DOI QR코드

DOI QR Code

Four multiplex PCR Sets of 11 LM Maize for LMO environmental monitoring in Korea

국내 LMO 자연환경 모니터링을 위한 11개 LM 옥수수의 동시검출기법 개발

  • Shin, Su Young (Division of Ecological Conservation, National Institute of Ecology (NIE)) ;
  • Lim, Hae-Song (Division of Ecological Conservation, National Institute of Ecology (NIE)) ;
  • Seol, Min-A (Division of Ecological Conservation, National Institute of Ecology (NIE)) ;
  • Jung, Young Jun (Division of Ecological Conservation, National Institute of Ecology (NIE)) ;
  • Kim, Il Ryong (Division of Ecological Conservation, National Institute of Ecology (NIE)) ;
  • Song, Hae Ryoung (Division of Ecological Conservation, National Institute of Ecology (NIE)) ;
  • Lee, Jung Ro (Division of Ecological Conservation, National Institute of Ecology (NIE)) ;
  • Choi, Wonkyun (Division of Ecological Conservation, National Institute of Ecology (NIE))
  • 신수영 (국립생태원 생태보전연구실) ;
  • 임혜송 (국립생태원 생태보전연구실) ;
  • 설민아 (국립생태원 생태보전연구실) ;
  • 정영준 (국립생태원 생태보전연구실) ;
  • 김일룡 (국립생태원 생태보전연구실) ;
  • 송해룡 (국립생태원 생태보전연구실) ;
  • 이중로 (국립생태원 생태보전연구실) ;
  • 최원균 (국립생태원 생태보전연구실)
  • Received : 2016.11.02
  • Accepted : 2016.12.15
  • Published : 2016.12.31

Abstract

With the increasing development and commercial use of genetically modified maize, it is essential to develop an appropriate method for detection of individual LMO (Living modified organism) events for monitoring the samples. In South Korea, commercial planting and accidental or unintentional releases of LMOs into the environment were not approved. In this study, to increase the efficiency of LMO detection, we developed simultaneous detection methods for 11 LM maize events. This multiplex PCR detection method is economical, as it saves time, cost and labor. We developed 11 individual LM maize events, and applied 4 multiplex PCR sets to the LM maize samples. These results are confirmed by applying the multiplex analysis of LMO environmental monitoring from 2012 to 2014, which represents the unintentionally released LM maize samples. The data were correlated with event specific PCR results. Our results indicate that the multiplex PCR method developed is suitable for detection of LM maize in LMO monitoring.

유전자변형 옥수수의 개발과 상업적 이용이 증가하면서 유전자변형 옥수수 의심시료의 LMO 이벤트를 확인 할 수 있는 적합한 방법 개발이 필요하다. 국내에서는 상업적 재배와 자연생태계의 의도적 비의도적 LMO의 유출이 허용되고 있지 않다. 본 연구에서는 국내 승인된 11개의 LM 옥수수 이벤트를 동시에 검출할 수 있는 동시검출기법을 개발하였다. 이 방법은 시간과 비용, 노동력을 절감할 수 있는 효율적인 방법으로 4종의 PCR set로 개발하였다. 2012년부터 2014년까지 국립환경과학원 및 국립생태원에서 실시한 LMO 자연환경 모니터링 의심시료를 이용하여 동시검출기법의 효율성을 검증하였다. 이러한 결과를 바탕으로 본 연구의 결과는 LMO 모니터링 시료의 분석에 적합한 방법임을 알 수 있었다.

Keywords

References

  1. BIOSAFETY SCANNER. Available from: http://en.biosafetyscanner.org/
  2. Center for Environmental Risk Assessment (CERA). Available from: http://www.cera-gmc.org/
  3. Cottenet G, Blancpain C, Sonnard V, Chuar PF (2013) Development and validation of a multiplex real-time PCR method to simultaneously detect 47 targets for the identification of genetically modified organisms. Anal Bioanal Chem 450:6831-6844
  4. Dorries HH, Remus I, Gronewald A, Gronewald C, Berghof-Jager K (2010) Development of a qualitative, multiplex real-time PCR kit for screening of genetically modified organisms (GMOs). Anal Bioanal Chem 936:2043-2054
  5. European Commission, Joint Research Centre (JRC). Available from: http://gmo-crl.jrc.ec.europa.eu
  6. GMO Detection Method Database (GMDD). Available from: http://gmdd.shgmo.org/index/search
  7. Heide BR, Dromtorp SM, Rudi K, Heir E, Holck AL (2008) Determination of eight genetically modified maize events by quantitative multiplex PCR and fluorescence capillary gel electrophoresis. European Food Research and Technology 227:1125-1137 https://doi.org/10.1007/s00217-008-0828-8
  8. Jo BH, Lee JR, Choi W, Moon JC, Shin SY, Eum SJ. Seol MA, Kim IR, Song HR (2015) Development of multiplex PCR-based detection method for five approved LM canola events in Korea. J Plant Biotechonol 42:117-122 https://doi.org/10.5010/JPB.2015.42.2.117
  9. Jo BH, Seol MA, Shin SY, Kim IR, Choi W, Eum SJ, Song HR, Lee JR (2016) Multiplex PCR method for environmental monitoring of approved LM cotton events in Korea. J Plant Biotechnol 43:91-98 https://doi.org/10.5010/JPB.2016.43.1.91
  10. KBCH: Korea Biosafety Clearing House. [Internet]. 2015. Status of risk assessment of GMO in Korea. Available from: http://www.biosafety.or.kr.
  11. Kim SY, Kim JH, Lee HJ, Kim HY (2010) Detection system of stacked genetically modified Maize using multiplex PCR. Food Sci Biotechnol 19(4):1029-1033 https://doi.org/10.1007/s10068-010-0144-5
  12. Meyer W, Caprioara-Buda M, Jeynov B, Corbisier P, Trapmann S, Emons H (2012) The impact of analytical quality criteria and data evaluation on the quantification of genetically modified organisms. European Food Research and Technology 235(4):597-610 https://doi.org/10.1007/s00217-012-1787-7
  13. National Insititute of Ecology (NIE) (2015) Detection methods for analysis of LMO. ISBN 979-11-86197-24-0. 10-43
  14. Oguchi T, Onishi M, Mano J, Akiyama H, Techima R, Futo S, Furui S, Kitta K (2010) Development of multiplex PCR method for simultaneous detectioin of four events of genetically modified maize: DAS-59122-7, MIR604, MON863 and MON88017. Food Hygiene and Safety Science 51(3):92-100 https://doi.org/10.3358/shokueishi.51.92
  15. Randhawa GJ, Singh M, Sood P, Bhoge RK (2014) Multitarget real-time PCR-based system: monitoring for unauthorized genetically modified events in india. J Agric Food Chem 62(29):7118-7130 https://doi.org/10.1021/jf502190e
  16. Shrestha HK, Hwu KK, Chang MC (2010) Advances in detection of genetically engineered crops by multiplex polymerase chain reaction methods. Trends in Food Science & Technology 21:442-454 https://doi.org/10.1016/j.tifs.2010.06.004
  17. Xu J, Zhu S, Miao H, Huang W, Qiu M, Huang Y, Fu X, Li Y (2007) Event-specific detection of seven genetically modified soybean and maizes using multiplex-PCR coupled with oligonucleotide microarray. J Agric Food Chem 55(14):5575-5579 https://doi.org/10.1021/jf070433m
  18. Yang L, Guo J, Pan A, Zhang H, Zhang K, Wang Z, Zhang D (2007) Event-specific quantitative detection of nine genetically modified maizes using one novel standard reference molecule. J Agric Food Chem 55(1):15-24 https://doi.org/10.1021/jf0615754