辣椒CaWRKY5启动子的分离及其调控元件分析  

Isolation and cis-Acting Analysis of the CaWRKY5 Promoter in Pepper

作　　者：刘志钦[1,2] 杨晟[1,2] 蔡金森[1,2] 黄雪盈[1,2] 马小玲[1,2] 石兰平[1,2] 王博[1,2] 郭吟枫[1,2] 陈桂信[3] 牟少亮[1,2] 官德义[1,4] 何水林[1,4] 

机构地区：[1]福建农林大学作物遗传育种与综合利用教育部重点实验室,福州350002 [2]福建农林大学生命科学学院,福州350002 [3]福建农林大学园艺学院,福州350002 [4]福建农林大学作物科学学院,福州350002

出　　处：《应用与环境生物学报》2013年第3期389-394,共6页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(30600391;30971718);福建省自然科学基金重点项目(2008J0003;2008J0049)资助~~

摘　　要：采用Genome walking方法从辣椒基因组DNA中分离获得辣椒CaWRKY5基因上游的启动子序列,命名为CaWRKY5p.为了分析CaWRKY5p对病原菌和机械损伤的应答以及可能的调控区域,将该基因5’端顺序4个不同缺失片段分别与GUS报告基因相融合构成表达载体,在农杆菌介导的烟草叶片瞬间表达系统中表达.结果表明:启动子TATA框为起始密码子ATG上游-140 bp至-136 bp之间的TATATAA,同时含有多种与逆境胁迫相关的顺式作用元件,如W-box、S-box、MeJA应答元件和ABRE类似元件等;CaWRKYK5基因上游-936 bp启动子可以应答青枯病菌、抗病信号分子以及机械损伤;CaWRKY5p应答青枯病菌和茉莉酸甲酯(MeJA)的核心元件可能位于启动子-886 bp至-489 bp之间,应答乙烯(ET)核心调控区域位于-936 bp和-886 bp之间,而应答机械损伤的核心元件位于启动子-336 bp以及ATG之间.A -936 bp upstream promoter sequence of the CaWRKY5 was isolated from the genomic DNA of pepper by genome walking technology and named CaWRKY5p. To understand the probable regulatory region of CaWRKY5p in response to pathogen and mechanical wounding, four 5＇ deletions of the promoter were fused to the reporter gene GUS and underwent an Agrobacterium-mediated transient expression assay in tobacco leaves. The results showed that the TATA-box of the promoter was located in -140 bp to -136 bp. Several cis-acting elements, including W-box, S-box, SA-responsive element, ABRE- like element, were found in the promoter. The -936 bp upstream of CaWRKY5 could be induced in response to Pseudomonas solanacearum infection, resistance signal molecules and mechanical wounding. The possible cis-acting elements of the promoter responding to P. solanacearum infection and MeJA induction were located from -886 bp to -489 bp. The ethylene responsive cis-acting element might locate within -936 bp to -886 bp, and the fragment from transcriptional start codon to -336 bp was sufficient for GUS expression in response to mechanical wounding. Fig 3, Ref 14

关 键 词：辣椒 CaWRKY5启动子 病原菌应答 机械损伤应答 烟草叶片瞬间表达 

分 类 号：S641.3[农业科学—蔬菜学] Q943.2[农业科学—园艺学]