《植物生理学报》 2014, 50(5): 665-674

大豆两个C2H2型转录因子基因序列特征及表达分析

侯思宇^1,2,3, 孙朝霞¹, 郭彬¹, 王玉国¹, 李贵全¹, 韩渊怀^1,2,3,*

山西农业大学¹农学院, ²农业生物工程研究所, 山西太谷030801; ³农业部黄土高原作物基因资源与种质创制重点实验室, 太原030031

通信作者：韩渊怀；E-mail: hanyuanhuai@163.com；Tel: 0354-6286399

摘　要：

干旱气候已严重影响需水作物大豆的产量, 而植物中C2H2型转录因子在应答非生物逆境中起到重要作用, 因此充分挖掘优异抗旱大豆种质的基因资源及功能研究, 为利用基因工程手段获得抗旱大豆种质奠定理论基础。本研究从大豆叶片中克隆到2个基因, 分别命名为GmZAT9-like和GmZAT5-like。序列特征分析表明二者都属于C2H2型转录因子, 均包含典型双锌指结构域和EAR保守基序, 且氨基酸同源性低于其他物种同源基因。分子进化树分析表明, 2个基因分别与拟南芥AtZAT9和ATZAT5基因划分为一类。基于荧光实时定量PCR技术检测到2个基因分别在叶片和根部组织特异性表达。通过半定量RT-PCR和荧光定量PCR分析大豆幼苗在PEG、SA、ABA、NaCl和4 ℃胁迫处理条件下2个基因的表达模式。结果表明, 在PEG和SA胁迫条件下, 叶片中GmZAT9-like基因表达在处理后期有升高趋势, 而根部GmZAT5-like基因在处理早期受到诱导表达; ABA胁迫条件下, 2个基因均在处理初期呈现表达升高趋势而后下降; 盐和冷胁迫条件下, 叶片中GmZAT9-like基因表达受到抑制, 而根部GmZAT5-like基因表达在冷胁迫处理初期呈现升高趋势。因此推测这两个基因与大豆非生物胁迫响应相关。

关键词：非生物胁迫; C2H2型; 转录因子; 基因表达分析

收稿：2013-12-06   修定：2014-04-10

资助：山西省青年科技基金(2011021032-1和2011021032-3)、山西农业大学引进人才基金(XB2009017)、山西省科技攻关项目(20120311005-3)和山西省留学基金(2010050)。

Cloning and Expression Analysis of Two C2H2 Transcription Factors in Soybean

HOU Si-Yu^1,2,3, SUN Zhao-Xia¹, GUO Bin¹, WANG Yu-Guo¹, LI Gui-Quan¹, HAN Yuan-Huai^1,2,3,*

¹School of Agriculture, ²Institute of Agricultural Biotechnology, Shanxi Agricultural University, Taigu, Shanxi 030801, China; ³Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau, Ministry of Agriculture, Taiyuan 030031, China

Corresponding author: HAN Yuan-Huai; E-mail: hanyuanhuai@163.com; Tel: 0354-6286399

Abstract:

Soybean yield can be greatly affected under dehydration condition. Since C2H2-type transcription factor plays an important role in the response to abiotic stresses, expecially drought stress, in plants, we cloned two C2H2-type genes, GmZAT9-like and GmZAT5-like, from soybean leaves. Their DNA and protein sequences were analyzed further. The results showed that GmZAT9-like and GmZAT5-like genes contained two conservative zinc finger domains and one EAR motif. Phylogenetic tree analysis showed that GmZAT9-like and GmZAT5-like genes, together with AtZAT9 and ATZAT5, were clustered to one group. The expression patterns of these two genes under abiotic stresses and hormone treatments were also investigated. It showed that under PEG and SA treatments GmZAT9-like gene was upregulated in leaves at the later time points, while the GmZAT5-like gene was induced at the early time points in roots. As for ABA treatment, the expression of these two genes were increased at the early time point and then decreased. Under the salt and cold stress condition, GmZAT9-like gene expression was suppressed in leaves, and the GmZAT5-like gene expression was increased at the beginning of cold treatment in root. Therefore, we speculate that these two genes may be involved in abiotic stress response.

Key words: abiotic stress; C2H2-type; transcription factor; gene expression analysis