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Developing a matrix reference material for screening of transgenic rice

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Abstract

Certified reference materials (CRMs) that are compatible with detection methods are needed to detect genetically modified organisms (GMOs). Screening is the first detection step in determining the possible presence of GMO ingredients in food or feed; however, screening has been hindered by the lack of GMO CRMs. In this study, transgenic rice materials were developed via the transformation of a construct harboring 11 commonly used screening elements. Digital PCR was utilized to identify a homozygous single-copy line termed SDrice. The qualitative detections of 11 elements in 21 transgenic materials demonstrated that the genomic DNA of the SDrice was suitable for use as a positive control in the screening of GMO ingredients. The suitability of SDrice as reference material was further checked by testing the sensitivity of 11 known conventional PCR assays, ranging from 10 to 50 copies of the SDrice genome. The standard curves that were created using SDrice DNA series as calibrators all exhibited good linearities in the relationships of the Ct values with the template copy numbers in these 11 real-time PCR assays. The LODs of the real-time PCR assays were estimated to be two to five copies of the SDrice genome. Comparisons of the SDrice with other GM rice revealed that significant differences existed in both the intercepts of the standard curves and the ΔCt values of the exogenous and reference genes for the P-35S, T-nos, HPT, T-35S, and Bar assays; the SDrice was not fit for quantification of other GM rice events. This study provided a matrix reference material (RM) that was suitable for screening GM rice, determination of sensitivity and a LOD of PCR assays, and overcame some of the drawbacks of plasmid DNA as reference material.

A matrix RM from transgenic rice harboring 11 commonly used screening targets

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References

  1. James C (2014) ISAAA Brief 49

  2. Chassy BM (2009) In: Kriz AL, Larkins BA (eds) Molecular genetic approaches to maize improvement, biotechnology in agriculture and forestry. Springer, Berlin, Heidelberg

    Google Scholar 

  3. Gruere GP, Rao SR (2007) Ag BioForum 10:51–64

    Google Scholar 

  4. Holst-Jensen A (2009) Biotechnol Adv 27:1071–1082

    Article  CAS  Google Scholar 

  5. Trapmann S, Emons H (2005) Anal Bioanal Chem 381:72–74

    Article  CAS  Google Scholar 

  6. Lauwaars M, Anklam E (2004) Accred Qual Assur 9:253–258

    Article  Google Scholar 

  7. Marmiroli N, Maestri E, Gullì M, Malcevschi A, Peano C, Bordoni R, De Bellis G (2008) Anal Bioanal Chem 392:369–384

    Article  CAS  Google Scholar 

  8. Holst-Jensen A, Rønning SB, Lovseth A, Berdal KG (2003) Anal Bioanal Chem 375:985–993

    CAS  Google Scholar 

  9. Gürtler PS, Meißner E, Busch U (2011) Nachweis von nicht zugelassenen gentechnisch-veränderten Organismen (GVO): weltweite Entwicklung; Importanalyse und Entwicklung von Nachweis-Methoden. 2011:LGL

  10. Trapmann S, Schimmel H, Kramer GN, Van den Eede G, Pauwels J (2002) J AOAC Int 85(3):775–779

    CAS  Google Scholar 

  11. European Reference materials (2010) ERM. http://www.erm-crm.org/Pages/ermcrmCatalogue.aspx. Accessed 15 May 2015

  12. Certified Reference Materials (2015) AOCS. http://www.aocs.org/LabServices/content.cfm?ItemNumber=19248. Accessed 15 May 2015

  13. CRL-GMFF (2011) Report on the application of P-35S, T-NOS and CryIAb/Ac methods for the detection of Genetically Modified Rice originating from China Using Real-Time PCR. http://gmo-crl.jrc.ec.europa.eu/doc/Report%20on%20application%20of%20P35_T_nos_CryLAb_Ac%20and%20rice%20PLD.pdf. Accessed 10 Dec. 2014

  14. IRMM (2009) Certification of reference materials of maize seed powder with different mass fractions of genetically modified 98140 maize. http://www.erm-crm.org/ERM_products/search/reports/BF427.pdf. Accessed 10 Dec. 2014

  15. Lievens A, Bellocchi G, De BD, Moens W, Savini C, Mazzara M, Van EG, Van BM (2010) Anal Bioanal Chem 396:2165–2173

    Article  CAS  Google Scholar 

  16. Taverniers I, Windels P, Vaitilingom. M, Milcamps A, Van B E, Van E G, De L M (2005) J Agric Food Chem 53:3041-3052

  17. Kuribara H, Shindo Y, Matsuoka T, Takubo K, Futo S, Aoki N, Hirao T, Akiyama H, Goda Y, Toyoda M, Hino A (2002) J AOAC Int 85:1077–1089

    CAS  Google Scholar 

  18. Shindo Y, Kuribara H, Matsuoka T, Futo S, Sawada C, Shono J, Akiyama H, Goda Y, Toyoda M, Hino A (2002) J AOAC Int 85:1119–1126

    CAS  Google Scholar 

  19. Cao Y, Wu G, Wu Y, Nie S, Zhang L, Lu C (2011) J Agric Food Chem 59:8550–8559

    Article  CAS  Google Scholar 

  20. Zhang L, Wu Y, Wu G, Cao Y, Lu C (2014) Anal Bioanal Chem 406:6385–6397

    Article  CAS  Google Scholar 

  21. Toyota A, Akiyama H, Sugimura M, Watanabe T, Kikuchi H, Kanamori H, Hino A, Esaka M, Maitani T (2006) Biosci Biotechnol Biochem 70:821–827

    Article  CAS  Google Scholar 

  22. Porebski S, Bailey LG, Baum BR (1997) Plant Mol Biol Rep 15:8–15

    Article  CAS  Google Scholar 

  23. Lin YJ, Zhang Q (2005) Plant Cell Rep 23:540–547

    Article  CAS  Google Scholar 

  24. Jin W, Shen P, Zhang X, Wan Y, Lin X, Yang L, Liu Y, Miao C, Wang X, Ma H (2012) Announcement by the Ministry of Agriculture No 1861-1-2012, 1-7. http://www.bjstandard.com/standard/271334.html. Accessed 15 Feb. 2015

  25. Zhang M, Song G, Li F, Li C, Zhao X, Dong L, Yan W, Xing Z, Song X, Liu N (2013) Announcement by the Ministry of Agriculture. No.2031-8-2013, 1-6. http://www.bjstandard.com/standard/282620.html. Accessed 15 Feb. 2015

  26. Yang L, Liu X, Zhang D, Shen P, Guo J, Jin W (2012) Announcement by the Ministry of Agriculture. No.1861-3-2012, 1-6. http://www.bjstandard.com/standard/271332.html. Accessed 15 Feb. 2015

  27. Lu C, Liu X, Wu Y, Wu G, Shen P, Yang L, Zhang D (2013) Announcement by the Ministry of Agriculture. No.2031-9-2013, 1-6. http://www.bjstandard.com/standard/282621.html. Accessed 15 Feb. 2015

  28. Xie J, Shen P, Peng Y, Li C, Song G, Sun Y (2013) Announcement by the Ministry of Agriculture. No.1943-1-2013, 1-6. http://www.bjstandard.com/standard/271328.html. Accessed 15 Feb. 2015

  29. Wu Y, WangY LJ, Li W, Zhang L, Li Y, Li X, Li J, Zhu L, Wu G (2014) Sci Rep. doi:10.1038/srep07358

    Google Scholar 

  30. Pan L, Zhang S, Yang L, Broll H, Tian F, Zhang D (2007) J AOAC Int 90:1639–1646

    CAS  Google Scholar 

  31. Lu C., Song G, Wu G, Wu Y, Cao Y, Li J, Luo J (2012) Announcement by the Ministry of Agriculture. No.1782-2-2012, 1-9. http://www.bjstandard.com/standard/271346.html. Accessed 5 Feb. 2015

  32. Xie J, Liu X, Su C, Yang L, Shen P, Zhang S, Sun Y (2012) Announcement by the Ministry of Agriculture. No.1782-3-2012, 1-9. http://www.bjstandard.com/standard/271347.html. Accessed 15 Feb. 2015

  33. Debode F, Janssen E, Berben G (2013) Eur Food Res Technol 236:659–669

    Article  CAS  Google Scholar 

  34. ISO. Foodstuffs—methods of analysis for the detection of genetically modified organisms and derived products - quantitative nucleic acid based methods. ISO 21570E:2005, 1-103

  35. JRC Compendium of Reference Methods for GMO Analysis. Quantitative PCR method for detection of rice event LLRICE62. http://gmo-crl.jrc.ec.europa.eu/gmomethods/docs/QT-EVE-OS-002.pdf. Accessed 16 Feb. 2015

  36. Grohmann L, Brunen-Nieweler C, Nemeth A, Waiblinger HU (2009) J Agric Food Chem 57:8913–8920

    Article  CAS  Google Scholar 

  37. Reiting R, Broll H, Waiblinger HU, Grohmann L (2007) J Verbr Lebensm 2:116–121

    Article  CAS  Google Scholar 

  38. JRC Compendium of Reference Methods for GMO Analysis. Qualitative PCR method for detection of Figwort Mosaic Virus 35S Promoter. http://gmo-crl.jrc.ec.europa.eu/gmomethods/docs/QL-ELE-00-015.pdf. Accessed 16 Feb. 2015

  39. Waiblinger HU, Ernst B, Anderson A, Pietsch K (2008) Eur Food Res Technol 226:1221–1228

    Article  CAS  Google Scholar 

  40. Weaver S, Dube S, Mir A, Qin J, Sun G, Ramakrishnan R (2010) Methods 50:271–276

    Article  CAS  Google Scholar 

  41. Dube S, Qin J, Ramakrishnan R (2008) Plos One 3:e2876

    Article  Google Scholar 

  42. ENGL (2008) Definition of Minimum Performance Requirements for Analytical Methods of GMO Testing European Network of GMO. Laboratories.http://gmo-crl.jrc.ec.europa.eu/doc/Min_Perf_Requirements_Analytical_methods.pdf. Accessed 20 July. 2015

  43. Caprioara-Buda M, Meyer W, Jeynov B, Corbisier P, Trapmann S, Emons H (2012) Anal Bioanal Chem 404:29–42

    Article  CAS  Google Scholar 

  44. Jiang Y, Yang H, Quan S, Liu Y, Shen P, Yang L (2015) Anal Bioanal Chem 407:6731–6739

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by grants from the National Major Special Project for the Development of Transgenic Organisms (grant nos. 2011ZX08012-003, 2014ZX0801202B and 2011ZX08012-005).

Conflict of interest

The authors declare no conflict of interest.

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Author notes

    Authors and Affiliations

    1. Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan, 430062, China

      Jun Li, Yuhua Wu, Xiaofei Li, Yulei Wang, Yunjing Li & Gang Wu

    2. Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China

      Jun Li, Yuhua Wu, Xiaofei Li, Yulei Wang, Yunjing Li & Gang Wu

    3. College of Life Science, South-Central University for Nationalities, 182 Min-Yuan Road, 430074, Wuhan, China

      Li Zhang

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    1. Jun Li
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    3. Xiaofei Li
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    Correspondence to Gang Wu.

    Additional information

    Jun Li and Yuhua Wu contributed equally to this work.

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    Li, J., Wu, Y., Li, X. et al. Developing a matrix reference material for screening of transgenic rice. Anal Bioanal Chem 407, 9153–9163 (2015). https://doi.org/10.1007/s00216-015-9081-8

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