海岛棉DELLA基因克隆及其功能分析-生物化学与分子生物学专业论文.docxVIP

III III Cloning and Functional Analysis of DELLA Gene in Gossypium Barbadense Abstract Object: Cotton is an important global economic crop. It is our countrys important economy and strategic materials too.The cotton fiber is the main product, which has become more and more important for its advantages. The Gossypium barbadense L. is the best Cultivated Variety .The world has paid more and more attention to the quality of the cotton fiber. Gibberellins (GAs) play important roles in cotton fiber development. DELLA proteins are negative regulators in the signaling pathway of GAs. In order to understand the role of GA signaling in fiber development, we initiated the characterization of this pathway in cotton plants. Methods: The gene GbGAI2 and GbGAI3 encoding the DELLA protein in G. barbadense was isolated and cloned through the method that the homology clones. Over-express GbGAI2 and GbGAI3 and their mutants in the wild type Arabidopsis by dipping the p35S:GAI2, p35S:gai2, p35S:GAI3, p35S:gai3 vector into the wild type Arabidopsis. Results and Conclusion: The gene GbGAI2 and GbGAI3 encoding the DELLA protein in G. barbadense was isolated and cloned. GbGAI2 include a 1,848 bp ORF which encoding 616 amino acid sequences，GbGAI3 include a 1,626 bp ORF which encoding 542 amino acid sequences. The typical domains of DELLA proteins were also found in the GbGAI2 and GbGAI3 proteins. Overexpression of GbGAI2, GbGAI3 and their mutants transgenic Arabidopsis were got by the floral dip (p35S:GAI2, p35S:GAI3, p35S:gai3), and these transgenic plantlets showed dwarf and late flowering phenotype, which were similar to that of GAs deficient or insensitive mutants. Key words: Gossypium barbadense; gibberellins; DELLA protein; Arabidopsis thaliana; transgenic plant 英文缩略词表 英文缩写 英文全称 中文名称 μl Minuteliter 微升 bp base pair 碱基对 DEPC Diethypyro carbonate 焦碳酸二乙酯 DNA Deoxyribonucleic acid 脱氧核糖核酸 d Day 天 ddH2O Distilled and deionized water 重蒸水 dNTP deoxyribonucleoside trephosphat 脱氧核苷三磷酸 E.coli Es