《植物生理学报》 2018, 54(5): 803-811

新疆野苹果MsCBF1基因的克隆及表达分析

张克闯^1,2, 李心悦³, 张一弓¹, 张道远^1,*, 王玉成¹

¹中国科学院新疆生态与地理研究所, 中国科学院干旱区生物地理与生物资源重点实验室, 乌鲁木齐830011; ²中国科学院大学, 北京100049; ³新疆师范大学生命科学学院, 乌鲁木齐830054

通信作者：张道远；E-mail: zhangdy@ms.xjb.ac.cn

摘　要：

新疆野苹果是现代栽培苹果的祖先, 对非生物胁迫具有很强的适应性, 是非常珍贵的天然基因资源库。本研究以新疆野苹果RNA为模板, RT-PCR扩增出CBF1基因cDNA序列, 命名为MsCBF1。MsCBF1基因完整开放阅读框ORF长660 bp, 编码219位氨基酸, 具有CBFs类转录因子典型的AP2保守结构域。亚细胞定位显示MsCBF1蛋白作用于细胞核; 转录激活活性分析显示MsCBF1蛋白具有转录激活活性, 其激活活性区域位于C端第166~219位氨基酸之间。基因表达分析显示, MsCBF1基因响应盐(200 mmol·L^-1 NaCl)和冷(4°C)胁迫诱导, 不响应干旱(300 mmol·L^-1甘露醇)胁迫诱导。

关键词：新疆野苹果; MsCBF1; 转录激活活性; 表达分析

收稿：2017-11-08   修定：2018-04-27

资助：国家重点研发计划(2016YFC0501505)和中国科学院特色研究所主要服务项目(TSS-2015-014-FW-4-3)。

Cloning and expression analysis of MsCBF1 gene from Malus sieversii

ZHANG Ke-Chuang^1,2, LI Xin-Yue³, ZHANG Yi-Gong¹, ZHANG Dao-Yuan^1,*, WANG Yu-Cheng¹

¹Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, Key Laboratory of Biogeography and Bioresource in Arid Land, Urumqi 830011, China; ²University of Chinese Academy of Sciences, Beijing 100049, China; ³School of Life Sciences, Xinjiang Normal University, Urumqi 830054, China

Corresponding author: ZHANG Dao-Yuan; E-mail: zhangdy@ms.xjb.ac.cn

Abstract:

Malus sieversii in Xinjiang of China is the ancestor of cultivated apples, showing a strong adaptability to abiotic stress, which is an extreme precious natural gene pool. In this study, the M. sieversii CBF1 (MsCBF1) gene were obtained by RT-PCR, the open reading frame (ORF) was 660 bp in length, encoding a amino acid sequence of 219 residues. The MsCBF1 protein contained a typical AP2 domain. The result of subcellular localization indicated that MsCBF1 was targeted to nucleus. The transcriptional activation activity analysis indicated that the C-terminal region (166–219 bp) of MsCBF1 had transcriptional activation ability. The result of qRT-PCR showed that MsCBF1 gene were up-regulated under salt stress (200 mmol.L^-1 NaCl) and cold treatment (4°C), but did not response to drought stress (300 mmol.L^-1 mannitol).

Key words: Malus sieversii; MsCBF1; transcriptional activation activity; expression analysis