二穗短柄草BdGATA2基因的表达及其功能分析  

Expression and Function Analysis of BdGATA2 Gene in Brachypodium distachyon

作　　者：严明雄 彭伟业[1] 李魏[1] 宋娜 段康乐 李佳欣 戴良英[1] 王冰 Yan Mingxiong;Peng Weiye;Li Wei;Song Na;Duan Kangle;Li Jiaxin;Dai Liangying;Wang Bing(College of Plant Protection,Hunan Agricultural University,Changsha,410128)

机构地区：[1]湖南农业大学植物保护学院,长沙410128

出　　处：《分子植物育种》2024年第9期2916-2922,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31672017);国家重点研发计划项目(2016YFD0200800,2016YFD0300707);湖南省教育厅科学研究项目优秀青年项目(19B247);湖南省大学生创新创业训练计划项目(s202010537051)共同资助。

摘　　要：植物GATA基因广泛调控生长发育、环境胁迫和次生代谢等生理功能,但是其作用机制尚不清晰。本研究克隆了二穗短柄草(Brachypodium distachyon)GATA转录因子BdGATA2基因,并进行亚细胞定位和表达分析,为深入研究GATA基因的功能机制提供理论参考。通过无缝克隆试剂盒从二穗短柄草cDNA中克隆BdGATA2基因;使用农杆菌介导烟草瞬时表达系统进行BdGATA2蛋白的亚细胞定位;利用酵母系统验证BdGATA2的转录激活作用;使用RT-qPCR检测在高温和低温胁迫下二穗短柄草体内的BdGATA2基因转录水平变化;利用STRING数据库构建BdGATA2蛋白互作网络。结果表明,BdGATA2与大麦(Hordeum vulgare)GATA基因AC256257.1相似度最高,达到98.32%,表明其具有高度的保守性;BdGATA2蛋白在细胞核中表达,且具有自激活作用;在低温胁迫下,BdGATA2基因表达上调;蛋白互作预测结果显示,BdGATA2蛋白分别与JAZ4(茉莉酸通路)和PYDK1(植物生长素合成途径)互作。结果说明,BdGATA2基因具有典型GATA结构域,发挥转录激活功能;在二穗短柄草受到高温和低温胁迫时,BdGATA2基因差异表达;互作网络分析推测BdGATA2蛋白可能参与抗病和生长等多种生理过程。Plant GATA genes extensively regulate physiological functions such as growth and development,environmental stress and secondary metabolism,but their mechanism is still unclear.To provide theoretical references for in-depth study of the function mechanism of GATA genes,the GATA transcription factor BdGATA2 gene was cloned from Brachypodium distachyon,and further analyzed its subcellular localization and expression in this study.BdGATA2 was cloned from the cDNA of B.distachyon using seamless cloning kit.Subcellular localization of BdGATA2 protein was conducted in tobacco according Agrobacterium-mediated transient expression system.Yeast system was used to verify transcriptional activation of BdGATA2.The transcription level of BdGATA2 gene was detected by RT-qPCR in B.distachyon under high temperature and low temperature stresses.The BdGATA2 protein interaction network was constructed using STRING database.The results showed GATA gene AC256257.1 in Hordeum vulgare shares highest sequence homology with BdGATA2,and the similarity reached to 98.04%,indicating the sequence conservation of BdGATA2.The results of subcellular localization showed that BdGATA2 protein was expressed in nucleus and had self-activation.BdGATA2 gene expression was up-regulated in low temperature stress.The results of protein interaction prediction showed that BdGATA2 protein interacted with JAZ4(Jasmonic acid pathway)and PYDK1(plant auxin synthesis pathway),respectively.The results indicated BdGATA2 gene has a typical GATA domain,which plays a role in transcriptional activation.When B.distachyon is subjected to high temperature and low temperature stresses,the BdGATA2 gene is differentially expressed;the interaction network analysis speculates that the BdGATA2 protein may be involved in various physiological processes such as plant disease resistance and plant growth.

关 键 词：二穗短柄草 BdGATA2 逆境胁迫 表达分析 

分 类 号：Q943.2[生物学—植物学]