尖孢镰刀菌黄瓜专化型NPS6和CPS1基因的克隆与致病机理研究

项目名称： 尖孢镰刀菌黄瓜专化型NPS6和CPS1基因的克隆与致病机理研究

项目编号： No.41301255

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 天文学、地球科学

项目作者： 叶旭红

作者单位： 沈阳农业大学

项目金额： 26万元

中文摘要： 由致病性尖孢镰刀菌侵染引起的枯萎病对作物生产造成了巨大的经济损失，对该菌的致病机理及其有效防治已经成为一个重大难题。本项目拟以尖孢镰刀菌可能存在的编码非核糖体多肽合成酶的NPS6基因和CPS1基因为切入点，采用PCR克隆、基因敲除突变株构建、生理生化特性分析、毒力检测以及实时定量PCR和454高通量转录组测序等现代分子生物学和传统微生物学结合的研究方法，对尖孢镰刀菌黄瓜专化型NPS6基因和CPS1基因进行克隆及功能分析，研究其致病力贡献，探索与这两个基因相关联的表达调控网络体系，寻找并筛选更多的与尖孢镰刀菌黄瓜专化型致病相关的基因。该项目有助于系统研究NPS6基因和CPS1基因对尖孢镰刀菌黄瓜专化型的致病贡献，为探索此两种基因对尖孢镰刀菌黄瓜专化型致病机理的功能、发现药物作用的靶位点、设计新型药物分子提供理论依据，为揭示尖孢镰刀菌的致病机理奠定理论基础。

中文关键词： 尖孢镰刀菌；枯萎病；NPS6基因；CPS1基因；转录组测序

英文摘要： The Fusarium wilt caused by pathogenic Fusarium oxysporum resulted in huge economic losses to crop production, so its pathogenic mechanism and effective prevention have become a major problem. In this work, we put the possible pathogenic gene NPS6 and CPS1 gene of Fusarium oxysporum f. sp. cucumerinum encoding non-ribosomal peptide synthaseas as entry point. By PCR cloning, gene knockout mutant construct, physiological and biochemical characteristics of virulence detection and real-time quantitative PCR and 454 high-throughput transcriptome sequencing of modern molecular biology research and traditional microbiology research methods, we clone the NPS6 and CPS1 gene and analyze their functions, study the virulence contributions, explore the regulation of expression of these two genes associated network system to find and screen more possible pathogenic genes. This work can show the pathogenic contribution to F. oxysporum of CPS1 and NPS6 genes, will give the theoretical basis of completely revealing F. oxysporum pathogenic mechanism, lay the theoretical foundation for exploring the pathogenic mechanism of the function of F. oxysporum and finding the target site of drug action, to design new drug molecules.

英文关键词： Fusarium oxysporum；Fusarium wilt；NPS6 gene；CPS1 gene；RNA-Seq