花生AhGPAT9基因启动子克隆及其功能分析  

Cloning and functional analysis of peanut AhGPAT9 promoter

作　　者：沈悦[1] 沈一[1] 刘永惠[1] 梁满 张旭尧 陈志德[1] SHEN Yue;SHEN Yi;LIU Yong-hui;LIANG Man;ZHANG Xu-yao;CHEN Zhi-de(Institute of Industrial Crops,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]江苏省农业科学院经济作物研究所,江苏南京210014

出　　处：《中国油料作物学报》2023年第3期533-541,共9页Chinese Journal of Oil Crop Sciences

基　　金：江苏省农业科技自主创新资金(CX(20)3121);国家自然科学基金(31701461);财政部和农业农村部:国家现代农业产业技术体系(CARS-13)。

摘　　要：为研究花生油脂合成功能基因AhGPAT9的调控机理,分析其启动子功能,从花生品种Tifrunner基因组中扩增获得AhGPAT9基因启动子序列,构建AhGPAT9全长启动子及其多个5’缺失启动子融合GUS基因的重组表达载体,利用农杆菌介导的植物转化体系,分析启动子的活性和表达模式,结果表明:AhGPAT9启动子序列全长1750 bp,核心区位于-257 bp~-128 bp,该启动子除了具备核心元件CAAT-box和TATA-box,还包含多种响应激素调控、胁迫诱导、光反应、胚乳特异表达和生长调控相关的顺式作用元件。此外,AhGPAT9主要在转基因拟南芥的幼苗中,以及抽薹期的茎和茎生叶、小花、9-12DAP角果和对应的成熟胚中表达。本研究结果将为进一步阐释AhGPAT9在花生油脂合成途径中的生物学功能提供新的理论依据。In order to explore the promoter function of AhGPAT9 gene involved in TAG de novo synthesis,a promoter sequence was cloned from the upstream of AhGPAT9 gene in peanut(Arachis hypogaea L.)cv Tifrunner genome,and bioinformatics analysis of it was carried out.After that,recombinant expression vectors of full-length promoter as well as several 5’-terminal deletion promoters fused with GUS reporter gene were constructed,respec-tively,and then transfected by Agrobacterium rhizogenes mediated plant transgenic systems to analyze the promoter activity and expression pattern of AhGPAT9.It was found that the full-length of AhGPAT9 promoter sequence was 1750 bp,and its core region was located in-257 bp to-128 bp.In addition to the core elements essential for eu-karyotic promoters,such as CAAT-box and TATA-box,the AhGPAT9 promoter also contained multiple cis-ele-ments that were responsive to hormone regulation,stress induction,light response,endosperm specific expression and growth regulation.Furthermore,AhGPAT9 was mainly expressed on seedlings,stems,cauline leaves,flowers,9-12DAP siliques and corresponding mature embryos of transgenic Arabidopsis by GUS histochemical staining test.Our findings provide a new theory for further revealing the biological functions of AhGPAT9 involved in the lipids biosynthetic pathway of peanut.

关 键 词：花生 AhGPAT9 启动子 顺式元件 GUS活性 

分 类 号：S565.2[农业科学—作物学]