Event-Specific Qualitative and Quantitative Detection of Genetically Modified Rice G6H1
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Event-Specific Qualitative and Quantitative Detection of Genetically Modified Rice G6H1 Xiaoli Xu 1 & Xiaoyun Chen 1 & Yongmin Lai 1 & Qing Zhang 2 & Yuhua Wu 3 & Jun Li 3 & Cheng Peng 1 & Xiaofu Wang 1 & Qingmei Miao 1 & Hui Liu 1 & Junfeng Xu 1 Received: 9 February 2018 / Accepted: 28 September 2018 / Published online: 6 October 2018 # Springer Science+Business Media, LLC, part of Springer Nature 2018
Abstract The genetically modified rice G6H1 expressing a fused protein of Cry1Ab/Vip3H and a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) G6 is a genetically modified event that has been approved for preproduction field testing in China. The purpose of this study was to establish an event-specific qualitative and quantitative detection method that could provide a stable, reliable system for monitoring this new transgenic event. In this study, event-specific qualitative and quantitative detection methods based on the 3′ integration junction sequence between host plant DNA and the integrated gene were developed. The limit of detection (LOD) of qualitative PCR was assessed to be 0.1%. The LOD of quantitative PCR was estimated to be ten haploid genome copies. The quantitative PCR detection method was verified with three mixed rice samples with known G6H1 contents, and the results agreed with the expected values. Evaluation of specificity and sensitivity indicated that the developed qualitative and quantitative PCR methods are reliable and can be used for the detection and quantification of G6H1. Keywords Genetically modified organism . G6H1 rice . Event-specific method . Real-time PCR
Introduction To meet the increasing food demand of the growing human population, genetically modified crops were developed to boost agricultural productivity despite the lack of arable land. Since 1989, with the government’s support, numerous transgenic rice lines were developed and tested at various stages based on the regulatory process for agricultural genetically modified organisms (GMOs) in China (Mäde et al. 2006). Most of these transgenic rice lines were genetically modified to express insecticidal genes with lepidopteran activity from bacterium Bacillus Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12161-018-1375-y) contains supplementary material, which is available to authorized users. * Junfeng Xu [email protected] 1
State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2
Planting Management Station of Ningbo City, Ningbo 315000, China
3
Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China
thuringiensis (Bt), with single or dual insecticidal genes or stacked with other types of transgenes (Li et al. 2015). In 2009, China’s Ministry of Agriculture issued biosafety certificates for the commercial production of two transgenic Bt rice lines, Huahui 1 and Bt Shanyou 63, in restricted areas, and the biosafety certificat
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