产黄青霉菌抗稻曲病菌活性物质的鉴定及其作用机制研究

项目名称： 产黄青霉菌抗稻曲病菌活性物质的鉴定及其作用机制研究

项目编号： No.31201560

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 植物保护学、园艺学与植物营养学

项目作者： 吴丹丹

作者单位： 中国科学院武汉病毒研究所

项目金额： 24万元

中文摘要： 随着杂交稻的大面积种植，稻曲病已成为影响水稻产量和稻米品质的重大危害之一。但到目前为止，微生物来源的抗稻曲病活性物质及其杀菌机制尚无报道。本课题组分离到一株高效抗稻曲病菌的产黄青霉KF-25，其抑菌活性物质可能为一种小分子抗真菌肽(antifungal peptide, AFP)，可抑制病原菌孢子萌发和使菌丝体细胞坏死或凋亡。本课题拟采用HPLC、MS、NMRI技术对AFP进行分离鉴定，阐明其化学组成和结构；采用蛋白质组学和生理生化检测技术，研究AFP作用下稻曲病菌的生理差异和蛋白表达差异；结合病原菌差异蛋白的功能、结构及其与AFP的虚拟互作，筛选AFP靶向的与病原菌生长和致病相关靶标；运用qPCR、GFP标记、X衍射技术，解析若干靶标的基因转录动态、蛋白的细胞定位和晶体结构。本研究不仅有助于揭示AFP抗稻曲病的作用机制，亦为防治其它真菌性植物病害提供靶标层面的有力借鉴。

中文关键词： 稻曲病菌；产黄青霉；活性物质；基因组；转录组

英文摘要： With the large-scale cultivation of hybrid rice, Ustilaginoidea virens has become one of the major hazards affecting rice yield and quality. But so far, active substances from microbial sources against Ustilaginoidea virens and their fungicidal mechanisms have not been reported. Our research group isolated a strain Penicillium chrysogenum KF-25, with high activity against Ustilaginoidea virens. Its active substance may be a small molecule anti-fungal peptide (AFP). AFP can inhibit spore germination and mycelium cell necrosis or apoptosis of the pathogen. This project intends to adopt technologies such as HPLC, MS, NMRI, for the separation and identification of AFP and to clarify its chemical composition and structure; to apply techniques such as proteomics, physiological observation and biochemical detection to study the differences of physiology and protein profile of pathogen after AFP processing; to consider about the functions and three-dimensional structure models of differentially expressed proteins, and the virtual interactions between their structure models and AFP, to screen AFP's targets involved in pathogen growth and pathogenicity; to use techniques of qPCR, GFP tag and X-ray diffraction to resolve the dynamics transcription of targeted genes, as well as cellular localization and crystal structure of

英文关键词： Ustilaginoidea virens；Penicillium chrysogenum；active substance；genome；transcriptome