APSES转录因子调控寡孢节丛孢菌丝发育及致病性的分子机理

APSES蛋白是真菌中特有的一类转录因子，参与真菌的生长、发育和致病性等多种生物学功能的调控。寡孢节丛孢是捕食线虫真菌研究的模式种，该真菌中存在五种APSES蛋白(StuA、Swi6和Mbp1等)，前期研究发现StuA参与该真菌的菌丝发育 (产孢和捕食器官形成等) 和杀线虫能力的调控。本项目拟在前期基础上对该真菌中APSES蛋白的功能及调控机制进行深入研究，通过实时定量PCR分析APSES基因在真菌不同发育阶段的转录模式，通过基因敲除等分子及生化技术比较APSES基因突变菌株与野生型菌株的表型差异，通过转录组分析APSES蛋白下游的靶基因及相关的调控途径，通过GFP标记和酵母双杂交等技术研究APSES与靶蛋白的表达定位及相互作用。总之，本项目拟通过阐明寡孢节丛孢APSES转录因子的功能及调控机制，为捕食线虫真菌的发育和致病机制研究提供重要的参考，也为高效线虫生防制剂的研究和开发奠定基础。

The APSES proteins belong to transcriptional factors that are unique to fungi, and they have been identified as key regulators of fungal growth, development and pathogenicity. Arthrobotrys oligospora is a typical strain to study the interaction between nematode-trapping fungi and nematodes. Recently, five APSES proteins (such as StuA, Swi6, and Mbp1) were found in the fungus A. oligospora, and the roles of StuA were identified by gene knockout, which involved in the regulation of multiple biological properties, such as mycelial growth, conidiation, trap formation and nematicidal activity. In this project, we tried to furtherly identify the roles and regulation mechanism of APSES in A. oligospora, analyze the transcription pattern of APSES proteins in different development stages using real time PCR, compare the phenotypic difference between the wild type strain and mutants using gene knockout and other molecular and biochemical techniques, analyze the downstream target genes of APSES proteins and their regulation network using RNA sequencing technology, analyze the location of target genes by fusion expression with the gene coding for green fluorescent protein, and the interactions between the APSES proteins and target proteins by yeast two-hybrid technology. In summary, we tried to illuminate the functions and regulation mechanism of APSES proteins in the development and pathogenicity of A. oligospora, our results would provide a basis for elucidating the molecular mechanism of mycelial development and pathogenicity of nematode-trapping fungi, and also help to develop more effective biocontrol agents.