海岛棉类黄酮代谢通路相关基因的克隆及序列分析

Cloning and Sequence Analysis of Related Genes Involved in Flavonoids Metabolism Pathway in Gossypium barbadense L

本研究以前期的转录组和荧光定量筛选为基础,以海岛棉品种06-146为材料,通过RT-PCR克隆海岛棉类黄酮代谢通路关键基因。利用在线软件分析相关蛋白的理化性质及功能结构域,并比较相关蛋白在不同物种之间的同源,构建相关蛋白的系统发育进化树。分析表明GbCHI、GbDFR和GbTT7的CDS全长分别为681 bp、1 020 bp和1 593 bp。除GbTT7外,GbCHI和GbDFR为酸性蛋白。除GbCHI外,GbTT7、GbDFR均为稳定蛋白。GbCHI、GbDFR和GbTT7均为亲水蛋白。GbDFR,GbTT7在N端存在两个和一个跨膜区。Gb CHI属于查尔酮合酶家族。GbTT7属于细胞色素P450蛋白酶家族,GbDFR属于SDR家族,且这两个基因与水稻亲缘关系较近,与亚洲棉、雷蒙德氏棉和陆地棉亲缘关系较远,推测与海岛棉枯萎病抗性相关,为类黄酮代谢通路关键基因在海岛棉抗病育种中的应用提供理论依据。

In this study, based on the early transcriptome and fluorescent quantitative screening, the key genes of flavonoids metabolism pathway in Gossypium barbadense L were cloned by RT-PCR with Gossypium barbadense L variety 06-146 as material. The physical, chemical properties and functional domains of related proteins were analyzed by online software, and the homology of related proteins among different species was compared, and phylogenetic tree of related proteins was constructed. The analysis showed that the CDS length of Gb CHI, GbDFR and GbTT7 were 681 bp, 1 020 bp and 1 593 bp respectively. In addition to GbTT7, GbCH1 and GbDFR are acidic proteins. In addition to GbCHI, GbTT7 and GbDFR were stable proteins. GbCHI, GbDFR and GbTT7 are hydrophilic proteins. GbDFR and GbTT7 have two and one transmembrane region at the N end. GbCHI belongs to the chalcone synthase family. GbTT7 belongs to the cytochrome P450 protein family, GbDFR belongs to the SDR family, and these two genes are closely related to the rice, but far from Gossypium arboreum L, Gossypium raimondii and Gossypium hirsutum L. genetic relationship, presumably associated with Gossypium barbadense L Fusarium wilt resistance. The results provided the theoretical basis for the application of key genes in the flavonoid metabolic pathway in the resistance breeding ofGossypium barbadense L.