A Multiplex PCR System for the Screening of Genetically Modified (GM) Maize and the Detection of 29 GM Maize Events Based on Capillary Electrophoresis
Hongmei Yi,
Ziyue Liang,
Jianrong Ge,
Haibo Zhang,
Fengze Liu,
Xuezhen Ren,
Jie Ren,
Haijie Wang,
Jiali Ren,
Xingxu Ren,
Ying Zhang,
Fang Jin,
Shiqiao Jin,
Yikun Zhao,
Fengge Wang
Affiliations
Hongmei Yi
Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agricultural and Forest Sciences (BAAFS), Beijing 100089, China
Ziyue Liang
Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agricultural and Forest Sciences (BAAFS), Beijing 100089, China
Jianrong Ge
Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agricultural and Forest Sciences (BAAFS), Beijing 100089, China
Haibo Zhang
Shaanxi Seed Administration Bureau, Xi’an 710018, China
Fengze Liu
National Agricultural Technology Extension and Service Center, Beijing 100026, China
Xuezhen Ren
National Agricultural Technology Extension and Service Center, Beijing 100026, China
Jie Ren
Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agricultural and Forest Sciences (BAAFS), Beijing 100089, China
Haijie Wang
Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agricultural and Forest Sciences (BAAFS), Beijing 100089, China
Jiali Ren
Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agricultural and Forest Sciences (BAAFS), Beijing 100089, China
Xingxu Ren
Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agricultural and Forest Sciences (BAAFS), Beijing 100089, China
Ying Zhang
Shaanxi Seed Administration Bureau, Xi’an 710018, China
Fang Jin
National Agricultural Technology Extension and Service Center, Beijing 100026, China
Shiqiao Jin
National Agricultural Technology Extension and Service Center, Beijing 100026, China
Yikun Zhao
Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agricultural and Forest Sciences (BAAFS), Beijing 100089, China
Fengge Wang
Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agricultural and Forest Sciences (BAAFS), Beijing 100089, China
The detection of genetically modified (GM) maize events is an inevitable necessity under the strict regulatory systems of many countries. To screen for GM maize events, we developed a multiplex PCR system to specifically detect 29 GM maize events as well as the cauliflower mosaic virus 35S promoter, the Agrobacterium tumefaciens nos terminator, the Streptomyces viridochromogenes pat gene, and the endogenous zSSIIb maize reference gene. These targets were divided into five panels for screening and event-specific detection by multiplex (10-plex, 7-plex, 7-plex, 4-plex, and 5-plex) PCR. All amplification products were separated and visualized by fluorescence capillary electrophoresis (CE). By taking advantage of the high resolution, multiple fluorescence detection, and high sensitivity of CE, our system was able to identify all targets simultaneously with a limit of detection of 0.1%. The accurate identification of specific amplification peaks from different GM maize materials by CE confirmed the specificity of the system. To verify the practical applicability of this system, we analyzed 20 blind samples. We successfully identified five MON810, four TC1507, and three MIR162 samples. The detection of concomitant elements also verified the accuracy of this approach. Our system can, therefore, be used for the screening and detection of GM maize events. The system, which is easy to use, facilitates high-throughput detection with the help of a high-throughput platform and automated identification software. Multiplex PCR coupled with CE is, thus, very suitable for the detection of genetically modified organisms (GMOs) with a large number of detection targets. Additional multiplexed electrophoretic targets can be easily incorporated as well, thereby increasing the usefulness of this system as the number of GMO events continues to increase.
