Expression and Functional Analysis of Genes Regulating Appressorium Formation in Magnaporthe Grisea

稻瘟病菌附着胞形成调控基因的表达及功能分析

• 批准号: 9874430
• 负责人: Ralph Dean
• 金额: $ 36万
• 依托单位: Clemson University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9874430&HistoricalAwards=false
• 关键词: Expression; Functional; Analysis; Genes; Regulating

The purpose of the proposed research is to elucidate the mechanismsregulating the development of the appressorium, a specialized cell requiredby the rice blast fungus Magnaporthe grisea (and other fungal pathogens)for successful host infection. Previous research focused on characterizingand cloning genes from appressorium developmental mutants. These data havebeen used to create a model of appressorium formation for further testing.In this project, advantage will be taken of progress to date as well as newresources available at the Clemson University Genomics Institute. It isnow possible to analyze the expression of many genes under a variety ofdevelopmental conditions. As many of those genes involved in appressoriumformation as possible will be identified through differential hybridizationanalysis. Gene expression in various appressorium deficient mutants willbe exploited to identify potentially key genes. The function of thesegenes will be investigated by gene disruption. Finally, to begin to definethe interplay of proteins regulating appressorium formation the interactionof gene products will be evaluated. Objectives include:1. Identify genes involved in the regulation and development ofappressorium formation. This objective covers the completion of ongoingresearch, specifically APP1 cloning and identifying genes tagged byinsertional mutagenesis. In new research, genes involved in appressoriumformation will be identified by differential hybridization of cDNAlibraries arrayed on high-density filters. Identified cDNAs will besequenced and deposited in public databases.2. Evaluate role of appressorium genes through gene disruption.3. Begin to define the regulatory networks governing appressorium formationby exploiting the 2-hybrid system.Results from this investigation have practical as well as conceptualsignificance. The identification of genes and pathways essential forinfection to occur can be exploited to create rational mechanism basedinhibitors.

本研究的目的是阐明稻瘟病菌（Magnaporthe grisea）（和其他真菌病原体）成功感染宿主所需的附着胞发育的调控机制。 以往的研究主要集中在附着胞发育突变体的特征分析和基因克隆上。这些数据已用于创建附着胞形成模型以进行进一步测试。在该项目中，将利用迄今为止的进展以及克莱姆森大学基因组学研究所可用的新资源。 现在可以分析许多基因在各种发育条件下的表达。 通过差异杂交分析，将尽可能多地鉴定出那些参与附着胞形成的基因。 将利用各种附着胞缺陷突变体的基因表达来鉴定潜在的关键基因。 这些基因的功能将通过基因破坏来研究。 最后，为了开始定义调节附着胞形成的蛋白质的相互作用，将评估基因产物的相互作用。目标包括：1.鉴定与附着胞形成的调控和发育有关的基因。 这一目标包括完成正在进行的研究，特别是APP 1克隆和鉴定插入突变标记的基因。 在新的研究中，参与附着胞形成的基因将通过排列在高密度滤膜上的cDNA差异杂交来鉴定。 鉴定的cDNA将被测序并保存在公共数据库中.通过基因破坏评价附着胞基因的作用.开始利用2-杂交系统来确定控制附着胞形成的调控网络，本研究的结果具有实际意义和概念意义。 对感染发生所必需的基因和途径的鉴定可以用来创造基于合理机制的抑制剂。