天女木兰MsARF5基因启动子克隆及功能分析  

Cloning and Functional Analysis of MsARF5 Gene Promoter from Magnolia sieboldii

作　　者：刘琳 郝昕 邱实 关士鑫 艾万峰 梅梅[1,2] 陆秀君 Liu Lin;Hao Xin;Qiu Shi;Guan Shixin;Ai Wanfeng;Mei Mei;Lu Xiujun(College of Forestry,Shenyang Agricultural University,Shenyang,110866;Liaoning Academy of Forest Science,Shenyang,110033)

机构地区：[1]沈阳农业大学林学院,沈阳110866 [2]辽宁省林业科学研究院,沈阳110033

出　　处：《分子植物育种》2024年第22期7378-7388,共11页Molecular Plant Breeding

基　　金：国家自然科学基金面上项目(No.31570621,No.31971647)资助。

摘　　要：天女木兰种子的形态和生理均具有休眠特性,其休眠解除程度与生长素的响应和信号转导有关,其中ARF基因家族起重要作用。因此,探究MsARF基因启动子区顺式作用元件及其活性结构,对解析ARF基因在天女木兰种子休眠解除中的作用机理有重要的意义。本研究利用染色体步移技术,以天女木兰种子DNA为模板,扩增MsARF5启动子区。通过Plant CARE和PLACE等在线工具共检测到50种221个作用元件,其中激素响应元件82个,分别参与到GA、MeJA、IAA、SA、ABA、ETH响应调控;光响应元件20个、代谢循环元件4个、胁迫响应元件21个,未知作用元件49个,确定了启动子的顺式作用元件TATA和CAAT作用位点。采用GUS染色的方法,通过构建不同长度的pMsARF5-pCAMBIA3301:GUS植物表达载体并侵染烟草叶片,确定MsARF5启动子对IAA和IBA有响应位点,响应区域主要在-2 544~-1 310区段和-859~-400区段。The seeds of Magnolia sieboldii have morphophysiological dormancy characteristics, and the degree of dormancy release is related to auxin response and signal transduction, in which the ARF gene family plays an important role. Therefore, to explore the cis-acting elements in the promoter region of MsARF gene and its active structure is of great significance for analyzing the mechanism of ARF gene in seed dormancy release of Magnolia sieboldii. In this study, the promoter region of MsARF5 was amplified using the seed DNA of Magnolia sieboldii by chromosome stepping technique. A total of 221 acting elements were detected by Plant CARE and PLACE, among which 82 hormone responsive elements were involved in the response regulation of GA, MeJA, IAA, SA, ABA and ETH, respectively. There were 20 photoresponsive elements, 4 metabolic cycle elements, 21 stress responsive elements and 49 unknown acting elements. The action sites of TATA and CAAT cis-acting elements of the promoter were determined. By using GUS staining method, pMsARF5-PcamBIA3301:GUS plant expression vectors with different lengths were constructed and infected tobacco leaves. The MsARF5 promoter had response sites to IAA and IBA, and the response regions were mainly in the-2 544~-1 310 and-859~-400 regions.

关 键 词：MsARF5基因 启动子克隆 顺式作用元件 GUS染色 天女木兰 

分 类 号：S792.99[农业科学—林木遗传育种]