A novel trait-specific real-time PCR method enables quantification of genetically modified (GM) maize content in ground grain samples containing stacked GM maize

详细信息    查看全文

• 作者：Akio Noguchi (1)
  Hiroshi Akiyama (1)
  Kosuke Nakamura (1)
  Kozue Sakata (1)
  Yasutaka Minegishi (2)
  Junichi Mano (3)
  Reona Takabatake (3)
  Satoshi Futo (4)
  Kazumi Kitta (3)
  Reiko Teshima (1)
  Kazunari Kondo (1)
  Tomoko Nishimaki-Mogami (1)
  
• 关键词：Genetically modified maize ; qPCR ; Trait ; specific method ; Stacked GM maize
• 刊名：European Food Research and Technology
• 出版年：2015
• 出版时间：February 2015
• 年：2015
• 卷：240
• 期：2
• 页码：413-422
• 全文大小：284 KB
• 参考文献：1. Davison J, Bertheau Y (2008) The theory and practice of European traceability regulations for GM food and feed. Cereal Foods World 53:186鈥?96
  2. Kuribara H, Shindo Y, Matsuoka T, Takubo K, Futo S, Aoki N, Hirao T, Akiyama H, Goda Y, Toyoda M, Hino A (2002) Novel reference molecules for quantitation of genetically modified maize and soybean. J AOAC Int 85:1077鈥?089
  3. Shindo Y, Kuribara H, Matsuoka T, Futo S, Sawada C, Shono J, Akiyama H, Goda Y, Toyoda M, Hino A (2002) Validation of real-time PCR analyses for line-specific quantitation of genetically modified maize and soybean using new reference molecules. J AOAC Int 85:1119鈥?126
  4. Permingeat HR, Reggiardo MI, Vallejos RH (2002) Detection and quantification of transgenes in grains by multiplex and real-time PCR. J Agric Food Chem 50:4431鈥?436 CrossRef
  5. Collonnier C, Schattner A, Berthier G, Boyer F, Cou茅-Philippe G, Diolez A, Duplan M-N, Fernandez S, Kebdani N, Kobilinsky A, Romaniuk M, de Beuckeleer M, de Loose M, Windels P, Bertheau Y (2005) Characterization and event specific-detection by quantitative real-time PCR of T25 maize insert. J AOAC Int 88:536鈥?46
  6. La Paz J-L, Garc铆a-Muniz N, Nadal A, Esteve T, Puigdom猫nech P, Pla M (2006) Interlaboratory transfer of a real-time polymerase chain reaction assay for quantitative detection of genetically modified maize event TC-1507. J AOAC Int 89:1347鈥?352
  7. Li X, Yang L, Zhang J, Wang S, Shen K, Pan L, Zhang D (2009) Simplex and duplex polymerase chain reaction analysis of Herculex RW (59122) maize based on one reference molecule including separated fragments of 5鈥?integration site and endogenous gene. J AOAC Int 92:1472鈥?483
  8. Kodama T, Kuribara H, Minegishi Y, Futo S, Watai M, Sawada C, Watanabe T, Akiyama H, Maitani T, Teshima R, Furui S, Hino A, Kitta K (2009) Evaluation of modified PCR quantitation of genetically modified maize and soybean using reference molecules: interlaboratory study. J AOAC Int 92:223鈥?33
  9. Berdal K, Holst-Jensen A (2001) Roundup Ready^庐 soybean event-specific real-time quantitative PCR assay and estimation of the practical detection and quantification limits in GMO analyses. Eur Food Res Technol 213:432鈥?38 CrossRef
  10. Taverniers I, Windels P, Van Bockstaele E, De Loose M (2001) Use of cloned DNA fragments for event-specific quantification of genetically modified organisms in pure and mixed food products. Eur Food Res Technol 213:417鈥?24 CrossRef
  11. Baeumler S, Wulff D, Tagliani L, Song P (2006) A real-time quantitative PCR detection method specific to widestrike transgenic cotton (event 281-24-236/3006-210-23). J Agric Food Chem 54:6527鈥?534 CrossRef
  12. Yang L, Pan A, Zhang K, Yin C, Qian B, Chen J, Huang C, Zhang D (2005) Qualitative and quantitative PCR methods for event-specific detection of genetically modified cotton Mon1445 and Mon531. Transgenic Res 14:817鈥?31 CrossRef
  13. Wu G, Wu Y, Xiao L, Lu C (2008) Event-specific qualitative and quantitative polymerase chain reaction methods for detection of genetically modified rapeseed Ms8xRf3 based on the right border junctions. J AOAC Int 91:143鈥?51
  14. James C (2012) Global Status of Commercialized Biotech/GM Crops: 2012, ISAAA Brie. ISAAA, Ithaca
  15. Akiyama H, Watanabe T, Wakabayashi K, Nakade S, Yasui S, Sakata K, Chiba R, Spiegelhalter F, Hino A, Maitani T (2005) Quantitative detection system for maize sample containing combined-trait genetically modified maize. Anal Chem 77:7421鈥?428 CrossRef
  16. Akiyama H, Sakata K, Makiyma D, Nakamura K, Teshima R, Nakashima A, Ogawa A, Yamagishi T, Futo S, Oguchi T, Mano J, Kitta K (2011) Interlaboratory study of DNA extraction from multiple ground samples, multiplex real-time PCR, and multiplex qualitative PCR for individual kernel detection system of genetically modified maize. J AOAC Int 94:1540鈥?547 CrossRef
  17. Onishi M, Matsuoka T, Kodama T, Kashiwaba K, Futo S, Akiyama H, Maitani T, Furui S, Oguchi T, Hino A (2005) Development of a multiplex polymerase chain reaction method for simultaneous detection of eight events of genetically modified maize. J Agric Food Chem 53:9713鈥?721 CrossRef
  18. Oguchi T, Onishi M, Mano J, Akiyama H, Teshima R, Futo S, Furui S, Kitta K (2010) Development of multiplex PCR method for simultaneous detection of four events of genetically modified maize: DAS-59122-7, MIR604, MON863 and MON88017. J Food Hyg Soc Jpn 51:92鈥?00 CrossRef
  19. Department of Food Safety Ministry of Health Labour and Welfare of Japan (2009) Notice No. 0803, Article 8
  20. Bilder CR, Tebbs JM (2009) Bias, efficiency, and agreement for group-testing regression models. J Stat Comput Simul 79:67鈥?0 CrossRef
  21. Chen CL, Swallow WH (1990) Using group testing to estimate a proportion, and to test the binomial model. Biometrics 46:1035鈥?046 CrossRef
  22. Yamamura K, Hino A (2007) Estimation of the proportion of defective units by using group testing under the existence of a threshold of detection. Commun Stat Simul Comput 36:949鈥?57 CrossRef
  23. Mano J, Yanaka Y, Ikezu Y, Onishi M, Futo S, Minegishi Y, Ninomiya K, Yotsuyanagi Y, Spiegelhalter F, Akiyama H, Teshima R, Hino A, Naito S, Koiwa T, Takabatake R, Furui S, Kitta K (2011) Practicable group testing method to evaluate weight/weight GMO content in maize grains. J Agric Food Chem 59:6856鈥?863 CrossRef
  24. Akiyama H, Sakata K, Kondo K, Tanaka A, Liu MS, Oguchi T, Furui S, Kitta K, Hino A, Teshima R (2008) Individual detection of genetically modified maize varieties in non-identity-preserved maize samples. J Agric Food Chem 56:1977鈥?983 CrossRef
  25. Akiyama H, Minegishi Y, Makiyama D, Mano J, Sakata K, Nakamura K, Noguchi A, Takabatake R, Futo S, Kondo K, Kitta K, Kato Y, Teshima R (2012) Quantification and identification of genetically modified maize events in non-identity preserved maize samples in 2009 using an individual kernel detection system. J Food Hyg Soc Jpn 53:157鈥?65 CrossRef
  26. Cavato T, Coombe T, Johnson S (2003) Patent No. US20060095986: Corn event pv-zmir13 (mon863) plants and compositions and methods for detection thereof
  27. Joint Research Centre-European Commission (2005) Event-specific method for the quantitation of maize line TC 1507 using real-time PCR Protocol. http://gmo-crl.jrc.ec.europa.eu/summaries/TC1507-WEB-Protocol-Validation.pdf
  28. Joint Research Centre-European Commission (2006) CRL assessment on the validation of an event specific method for the relative quantitation of maize line MON 810 DNA using real-time PCR as carried out by Federal Institute for Risk Assessment (BfR). http://gmo-crl.jrc.ec.europa.eu/summaries/Mon810_validation_report.pdf
  29. Joint Research Centre-European Commission (2007) Event-specific method for the quantitation of maize 59122 using real-time PCR Protocol. http://gmo-crl.jrc.ec.europa.eu/summaries/59122-ProtocolValidation.pdf
  30. Joint Research Centre-European Commission (2010) Event-specific method for the quantitation of maize line MON 88017 using real-time PCR Protocol. http://gmo-crl.jrc.ec.europa.eu/summaries/MON88017_validated_Method_correctedversion1.pdf
  31. Yoshimura T, Kuribara H, Matsuoka T, Kodama T, Iida M, Watanabe T, Akiyama H, Maitani T, Furui S, Hino A (2005) Applicability of the quantification of genetically modified organisms to foods processed from maize and soy. J Agric Food Chem 53:2052鈥?059 CrossRef
  32. Hern谩ndez M, Duplan M-N, Berthier G, Va茂tilingom M, Hauser W, Freyer R, Pla M, Bertheau Y (2004) Development and comparison of four real-time polymerase chain reaction systems for specific detection and quantification of / Zea mays L. J Agric Food Chem 52:4632鈥?637 CrossRef
  33. Joint Research Centre-European Commission (2005) Event-specific method for the quantitation of maize line MON 863 using real-time PCR Protocol. http://gmo-crl.jrc.ec.europa.eu/summaries/MON863-WEB-Protocol-Validation.pdf
  34. Joint Research Centre-European Commission (2005) Event-specific method for the quantitation of maize line NK 603 using real-time PCR Protocol. http://gmo-crl.jrc.ec.europa.eu/summaries/NK603report_mm.pdf
  35. Arumuganathan K, Earle ED (1991) Nuclear DNA content of some important plant species. Plant Mol Biol Rep 9:208鈥?18 CrossRef
  36. Trifa Y, Zhang D (2004) DNA content in embryo and endosperm of maize kernel ( / Zea mays L.): impact on GMO quantification. J Agric Food Chem 52:1044鈥?048 CrossRef
  37. Heck GR, Armstrong CL, Astwood JD, Behr CF, Bookout JT, Brown SM, Cavato TA, DeBoer DL, Deng MY, George C, Hillyard JR, Hironaka CM, Howe AR, Jakse EH, Ledesma BE, Lee TC, Lirette RP, Mangano ML, Mutz JN, Qi Y, Rodriguez RE, Sidhu SR, Silvanovich A, Stoecker MA, Yingling RA, You J (2005) Development and characterization of a CP4 EPSPS-Based, Glyphosate-Tolerant Corn Event. Crop Sci 45:329鈥?39 CrossRef
  38. Joint Research Centre-European Commission (2010) Event-specific method for the quantitation of maize line MON 88017 using real-time PCR Validation Report. http://gmo-crl.jrc.ec.europa.eu/summaries/MON88017_val_report_correctedVersion1.pdf
  39. Joint Research Centre-European Commission (2006) Event-specific method for the quantitation of maize line NK 603 using real-time PCR Validation Report. http://gmo-crl.jrc.ec.europa.eu/summaries/NK603report_mm.pdf
  40. Joint Research Centre-European Commission (2005) Event-specific method for the quantitation of maize line TC 1507 using real-time PCR Validation Report. http://gmo-crl.jrc.ec.europa.eu/summaries/TC1507-report_mm.pdf
  41. Joint Research Centre-European Commission (2006) Event-specific method for the quantitation of maize line DAS-59122-7 using real-time PCR Validation Report. http://gmo-crl.jrc.ec.europa.eu/summaries/59122_val_report.pdf
  42. Joint Research Centre-European Commission (2005) Event-specific method for the quantitation of maize line MON 863 using real-time PCR Validation Report. http://gmo-crl.jrc.ec.europa.eu/summaries/MON863-Val-report_mm.pdf
  43. United States Department of Agriculture (2013) USDA鈥檚 National Agricultural Statistics Service. Genetically engineered varieties of corn, upland cotton, and soybeans, by State and for the Unites States, 2000-13. http://www.ers.usda.gov/datafiles/Adoption_of_Genetically_Engineered_Crops_in_the_US/alltables.xls
  44. Mano J, Shigemitsu N, Futo S, Akiyama H, Teshima R, Hino A, Furui S, Kitta K (2009) Real-time PCR array as a universal platform for the detection of genetically modified crops and its application in identifying unapproved genetically modified crops in Japan. J Agric Food Chem 57:26鈥?7 CrossRef
  
• 作者单位：Akio Noguchi (1)
  Hiroshi Akiyama (1)
  Kosuke Nakamura (1)
  Kozue Sakata (1)
  Yasutaka Minegishi (2)
  Junichi Mano (3)
  Reona Takabatake (3)
  Satoshi Futo (4)
  Kazumi Kitta (3)
  Reiko Teshima (1)
  Kazunari Kondo (1)
  Tomoko Nishimaki-Mogami (1)
  
  1. National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan
  2. Nippon Gene Co. Ltd., 1-5 Kandanishiki-cho, Chiyoda-ku, Tokyo, 101-0054, Japan
  3. National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki, 305-8642, Japan
  4. Fasmac Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa, 243-0041, Japan
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Food Science
  Analytical Chemistry
  Biotechnology
  Agriculture
  Forestry
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1438-2385

文摘

Stacked genetically modified (GM) maize is increasingly produced; thereby, current event-specific quantitative real-time polymerase chain reaction (qPCR) methods have led to the overestimation of GM organism (GMO) content compared with the actual weight/weight percentage of GM organism in maize samples. We developed a feasible qPCR method in which the GMO content is calculated based on the quantification of two herbicide-tolerant trait genes, 5-enolpyruvylshikimate-3-phosphate synthase from Agrobacterium sp. strain CP4 (cp4epsps) and phosphinothricin N-acetyl-transferase from Streptomyces viridochromogenes (pat) to quantify the GMO content in ground grain samples containing stacked GM maize. The GMO contents of two genes were quantified using a plasmid calibrant and summed for quantification of total GMO content. The trait-specific method revealed lower biases for examination of test samples containing stacked GM maize compared with the event-specific method. Our results clearly show that the trait-specific method is not only simple and cost-effective, but also useful in quantifying the GMO content in ground grain samples containing stacked GM maize, which are expected to be major events in the near future. The developed method would be the only feasible way to conduct the quantification of GMO content in the ground maize samples containing stacked GM maize for the verification of the labeling regulation.