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Event-specific qualitative and quantitative detection of transgenic rice Kefeng-6 by characterization of the transgene flanking sequence

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Abstract

Using TAIL-PCR (thermal asymmetric interlaced PCR), we analyzed the flanking sequences of transfer DNA (T-DNA) in transgenic rice Kefeng-6, which has insect-resistant genes, Cry1Ac and CpTi. Two junction sequences of T-DNA were identified in the Kefeng-6 genome: one integrated in chromosome 6 with an additional 22-bp insertion and another one in chromosome 9 with a 4-bp insertion between rice genomic DNA and T-DNA. Based on these inserts and border sequences, the event-specific qualitative and quantitative PCR system was established for this line. The relative limit of qualitative detection was assessed to be between 0.1 and 0.05%, and the absolute limit of detection in the quantitative PCR was approximately five initial copies. Two mixed rice samples with known Kefeng-6 contents were used to verify the quantitative method, from which the expected results were observed. This study provides a reliable method and information for detection, identification, and quantification of the presence of non-authorized GM rice Kefeng-6.

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Acknowledgments

We thank Feng Wang (Fujian Academy of Agricultural Sciences) for providing the transgenic rice Kefeng-6. This research was funded by the following programs: Safety Administration Progresses of Agricultural Transgenic Organisms projects, Agricultural Ministry of the People’s Republic of China, and Important National Science & Technology Specific Projects (2008ZX08012-001), Central Public-interest Scientific Institution Basal Research Fund, China National Rice Institute.

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Authors and Affiliations

  1. China National Rice Research Institute, Tiyuchang Road 359, 310006, Hangzhou, Zhejiang, People’s Republic of China

    Wei-Xia Wang, Feng-Xiang Lai & Qiang Fu

  2. College of Biological and Environmental Engineering, Zhejiang University of Technology, Chaowang Road, 310014, Hangzhou, Zhejiang, People’s Republic of China

    Ting-Heng Zhu

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  1. Wei-Xia Wang
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Correspondence to Wei-Xia Wang or Qiang Fu.

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Wang, WX., Zhu, TH., Lai, FX. et al. Event-specific qualitative and quantitative detection of transgenic rice Kefeng-6 by characterization of the transgene flanking sequence. Eur Food Res Technol 232, 297–305 (2011). https://doi.org/10.1007/s00217-010-1389-1

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  • DOI: https://doi.org/10.1007/s00217-010-1389-1

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