项目的完成为阐述盾壳霉-核盘菌互作的分子机理奠定基础，深度挖掘盾壳霉资源提供新的思路，并拓宽人们对广泛存在NLP蛋白编码基因功能的认识。

Coniothyrium minitans is a specific mycoparasitic fungus of Sclerotinia. It parasitizes sclerotia and mycelium and has a significant control effect on the disease of stem rot caused by Sclerotinia sclerotiorum. Ethylene- and necrosis-inducing proteins (NLPs) are a class of secreted proteins widely existing in oomycetes, bacteria and fungi. In the previous research of the applicant, it was found that the gene expression of SsNep2 was significantly up-regulated (log2fold = 4.91) after contact with C. minitans. Interestingly, silencing the gene significantly enhanced the resistance of S. sclerotiorum to parasitism. The applicant speculated that the SsNep2 played a role of "susceptible gene" in the process of parasitizing S. sclerotiorum, and had potential application value. This project intends to use gene knockout, complementary technology to analyze the role of SsNep2 in the interaction between S. sclerotiorum and C. minitans; through protein subcellular localization and yeast two-hybrid to clarify the molecular mechanism of SsNep2 promoting C. minitans parasitization; overexpression of SsNep2 in C. minitans to determine whether it can improve the mycoparasitic ability of C. minitans. The completion of the project will lay a foundation for elaborating the molecular mechanism of the interaction between S. sclerotiorum and C. minitans, provide new ideas for further mining the resources of C. minitans, and broaden people's understanding of the functions of NLP coding genes.