玉米热激转录因子基因ZmHSF05的克隆与功能分析  

Cloning and Functional Analysis of Maize Heat Shock Transcription Factor Gene ZmHSF05

作　　者：董立红 范瑞 陈永欣 翟广谦 张璐[1] 李文和 DONG Lihong;FAN Rui;CHEN Yongxin;ZHAI Guangqian;ZHANG Lu;LI Wenhe(Maize Research Institute of Shanxi Agricultural University,Xinzhou,Shanxi 034000,China;Dryland Agricultural Research Center of Shanxi Agricultural University,Taiyuan 030031,China)

机构地区：[1]山西农业大学玉米研究所,山西忻州034000 [2]山西农业大学旱地农业研究中心,太原030031

出　　处：《西北植物学报》2020年第8期1294-1302,共9页Acta Botanica Boreali-Occidentalia Sinica

基　　金：山西省农业科学院优秀青年项目(ycx2020YQ60);科技部七大作物育种项目(2018YFD0100204)。

摘　　要：该研究通过RT-PCR扩增和测序从玉米幼苗中克隆到一个高温胁迫诱导的热激转录因子(heat shock transcription factors,HSFs)基因,命名为ZmHSF05,其开放阅读框(ORF)长度为1080 bp,编码359个氨基酸。序列分析发现,ZmHSF05具有热激转录因子蛋白的典型结构特征,含有一个DNA结合域(BD)和一个AHA激活功能结构域(AD/AHA),并且具有核定位信号(NLS)。亚细胞定位分析发现,pEGAD-ZmHSF05融合蛋白仅分布在细胞核中,表明ZmHSF05基因定位于细胞核。对ZmHSF05基因起始密码子ATG上游2102 bp序列进行顺式作用元件分析发现,该启动子区域包含多个激素和逆境胁迫响应元件。定量RT-PCR结果表明,ZmHSF05基因在根中表达水平较高,并且受高温、干旱和盐胁迫上调表达。转基因功能分析发现,ZmHSF05基因的拟南芥超表达株系耐热性显著提高,说明ZmHSF05对植物的耐热性具有正向调控作用,该研究为深入理解植物耐热性分子机制提供了新依据。In this study,we cloned a heat shock transcription factors(HSFs)gene,ZmHSF05 from maize(Zea mays)seedlings by RT-PCR amplification and sequencing.The open reading frame of ZmHSF05 is 1080 bp in length,which encodes 359 amino acid residues.Sequence analysis indicated that ZmHSF05 possessed a conserved DNA binding domain(BD),an activator domain(AD/AHA)and a nuclear localization signal(NLS)site.Subcellular localization analysis showed that pEGAD-ZmHSF05 fusion protein only distributed in the nucleus,indicating that ZmHSF05 gene localized in the nucleus.Cis-acting element analysis of the 2102 bp sequence upstream of the ATG of ZmHSF05 gene initiation codon revealed that the promoter region of ZmHSF05 contained several hormone and stress response related cis-elements.qRT-PCR result indicated that ZmHSF05 was highly expressed in root,and responsive to heat,drought and salt stresses.ZmHSF05 overexpress transgenic lines of Arabidopsis exhibited more thermo-tolerant phenotype under heat stress,including higher survival rate and cell membrane thermostability than wild type plants.These results indicated ZmHSF05 could be a positive regulator in heat stress response pathway.

关 键 词：玉米 热激转录因子 基因表达 热胁迫 功能分析 

分 类 号：Q785[生物学—分子生物学] Q786