杨树PtoCIPK2基因的克隆和表达分析  

Cloning and expression analysis of PtoCIPK2 in Populus tomentosa

作　　者：张杰伟[1] 管阳[1] 李瑞芬[1] 陈亚娟[1] 朱丹[1] 魏建华[1] 

机构地区：[1]北京市农林科学院北京农业生物技术研究中心/农业基因资源与生物技术北京市重点实验室,北京100097

出　　处：《广东农业科学》2013年第20期132-136,共5页Guangdong Agricultural Sciences

基　　金：国家"863"计划项目(2011AA100201);北京市自然科学基金(5132010);国家自然科学基金(30972392);国家"973"计划项目(2012CB114501)

摘　　要：钙调磷酸酶B类似蛋白互作蛋白激酶(CIPKs)在植物生长发育和抗逆过程中发挥着重要作用。以2个月龄的杨树(Populus tomentosa Carr.cv‘BJHR01’)幼苗总RNA反转录的cDNA为模板,克隆得到PtoCIPK2(GeneBank登录号:KF225478)基因全长cDNA序列。该基因包含1 395 bp开放读码框,编码464个氨基酸,预测蛋白分子量为52.8 ku,等电点为9.19。蛋白质结构分析表明,PtoCIPK2具有保守的蛋白激酶结构域和NAF结构域。Real-Time qRT-PCR分析表明,PtoCIPK2在杨树幼苗主根、茎和叶片中均有表达,在叶片中的表达量最高,约为其在茎中表达量的4倍。PtoCIPK2响应了NaCl(300 mmol/L)、干旱和ABA(200μmol/L)的胁迫。在NaCl和干旱胁迫12 h后,PtoCIPK2的表达量分别上调了28和8倍,推测该基因在响应高盐和干旱胁迫时起重要作用。CBL-interacting protein kinases （CIPKs） play an important role in development and stress-resistant of plant. In this study, the full length cDNA of PtoC1PK2 （GeneBank Accession No. KF225478） was cloned from the cDNA library prepared from 2-month-old Populus tomentosa trees using RT-PCR technique. The CDS of PtoCIPK2 contained 1 395 nucleotide, encoding 464 amino acid with the molecular weight about 52.8 ku and PI of 9.19. Protein structure analysis showed that PtoCIPK2 contained conservative P-kinase and NAF domain. Real time quantitative RT-PCR showed that the expression of PtoCIPK2 was higher in leaves in 6-month-old trees, about 4 times than that of in stem. Meanwhile, the results suggested that expression of PtoCIPK2 possibly responded to the stress of dehydration, ABA and NaCl in P. tomentosa. After 12 h under NaCl and dehydration stressed, expression of PtoCIPK2 was 28 and 8 times than control respectively, indicating that PtoCIPK2 would be mainly involved in dehydration and salt stresses.

关 键 词：蛋白激酶 杨树 PtoCIPK2 盐胁迫 干旱胁迫 

分 类 号：S792.11[农业科学—林木遗传育种] Q943.2[农业科学—林学]