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本研究依据已公布的雷蒙德氏棉基因组测序结果,采用同源克隆的方法,以陆地棉TM-1幼苗为材料,利用RT-PCR技术克隆得到陆地棉光敏色素B基因(Gh PHYB)的c DNA序列。结果显示:该基因ORF全长3 582 bp,编码1 194个氨基酸;通过与已知的拟南芥光敏色素B氨基酸序列比对及蛋白结构预测,发现该基因包含植物光敏色素完整的结构,与烟草、拟南芥、水稻、小麦和玉米的氨基酸序列相似性分别为87.2%、77.9%、74.5%、74.5%和54.6%。同时,本研究还构建了Gh PHYB基因的RNAi干涉载体用于转化棉花,并通过构建棉花PHYB RNAi突变体了解陆地棉PHYB基因对棉花株型、开花期、产量、棉纤维品质、抗逆性等方面的调控作用。研究结果可为该基因今后应用于棉花生产奠定基础。
In this study, based on published results of genome sequencing of Raymond cotton, homologous cloning method was used to clone Gh PHYB from Gossypium hirsutum TM-1 seedlings using RT-PCR technique c DNA sequence. The results showed that ORF of this gene was 3 582 bp in length and encoded 1 194 amino acids. The amino acid sequence of Arabidopsis phytohemagglutinin B was compared with that of Arabidopsis thaliana, and the protein structure of the gene was predicted. The ORF of the gene contained the complete structure of phytochrome, The amino acid sequence identities of tobacco, Arabidopsis, rice, wheat and maize were 87.2%, 77.9%, 74.5%, 74.5% and 54.6%, respectively. At the same time, RNAi interference vector of Gh PHYB gene was also constructed to transform cotton, and PHYB RNAi mutant of cotton was used to understand the cotton plant type, flowering period, yield, cotton fiber quality, resistance, etc. The regulatory role. The results can lay a foundation for future application of this gene in cotton production.