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园艺学报 ›› 2017, Vol. 44 ›› Issue (4): 712-722.doi: 10.16420/j.issn.0513-353x.2016-0758

• 研究论文 • 上一篇    下一篇

沙冬青AmCSDP基因特性及转基因烟草的抗寒性分析

王红蕾,王艳萍,于婷乔,刘胜利,陈玉珍,卢存福*   

  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)
  • 出版日期:2017-04-25 发布日期:2017-04-25

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

WANG Honglei,WANG Yanping,YU Tingqiao,LIU Shengli,CHEN Yuzhen,and LU Cunfu*   

  1. (College of Biological Sciences and Biotechnology,National Engineering Laboratory for Tree Breeding,Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Education Ministry,Beijing Forestry University,Beijing 100083,China)
  • Online:2017-04-25 Published:2017-04-25

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(GenBank登录号:KX756575)全长540 bp,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。

关键词: 沙冬青, 冷休克蛋白基因AmCSDP, 转基因, 低温抗性

Abstract:

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(GenBank 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(GenBank accession number XP_016170210.1)from Arachis ipaensis. To further confirm the functionof 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.

Key words: Ammopiptanthus mongolicus, cold shock domain protein gene AmCSDP, transgene, cold tolerance

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