橡胶树蛋白激酶基因SnRK2.7的克隆及表达  

Cloning and expression analysis of sucrose non-fermenting 1-related protein kinase 2.7(SnRK2.7)gene in Hevea brasiliensis

作　　者：刘云飞 李言[2] 田维敏[2] LIU Yunfei;LI Yan;TIAN Weimin(College of Tropical Crops,Hainan university,Haikou,Hainan 570228,China;Key Laboratory of Biology and Genetic Resources of Rubber Tree,Ministry of Agriculture/State Key Laboratory Incubation Base for Cultivation&Physiology of Tropical Crops,Rubber Research Institute,CATAS,Haikou,Hainan 571101,China)

机构地区：[1]海南大学热带作物学院,海口570228 [2]中国热带农业科学院橡胶研究所/农业农村部橡胶树生物学与遗传资源利用重点实验室/海南省热带作物栽培生理学重点实验室,海口571101

出　　处：《热带生物学报》2024年第1期1-9,共9页Journal of Tropical Biology

基　　金：广东省重点领域研发计划项目(2020b020217002);海南省科技专项(ZDKJ2021012);国家天然橡胶产业技术体系项目(CARS-33-YZ1)。

摘　　要：为了探索蔗糖非发酵相关蛋白激酶2(Sucrose non-fermentating1-related protein kinase 2,SnRK2)在巴西橡胶树响应低温胁迫应答中的作用,采用RT-PCR技术,从橡胶树抗寒无性系93-114的叶片中克隆了SnRK2亚家族的一个成员,命名为HbSnRK2.7。该基因包含一个1095 bp的开放阅读框,编码364个氨基酸。HbSnRK2.7的分子量为41.39 kD,等电点为4.70,具有保守丝氨酸/苏氨酸蛋白激酶结构域,预测其能催化磷酸基从ATP转移到蛋白质底物上的丝氨酸/苏氨酸残基上,也是磷酸化位点的集中区域,还具有非生物胁迫所需的结构域和ABA(脱落酸)依赖的HbSnRK2.7激活所需结构域。q-PCR结果显示,HbSnRK2.7基因响应ABA诱导,呈下调表达模式。在ABA处理8 h时,表达量下调到最低点。在低温胁迫下,HbSnRK2.7的表达量极显著降低。低温胁迫4 h和8 h时,HbSnRK2.7持续下调表达,低温胁迫24 h时,HbSnRK2.7表达量下调到最低,大约为非胁迫表达量的1/2。而且,HbSnRK2.7在不抗寒种质中的表达量显著高于抗寒种质。这些结果表明,HbSnRK2.7可能作为负调控因子介导橡胶树依赖ABA的低温胁迫抗性形成。To explore the role of sucrose non-fermentation associated protein kinase 2(SnRK2)in the response of Hevea brasiliensis to low temperature stress,a member of the SnRK2 subfamily,named HbSnRK2.7,was cloned from the leaves of the cold-resistant clone Reyan‘93-114’of Hevea brasiliensis by using RT-PCR.The gene cloned contains an open reading frame of 1095 bp that encodes a protein of 364 amino acids.HbSnRK2.7 has a conserved serine/threonine protein kinase domain with a molecular weight of 41.39 kD and an isoelectric point of 4.70.It is predicted that it can catalyze the transfer of phosphate groups from ATP to the serine/threonine residues on the protein substrate,which is also the concentrated region of phosphorylation sites.It also has domains needed for abiotic stress and ABA-dependent domains for HbSnRK2.7 activation.Quantitative PCR(qPCR)results showed that HbSnRK2.7 gene was down-regulated in response to ABA induction.When treated with ABA for 8 h,the expression was down-regulated to the lowest point.Under low temperature stress,the expression of HbSnRK2.7 decreased significantly.The expression of HbSnRK2.7 was down-regulated continuously under low temperature stress for 4 h and 8 h,and the expression of HbSnRK2.7 was down-regulated to the lowest level at 24 h under low temperature stress,which was about 1/2 of non-stress expression.Moreover,the expression of HbSnRK2.7 was significantly higher in the cold-susceptible clones than in the cold-resistant clones.These results suggest that HbSnRK2.7 may act as a negative regulatory factor to mediate the formation of low temperature stress resistance of Hevea brasiliensis dependent on ABA.

关 键 词：巴西橡胶树 蔗糖非发酵相关蛋白激酶 荧光定量PCR 低温胁迫抗性 脱落酸信号途径 

分 类 号：S794.1[农业科学—林木遗传育种]