复苏植物旋蒴苣苔J结构域蛋白编码基因BhDNAJC2的克隆、表达与功能  

Cloning,Expression and Functional Analysis of a J-domain Proteincoding Gene,BhDNAJC2,from the Resurrection Plant Boea hygrometrica

作　　者：陈世璇 张振南[2,3] 王波[2] 朱燕[2] 龚月桦[1,4] 孙冬梅[3] 邓馨[2] 

机构地区：[1]西北农林科技大学生命科学学院,杨凌712100 [2]中国科学院植物研究所北方资源植物重点实验室,北京100093 [3]黑龙江八一农垦大学生命科学技术学院,大庆163319 [4]宜宾学院生命科学与食品工程学院,宜宾644000

出　　处：《植物学报》2015年第2期180-190,共11页Chinese Bulletin of Botany

基　　金：国家高技术研究发展计划(No.2007AA021403);国家转基因生物新品种培育科技重大专项(No.2009ZX08009-060B);国家自然科学基金委员会青年科学基金(No.31100195);宜宾学院重点科研项目(No.2013QD07)

摘　　要：热激蛋白(HSP)是一类在受到逆境刺激后大量表达的蛋白质,能够帮助蛋白质正确折叠,促使变性蛋白质降解,缓解逆境胁迫对生物体的损伤。为揭示热激蛋白在耐旱的复苏植物中的保护作用,该研究对复苏植物旋蒴苣苔(Boea hygrometrica)HSP40家族中J结构域蛋白Bh DNAJC2的编码基因进行了克隆、表达与功能分析。Real-time PCR检测表明,该基因受脱水、低温、热激等多种逆境条件和脱落酸(ABA)诱导表达。Bh DNAJC2-YFP定位于细胞质、内质网和细胞核。过表达Bh DNAJC2的拟南芥(Arabidopsis thaliana)株系在干旱、热激、盐胁迫和碱胁迫下均表现出明显的抗逆性。综上所述,Bh DNAJC2可能在旋蒴苣苔抗旱、耐热及耐盐碱等胁迫反应中起关键作用。Heat shock proteins （Hsps） can relieve environmental stress damage by aiding proper folding or degradation of denatured proteins. The resurrection plant Boea hygrometrica can tolerate desiccation and revive after rehydration. In desiccated leaves of B. hygrometrica, an Hsp-coding gene was cloned and designated BhDNAJC2. Sequence analysis revealed that BhDNAJC2 contained a J-domain and thus was a novel member of the Hsp40 family. Real-time PCR re- vealed that BhDNAJC2 was induced by dehydration, cold, heat, oxidative, alkaline stresses and abscisic acid treatment. Confocal microscopy revealed the BhDNAJC2-YFP fusion protein in the endoplasmic reticulum, nucleus and cytoplasm. Compared to the wild type, Arabidopsis plants overexpressing BhDNAJC2 showed improved stress tolerance under drought, heat, salt and alkaline stresses, as indicated by better growth, higher FvlFrn, and lower electrolytic leakage. BhDNAJC2 likely plays a key role in resistance to osmotic, saline, alkaline and heat stresses in B. hygrometrica.

关 键 词：热激蛋白 结构域 逆境胁迫 亚细胞定位 转基因植物 

分 类 号：Q943.2[生物学—植物学]