陆地棉两个同源基因GhBlind的克隆与表达分析

Blind同源基因对植物株型调控起着重要作用。本研究用同源克隆策略，在陆地棉(Gossypium hirsutum L.) 腋芽部位cDNA中克隆出2个Blind同源基因GhBlind1(GenBank登录号为HQ115643)和GhBlind2(登录号为HQ115644)。GhBlind1和GhBlind2由3个外显子和2个内含子组成，分别编码359个和262个氨基酸残基。组织特异性表达分析表明, GhBlind1在腋芽部位和茎尖分生组织部位优势表达，在根、叶、茎尖、幼嫩纤维表达，而在茎部不表达；GhBlind2在腋芽部位以及根、茎、叶片、茎尖表达，而在纤维不表达。通过重组PCR技术，构建表达载体Psuper-gus-b1和Psuper-gus-b2；应用根癌农杆菌EHA105介导转化棉花胚性愈伤组织进行瞬时表达分析。将棉花胚性愈伤组织共培养4 d后，2个表达载体转化的胚性愈伤组织中均能检测到GUS活性，但Psuper-gus-b2表达活性强于Psuper-gus-b1。对Blind同源蛋白的保守结构域比对发现，GhBlind1和GhBlind2与Blind同源蛋白的一致性分别达94%和91%。以Blind同源蛋白作为外参，结合NCBI已公开的23个棉花MYB蛋白构建系统发生树，发现GhBlind1和GhBlind2与棉花大多数MYB蛋白遗传距离较远，和Blind同源蛋白组成一个较小的分支。

Isolation and Expression Analysis of Two GhBlind Homologs in Upland Cotton (Gossypium hirsutum L.)

In higher plants,Blind homologs play an important role in plant architectures.Two Blind orthologs,GhBlind1 and GhBlind2,were isolated from cDNA of the shoot apex of upland cotton(Gossypium hirsutum L.) by homologous cloning strategy.GenBank accession number is HQ115643 for GhBlind1,and HQ115644 for GhBlind2.Similar to other Blind homologs,GhBlind1 and GhBlind2 were consisted of three exons and two introns,and encoded 359 and 262 amino acids,respectively.Based on the RT-PCR analysis,it was found that GhBlind...