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克隆海岛棉木质素合成关键酶基因GbHCT全长cDNA序列,分析其在纤维不同发育时期的表达情况。根据海岛棉转录组数据中的HCT序列设计引物,采用RT-PCR技术克隆基因,通过生物信息学方法分析GbHCT的cDNA序列和氨基酸序列。通过分析转录组数据研究GbHCT在纤维不同发育时期的表达。从海岛棉中克隆了1个编码HCT的基因GbHCT,开放阅读框长度为1311bp,编码436个氨基酸,蛋白质分子量为48.58kD,等电点为6.03。GbHCT氨基酸序列中含有HTLGD和DFGWG2个保守基序。GbHCT氨基酸与陆地棉、亚洲棉和雷蒙德氏棉HCT一致性较高,与其他植物中的HCT氨基酸的一致性为55.7%-60.7%。转录组数据分析发现GbHCT在纤维不同发育阶段都有表达,在5DPA的纤维中表达量最高,表明该基因可能参与棉花纤维的发育。
The full length cDNA of GbHCT, a key enzyme in cotton lignin synthesis, was cloned and the expression of GbHCT at different developmental stages was analyzed. The primers were designed according to the HCT sequence in the transcriptome of island cotton. The gene was cloned by RT-PCR and the cDNA sequence and amino acid sequence of GbHCT were analyzed by bioinformatics method. The expression of GbHCT in different developmental stages of fiber was studied by analyzing transcriptome data. One HCT-encoding gene, GbHCT, was cloned from island cotton and contained an open reading frame of 1311bp encoding a protein of 436 amino acids with a molecular weight of 48.58kD and an isoelectric point of 6.03. GbHCT amino acid sequence contains HTLGD and DFGWG2 conserved motifs. The homologies of GbHCT amino acids with HCTs of G. hirsutum, A. purpurea and G. mongolica were high, which were 55.7% -60.7% identical to those of other plant HCT amino acids. Transcriptome data analysis showed that GbHCT was expressed in different developmental stages of fiber, and the highest expression was found in 5DPA fiber, which indicated that GbHCT may be involved in the development of cotton fiber.