《植物生理学报》 2014, 50(10): 1563-1573

山葡萄转录因子基因VaDREB的克隆及其抗逆功能分析

周伟^1,2,*, 吴宪^1,3,*, 贾承国¹, 胡瑞雪², 杜茜³, 李启云³, 潘洪玉^1,**

¹吉林大学植物科学学院, 吉林长春130062; ²大庆市农业技术推广中心, 黑龙江大庆163311; ³吉林省农业科学院, 吉林长春130124

通信作者：周；E-mail: panhongyu@jlu.edu.cn；Tel: 0431-87836251

摘　要：

DREB/CBF (dehydration-responsive element binding protein/C-repeat binding factor)转录因子能特异的与DRE/CRT (dehydration-responsive element/C-repeat)顺式作用元件结合, 在植物干旱、高盐和低温等非生物胁迫应答中起着重要的调控作用。本研究从山葡萄中克隆得到了一个VaDREB转录因子基因(登录号XM_002283076.1), 并对其进行了序列分析、亚细胞定位分析以及酵母和拟南芥中的功能鉴定。序列分析表明, VaDREB开放阅读框全长459 bp, 编码152个氨基酸, 预测蛋白质分子量为17 kDa, 等电点为8.67, 包含一个AP2/ERF结合域。氨基酸序列比对和系统进化分析结果表明, 该基因属于DREB/CBF转录因子A-5亚类。亚细胞定位结果表明VaDREB蛋白定位于细胞核中。重组酵母菌株与表达VaDREB基因的转基因拟南芥株系均表现出明显增强的盐、干旱、低温和高温胁迫耐受性表型, 基因表达分析结果表明, 转基因拟南芥株系中抗逆相关功能基因的表达量出现了明显上调, 这些结果证明VaDREB转录因子在植物抗逆调控过程中起着重要作用。

关键词：DREB/CBF转录因子; 逆境胁迫; 亚细胞定位; 转基因拟南芥; 山葡萄

收稿：2014-05-28   修定：2014-08-23

资助：农业部转基因生物新品种培育重大项目(2013ZX08004004)、“十二五”国家科技支撑计划课题(2012BAD19B04)和吉林省玉米产业技术体系岗位科学家(201203-1)。

Cloning and Analysis of Resistance Function of VaDREB Transcription Factor from Vitis amurensis

ZHOU Wei^1,2,*, WU Xian^1,3,*, JIA Cheng-Guo¹, HU Rui-Xue², DU Qian³, LI Qi-Yun³, PAN Hong-Yu^1,**

¹College of Plant Science, Jilin University, Changchun, Jilin 130062, China; ²The Daqing City Agro-Tech Extension and Service Center, Daqing, Heilongjiang 163311, China; ³Jilin Academy of Agricultural Sciences, Changchun, Jilin 130124, China

Corresponding author: ZHOU Wei; E-mail: panhongyu@jlu.edu.cn; Tel: 0431-87836251

Abstract:

The transcription factors DREBs/CBFs specifically interact with the DRE/CRT cis-acting element and play important roles in tolerance response to drought, high-salinity and low temperature in plant. In this study, we isolated a gene encoding DREB transcription factor from Vitis amurensis (GenBank No. XM_002283076.1) and analyzed its sequence, subcellular localization and function in recombinant yeast cells and T₃ generation transgenic Arabidopsis. The DNA sequences of VaDREB comprised a 459 bp ORF, encoding a putative protein of 152 amino acids with a predicted molecular mass of 17 kDa and isoelectric point (pI) of 8.67. The deduced amino acid sequences of VaDREB contained a highly conserved AP2/ERF domain. Sequence comparison and phylogenetic analysis revealed that VaDREB belonged to the A-5 subgroup of the DREB/CBF subfamily. The results of subcellular localization analysis suggested that the VaDREB protein was targeted to the nucleus. The recombinant yeast cells and transgenic Arabidopsis gained higher tolerance to salt, drought and cold stress. But also, the expression level of some stress responsive genes were significantly upregulated in transgenic Arabidopsis compared with those in the wild-type Arabidopsis. These results suggest that VaDREB may play a vital regulatory role in abiotic stress responses in Vitis amurensis.

Key words: DREB/CBF transcription factor; stress tolerance; subcellular localization; transgenic Arabidopsis; Vitis amurensis