沙冬青AmCSDP基因特性及转基因烟草的抗寒性分析  

Characterization of AmCSDP Gene from Ammopiptanthus mongolicus and the Analysis of Cold Resistance in Transgenic Tobacco Plants

作　　者：王红蕾[1] 王艳萍[1] 于婷乔[1] 刘胜利[1] 陈玉珍[1] 卢存福[1] 

机构地区：[1]北京林业大学生物科学与技术学院林木育种国家工程实验室教育部林木花卉遗传育种重点实验室,北京100083

出　　处：《园艺学报》2017年第4期712-722,共11页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31270737);高等学校学科创新引智计划项目(B13007);长江学者和创新团队发展计划项目(IRT13047);北京市自然科学基金项目(6112016)

摘　　要：冷休克结构域蛋白(CSPs)是含有不同数量冷休克结构域(cold shock domain,CSD)的低温响应蛋白,在低温胁迫中发挥重要作用。以沙冬青(Ammopiptanthus mongolicus)为材料,克隆得到了冷休克结构域蛋白基因AmCSDP。生物信息学分析显示AmCSDP(Gen Bank登录号:KX756575)全长540bp,编码180个氨基酸,二级结构由8.9%的α–螺旋、47.8%的β–折叠和43.3%的无规则卷曲组成,其N端含有冷休克结构域。进化分析表明沙冬青与花生同源性最高。构建了AmCSDP真核表达载体Pcambia2300-35S-AmCSDP-OCS,经农杆菌介导转入本氏烟中。对转基因T1代和野生型烟草进行4℃低温胁迫处理,结果证实转基因本氏烟比野生型的耐冷能力强。Cold shock domain proteins（CSPs）are cold induced proteins which play a critical role for the adaptation of plant to low-temperature stress. They are widely distributed in bacteria,plants and animals. In this study,we have cloned a Cold shock domain protein gene AmCSDP（Gen Bank accession number KU744005）from Ammopiptanthus mongolicus. Bioinformatics analysis showed that the ORF of AmCSDP is 540 bp,encoding 180 amino acids. The AmCSDP secondary structure consists of 8.9% α-helix,47.8% β-floding and 43.3% random coil and contains an N-terminal CSD domain. Phylogenetic analysis showed that AmCSDP shared high similarity to the homologous gene（Gen Bank accession number XP016170210.1）from Arachis ipaensis. To further confirm the function of AmCSDP,eukaryotic expression vector Pcambia2300-35S-AmCSDP-OCS was constructed and transformed into Nicotiana benthamiana by Agrobacterium. Moreover,the transgenic tobacco plants exhibited higher germination rates than the wild types under cold stress. The results suggested that AmCSDP could be a potential candidate for engineering cold tolerance in plants.

关 键 词：沙冬青 冷休克蛋白基因AmCSDP 转基因 低温抗性 

分 类 号：S68[农业科学—观赏园艺]