棉花黄萎病蛋白质组学分析及GbCHI基因功能分析-遗传学专业毕业论文.docxVIP

山东农业大学硕士学位论文 山东农业大学硕士学位论文 PAGE PAGE 1 中 文 摘 要 棉花是我国重要的经济作物，棉花黄萎病主要是由大丽轮枝菌引起的土传维管束病 害。黄萎病已成为严重影响棉花产量与棉纤维品质的重要真菌性病害，目前国内外在黄 萎病防治以及抗病育种方面尚未取得显著进展。本实验以海岛棉 7124 为材料，利用双 向电泳和质谱分析技术比较侵菌和未侵菌的棉花根部蛋白质组差异来研究海岛棉与大 丽轮枝菌之间的防卫反应机制。得到以下研究结果： 1、从抗黄萎病品种海岛棉 7124 中鉴定出 51 个上调和 18 个下调的蛋白。这些蛋白主要 包括防卫应答相关蛋白，初级与次级代谢相关蛋白，脂质运输与细胞骨架相关蛋白。 2、利用比较蛋白质组学方法从海岛棉7124根部蛋白中分离到受棉花黄萎病病原菌诱导 的几丁质酶，克隆了GbCHI基因的cDNA序列。 3、对GbCHI基因的表达特征及其蛋白的抑菌功能等进行了分析鉴定。qRT-PCR分析表 明GbCHI基因在棉花根、茎、叶、花和胚珠中均有表达。 4、GbCHI基因表达受大丽轮枝菌、水杨酸(SA)、乙烯(ACC)和茉莉酸(JA)诱导；亚细胞 定位分析显示GbCHI蛋白主要分布在细胞膜上。 5、体外抑菌实验证明GbCHI蛋白能显著抑制大丽轮枝菌孢子的萌发和菌丝的生长。这 些研究结果为了解GbCHI的功能及其在棉花黄萎病抗性分子育种中的应用提供了实验 依据和思路。 关键词：海岛棉；蛋白质组；黄萎病；几丁质酶；抑菌活性 棉花黄萎病蛋白质组学研究及 棉花黄萎病蛋白质组学研究及 GbCHI 基因功能分析 Abstract Cotton is one of the most important economic crops in our country, verticillium wilt of cotton is a vascular disease mainly caused by the soil-born filamentous fungus Verticillium dahliae and gives serious wilt phenotype for cotton and results in severe loss of cotton production as well as downgrade of fiber quality. At present few remarkable research progress was made in the control and breeding against the pathogen. In this study, to study the mechanisms associated with defense responses in wilt-resistant sea-island cotton (Gossypium barbadense) upon V. dahliae infection, a comparative proteomic analysis between infected and mock-inoculated roots was performed by 2-DE combined with local EST database-assisted MS/MS analysis. The results are as follows： 1、A total of 51 up-regulated and 18 down-regulated proteins were identified from G. barbadense var. Hai 7124, a cultivar showing resistance against V. dahliae. These proteins are mainly involved in defense and stress responses, primary and secondary metabolisms, lipid transport and cytoskeleton reorganization. 2、Previous study identified a pathogen-responsive chitinase (named GbCHI) from sea-island cotton (Gossypium barbadense) by comparative proteomics. The GbCHI cDNA was cloned 3、The anti-fungus function of the gene products w