捕食线虫真菌形成捕食器官的信号调控机制研究

项目名称： 捕食线虫真菌形成捕食器官的信号调控机制研究

项目编号： No.U1402265

项目类型： 联合基金项目

立项/批准年度： 2015

项目学科： 管理科学

项目作者： 杨金奎

作者单位： 云南大学

项目金额： 220万元

中文摘要： 全球每年由于作物病原线虫引起巨大的经济损失，捕食线虫真菌是线虫生防的重要研究材料。捕食器官（捕器）是真菌捕杀线虫的重要武器，它的产生和数量与真菌的捕杀效率呈正相关，然而捕器形成的信号调控机制还不清楚。本项目以产生不同捕器的三种代表性捕食线虫真菌为研究对象，采用同源性分析筛选真菌基因组中的信号蛋白基因，通过定量PCR和基因敲除等方法鉴定参与捕器形成调控的信号蛋白；采用表达谱和蛋白组技术筛选效应蛋白下游的转录因子和转录因子调控的靶基因，通过定量PCR等技术鉴定信号通路下游的转录因子和靶基因；采用绿色荧光蛋白标记、酵母双杂交和表达谱等技术确定关键蛋白的亚细胞定位、蛋白之间的相互作用和信号通路之间的交互作用。本项目针对线虫生防制剂毒力不高和防效不稳的关键问题，通过揭示捕器形成的信号调控机制，为提高捕食线虫真菌捕杀线虫的能力和开发高效线虫生防制剂奠定基础，也有助于诠释病原真菌生活模式转变的分子机制。

中文关键词： 捕食线虫真菌；捕食器官；信号蛋白；；信号途径；调控机制

英文摘要： Plant-parasitic nematodes cause severe damages to world agriculture every year, nematode-trapping fungi (NTF) are important materials for biological control of animal and plant disease caused by nematodes. Trapping devices (traps) are the important tools for NTF to capture nematodes, and the formation and number of traps are closely related to the nematicidal efficiency, while the signaling mechanism regulating the trap formation in NTF is still unclear. In this project, three NTF producing different traps were used as target strains. The potential proteins (genes) involved in signaling pathway in these NTF were screened out from their genomes by homologous analysis, and those proteins involved in singaling regulation of trap formation were furtherly identified by real time PCR (QPCR), gene knockout and complementary experiments. The potential target proteins of the effectors and target genes under the downstream of transcription factors were characterized by gene expression profile and comparative phosphoproteomic analyses between the wild type and knockout strains, and the roles of transcription factors and target genes were identified by QPCR and other molecular biological techniques. Moreover, the subcellular localization of these key signaling proteins, interactions between different proteins in signaling pathway and the crosstalk among different signaling pathways were confirmed by green fluorescence protein fusion gene technique, QPCR and the yeast two-hybrid system, respectively. Our project focused on the key questions of nematode bio-control agents, such as low virulence and unstable nematocidal effect, our results will provide a basis to improve the nematicidal activity of NTF and develop more targeted anti-nematode agents by revealing the singaling mechanisms regulating the trap formation in NTF, and also help to elucidate the switch of nematophagous fungi from the saprophytic to the predacious lifestyles.

英文关键词： Nematode-trapping fungi;Trapping devices;Signaling proteins;Signaling pathway;Regulatory mechanism