雪菜BjSnRK2E基因的克隆及表达分析  

Cloning and Expression Analysis of BjSnRK2E Gene from Potherb Mustard(Brassica juncea)

作　　者：张春兰[1] 满丽莉 包乌日娜 向殿军 刘鹏[2] Zhang Chunlan;Man Lili;Bao Wurina;Xiang Dianjun;Liu Peng(College of Life Science, Inner Mongolia University for Nationalities, Tongliao, 028042;College of Agriculture, Inner Mongolia University for Nationalities, Tongliao, 028042)

机构地区：[1]内蒙古民族大学生命科学学院,通辽028042 [2]内蒙古民族大学农学院,通辽028042

出　　处：《分子植物育种》2018年第6期1758-1768,共11页Molecular Plant Breeding

基　　金：牡丹江市科学技术计划项目(G2013n0012)资助

摘　　要：环境胁迫是造成作物品质和产量损失的重要因子。Sn RK2家族基因编码植物体内一类丝氨酸/苏氨酸蛋白激酶,是影响植物抗逆境胁迫能力的关键调控基因。采用电子克隆与RT-PCR相结合的技术,从雪菜(Brassica juncea Coss.Var.Multiceps Tsen et Lee)克隆了一个Sn RK2基因,命名为Bj Sn RK2E(Gen Bank登录号为MF423121)。序列分析显示,该基因全长1 218 bp,编码区为1 089 bp,编码362个氨基酸。Scan Prosite预测显示,Bj Sn RK2E蛋白包含一个ATP结合位点和一个丝氨酸/苏氨酸激酶结构域。多序列比对和进化树分析表明,Bj Sn RK2E蛋白与其他植物的Sn RK2蛋白分享较高的相似性,与拟南芥的At Sn RK2.6和马铃薯St Sn RK2.3蛋白亲缘关系最近,处在同一进化分枝。实时定量RT-PCR检测结果显示,Bj Sn RK2E基因在根部的表达量最高,其次是茎,叶片的表达量最低;Bj Sn RK2E基因积极参与高盐、干旱和ABA胁迫应答反应,推测该基因功能与植物抗逆境胁迫有关。Environmental stresses are important factors leading to decrease in crop yield and quality loss. The SnRK2 family genes, which are key regulators of plant resistance to abiotic stresses, encode a class of serine/threonine protein kinases in plants. The cDNA sequence of a SnRK2 gene designated as BjSnRK2E （GenBank Accession No. MF423121） was successfully isolated from potherb mustard by the combination technology of RT-PCR and electric cloning. Sequence analysis showed that the full-length cDNA of BjSnRK2E was 1 218 bp and contained a complete open reading frame （ORF） of 1 089 bp, which encoded 362 amino acid residues. ScanProsite prediction result indicated that the BjSnRK2E protein contained a ATP binding site and a serine/threonine enzyme activity domain. Multiple sequence alignment and phylogenetic tree analysis suggested that BjSnRK2E protein shared high similarity with SnRK2 proteins in other plant, and it was at the same evolutionary branch with AtSnRK2.6 in Arabidopsis thaliana as well as StSnRK2.3 in potato. Real time quantitative RT-PCR results showed that BjSnRK2E gene expressed most strongly in roots of potherb mustard,followed by that in stems, and the expression in leaves was the lowest. BjSnRK2E gene was involved in high salt, drought and ABA stress response, suggesting that the function of this gene was related to plant stress resistance.

关 键 词：雪菜 BjSnRK2E基因 克隆 特性分析 

分 类 号：S637.2[农业科学—蔬菜学]