CP4-EPSPS转基因棉花植株鉴定方法比较分析

doi:10.13560/j.cnki.biotech.bull.1985.2017-0250

生物技术通报 ›› 2017, Vol. 33 ›› Issue (4): 114-118.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0250

CP4-EPSPS转基因棉花植株鉴定方法比较分析

郭文芳, 王楠, 李刚强, 许芳芳, 杨彩峰, 刘德虎

中国农业科学院生物技术研究所,北京 100081

收稿日期:2017-02-18 出版日期:2017-04-25 发布日期:2017-04-25
作者简介:郭文芳,女,博士研究生,研究方向：抗病、耐盐碱转基因棉花培育;E-mail：guowf87@163.com
基金资助:
国家转基因生物新品种培育科技重大专项资助项目（2016ZX08005-004）

Comparative Analysis of Identification Methods of CP4-EPSPS Transgenic Cotton Plants

GUO Wen-fang, WANG Nan, LI Gang-qiang, XU Fang-fang, YANG Cai-feng, LIU De-hu

Biotechnology Research Institute,Chinese Academy of Agricultural Science,Beijing 100081

Received:2017-02-18 Published:2017-04-25 Online:2017-04-25

摘要/Abstract

摘要： CP4-EPSPS是从土壤农杆菌CP4菌株中分离得到的抗除草剂草甘膦基因,是转基因植物研究中最为常用的一种筛选标记基因。但是在CP4-EPSPS转基因植物后期筛选过程中仅凭简单的草甘膦抗性筛选并不能得到完全准确的鉴定结果,常会出现假阳性或漏选的情况。本研究以抗草甘膦转CP4-EPSPS基因棉花为材料,对PCR扩增、ELISA、CP4-EPSPS蛋白检测试纸条及草甘膦抗性四种转基因阳性植株筛选方法进行比较,从筛选结果的准确性、时效性、易用性及实惠性等不同方面综合分析了各种筛选方法的优缺点,结果表明ELISA、试纸条检测方法的准确性显著高于PCR检测与0.3%浓度草甘膦抗性检测,而试纸条检测操作更加便捷,因此建议将草甘膦抗性与试纸条检测方法结合起来使用。本研究结果为以CP4-EPSPS基因为标记筛选的转基因植物的室内及大田筛选提供实验依据。

关键词: 转基因棉花, CP4-EPSPS, 草甘膦抗性, 转基因植株筛选

Abstract: CP4-EPSPS isolated from Agrobacterium strain CP4 is a glyphosate resistant gene and one of the most commonly used marker genes in transgenic plant research. However,we found that investigated results of the transgenic plants were not always completely accurate only using simple glyphosate resistance assay in the identification of CP4-EPSPS transgenic plants,which resulted in false-positive plants or missing positive transgenic plants. In this study,by use CP4-EPSPS transgenic cottons that we had gained,the four screening methods of CP4-EPSPS transgenic plants were compared,which included PCR,ELISA,resistant test strips and glyphosate resistance assay. The results show that the accuracy of ELISA and test strip are significantly higher than PCR and 0.3% glyphosate,but the test strip test is more convenient than ELISA basing on the comprehensive analysis,accuracy,timeliness,ease of use and price in identification of transgenic plants. Therefore we think combination glyphosate resistance assay with resistant strip test is the best choice. Our finding provides significant guidance for people engaging in the identification of CP4-EPSPS transgenic plants both in the laboratory and field.

Key words: transgenic cotton, CP4-EPSPS, glyphosate resistance, identification of transgenic plants

郭文芳, 王楠, 李刚强, 许芳芳, 杨彩峰, 刘德虎. CP4-EPSPS转基因棉花植株鉴定方法比较分析[J]. 生物技术通报, 2017, 33(4): 114-118.

GUO Wen-fang, WANG Nan, LI Gang-qiang, XU Fang-fang, YANG Cai-feng, LIU De-hu. Comparative Analysis of Identification Methods of CP4-EPSPS Transgenic Cotton Plants[J]. Biotechnology Bulletin, 2017, 33(4): 114-118.

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CP4-EPSPS转基因棉花植株鉴定方法比较分析

Comparative Analysis of Identification Methods of CP4-EPSPS Transgenic Cotton Plants

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