玉米早期籽粒中强表达启动子的筛选  

Screening of Strongly Expressed Promoters in Immature Maize Kernels

作　　者：王莉[1,2] 王作平 张中保 白玲[1] 吴忠义[2] Wang Li;Wang Zuoping;Zhang Zhongbao;Bai Ling;Wu Zhongyi(State Key Laboratory of Crop Stress Adaption and Improvement,Henan University,Kaifeng 475004,Henan,China;Beijing Agro-Biotechnology Research Center,Beijing Academy of Agriculture and Forestry Sciences/Beijing Key Laboratory of Agricultural Gene Resources and Biotechnology,Beijing 100097,China)

机构地区：[1]河南大学省部共建作物逆境适应与改良国家重点实验室,河南开封475004 [2]北京市农林科学院北京农业生物技术研究中心/北京市农业基因资源和生物技术重点实验室,北京100097

出　　处：《作物杂志》2020年第4期114-120,共7页Crops

基　　金：北京市科委项目(Z171100001517001);北京市农林科学院储备性项目(KJCX20200407和KJCX20200104)。

摘　　要：在植物基因表达调控的过程中,启动子作为调控基因表达的顺式元件起着重要作用。为了筛选在玉米早期籽粒中强表达的启动子,选取6个启动子pCaMV35SD、pUbiquitin、pZmActin1、pZmSTK2、pZm66589和pZmbHLH148,分别构建EGFP表达载体并转染玉米原生质体,快速验证载体功能构建的正确性;同时采用花粉磁转染法将6个表达载体导入玉米自交系郑58中,并对不同启动子驱动的EGFP载体在授粉后48h玉米籽粒中的荧光强度和荧光检出率进行观察和统计分析。结果表明,6个启动子驱动EGFP表达载体构建正确;pCaMV35SD启动子驱动EGFP表达载体的荧光最强,6个启动子驱动EGFP表达载体由强到弱依次为pCaMV35SD>pZmSTK2>pZm66589>pZmbHLH148>pUbiquitin>pZmActin1,其荧光检出率分别为27.17%、27.17%、29.83%、23.84%、13.40%和30.57%。Promoters play an important role in regulating gene expression.To screen strongly expressed promoters in immature maize kernels,six promoters,pCaMV35SD,pUbiquitin,pZmActin1,pZmSTK2,pZm66589,and pZmbHLH148,were for EGFP expression in this study.Maize protoplasts were transfected to quickly verify the vector function.At the same time,six expression vectors were introduced into Zheng 58 by pollen magnetic transfection.Later,the expression of EGFP driven by different promoters in immature maize kernels were observed 48 h after pollination.The results showed that the expression of EGFP was detected in case of all six promoters but the fluorescence of EGFP driven by pCaMV35SD promoter was the strongest,and the order of EGFP fluorescence strength driven by six promoters was pCaMV35SD>pZmSTK2>pZm66589>pZmbHLH148>pUbiquitin>pZmActin1,with the fluorescence detection rate of 27.17%,27.17%,29.83%,23.84%,13.40%and 30.57%,respectively.

关 键 词：启动子 花粉磁转染法 玉米籽粒 绿色荧光蛋白 玉米原生质体 

分 类 号：S513[农业科学—作物学]