MAPK基因调控粉红螺旋聚孢霉寄生核盘菌的机制研究

粉红螺旋聚孢霉可寄生多种植物病原真菌，具有巨大的生防潜力。项目组前期通过对高效菌株67-1寄生核盘菌转录组测序，获得显著上调表达的促分裂原活化蛋白激酶（MAPK）编码基因crmapk，并证明其在寄生核盘菌过程中发挥了重要作用。MAPK是真菌细胞信号转导过程中一类重要的蛋白激酶，可通过磷酸化级联反应调控真菌生长、产孢、致病和分化等过程，但MAPK参与菌寄生作用的分子机制尚不明确。本项目拟在67-1野生菌株和crmapk敲除突变株寄生转录组测序基础上，研究差异基因和相关的代谢途径及代谢产物与菌寄生的关系；利用酵母双杂交、Pull-down和Co-IP等技术筛选鉴定Crmapk互作蛋白，研究其对菌寄生和相关代谢产物的影响，最终阐明crmapk调控菌寄生的分子机制。本研究对揭示粉红螺旋聚孢霉生防作用机制具有重要的意义，同时对进一步挖掘利用高效生防功能基因、提高病害防治水平具有重要的指导意义。

Clonostachys rosea (syn. Gliocladium roseum) is a widely distributed mycoparasite that has shown great potential against various plant fungal diseases. In our previous study, the transcriptome of a highly efficient C. rosea isolate 67-1 parasitizing Sclerotinia sclerotiorum sclerotia was constructed, from which a mitogen-activated protein kinase (MAPK) encoding gene crmapk that significantly upregulated expressed under the induction of sclerotia was obtained. The functions of crmapk in C. rosea mycoparasitism and biocontrol activity were confirmed by gene knockout and complementation. MAPK is a kind of important protein kinase that involves in signal transduction in fungi, and in turn regulates fungal growth, sporulation, pathogenicity and differentiation through phosphorylation cascade. However, the mechanisms of MAPK in mycoparasitism are not clear yet. In this study, the roles of the differentially expressed genes and the metabolic pathways related will be investigated and correlation among metabolites and mycoparasitism will be analyzed basing on the transcriptome of the wild type 67-1 and crmapk-deficient mutant. The interaction proteins of Crmapk will be caught and identified by using the techniques of yeast two-hybrid system, pull-down and Co-IP, their functions to mycoparasitism and relevant metabolites will be demonstrated, and ultimately, to reveal the molecular mechanism of crmapk regulating fungal parasitism. This research will provide insight into the mechanisms underlying biocontrol of C. rosea and facilitate to explore and utilize highly efficient biocontrol-related genes and increase control efficacy to plant diseases.