Rapid Visual Detection of Four Specific Transgenic Events in GM Soybean Using Loop-Mediated Isothermal Amplification Method
- 20 Downloads
Many specific detection assays have been developed in order to efficiently detect GMOs (genetically modified organisms) in feed and food. Existing approaches to screen or detect GM crops always rely on thermal cycling device. Therefore, these rapid visual detection assays were developed and characterized for the four transgenic soybean (Glycine max (L.) Merr.) events MON87701, MON87705, MON87708, and MON87769 using loop-mediated isothermal amplification (LAMP) method. Four sets of specific primers were successfully designed to recognize the junction sequences of specific soybean events. The optimum primers and reaction temperature of each assay were tested respectively. Different kinds of genomic DNAs of transgenic events and non-transgenic crop samples were examined with the methods that indicated the good specificity of GM soybean species. The detection limit of each LAMP assays could reach 0.05%, showing sensitivity 10 times higher than that of the regular PCR method. Due to their simplicity, rapidness and low cost, the established LAMP assays would be practical for detecting the GM soybeans MON87701, MON87705, MON87708, MON87769 and their derivatives.
Keywords:Glycine max GM soybean specific events LAMP methods
This study was supported by the National Transgenic Plant Special Fund, China (2014ZX0801202B), Scientific and Technological development Plan Project, JiLin, China (20170204012NY), and Jilin Agricultural Technological Innovation Project, China (CXGC2017JQ017). We do appreciate for the financial support.
COMPLIANCE WITH ETHICAL STANDARDS
The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.
- 1.Global Status of Commercialized Biotech/GM Crops in 2017: Biotech Crop Adoption Surges as Economic Benefits Accumulate in 22 Years, Ithaca: ISAAA Brief, 2017, no. 53.Google Scholar
- 12.Mori, Y., Kanda, H., and Notomi, T., Loop-mediated isothermal amplification (LAMP): recent progress in research and development, J. Infect. Chemother., 2013, vol. 3, pp. 401–411.Google Scholar
- 13.Köppel, R., Bucher, T., Meuwly, A., and Moor, D., Multiplex real-time PCR method for the detection and quantification of DNA from the four transgenic soy traits DP-356043-5, DP-305423-1, MON 87701, and BPS-CV127-9 and lectin, Eur. Food Res. Technol., 2014, vol. 2, pp. 347–355.Google Scholar
- 14.Mazzara, M., Delobel, C., Pinski, G., Savini, C., and Van den Eede, G., Event-Specific Method for the Quantification of Soybean MON87769 Using Real-Time PCR, Luxembourg: Joint Res. Center, EURL, 2012. https://doi.org/10.2788/4544
- 15.Savini, C., Mazzara, M., Munaro, J., and Kreysa, J., Event-Specific Method for the Quantification of Soybean MON87769 Using Real-Time PCR, Luxembourg: Joint Res. Center, EURL, 2013. https://doi.org/10.2788/90943
- 16.Savini, C., Mazzara, M., Delobel, C., Pinski, G., and Van den Eede, G., Event-Specific Method for the Quantification of Soybean MON87705 Using Real-Time PCR, Luxembourg: Joint Res. Center, EURL, 2012. https://doi.org/10.2788/48383
- 17.Gao, A.G., Kolacz, K.H., MacRae, T.C., Miklos, J.A., Paradise, M.S., Perlak, F.J., and Dressel, T., Soybean Plant and Seed Corresponding to Transgenic Event MON87701 and Methods for Detection Thereof, US Patent Application 8455198, June 4, 2013.Google Scholar
- 18.Wagner, N., Burns, W.C., Godsy, E.J., and Roberts, P.D., Soybean Transgenic Event MON87705 and Methods for Detection Thereof, US Patent Application 8692080, April 8, 2014.Google Scholar
- 19.Froman, B., Duong, C., and Listello, J., Soybean Transgenic Event MON87769 and Methods for Detection Thereof, US Patent Application 8692076, April 8, 2014.Google Scholar
- 20.Brinker, R.J., Burns, W.C., Feng, P., Gupta, A., Hoi, S.W., Malven, M., and Wu, K., Soybean Transgenic Event MON87708 and Methods for Detection Thereof, US Patent Application 8501407, August 6, 2013.Google Scholar
- 22.Fukuta, S., Mizukami, Y., Ishida, A., Ueda, J., Hasegawa, M., Hayashi, I., Hashimoto, M., and Kanbe, M., Real-time loop-mediated isothermal amplification for the CaMV-35 promoter as a screening method for genetically modified organisms, Eur. Food Res. Technol., 2004, vol. 218, pp. 496–500.CrossRefGoogle Scholar
- 25.Tang, D., Zhang, J., Gao, S., Fan, Y., Xiao, Y., and Shi, L., LAMP (Loop-Mediated Isothermal Amplification) Detection Primer Group, Kit and Method for Transgenic Soybean DP-305423 and Derived Varieties Thereof, China Patent Application 102634589, August 21, 2013.Google Scholar