转基因抗虫耐除草剂玉米MON87411精准定量检测方法的建立  

Establishment of accurate quantitative detection method for insect-resistant and herbicide-tolerant maize MON87411

作　　者：李凌燕 张旭冬[1] 陈子言 王颢潜 乌兰 田锦[2] 张秀杰[1] 陈红 梁晋刚 LI Lingyan;ZHANG Xudong;CHEN Ziyan;WANG Haoqian;WU Lan;TIAN Jin;ZHANG XiujieC;CHEN Hong;LIANG Jingang(Development Center of Science and Technology,Ministry of Agriculture and Rural Affairs,Beijing 100176,China;Beijing Vocational College of Agriculture,Beijing 102442,China)

机构地区：[1]农业农村部科技发展中心,北京100176 [2]北京农业职业学院,北京102442

出　　处：《生物安全学报》2023年第1期38-45,共8页Journal of biosafety

基　　金：农业科研杰出人才培养计划(2021年);转基因生物新品种培育重大专项(2019ZX08013011、2016ZX08012003)。

摘　　要：【目的】抗虫耐除草剂玉米MON87411是孟山都远东有限公司利用农杆菌介导方法研发的玉米转化体,已获得我国进口用作加工原料的农业转基因生物安全证书。为满足生物安全监管的要求,亟需建立该转化体的定量检测方法。【方法】根据抗虫耐除草剂玉米MON87411的两端旁侧序列信息设计引物和Taqman探针,进行引物筛选、特异性检测、PCR体系优化、标准曲线建立、正确度及精密度检测、检出限及定量限测试、微滴数字PCR方法验证等。【结果】该方法能特异、定量地检测出抗虫耐除草剂玉米MON87411转化体成分,检出限低至10拷贝,定量限为40拷贝。对测试样品定值准确,经微滴数字PCR方法验证结果一致。【结论】本方法为该新品种转基因玉米品系的精准定量提供了一种新的检测方法,为生物安全监管提供了有效的技术支撑。【Aim】Insect-resistant and herbicide-tolerant maize MON87411 is a maize event developed by Monsanto Far East Co.Ltd.using agrobacterium mediated transformation through recombinant DNA techniques.It has obtained a safety certificate for genetically modified agricultural organisms imported to China as raw materials for processing.Establishing a quantitative detection method for the event to meet the requirements of biosafety compliance is necessary.【Method】Primers and TaqMan probes were designed based on the flanking sequence information of maize MON87411.After primer screening,specificity detection,PCR system optimization,standard curve establishment,accuracy and precision detection,limit of detection(LOD),limit of quantification(LOQ)detection,and droplet digital PCR validation were performed.【Result】The results showed that the method could specifically and quantitatively detect the components of maize MON87411,with a LOD of 10 copies and a LOQ of 40 copies.The values of the tested samples were accurate,and the results were consistent with those of droplet digital PCR.【Conclusion】The method provides a new detection method for accurately quantifying the new genetically modified maize MON87411 and provides effective technical support for biosafety supervision.

关 键 词：转基因玉米 MON87411 抗虫耐除草剂 实时荧光PCR 微滴数字PCR 精准定量 

分 类 号：S513[农业科学—作物学]