小麦酪蛋白激酶TaCK2α启动子的克隆与启动活性分析  

Cloning and activity analysis of the promoter of wheat casein kinase TaCK2α

作　　者：冯燕 吴志会[2] 曹雨欣 申松松 崔钟池 丁大烁 王海燕[1] 刘大群[1] FENG Yan;WU Zhihui;CAO Yuxin;SHEN Songsong;CUI Zhongchi;DING Dashuo;WANG Haiyan;LIU Daqun(College of Plant Protection,Hebei Agricultural University/Biological Control Technological Innovation Center of Plant Diseases and Plant Pests of Hebei Province,Baoding 071000,China;Tangshan Academy of Agricultural Sciences,Tangshan 063001,China)

机构地区：[1]河北农业大学植物保护学院/河北省农作物病虫害生物防治技术创新中心,河北保定071000 [2]河北省唐山市农业科学研究院,河北唐山063001

出　　处：《河北农业大学学报》2025年第2期54-61,共8页Journal of Hebei Agricultural University

基　　金：国家自然科学基金(32172384;31501623);河北省自然科学基金(C2020204028);中央引导地方科技发展资金项目(246Z6504G).

摘　　要：酪蛋白激酶2(Casein kinase 2,CK2)是在真核生物中普遍存在的组成型活性酶,已被证明广泛参与植物生长发育和环境胁迫反应。然而,CK2在小麦与叶锈菌互作过程中发挥的功能尚不清楚。本研究前期基于小麦TcLr19接种叶锈菌的转录组测序,筛选获得1个酪蛋白激酶基因TaCK2α,并证明其参与小麦抗叶锈病防御反应。为了进一步探究TaCK2α在小麦与叶锈菌互作过程中的转录调控机制,本研究基于中国春小麦数据库,获取TaCK2α基因上游启动子序列,克隆获得了TaCK2α启动子,命名为pTaCK2α。利用PlantCARE在线网站预测pTaCK2α包含的顺式作用元件,发现pTaCK2α序列含有启动子核心元件、逆境胁迫响应元件、激素响应元件、光响应元件;构建了pTaCK2α融合β-葡萄糖苷酸酶(β-Glucuronidase,GUS)、绿色荧光蛋白(Green fluorescent protein,GFP)报告基因的植物表达载体pTaCK2α-pBI121、pTaCK2α-pCamA,借助农杆菌转化本氏烟草,GUS组织化学染色及GFP荧光观察证明TaCK2α基因启动子具有启动活性。这些结果为探究TaCK2α在小麦抗叶锈病防御反应中的转录调控机制奠定了坚实的基础。Casein kinase 2 is a ubiquitous constitutively active enzyme that is consistently involved in plant growth,development,and responses to environmental stresses.However,the function of CK2 playing in the interaction between wheat Puccinia triticina(Pt)is unclear.A casein kinase gene,TaCK2α,was identified based on transcriptome sequencing of TcLr19 wheat inoculated with Pt,indicating its involvement in wheat defense response against Pt.In order to further explore its regulatory mechanism,the promoter sequence of TaCK2αwas cloned according to the sequence of China Spring wheat database which was named as pTaCK2α.The cis-regulatory elements of TaCK2αpromoter were predicted by PlantCARE,and the results showed that TaCK2αpromoter contained core elements,stress response elements,hormone response elements and light response elements.The fusion vectors pTaCK2α-pBI121,pTaCK2α-pCamA carryingβ-Glucuronidase and Green fluorescent protein reporter gene driven by pTaCK2αwere constructed and transferred into tobacco via Agrobacterium-mediated method.GUS histochemical staining and GFP fluorescence observation showed that the promoter of TaCK2αgene has promoter activity.Taken together,these results lay a solid foundation for exploring the regulation mechanism of TaCK2αinvolved in wheat defense response against Pt.

关 键 词：小麦叶锈菌 酪蛋白激酶2α 顺式作用元件 启动子 启动活性 

分 类 号：S432.1[农业科学—植物病理学]