大豆GmWRKY86基因的克隆及表达  

Cloning and expression analysis of GmWRKY86 in soybean

作　　者：王静[1] 付晶[1] 李晓伟 WANG Jing;FU Jing;LI Xiao-wei(Cangzhou Technical College,Cangzhou,Hebei 061001,China;Hebei University,Baoding,Hebei 071002,China)

机构地区：[1]沧州职业技术学院,河北沧州061001 [2]河北大学,河北保定071002

出　　处：《食品与机械》2020年第2期69-72,共4页Food and Machinery

基　　金：河北省教育厅自然科学基金项目(编号:17ZA0105)。

摘　　要：为了研究大豆植物中GmWRKY86基因在植株发育和抵御非生物胁迫中的生物学功能,利用其开放阅读框(opening reading frame,ORF)的PCR产物进行基因克隆,并运用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)技术检测该基因在不同组织中的表达。序列分析显示:GmWRKY86基因包含3个内含子和4个外显子,其开放阅读框长度为1 572bp,编码523个氨基酸,所编码蛋白质的等电点为8.15,预测分子量为56.82kDa,并且含有2个高度保守的WRKY结构域。进化树分析表明,GmWRKY86蛋白与来自草莓的FvWRKY42和葡萄的VvWRKY2聚为一支。荧光定量RT-PCR分析表明,在检测的所有组织中GmWRKY86都有表达,其中花中表达量最高,种子中表达量最低,说明GmWRKY86基因参与了大豆花器官的发育。In order to study the biological function of GmWRKY86 gene in soybean on development and resistance against abiotic stress,PCR products of its opening reading frame,ORF were used to clone GmWRKY86 gene,and the gene expressions in different tissues were detected by using realtime fluorescence quantitative PCR(qRT-PCR).Sequence analysis showed that the GmWRKY86 gene can encode 523 amino acids with an isoelectric point of 8.15,a molecular weight predicted to be 56.82 kDa,and an open reading frame of 1 572 bp.GmWRKY86 protein has two highly conserved domains,and the GmWRKY86 gene contains three introns and four exons.Phylogenetic tree analysis indicated that the GmWRKY86 protein belongs to the same branch as the FvWRKY42(Fragaria vesca)and VvWRKY2(Vitis vinifera).qRT-PCR analysis showed that GmWRKY86 gene was expressed in all tissues examined,with the highest expression in flowers and the lowest expression in seeds,indicating that GmWRKY86 gene involves in the development of soybean flower organs.

关 键 词：大豆 GmWRKY86 克隆 表达分析 

分 类 号：S565.1[农业科学—作物学]