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通过海岛棉基因组数据库和GSDS 2.0在线生物信息学软件,分析GbHCT基因的结构,利用实时荧光定量PCR技术,检测GbHCT基因在海岛棉不同组织和棉纤维不同发育时间的表达量。结果表明,该基因DNA序列长度为1 953bp,开放阅读框长度为1 311bp,编码436个氨基酸,在海岛棉基因组中对应scaffold3453序列,含有1个内含子和2个外显子。GbHCT基因在不同组织和纤维发育不同时期中均有表达,在各个组织中,苞叶和花中表达量最高,在棉纤维不同发育时期开花后5d和25d表达量最强。该基因可能参与棉纤维发育伸长期和次生壁增厚期。为深入研究该基因的功能,构建植物表达载体pEGAD-GbHCT并转入根癌农杆菌,为下一步研究奠定试验基础。
The structure of GbHCT gene was analyzed by the Gossypium barbadense genome database and GSDS 2.0 online bioinformatics software, and the expression level of GbHCT gene in different tissues of island island cotton and different developmental stages of cotton fiber were detected by real-time fluorescence quantitative PCR. The results showed that the DNA sequence of this gene was 1 953 bp in length and contained an open reading frame of 1131 bp, encoding a protein of 436 amino acids. The DNA sequence of this gene corresponded to the sequence of scaffold3453 and contained 1 intron and 2 exons. GbHCT gene was expressed in different tissues and different stages of fiber development. In all tissues, the expression level of GbHCT gene was highest in husk and flower, and was the highest at 5d and 25d after flowering at different developmental stages of cotton fiber. The gene may be involved in the development of cotton fiber elongation and secondary wall thickening period. In order to further study the function of this gene, the plant expression vector pEGAD-GbHCT was constructed and transformed into Agrobacterium tumefaciens, which laid the experimental foundation for further research.