茉莉JsMYB108和JsMYB305转录因子对萜烯合成酶基因TPS启动子的调控研究  

Regulating JsTPS Promoters by JsMYB108 and JsMYB305 in Jasminum sambac

作　　者：张月 胡莉 万超 胡双玲 尹秦婠 袁媛[1] ZHANG Yue;HU Li;WAN Chao;HU Shuangling;YIN Qinwan;YUAN Yuan(College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)

机构地区：[1]福建农林大学园艺学院,福建福州350002

出　　处：《福建农业学报》2024年第8期927-937,共11页Fujian Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31902050);福建省自然科学基金项目(2022J01588);福建农林大学科技创新专项基金(KFB23033)。

摘　　要：【目的】探明茉莉香气合成相关转录因子JsMYB108和JsMYB305对3个萜烯合成酶基因JsTPS启动子的调控机制。【方法】以茉莉叶片DNA为模板,采用染色体步移法克隆3个JsTPS基因的启动子片段,预测启动子序列顺式作用元件,并在此基础上将3个JsTPS基因启动子片段分别构建到报告载体pGWB433,单独转化烟草叶片或与pK7FWG2.0-JsMYB108、pK7FWG2.0-JsMYB305效应载体共同转化烟草叶片以检测JsMYB108和JsMYB305对3个TPS基因启动子的激活作用;同时,通过酵母单杂交实验进一步检验JsMYB108和JsMYB305与3个TPS基因启动子的结合作用。【结果】克隆了3个JsTPS基因启动子片段,长度分别为1357、1849、1005 bp。这些序列中均含MYB识别位点,不同启动子序列包含不同顺式作用元件,如光反应元件、损伤响应元件、脱落酸诱导顺式作用元件等;叶片GUS染色和GUS活性检测结果显示3个JsTPS基因启动子片段均具有启动活性且JsMYB108和JsMYB305能不同程度地激活3个TPS基因启动子活性。JsMYB108使JsTPS1、JsTPS3、JsTPS4启动子活性增强至对照的1.96倍、6.47倍和4.15倍;JsMYB305使JsTPS1、JsTPS3、JsTPS4启动子活性增强至对照的1.57倍、15.18倍和3.12倍;酵母单杂交结果显示JsMYB108和JsMYB305能与JsTPS1、JsTPS3和JsTPS4启动子结合。【结论】JsMYB108和JsMYB305可以不同程度激活JsTPS基因启动子活性,推测这2个转录因子在茉莉花萜烯香气合成和代谢过程具有一定作用。【Objective】Regulation functions of JsMYB108 and JsMYB305 on the promoters of three terpene synthase genes(TPSs)relating to the aroma synthesis of jasmine were analyzed.【Method】The promoter fragments of JsTPSs were cloned by genome walking with the DNA of jasmine leaves as template to determine the sequences of the cis-acting elements in them.The fragments were constructed separately in the reporter vector pGWB433.Then,tobacco leaves were transformed with the reporter vector alone or with pK7FWG2.0-JsMYB108 and pK7FWG2.0-JsMYB305 effect vectors to detect the activation of JsMYB108 and JsMYB305 on the promoters.Binding of JsMYB108 and JsMYB305 to the promoters was verified by the yeast one-hybrid assay.【Result】The cloned promoter fragments of the three JsTPSs were 1357 bp,1849 bp,and 1005 bp with MYB recognition sites.The elements relating to light response,damage response,and abscisic acid induced cis-acting were predicted in different promoter sequences.The GUS staining and activity detection in tobacco leaves confirmed varying degrees of activity of the fragments by introducing JsMYB108 and JsMYB305.Comparing to control,JsMYB108 expanded the activities 1.96-fold on the promoter of JsTPS1,6.47-fold on that of JsTPS3,and 4.15-fold on that of JsTPS4,while JsMYB305 did 1.57-fold,15.18-fold,and 3.12-fold,respectively.The yeast one-hybrid assay further verified the bindings of JsMYB108 and JsMYB305 to JsTPS1,JsTPS3,and JsTPS4.【Conclusion】JsMYB108 and JsMYB305 could activate the promoters of JsTPS1,JsTPS3,and JsTPS4.These two transcription factors might play a key role in the synthesis and metabolism of aromatic terpenes in jasmine flowers.

关 键 词：茉莉 转录因子 TPS 启动子 调控 

分 类 号：S685.16[农业科学—观赏园艺]