普通小麦中TaMYC1基因启动子的克隆和功能分析  

Cloning and Functional Analysis of Promoter of Ta MYC1 Gene in Common Wheat

作　　者：罗香怡 宗渊 曹东[3] 甘晓龙 李云[3] 刘宝龙[3] 魏乐 Luo Xiangyi;Zong Yuan;Cao Dong;Gan Xiaolong;Li Yun;Liu Baolong;Wei Le(College of Life Sciences,Qinghai Normal University,Xining,810008;College of Agriculture and Animal Husbandry,Qinghai University,Xining,810016;Key Laboratory of Crop Molecular Breeding of Qinghai Province,Chinese Academy of Sciences,Xining,810001)

机构地区：[1]青海师范大学生命科学学院,西宁810008 [2]青海大学农牧学院,西宁810016 [3]中国科学院青海省作物分子育种重点实验室,西宁810001

出　　处：《分子植物育种》2022年第8期2456-2462,共7页Molecular Plant Breeding

基　　金：青海省应用基础研究项目(2018-ZJ-762);青海省科技成果转化专项-前补助(2018-NK-133)共同资助。

摘　　要：花青素在植物生长发育和调节外界胁迫中发挥重要作用,TaMYC1是调控小麦四倍体紫色果皮花青素合成主效基因Pp3的候选基因。本研究对TaMYC1启动子的活性进行差异分析,结果表明TaMYC1w的弱活性是导致TaMYC1基因在白粒小麦中无法诱导花青素的原因之一。从紫粒小麦GY115和白粒小麦Opata分离到TaMYC1p启动子和TaMYC1w启动子,分别为3198和2186 bp。构建TaMYC1启动子植物表达载体TaMYC1p:GUS和TaMYC1w:GUS并转入本氏烟草发现TaMYC1启动子能够驱动GUS基因在烟草表皮中的表达,且TaMYC1p启动子在烟草表皮中表达量明显高于TaMYC1w,说明TaMYC1p启动子的活性要远高于TaMYC1w,这可能就是紫粒小麦中TaMYC1转录水平较高,并能够诱导激活花青素合成代谢通路的原因之一。本研究对TaMYC1p和TaMYC1w启动子进行生物信息学分析以及转基因过表达验证等分子生物学实验,为后续有色小麦花青素生物合成代谢研究提供可靠的分子理论基础。Anthocyanins play an important role in plant growth and development and regulation of external stresses,TaMYC1 is candidate gene of major gene Pp3 that regulates anthocyanin synthesis in purple pericarp of wheat.In this experiment mainly analyzed the difference in the activity of the TaMYC1 promoter,proving that the weak activity of TaMYC1w is one of the reasons why the TaMYC1 gene cannot induce anthocyanin in white wheat.TaMYC1p promoter and TaMYC1w promoter were isolated from purple wheat GY115 and white wheat Opata,which are 3198and 2186 bp respectively.Construction of TaMYC1 promoter plant expression vectors TaMYC1p:GUS and TaMYC1w:GUS,transferred into Nicotiana benthamiana,it was found that the TaMYC1 promoter can drive the expression of the GUS gene in the tobacco epidermis,and the expression level of the TaMYC1p promoter in the tobacco epidermis was significantly higher than that of TaMYC1w,indicating that the activity of the TaMYC1p promoter is much higher than that of TaMYC1w,which may be one of the reasons why the transcription level of TaMYC1 is higher in purple wheat and it can induce the activation of anthocyanin anabolic pathway.In this study,molecular biology experiments such as bioinformatics analysis and transgene overexpression verification of TaMYC1p and TaMYC1w promoters were carried out to provide a reliable molecular theoretical basis for subsequent studies of colored wheat anthocyanin biosynthesis and metabolism.

关 键 词：紫粒小麦 花青素 TaMYC1 启动子 

分 类 号：S512.1[农业科学—作物学]