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Identification and Quantification of Genetically Modified Moonshade Carnation Lines Using Conventional and TaqMan Real-Time Polymerase Chain Reaction Methods

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Abstract

Genetically modified carnation (Dianthus caryophyllus L.) Moonshade was approved for planting and commercialization in several countries from 2004. Developing methods for analyzing Moonshade is necessary for implementing genetically modified organism labeling regulations. In this study, the 5'-transgene integration sequence was isolated using thermal asymmetric interlaced (TAIL)-PCR. Based upon the 5'-transgene integration sequence, conventional and TaqMan real-time PCR assays were established. The relative limit of detection for the conventional PCR assay was 0.05 % for Moonshade using 100 ng total carnation genomic DNA, corresponding to approximately 79 copies of the carnation haploid genome, and the limits of detection and quantification of the TaqMan real-time PCR assay were estimated to be 51 and 254 copies of haploid carnation genomic DNA, respectively. These results are useful for identifying and quantifying Moonshade and its derivatives.

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Abbreviations

GMO:

Genetically modified organism

PCR:

Polymerase chain reaction

TAIL-PCR:

Thermal asymmetric interlaced-PCR

LOD:

Limit of detection

LOQ:

Limit of quantification

f3'5'h :

Flavonoid 3′, 5′ hydroxylase

dfr :

Dihydroflavonol-4-reductase

SuRB :

Sulfonylurea resistance gene B

SD:

Standard deviation

References

  1. Kahl, L. S. (1994). Summary of consultation with Calgene, Inc. Concerning Flavr Savr Tomoto. FDA. 1994, Docket No. 91A20330.

  2. Clive, J. (2011). ISAAA Brief 43-2011//Global Status of Commercialized Biotech/GM Crops: 2011.

  3. Losey, J. E., Rayor, L. S., & Carter, M. E. (1999). Nature, 399, 214.

    Article  CAS  Google Scholar 

  4. Clark, E. A. (2006). Euphytica, 148, 47–60.

    Article  CAS  Google Scholar 

  5. Xiong, H. B., Cheng, Z. Q., & Wang, L. X. (2007). Southwest China Journal of Agricultural Sciences, 17, 340–346.

    Google Scholar 

  6. Chin, Y. L., Greg, N., Terese, W. R., Stephen, F. C., Richard, Y., & Michael, J. D. (1991). Nature Biotechnology, 9, 864–868.

    Article  Google Scholar 

  7. Marchant, R., Davey, M. R., Lucas, J. A., Lamb, C. J., Dixon, R. A., & Power, J. B. (1998). Molecular Breeding, 4(3), 187–194.

    Article  CAS  Google Scholar 

  8. Elomaa, P., Helariutta, Y., Kotilainen, M., & Teeri, T. H. (1996). Molecular Breeding, 2(1), 41–50.

    Article  CAS  Google Scholar 

  9. Wilmink, A., Van de Ven, B. C. E., & Dons, J. J. M. (1992). Plant Cell Reports, 11(2), 76–80.

    Article  Google Scholar 

  10. Liau, C. H., You, S. J., Prasad, V., Hsiao, H. H., Lu, J. C., Yang, N. S., et al. (2003). Plant Cell Reports, 21, 993–998.

    Article  CAS  Google Scholar 

  11. Kiyokawa, S., Kikuchi, Y., Kamada, H., & Harada, H. (1996). Plant Cell Reports, 15(8), 606–609.

    Article  CAS  Google Scholar 

  12. Aida, R., Hirose, Y., Kishimoto, S., & Shibata, M. (1999). Plant Science, 148, 1–7.

    Article  CAS  Google Scholar 

  13. Pellegrineschi, A., & Davolio, M. O. (1996). Plant Cell. Tissue and Organ Culture, 47(1), 79–86.

    Article  Google Scholar 

  14. Dolgov, S. V., Mitiouchkina, T. Y., & Skryabin, K. G. (1997). Acta Horticulturae, 447, 329–334.

    Google Scholar 

  15. Hosokawa, K., Matsuki, R., Oikawa, Y., & Yamamura, S. (2000). Plant Cell Reports, 19(5), 454–458.

    Article  CAS  Google Scholar 

  16. Nishihara, M., Ito, M., Tanaka, I., Kyo, M., Ono, K., Irifune, K., et al. (1993). Plant Physiology, 102(2), 357–361.

    CAS  Google Scholar 

  17. Hu, J., Gu, Q. X., & Chi, J. (2004). Carnation production technology. Beijing: China Agricultural Press.

    Google Scholar 

  18. Available from : http://www.isaaa.org/gmapprovaldatabase/events/default. asp? EventID = 132. Accessed April 11, 2011.

  19. Commission, E. (2003). Commission regulation (EC) no. 1829/2003 of September 22, 2003, concerning on genetically modified food and feed. Official Journal of Europian Community, L268, 1–23.

    Google Scholar 

  20. Commission, E. (2003). Commission regulation (EC) no. 1830/2003 of September 22, 2003, concerning the traceability of food and feed products produced from genetically modified organisms and amending directive2001/18 /EC. Official Journal of Europian Community, L268, 24–28.

    Google Scholar 

  21. Ministry of Agriculture and Forestry. (2000). Guidelines for labeling of genetically modified agriculture products. MAF notification 2000–31. Ministry of Agriculture and Forestry, Korea.

  22. Matsuoka, T. (2001). GMO Labeling and Detection Methods in Japan. APEC-JIRCAS Joint Symposium and Workshop on Agriculture Biotechnology

  23. Yang, L. T., Xu, S. C., Pan, A. H., Yin, C. S., Zhang, K. W., Wang, Z. Y., et al. (2005). Journal of Agricultural and Food Chemistry, 53, 9312–9318.

    Article  CAS  Google Scholar 

  24. Liu, Y. G., Mitsukawa, N., Oosumi, T., & Whittier, R. F. (1995). The Plant Journal, 8, 457–463.

    Article  CAS  Google Scholar 

  25. Royal botanic Gardens, Kew. (2005). Plant c-DNA value database (release 4.0, October 2005). Available from : http://www.rbgkew.org.uk/cval/homepage. html. Accessed April 11, 2011

  26. Zhu, H., Jiang, L., Tao, S., Lin, H., Wang, J., Tan, F., et al. (2011). Journal of AOAC International, 94(4), 1227–1232.

    CAS  Google Scholar 

  27. Available from: http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/dir030-2002. Accessed April 11, 2011.

  28. Codex Committee on Methods of Analysis and Sampling (1998). In House method validation. In House validated methods and their role in methods of analysis for codex purposes. Codex Alimentarius Commission. CX/MAS98/8:23–27.

  29. Peccoud, J., & Jacob, C. (1996). Biophysical Journal, 71, 101–108.

    Article  CAS  Google Scholar 

  30. Wu, G., Wu, Y. H., Xiao, L., & Lu, C. M. (2008). Transgenic Research, 17(5), 851–862.

    Article  CAS  Google Scholar 

  31. Jia, J., Sun, J., Bai, L., Peng, L., & Pan, A. (2010). Chinese Agricultural Science Bulletin, 26(12), 35–39.

    Google Scholar 

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Acknowledgments

This work was financially supported by Shanghai Agricultural Science Committee foundation of China, grant no. 6-4 (2009), Shanghai Agricultural Science Key Research Foundation, grant no. 1-8 (2011), the Public Research Platform Foundation of SSTC, grant no. 10DZ2294103, the Fund of Key Project of Science and Technology of the Shanghai Committee of Agriculture (2008-8-9), and the Youth Fund of Shanghai Academy of Agricultural Sciences, PR China (2010–15).

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Authors and Affiliations

  1. Biotech Research Institute of Shanghai Academy of Agricultural Sciences, Shanghai, China

    Peng Li, Junwei Jia, Lan Bai, Aihu Pan & Xueming Tang

  2. Shanghai Key Laboratory of Agricultural Genetics and Breeding, Shanghai, China

    Peng Li, Junwei Jia, Lan Bai, Aihu Pan & Xueming Tang

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  1. Peng Li
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  2. Junwei Jia
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  3. Lan Bai
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  4. Aihu Pan
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  5. Xueming Tang
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Correspondence to Aihu Pan or Xueming Tang.

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Li, P., Jia, J., Bai, L. et al. Identification and Quantification of Genetically Modified Moonshade Carnation Lines Using Conventional and TaqMan Real-Time Polymerase Chain Reaction Methods. Appl Biochem Biotechnol 170, 1151–1162 (2013). https://doi.org/10.1007/s12010-013-0254-7

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