棉花转录因子基因(GhMYB11)的克隆与表达分析  

Cloning and Expression Analysis of the Transcription Factor Gene(Gh MYB11) in Gossypium hirsutum L.

作　　者：李菲[1] 柳展基[1] 王立国[1] 刘勤红[1] 刘任重[1] 

机构地区：[1]山东棉花研究中心,济南250100

出　　处：《农业生物技术学报》2015年第2期161-169,共9页Journal of Agricultural Biotechnology

基　　金：山东省农业生物资源创新利用研究项目和国家棉花产业技术体系(No.CARS-18-10)

摘　　要：MYB基因家族在植物应对外界生物和非生物胁迫的过程中有重要的调控作用。本研究利用二倍体雷蒙德氏棉(Gossypium raimondii)基因组数据库,从陆地棉(G.hirsutum)品种鲁棉研32号中克隆到一个新的MYB转录因子Gh MYB11(Gen Bank登录号:HQ234875.1),c DNA全长1 001 bp,开放阅读框828bp。通过与模式植物拟南芥(Arabidopsis thaliana)和主要作物小麦(Triticum aestivum L.)、玉米(Zea mays L.)、水稻(Oryza sativa L.)MYB蛋白的氨基酸序列比对发现,Gh MYB11蛋白与拟南芥中脱落酸(abscisic acid,ABA)合成和信号传导的重要基因At MYB13/14编码的蛋白高度相似(E值分别为7e-83和1e-90),含两个高度保守的MYB结构域R2R3及一个转录激活结构域。实时定量PCR分析发现,Gh MYB11基因在黄萎病菌(Verticillium dahliae)侵染、干旱、盐和氧化胁迫处理后的棉花幼苗叶片中表达显著上调。Gh MYB11基因可能在棉花生物和非生物胁迫反应中起重要调控作用,是陆地棉品种抗逆性遗传改良的重要候选基因。MYB gene family are important transcription factors involving in plant responses to biotic and abiotic stresses by controlling the expression of diverse functional genes. Utilizing genomic sequences of diploid Gossypium raimondii, a new cotton MYB transcription factor gene GhMYB11 （GenBank accession No. HQ234875.1） was cloned from upland cotton（G, hirsutum） cultivar Lumianyan 32. The full length of GhMYBll cDNA was 1 001 bp coding an open reading frame （ORF） of 828 bp. There were 11 SNPs （single nucleotide polymorphism） between the ORF sequences of GhMYB11 and G. raimondii 001G177100.1, of which 7 SNPs caused amino acid alternation. Phylogenetic analysis showed that GhMYBll was highly similar （E value=7e-83 and le-90, respectively） to AtMYB13/14, which were 2 important factors involving in ABA synthesis and signal transduction in Arabidopsis thaliana, with 2 highly conservative MYB DNA binding domains R2R3 and a transactivation domain. Real-time quantitative PCR analysis revealed that after Verticilliura dahliae strain VD8 inoculated, GhMYB11 expression of cotton seedlings very significantly up- regulated by 14.49 times（P〈0.01） in cotton leaves and by 5-6 times significantly（P〈0.05） after 10% PEG6000, 150 mmol/L NaCl and 200 mmol/L H2O2 treated, respectively. The result suggested that GhMYB11 maight play a key role in cotton responding to biotic and abiotic stresses and could be treated as an important candidate gene in genetic improvement for stress resistance of upland cotton cultivars. This work provides basic materials for cotton stress tolerance enhancement through genetic engineering.

关 键 词：棉花 抗逆 MYB转录因子 基因克隆 

分 类 号：S562[农业科学—作物学]