棉花GhMYB0基因的克隆、表达分析及功能鉴定  

Cloning, Expression, and Functional Analysis of GhMYB0 Gene from Cotton(Gossypium hirsumtum L.)

作　　者：王诺菡[1,2] 于霁雯[2] 吴嫚[2] 马启峰[1] 李兴丽[2] 裴文锋[2] 李海晶[2] 黄双领[2] 张金发[2] 喻树迅[1,2] 

机构地区：[1]西北农林科技大学农学院,陕西杨凌712100 [2]中国农业科学院棉花研究所/棉花生物学国家重点实验室,河南安阳455000

出　　处：《作物学报》2014年第9期1540-1548,共9页Acta Agronomica Sinica

基　　金：国家重点基础研究发展计划(973计划)项目(2010CB126006)资助

摘　　要：MYB类转录因子是植物转录因子最大的家族之一,参与控制植物腺毛细胞的模式和形态建成。本研究利用雷蒙德氏棉(Gossypium raimondii)D5基因组数据库以AtMYB0(GL1,NM_113708)蛋白为参比序列获得同源基因GrMYB0,从徐州142中克隆了陆地棉的GhMYB0,其开放阅读框长度为843 bp,编码280个氨基酸。经过保守结构域分析和亚细胞定位确定GhMYB0为R2R3-MYB转录因子。qRT-PCR的结果表明,GhMYB0在徐州142开花当天开始高调表达,开花后20 d表达量达高峰;在所有的组织器官中,花中表达量最高,其次为胚珠。转基因功能分析结果表明,在野生型拟南芥(Columbia)中过表达GhMYB0,使其叶片表皮毛与野生型相比明显减少;该基因在拟南芥突变体gl-1中过表达,能恢复表皮毛缺失型突变体的表型,说明该基因可能对拟南芥表皮毛的形态建成发挥一定作用,本试验为研究R2R3-MYB转录因子在棉纤维起始和伸长过程中的调控作用提供有力证据。MYB transcription factor, one of the most important protein families in plants, is involved in the regulation of secon-dary metabolism, morphogenesis of plant, responding to environment stress and plant hormone. In this study, we used D5 genomic bank of Gossypium raimondii as the reference to AtMYB0 （GL1, NM_113708） protein, and cloned the full-length cDNA of a new MYB transcription factor gene GhMYB0 from cotton （Gossypium hirsutum L.）. The open reading frame of GhMYB0 is 843 bp in length, which encodes 280 amino acid residues. GhMYB0 was confirmed as R2R3-MYB transcription factor via conserved struc-ture analysis and subcellular localization. The qRT-PCR result indicated that GhMYB0 was highly expressed at the blossom day, its expresssion amount reached the peak after 20 days, with the most amount in flower, then in ovules. Transgenic funtion analysis indicated that GhMYB0 over-expressed in Arabidopsis lines, showing fewer trichomes in leaf epiderm than in that of the wild type, which suggests that the gene can restore the notrichome phenotype of gl-1 mutant of Arabidopsis. In addition, the transgenic lines had shorter plant heights, longer vegetative growth time, and pollen abortion.

关 键 词：棉花 MYB转录因子 植物表达载体构建 遗传转化 

分 类 号：S562[农业科学—作物学]