棉花MYB转录因子基因GbMYB5的克隆及表达分析  

Cloning and expression analysis of cotton MYB transcription factor gene GbMYB5

作　　者：胡雪虹[1,2] 陈天子[1] 杨郁文[1] 张保龙[1] 王荣富[2] 

机构地区：[1]江苏省农业科学院农业生物技术研究所,江苏南京210014 [2]安徽农业大学生命科学院,安徽合肥230061

出　　处：《江苏农业学报》2012年第1期12-17,共6页Jiangsu Journal of Agricultural Sciences

基　　金：转基因专项(2009ZX08005-003B);江苏省自主创新项目[CX(09)112]

摘　　要：在对陆地棉Maxxa BAC文库所获得的一个BAC克隆进行测序分析过程中,发现一个新的MYB类转录因子,为探明其在棉花中的表达模式,采用RT-PCR技术从海岛棉品种H7124中克隆出该基因,命名为GbMYB5(GenBank登录号为:JF820389)。GbMYB5基因全长1 105 bp,编码277个氨基酸。RT-PCR结果表明GbMYB5基因在棉花的茎、叶、蕾、絮和未成熟的种子中均有表达,尤以叶片中的表达水平最高。重金属胁迫可短暂抑制GbMYB5基因表达,但表达量在处理48 h后回升到正常水平。PEG、脱落酸和赤霉酸诱导均可增强GbMYB5基因的表达。另外,构建了含有GbMYB5基因全长的植物过量表达载体,转化烟草。经PCR检测获得目的基因正常表达的转基因烟草9株,为研究该基因的抗逆作用奠定了基础。MYB transcription factor,one of the most important protein families in plants,is involved in the regulation of secondary metabolism,morphogenesis of plant,responding to environment stress and plant hormone.A new MYB transcription factor was found in a BAC library of Gossypium hirsutum L.cv.Maxxa..In order to explore its expression pattern in cotton,it was cloned from Gossypium barbadense 7124 by RT-PCR,and named GbMYB5（GenBank No.：JF820389）.The sequencing analysis showed that GbMYB5 cDNA covered 1 105 bp,and encoded a polypeptide of 277 amino acids.It was expressed in the stems,leaves,buds,lints and immature seeds of cotton,but the highest expression was in the leaves.The expression of GbMYB5 gene was inhibited in 24 h after heavy metals stress,but recovered to normal level in 48 h.The expression level of GbMYB5 increased significantly with PEG,abscisic acid（ABA） or gibberellic acid（GA） treatment.In addition,a plant expression vector pCAMBIA2301-GbMYB5 was constructed,and was transformed into tobacco.Nine individuals were obtained successfully,which showed the expression of GbMYB5 gene at transcription level.This would facilitate the functional analysis of GbMYB5 gene in plant tolerance to abiotic stress.

关 键 词：棉花 MYB转录因子 抗逆 烟草转化 

分 类 号：Q78[生物学—分子生物学]