Skip to main content
SpringerLink
Log in

Detection of genetically modified rice: a construct-specific real-time PCR method based on DNA sequences from transgenic Bt rice

  • Original Paper
  • Published:
European Food Research and Technology Aims and scope Submit manuscript

Abstract

Genetically modified rice varieties developed in China are close to approval for agricultural cultivation and production. However, so far no method has been reported for specific detection of transgenic varieties of this crop. In the present study, rice seeds assumed to consist of field-tested Bt rice (‘Anti-pest Shanyou 63’ and ‘Anti-pest Jinyou 63’) were used as reference material to determine transgenic DNA sequences. The transition between the cryIA(b) and cryIA(c) fusion gene and the nopaline synthase terminator (nos) sequence was used to develop a construct-specific real-time PCR based detection method. This Bt rice specific detection system was combined with a recently published quantitative real-time PCR method for the rice-specific (Oryza sativa L.) reference gene gos9. The complete PCR assay for detection of transgenic Bt rice was in-house validated and the limit of quantification was found to be below 0.1% Bt rice relative to the rice content. Application of the PCR assay should allow more precise detection of transgenic rice varieties in imported food products which are so far not approved in the EU.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Regulation (EC) No. 1829/2003 of the European Parliament and of the Council of 22 September 2003 on genetically modified food and feed. Off J Eur Commun L268:1–23

  2. Regulation (EC) No. 1830/2003 of the European Parliament and of the Council of 22 September 2003 concerning the traceability and labelling of genetically modified organisms and the traceability of food and feed products produced from genetically modified organisms and amending Directive 2001/18/EC. Off J Eur Commun L268:24–28

  3. Huang J, Hu R, Rozelle S, Pray C (2005) Science 308:688–690

    Article  CAS  Google Scholar 

  4. Bajaj S, Mohanty A (2005) Plant Biotechnol J 3:275–307

    Article  CAS  Google Scholar 

  5. Anonymous (2006) AGBIOS GM database. http://www.agbios.com/dbase.php

  6. Anonymous (2003) GMO-Watch Report. Genetically modified crops: molecular and regulatory facts. Version 2. http://www.bats.ch/gmo-watch/GVO-report140703.pdf

  7. Tu J, Zhang G, Datta K, Xu C, He Y, Zhang Q, Khush GS, Datta SK (2000) Nature Biotechnol 18:1101–1104

    Article  CAS  Google Scholar 

  8. Pray CE, Ramaswami B, Huang J, Hu R, Bengali P, Zhang H (2006) Int J Technol Globalisation 2:137–157

    Google Scholar 

  9. Sze PC, Cotter J (2005) Science 310:232

    Google Scholar 

  10. Anonymous (2005) Testing results for GE rice in China. http://www.greenpeace.org/raw/content/international/press/reports/RiceatRiskTestResults.pdf

  11. Holland PM, Abramson RD, Watson R, Gelfand DH (1991) Proc Natl Acad Sci USA 88:357–362

    Google Scholar 

  12. Commission Recommendation 2004/787 (EC) of 4 October 2004 on technical guidance for sampling and detection of genetically modified organisms and material produced from genetically modified organisms as or in products in the context of Regulation (EC) No. 1830/2003

  13. ISO 21571:2005 Foodstuffs. Methods of analysis for the detection of genetically modified organisms and derived products – nucleic acid extraction. International Standardization Organisation, Geneva, Switzerland

  14. Official Collection of Test Methods (2001) Screening for detection of genetically modified DNA sequences in foodstuffs by detection of DNA sequences which are frequently present in genetically modified organisms German Federal Foodstuffs Act – Food Analysis, Article 35, L 00.00-31, Beuth-Verlag, Berlin

  15. ISO 21569:2005. Foodstuffs – methods of analysis for the detection of genetically modified organisms and derived products – qualitative nucleic acid based methods. International Standardization Organisation, Geneva, Switzerland

  16. ISO 21570:2006. Foodstuffs – methods of analysis for the detection of genetically modified organisms and derived products – quantitative nucleic acid based methods. International Standardization Organisation, Geneva, Switzerland

  17. Pietsch K, Waiblinger HU, Brodmann P, Wurz A (1997) Dtsch Lebensm Rundsch 93:35–38

    CAS  Google Scholar 

  18. Ehlers B, Strauch E, Goltz M, Kubsch D, Wagner H, Maidhof H, Bendiek J, Appel B, Buhk HJ (1997) Bundesgesundheitsbl 4:118–121

    Article  Google Scholar 

  19. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) J Mol Biol 215:403–410

    Article  CAS  Google Scholar 

  20. Ding J, Jia J, Yang L, Wen H, Zhang C, Liu W, Zhang D (2004) J Agric Food Chem 52:3372–3377

    Article  CAS  Google Scholar 

  21. Hernandez M, Esteve T, Pla M (2005) J Agric Food Chem 53:7003–7009

    Article  CAS  Google Scholar 

  22. Tu J, Datta K, Oliva N, Zhang G, Xu C, Khush GS, Zhang Q, Datta SK (2003) Plant Biotechnol J 1:155–165

    Article  CAS  Google Scholar 

  23. Tu J, Datta K, Alam MF, Fan Y, Khush GS, Datta SK (1998) Plant Biotechnol 15:195–203

    CAS  Google Scholar 

  24. Arumuganathan K, Earle ED (1991) Plant Mol Biol Rep 9:208–218

    CAS  Google Scholar 

  25. Huang J, Ruifa H, Scott R, Pray C (2005) Science 310:231–233

    Article  Google Scholar 

  26. Community Reference Laboratory for GM Food and Feed (2006) Event-specific method for the quantification of rice line LLRICE62 using real-time PCR. http://gmo-crl.jrc.it/summaries/LLRICE62_validated_Protocol.pdf

  27. Xue Q, Duan X, Xu D, Wu R (1996) Production and testing of insect-resistant transgenic rice plants. In: Proceedings of the Third International Rice Genetics Symposium on rice genetics III, Manila, 16–20 October 1995, pp 239–246

  28. Tu J, Ona I, Zhang Q, Mew TW, Khush GS, Datta SK (1998) Theor Appl Genet 97:31–36

    Article  CAS  Google Scholar 

  29. Tu J, Datta K, Khush GS, Zhang Q, Datta SK (2000) Theor Appl Genet 101:15–20

    Article  CAS  Google Scholar 

  30. Paine JA, Shipton CA, Chaggar S, Howells RM, Kennedy MJ, Vernon G, Wright SY, Hinchliffe E, Adams JL, Silverstone AL, Drake R (2005) Nat Biotechnol 23:482–487

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr. Janet Cotter (University of Exeter, Exeter, UK) for providing us with the rice sample materials. We also thank Dr. Joachim Bendiek (Bundesamt für Verbraucherschutz und Lebensmittelsicherheit, Berlin) for critical reading of the manuscript.

Author information

Authors and Affiliations

  1. Landesamt für Verbraucherschutz Sachsen-Anhalt, Dezernat 33, Freiimfelder Str. 66/68, 06112, Halle, Germany

    Dietrich Mäde & Christine Degner

  2. Bundesamt für Verbraucherschutz und Lebensmittelsicherheit, Abteilung Gentechnik, Referat 405, Mauerstr. 39-42, 10117, Berlin, Germany

    Lutz Grohmann

Authors
  1. Dietrich Mäde
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christine Degner
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lutz Grohmann
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lutz Grohmann.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mäde, D., Degner, C. & Grohmann, L. Detection of genetically modified rice: a construct-specific real-time PCR method based on DNA sequences from transgenic Bt rice. Eur Food Res Technol 224, 271–278 (2006). https://doi.org/10.1007/s00217-006-0467-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00217-006-0467-x

Keywords

Search

Navigation