Differential Detection and Identification of Transgenic Tomato Solanum Lycopersicum by Multiplex PCR Technique in Tehran
Subject Areas : Microbiology
A. Azizi
1
(M. Sc. of Food Science and Technology, Science and Research Branch, Islamic Azad University, Tehran, Iran.)
A. Sarifan
2
(Associate Professor of the Department of Food Science and Technology, Science and Research Branch, Islamic Azad University, Tehran, Iran.)
M. Emtiazjoo
3
(Associate Professor of Faculty of Life Sciences, North Branch, Islamic Azad University, Tehran, Iran.)
Keywords: PCR, tomato, Transgenic Identification,
Abstract :
Introduction: Food industry has an important value all over the would. Tomatoes and it’s other products are one of the important agricultural products in food industry. Transgenic plants and seeds are one of the important achievements of modern biotechnology in field of agriculture that have captured a part of food marketing in recent years. Therefore, identify and examine whether transgenic agricultural products are transgenic or not, it is necessary because there are still many opinions for and against the use of these products in the world.Materials and Methods: In this study, 10 types of tomatoes along with sepals were collected by simple sampling method from Tajrish bazaar, Shahrvand-e farmaniyeh, vegetable market East and West of Tehran and were examined. At first, P35S F/R, NOS-1/NOS-3, RBCLF/R and NPTǁ F/R primers were used. Nucleotide fragments were amplified respectively at 195bp and 180bp.Results: Amplified nucleotide fragments confirmed 100% similarity of the nucleotide sequence to the samples in the gene bank that the studied products were genetically engineered. Also, in order to identify transgenic tomatoes, aspecific PG F/R primer pair was used to amplify the antigens of polygalacturonase gene. All specimens that had P35S promoter band or NOS terminator, or both, amplified the 384bp band associated with part of the PG gene antisense, and found a 100% similarity in the gene sequence registered in the gene bank.Conclusion: The results of this investigation showed that transgenic tomatoes exist in Iran’s market without consumer awareness.
Ahmed, F. E. (2002). Detection of genetically modified organisms in foods. TRENDS in Biotechnology, 20(5), 215-223.
Amin, L., Azad, M. A. K., Gausmian, M. H. & Zulkifli, F. (2014). Determinants of public attitudes to genetically modified salmon. PloS one, 9(1), e86174.
Arun, Ö. Ö., Yılmaz, F. & Muratoğlu, K. (2013). PCR detection of genetically modified maize and soy in mildly and highly processed foods. Food Control, 32(2), 525-531.
Ashraf, M. & Akram, N.A. (2009). Improving salinity tolerance of plants through conventional breeding and genetic engineering: an analytical comparison. Biotechnology advances, 27(6), 744-752. [In Persian]
Briefs, I. S. A. A. (2016). Global status of commercialized biotech/GM crops 2016.
Bruderer, S., Leitner, K. E. & Lindenmeyer, J. )2003(. Genetically Modified (GM) crops: molecular and regulatory details. Basel, Švica, BATS, Centre for biosafety and sustainability (http://www. bats. ch/gmo-watch/).
Burton‐Freeman, B., Talbot, J., Park, E., Krishnankutty, S. & Edirisinghe, I. (2012). Protective activity of processed tomato products on postprandial oxidation and inflammation: a clinical trial in healthy weight men and women. Molecular Nutrition & Food Research, 56(4), 622-631.
Constable, A., Jonas, D., Cockburn, A., Davi, A., Edwards, G., Hepburn, P., Herouet-Guicheney, C., Knowles, M., Moseley, B., Oberdörfer, R. and Samuels, F. (2007). History of safe use as applied to the safety assessment of novel foods and foods derived from genetically modified organisms. Food and Chemical Toxicology, 45(12), 2513-2525.
Demeke, T. & Ratnayaka, I. (2008). Multiplex qualitative PCR assay for identification of genetically modified canola events and real-time event-specific PCR assay for quantification of the GT73 canola event. Food Control, 19(9), 893-897.
Demyttenaere, J. C. (2018). Natural or Synthetic? The Legal Framework in the EU for the Production of Natural Flavouring
Ingredients. In Biotechnology of Natural Products, Springer, Cham 281-305.
Faostat, F. A. O. (2012). Disponível em:< http://faostat. fao. org>. Acesso em, 14.
Foley, J. A., Ramankutty, N., Brauman, K. A., Cassidy, E. S., Gerber, J. S., Johnston, M., Mueller, N. D., O’Connell, C., Ray, D. K., West, P. C. & Balzer, C. (2011). Solutions for a cultivated planet. Nature, 478(7369), 337-342.
Fooladgar, F., Talebi, M. & Bahar, M. (2019). Detection and identification of genetically-modified tomatoes using PCR technique and sequencing. Journal of Science and Technology of Greenhouse Culture-Isfahan University of Technology, 9(4), 71-80. [In Persian]
Fraiture, M. A., Herman, P., Taverniers, I., De Loose, M., Deforce, D. & Roosens, N. H. (2015). Current and new approaches in GMO detection: challenges and solutions. BioMed research international, 2015, 1-22.
Gaudron, T., Peters, C., Boland, E., Steinmetz, A. & Moris, G. (2009). Development of a quadruplex-real-time-PCR for screening food for genetically modified organisms. European Food Research and Technology, 229(2), 295-305.
Ghanian, M., Ghoochani, O. M., Kitterlin, M., Jahangiry, S., Zarafshani, K., Van Passel, S. & Azadi, H. (2016). Attitudes of agricultural experts toward genetically modified crops: A case study in Southwest Iran. Science and engineering ethics, 22(2), 509-524. [In Persian]
Godfray, H. C. J., Beddington, J.R., Crute, I. R., Haddad, L., Lawrence, D., Muir, J. F., Pretty, J., Robinson, S., Thomas, S. M. & Toulmin, C. (2010). Food security: the challenge of feeding 9 billion people. science, 327(5967), 812-818.
Hashemi, M. & Shojae Al-sadati, S. A. (2012). Genetically Modified food: opportunities and challenges. Iranian Journal of Food science and Technology, 7(24).89-102. [In Persian]
Hemmer, W. (1997). Foods derived from genetically modified organisms and detection methods, 1-3.
Holst-Jensen, A., Bertheau, Y., de Loose, M., Grohmann, L., Hamels, S., Hougs, L., Morisset, D., Pecoraro, S., Pla, M., Van den Bulcke, M. & Wulff, D. (2012). Detecting un-authorized genetically modified organisms (GMOs) and derived materials. Biotechnology Advances, 30(6), 1318-1335.
ISAAA (2012). International Service for the Acquistion of Agri-biotech Applications. ISAAA Briefs 42. Global statuse of commercialized Biotech/GM Crops: 2012: International Service for the Acquisition of Agri-biotech Applications.
Jia, H. (2003). GM Labeling in china beset by problems. Nature Biotechnology 21(8), 835-836.
Jinxia, A., Qingzhang, L., Xuejun, G., Yanbo, Y., Lu, L. & Minghui, Z. (2011). A multiplex nested PCR assay for the simultaneous detection of genetically modified soybean, maize and rice in highly processed products. Food Control, 22(10), 1617-1623.
Jobes, D. V., Hurley, D. L. & Thien, L. B. (1995). Plant DNA isolation: a method to efficiently remove polyphenolics, polysaccharides, and RNA. Taxon, 44(3), 379-386.
Kay, S. & Van den Eede, G. (2001). The limits of GMO detection. Nature Biotechnology, 19(5), 405-405.
Kaya, I. H., Poyrazoglu, E. S., Artik, N. & Konar, N. (2013). Academicans’ Perceptions and Attitudes toward GM-Organisms and–Foods. International Journal of Biological, Ecological and Environmental Sciences IJBEES, 2(2), 20-24.
Kuiper, H. A. (1999). Summary report of the ILSI Europe workshop on detection methods for novel foods derived from genetically modified organisms. Food Control, 10, 339-349.
Lin, H. Y., CHIUEH, L. C. & SHIH, D. Y. C. (2000). Detection of genetically modified soybeans and maize by the polymerase chain reaction method. Journal of Food and Drug Analysis, 8(3).
Lipp, M., Brodmann, P., Pietsch, K., Pauwels, J. & Anklam, E. (1999). IUPAC collaborative trial study of a method to detect genetically modified soy beans and maize in dried powder. Journal of AOAC International, 82(4), 923-928.
Liu, J. Y., Deng, X., Kang, L. & Chen, D. M. (2006). Detection of transgenic papaya by SYBR-Green real time PCR. Journal of HUNAN Agriculture University, 32(4), 371-374.
Lübeck, M. (2002). Detection of genetically modified plants: Methods to sample and analyse GMO content in plants and plant products. Danish Forest and Nature Agency.
Marmiroli, N., Maestri, E., Gulli, M., Malcevschi, A., Peano, C., Bordoni, R. & De Bellis, G. (2008). Methods for detection of GMOs in food and feed. Analytical and Bioanalytical Chemistry, 392(3), 369.
Matsuoka, T., Kuribara, H., Takubo, K., Akiyama, H., Miura, H., Goda, Y., Kusakabe, Y., Isshiki, K., Toyoda, M. & Hino, A. (2002). Detection of recombinant DNA segments introduced to genetically modified maize (Zea mays). Journal of Agricultural and Food Chemistry, 50(7), 2100-2109.
Meyer, R. (1999). Development and application of DNA analytical methods for the detection of GMOs in food. Food control, 10(6), 391-399.
Nazarenko, I., Lowe, B., Darfler, M., Ikonomi, P., Schuster, D. & Rashtchian, A. (2002). Multiplex quantitative PCR using self-quenched primers labeled with a single fluorophore. Nucleic acids Research, 30(9), e37-e37.
Nourozi, P. (2003). Biotechnology perspectives and issues in sustainable development. The first food and sustainable development conference. [In Persian]
Norouzi, P., Mahmoudi, S. & Jafari, R. (2016). Priorities of Transformation for Field and Horticultire Crops in Iran Biotechnology committee of the ministry of Agriculture Jihad. Mashhad University Jihad Publications. 78 pages Ministry of Agriculture Jihad, 2016. From (www.pr.maj.ir). [In Persian]
Park, S. H. (2005). Current status of regulation on GM food in Korea. Shokuhin eiseigaku zasshi. Journal of the Food Hygienic Society of Japan, 46(1), 4-J7.
Rani, S. J. & Usha, R. (2013). Transgenic plants: Types, benefits, public concerns and future. journal of pharmacy research, 6(8), 879-883.
Rizwan, M., Rodriguez‐Blanco, I., Harbottle, A., Birch‐Machin, M.A., Watson, R.E.B. & Rhodes, L.E. (2011). Tomato paste rich in lycopene protects against cutaneous photodamage in humans in vivo: a randomized controlled trial. British Journal of Dermatology, 164(1), 154-162.
Sonmezalp, C. Z. (2004). Detection of genetically modified insect resistant tomato via polymerase chain reaction (Doctoral dissertation, Doctoral dissertation, Middle East Technical University).
Tan, W., J. Dong, H. L. Deng, H. Z. Wu & Xu, B. L. (2003). Protocol of the qualitative polymerase chain reaction for detection of genetically modified plant components in food, industry standard of entry exit inspection and quarantine of the People’s Republic of China. SN/T 1202, China: Industry Standard.
Tas, M., Balci, M., Yüksel, A. & Yesilçubuk, N. S. (2015). Consumer awareness, perception and attitudes towards genetically modified foods in Turkey. British Food Journal.
Tengel, C., Schüßler, P., Setzke, E., Balles, J. & Sprenger-Haussels, M. (2001). PCR-based detection of genetically modified soybean and maize in raw and highly processed foodstuffs. BioTechniques, 31(2), 426-429.
Vidhya, S. S., Gowda, P.H., Yogendra, K.N., Ningaraju, T. M. & Salome, T. (2012). Detection of genetically modified cotton seeds using PCR and real-time PCR.
Wen-Tao, X., Kun-Lun, H., Ai-Ke, D. & Yun-Bo, L. (2006). PCR for the detection of the anti-herbicide genes in genetically modified organisms. Chinese Journal of Agricultural Biotechnology, 3(1), 53-57.
Zhai, W. X., Chen, C. Y., Zhu, X. F., Chen, X. W., Zhang, D. H., Li, X. B. & Zhu, L. H. (2004). Analysis of tDNA-Xa21 loci and bacterial blight resistance effects of the transgene Xa21 in transgenic rice. Theoretical and Applied Genetics, 109, 534–542.
_||_Ahmed, F. E. (2002). Detection of genetically modified organisms in foods. TRENDS in Biotechnology, 20(5), 215-223.
Amin, L., Azad, M. A. K., Gausmian, M. H. & Zulkifli, F. (2014). Determinants of public attitudes to genetically modified salmon. PloS one, 9(1), e86174.
Arun, Ö. Ö., Yılmaz, F. & Muratoğlu, K. (2013). PCR detection of genetically modified maize and soy in mildly and highly processed foods. Food Control, 32(2), 525-531.
Ashraf, M. & Akram, N.A. (2009). Improving salinity tolerance of plants through conventional breeding and genetic engineering: an analytical comparison. Biotechnology advances, 27(6), 744-752. [In Persian]
Briefs, I. S. A. A. (2016). Global status of commercialized biotech/GM crops 2016.
Bruderer, S., Leitner, K. E. & Lindenmeyer, J. )2003(. Genetically Modified (GM) crops: molecular and regulatory details. Basel, Švica, BATS, Centre for biosafety and sustainability (http://www. bats. ch/gmo-watch/).
Burton‐Freeman, B., Talbot, J., Park, E., Krishnankutty, S. & Edirisinghe, I. (2012). Protective activity of processed tomato products on postprandial oxidation and inflammation: a clinical trial in healthy weight men and women. Molecular Nutrition & Food Research, 56(4), 622-631.
Constable, A., Jonas, D., Cockburn, A., Davi, A., Edwards, G., Hepburn, P., Herouet-Guicheney, C., Knowles, M., Moseley, B., Oberdörfer, R. and Samuels, F. (2007). History of safe use as applied to the safety assessment of novel foods and foods derived from genetically modified organisms. Food and Chemical Toxicology, 45(12), 2513-2525.
Demeke, T. & Ratnayaka, I. (2008). Multiplex qualitative PCR assay for identification of genetically modified canola events and real-time event-specific PCR assay for quantification of the GT73 canola event. Food Control, 19(9), 893-897.
Demyttenaere, J. C. (2018). Natural or Synthetic? The Legal Framework in the EU for the Production of Natural Flavouring
Ingredients. In Biotechnology of Natural Products, Springer, Cham 281-305.
Faostat, F. A. O. (2012). Disponível em:< http://faostat. fao. org>. Acesso em, 14.
Foley, J. A., Ramankutty, N., Brauman, K. A., Cassidy, E. S., Gerber, J. S., Johnston, M., Mueller, N. D., O’Connell, C., Ray, D. K., West, P. C. & Balzer, C. (2011). Solutions for a cultivated planet. Nature, 478(7369), 337-342.
Fooladgar, F., Talebi, M. & Bahar, M. (2019). Detection and identification of genetically-modified tomatoes using PCR technique and sequencing. Journal of Science and Technology of Greenhouse Culture-Isfahan University of Technology, 9(4), 71-80. [In Persian]
Fraiture, M. A., Herman, P., Taverniers, I., De Loose, M., Deforce, D. & Roosens, N. H. (2015). Current and new approaches in GMO detection: challenges and solutions. BioMed research international, 2015, 1-22.
Gaudron, T., Peters, C., Boland, E., Steinmetz, A. & Moris, G. (2009). Development of a quadruplex-real-time-PCR for screening food for genetically modified organisms. European Food Research and Technology, 229(2), 295-305.
Ghanian, M., Ghoochani, O. M., Kitterlin, M., Jahangiry, S., Zarafshani, K., Van Passel, S. & Azadi, H. (2016). Attitudes of agricultural experts toward genetically modified crops: A case study in Southwest Iran. Science and engineering ethics, 22(2), 509-524. [In Persian]
Godfray, H. C. J., Beddington, J.R., Crute, I. R., Haddad, L., Lawrence, D., Muir, J. F., Pretty, J., Robinson, S., Thomas, S. M. & Toulmin, C. (2010). Food security: the challenge of feeding 9 billion people. science, 327(5967), 812-818.
Hashemi, M. & Shojae Al-sadati, S. A. (2012). Genetically Modified food: opportunities and challenges. Iranian Journal of Food science and Technology, 7(24).89-102. [In Persian]
Hemmer, W. (1997). Foods derived from genetically modified organisms and detection methods, 1-3.
Holst-Jensen, A., Bertheau, Y., de Loose, M., Grohmann, L., Hamels, S., Hougs, L., Morisset, D., Pecoraro, S., Pla, M., Van den Bulcke, M. & Wulff, D. (2012). Detecting un-authorized genetically modified organisms (GMOs) and derived materials. Biotechnology Advances, 30(6), 1318-1335.
ISAAA (2012). International Service for the Acquistion of Agri-biotech Applications. ISAAA Briefs 42. Global statuse of commercialized Biotech/GM Crops: 2012: International Service for the Acquisition of Agri-biotech Applications.
Jia, H. (2003). GM Labeling in china beset by problems. Nature Biotechnology 21(8), 835-836.
Jinxia, A., Qingzhang, L., Xuejun, G., Yanbo, Y., Lu, L. & Minghui, Z. (2011). A multiplex nested PCR assay for the simultaneous detection of genetically modified soybean, maize and rice in highly processed products. Food Control, 22(10), 1617-1623.
Jobes, D. V., Hurley, D. L. & Thien, L. B. (1995). Plant DNA isolation: a method to efficiently remove polyphenolics, polysaccharides, and RNA. Taxon, 44(3), 379-386.
Kay, S. & Van den Eede, G. (2001). The limits of GMO detection. Nature Biotechnology, 19(5), 405-405.
Kaya, I. H., Poyrazoglu, E. S., Artik, N. & Konar, N. (2013). Academicans’ Perceptions and Attitudes toward GM-Organisms and–Foods. International Journal of Biological, Ecological and Environmental Sciences IJBEES, 2(2), 20-24.
Kuiper, H. A. (1999). Summary report of the ILSI Europe workshop on detection methods for novel foods derived from genetically modified organisms. Food Control, 10, 339-349.
Lin, H. Y., CHIUEH, L. C. & SHIH, D. Y. C. (2000). Detection of genetically modified soybeans and maize by the polymerase chain reaction method. Journal of Food and Drug Analysis, 8(3).
Lipp, M., Brodmann, P., Pietsch, K., Pauwels, J. & Anklam, E. (1999). IUPAC collaborative trial study of a method to detect genetically modified soy beans and maize in dried powder. Journal of AOAC International, 82(4), 923-928.
Liu, J. Y., Deng, X., Kang, L. & Chen, D. M. (2006). Detection of transgenic papaya by SYBR-Green real time PCR. Journal of HUNAN Agriculture University, 32(4), 371-374.
Lübeck, M. (2002). Detection of genetically modified plants: Methods to sample and analyse GMO content in plants and plant products. Danish Forest and Nature Agency.
Marmiroli, N., Maestri, E., Gulli, M., Malcevschi, A., Peano, C., Bordoni, R. & De Bellis, G. (2008). Methods for detection of GMOs in food and feed. Analytical and Bioanalytical Chemistry, 392(3), 369.
Matsuoka, T., Kuribara, H., Takubo, K., Akiyama, H., Miura, H., Goda, Y., Kusakabe, Y., Isshiki, K., Toyoda, M. & Hino, A. (2002). Detection of recombinant DNA segments introduced to genetically modified maize (Zea mays). Journal of Agricultural and Food Chemistry, 50(7), 2100-2109.
Meyer, R. (1999). Development and application of DNA analytical methods for the detection of GMOs in food. Food control, 10(6), 391-399.
Nazarenko, I., Lowe, B., Darfler, M., Ikonomi, P., Schuster, D. & Rashtchian, A. (2002). Multiplex quantitative PCR using self-quenched primers labeled with a single fluorophore. Nucleic acids Research, 30(9), e37-e37.
Nourozi, P. (2003). Biotechnology perspectives and issues in sustainable development. The first food and sustainable development conference. [In Persian]
Norouzi, P., Mahmoudi, S. & Jafari, R. (2016). Priorities of Transformation for Field and Horticultire Crops in Iran Biotechnology committee of the ministry of Agriculture Jihad. Mashhad University Jihad Publications. 78 pages Ministry of Agriculture Jihad, 2016. From (www.pr.maj.ir). [In Persian]
Park, S. H. (2005). Current status of regulation on GM food in Korea. Shokuhin eiseigaku zasshi. Journal of the Food Hygienic Society of Japan, 46(1), 4-J7.
Rani, S. J. & Usha, R. (2013). Transgenic plants: Types, benefits, public concerns and future. journal of pharmacy research, 6(8), 879-883.
Rizwan, M., Rodriguez‐Blanco, I., Harbottle, A., Birch‐Machin, M.A., Watson, R.E.B. & Rhodes, L.E. (2011). Tomato paste rich in lycopene protects against cutaneous photodamage in humans in vivo: a randomized controlled trial. British Journal of Dermatology, 164(1), 154-162.
Sonmezalp, C. Z. (2004). Detection of genetically modified insect resistant tomato via polymerase chain reaction (Doctoral dissertation, Doctoral dissertation, Middle East Technical University).
Tan, W., J. Dong, H. L. Deng, H. Z. Wu & Xu, B. L. (2003). Protocol of the qualitative polymerase chain reaction for detection of genetically modified plant components in food, industry standard of entry exit inspection and quarantine of the People’s Republic of China. SN/T 1202, China: Industry Standard.
Tas, M., Balci, M., Yüksel, A. & Yesilçubuk, N. S. (2015). Consumer awareness, perception and attitudes towards genetically modified foods in Turkey. British Food Journal.
Tengel, C., Schüßler, P., Setzke, E., Balles, J. & Sprenger-Haussels, M. (2001). PCR-based detection of genetically modified soybean and maize in raw and highly processed foodstuffs. BioTechniques, 31(2), 426-429.
Vidhya, S. S., Gowda, P.H., Yogendra, K.N., Ningaraju, T. M. & Salome, T. (2012). Detection of genetically modified cotton seeds using PCR and real-time PCR.
Wen-Tao, X., Kun-Lun, H., Ai-Ke, D. & Yun-Bo, L. (2006). PCR for the detection of the anti-herbicide genes in genetically modified organisms. Chinese Journal of Agricultural Biotechnology, 3(1), 53-57.
Zhai, W. X., Chen, C. Y., Zhu, X. F., Chen, X. W., Zhang, D. H., Li, X. B. & Zhu, L. H. (2004). Analysis of tDNA-Xa21 loci and bacterial blight resistance effects of the transgene Xa21 in transgenic rice. Theoretical and Applied Genetics, 109, 534–542.