一种可视化快速筛选转基因玉米的方法研究  

A Visual Method for Rapid Screening of Transgenic Maize

作　　者：柳青[1,2] 刘洋 胡阳[1,2] 叶欣男 刘洁 刘相国 郝东云 郭嘉[1,2] 尹悦佳 LIU Qing;LIU Yang;HU Yang;YE Xin-nan;LIU Jie;LIU Xiang-guo;HAO Dong-yun;GUO Jia;YIN Yue-jia(Institute of Agricultural Biotechnology,Jilin Academy of Agricultural Sciences,Changchun 130033;Jilin Provincial Key Laboratory of Agricultural Biotechnology,Changchun 130033;College of Agronomy,Jilin Agricultural University,Changchun 130118,China)

机构地区：[1]吉林省农业科学院农业生物技术研究所,长春130033 [2]吉林省农业生物技术重点实验室,长春130033 [3]吉林农业大学农学院,长春130118

出　　处：《玉米科学》2021年第2期50-55,59,共7页Journal of Maize Sciences

基　　金：吉林省农业科技创新工程自由创新项目(CXGC2018ZY031);吉林省科技发展计划项目自然科学基金项目(20170101014JC);吉林省科技发展计划项目自然科学基金项目(20190201290JC)。

摘　　要：利用可视化表型分析快速筛选转基因后代可以有效提高遗传转化和基因功能的验证效率。本文研究一种高效、低成本的筛选转基因玉米的新方法,该方法利用愈伤组织和糊粉层特异性表达启动子Ltp2和可视化红色荧光蛋白编码基因DsRed构建植物表达载体pCAMBIA3300-DsRed,通过农杆菌介导法侵染玉米HiII幼胚,诱导出分子检测为阳性的红色愈伤组织,肉眼可视化筛选使得组培筛选更加直观,缩减了筛选时间。T_(0)代玉米子粒中呈现出阳性的红色种子和阴性的白色种子,肉眼快速分离鉴定转基因后代材料,极大地减少了分子检测工作量,降低后续基因功能验证的研究成本。Rapid screening of transgenic events by visual phenotype analysis can improve the efficiency of transformation and gene function verification.In this study,a new method of screening transgenic maize was estab⁃lished.A new plant expression vector pcambia3300-DsRed was constructed by using the specific expression promot⁃er LTP2 of callus and aleurone layer and the visual red fluorescent protein coding gene DsRed.After infection of maize HiII immature embryos by Agrobacterium mediated method,the red callus was induced,and identified as pos⁃itive transgenic event by PCR.The visual screening method made tissue culture screening more effectively and re⁃duced the screening time.The positive red seeds and negative white seeds were visible in T_(0) generation improved the efficiency of transformation while reducing the molecular identification cost of the transgenic events.

关 键 词：玉米 转基因 红色荧光蛋白 可视化 快速筛选 

分 类 号：S513.035.3[农业科学—作物学]