番茄SlNAC1基因启动子的盐应答功能分析  

Salt response analysis of tomato SlNAC1 promoter

作　　者：古袁扬 乐露 马欣荣[2] 涂升斌[2] 李辉[2] 庄勇 廖海[1] 黄维藻[2] GU Yuanyang;YUE Lu;MA Xinrong;TU Shengbin;LI Hui;ZHUANG Yong;LIAO Hai;HUANG Weizao(School of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610031,China;Chengdu Institute of Biology,Chinese Academy of Sciences,Chengdu 610041,China;College of Ecology and Environment,Chengdu University of Technology,Chengdu 610059,China)

机构地区：[1]西南交通大学生命科学与工程学院,成都610031 [2]中国科学院成都生物研究所,成都610041 [3]成都理工大学生态环境学院,成都610059

出　　处：《应用与环境生物学报》2021年第4期988-993,共6页Chinese Journal of Applied and Environmental Biology

基　　金：中国科学院国际合作局国际伙伴计划“一带一路”专项(151751K YSB20180006);四川省国际科技创新项目(2019YFH0043)资助。

摘　　要：番茄NAC转录因子编码基因SlNAC1受假单胞菌、盐、干旱和低温等多种生物和非生物胁迫诱导表达,但其转录调控机制仍不清楚.为研究SlNAC1的盐应答转录调控机制,分离SlNAC1基因的启动子并分析其盐应答功能.构建4个5′-缺失的SlNAC1启动子(起始密码子上游2039 bp、1508 bp、1373 bp和777 bp)驱动的GUS基因表达载体,并利用农杆菌介导法分别转化烟草(Nicotiana benthamiana),随后对转基因植株进行NaCl处理和GUS染色分析.结果显示,未经NaCl处理的转基因植株均不被明显染色,而NaCl处理后,除777 bp启动子转基因植株外,2039 bp、1508 bp和1373 bp启动子转基因植株都被明显染色.这说明SlNAC1启动子中盐应答元件位于-1373 bp和-777 bp之间.结合该区间有4个盐应答元件——GT1GMSCAM4元件(核心序列为GAAAAA)的预测分析结果,推断这4个GT1GMSCAM4元件中的一个或者多个协同负责SlNAC1基因的盐应答转录调控.这4个GT1GMSCAM4元件将用于筛选SlNAC1盐应答的转录调控因子.The SlNAC1 gene encodes the transcription factor NAC in Solanum lycopersicum.It is induced by various biotic and abiotic stresses;however,the mechanism of its transcriptional regulation remains unknown.To study the regulation of SlNAC1 transcription under salt stress,we have isolated the SlNAC1 promoter and analyzed its response to salt.Four 5-deleted SlNAC1 promoter fragments,measuring 2039,1508,1373,and 777 bp upstream from the start codon,were generated and fused to the GUS reporter gene,respectively.Transgenic Nicotiana benthamiana plants,harboring each GUS construct,were obtained by Agrobacterium tumefaciens-mediated transformation,and then used to analyze GUS expression by NaCl induction and GUS staining.In the absence of NaCl treatment,all stainings of transgenic plant samples were not significant.After NaCl treatment,the 2039 bp,1508 bp and 1373 bp promoter transgenic plants were significantly stained,except for the 777 bp promoter transgenic plants,indicating that cis-element(s),responsible for NaCl response,exist in the fragment between-1373 and-777 bp.In combination with the prediction that four salt-responsive elements(GT1 GMSCAM4,with core sequence of GAAAAA)exist in this region,we infer that one or more of these elements are responsible for the transcriptional regulation of SlNAC1 genes under salt stress.These four GT1 GMSCAM4 elements will be used for further screening of transcription factors regulating SlNAC1 expression in salt stress conditions.

关 键 词：转录因子 SlNAC1 启动子 顺式元件 盐应答 

分 类 号：S641.2[农业科学—蔬菜学]