球孢白僵菌AMPK激酶Snf1调控生长发育与致病力的分子机制

Beauveria bassiana, an important Hyphomycete entomopathogenic fungus, has great potential in the biological control of insect pests. The preliminary research suggested that the Snf1 kinase has an important role in regulating fungal development and virulence. This study is sought to find alternative strategy for enhancing the fungal virulence by elucidating the function of Snf1 kinase in B. bassiana. First, it is to characterize the subunit of Snf1 kinase complex by immunoprecipitation, and elucidate the role of each subunit in fungal development and pathogenicity. Second, the target proteins downstream of the Snf1 kinase were characterized by the comparative phosphoproteome between wild type and knockout strain. Third, the function and the mechanism mediated by the target protein of Snf1 kinase were characterized with the strategy of gene replacement. Fourth, the functional genes controlled by Snf1 kinase was used to construct the recombinant strain with improved virulence and biocontrol potential. These results would be the first finding elucidating the Snf1-mediated function and mechanism in entomopathogenic fungi, and add the new knowledge to what are known about the virulence of entomopathogen. Such theory and technique would be helpful for improving the virulence of strain, and propelling the commercialization of mycoinsecticides forward.

以球孢白僵菌为代表的丝孢类杀虫真菌在害虫生物防治中具有广阔的应用前景。前期工作显示球孢白僵菌Snf1激酶(BbSnf1)控制菌株生长发育和致病力。本项目拟通过阐明Snf1激酶的调控机制，探求增强生防真菌毒力的全新思路。第一，通过免疫共沉淀技术，解析激酶复合物各亚基，并阐明各亚基在菌株生长发育和侵染过程中的功能；第二，通过分析野生菌株和敲除菌株在特定条件下的差异磷酸化蛋白质组，鉴定蛋白激酶的下游靶蛋白；第三，利用基因敲除技术并结合生化技术，阐明蛋白激酶下游关键靶蛋白的功能和调控机制；最后，利用Snf1调控的关键基因构建毒力显著增强的重组球孢白僵菌。本项目的实施和完成将首次明确Snf1激酶在生防真菌中的调控机制，有助于完善杀虫真菌侵染毒力的理论体系，有利于形成增强生防真菌毒力的全新技术方案，从而推动生防真菌制剂的技术进步和产业化进程。

以球孢白僵菌为代表的丝孢类杀虫真菌在害虫生物防治中具有广阔的应用前景。研究工作显示球孢白僵菌Snf1激酶(BbSnf1)控制菌株生长发育和致病力。本项目通过阐明Snf1激酶的功能及其调控机制，探求增强生防真菌毒力的全新思路。第一，通过基因敲除技术，鉴定BbSNF1的生物学功能；第二，通过免疫共沉淀技术，解析激酶复合物各亚基，解析与BbSNF1相互作用的蛋白；第三，通过分析野生菌株和敲除菌株在特定条件下的差异磷酸化蛋白质组，鉴定蛋白激酶的下游靶蛋白；第四，利用基因敲除技术并结合生化技术，阐明蛋白激酶下游关键靶蛋白的功能和调控机制。本项目的实施和完成首次明确Snf1激酶在生防真菌中的生物学功能和调控机制，有助于完善杀虫真菌侵染毒力的理论体系，有利于形成增强生防真菌毒力的全新技术方案，从而推动生防真菌制剂的技术进步和产业化进程。