Detection of the 35S promoter by real-time PCR as a transgenicity indicator in food and Gossypium sp.
DOI:
https://doi.org/10.15517/am.v31i1.37151Keywords:
genetically modified organisms, SYBR green, transgenic food, Costa RicaAbstract
Introduction. Genetically modified crops (GMC) are of particular interest due to their impact on the global economy. Therefore, as a general concern, many countries have established some regulations in regards to genetically modified organisms (GMOs). In Costa Rica, the cultivation of GMOs has been practiced since 1991; however, there´s a lack of studies that monitor the execution and compliance with the biosafety regulations. Objective. The objective of the present study was to identify the presence or absence of transgenicity in processed foods for human and animal consumption, as well as in cotton seeds. Material and methods. The real-time PCR technique was used to target the 35S promoter sequence, derived from the cauliflower mosaic virus (CaMV), as a marker to detect the presence of transgenes in processed foods for human and animal consumption as well as in wild or cultivated cotton seeds collected nearby a GM cotton farm in May 2017. Results. In the analyzed samples there was a high incidence of an 82 bp fragment, corresponding to the 35S promoter sequence, being absent only in organic corn crops and their derivatives (tortillas, corn powder). Results suggest the presence of GMO traces in the Costa Rican food market, additionally it reveals the urgency of implementing adequate labeling for food traceability. Furthermore, the presence of transgenic cotton in the vicinity of a GM cotton farm was identified, suggesting the relevance of surveillance in aspects of biosafety and genetic manipulation of crops. Conclusion. The presence of traces for GMOs in Costa Rican processed food, demonstrates the importance of continuing this monitoring to provide enough elements for a critic discussion about food traceability and potential transgene flow into wild plant material.
Downloads
References
Barbau-Piednoir, E., A. Lievens, G. Mbongolo-Mbella, N. Roosens, M. Sneyers, A. Leunda-Casi, and M. Van-den-Bulcke. 2010. SYBR®Green qPCR screening methods for the presence of “35S promoter” and “NOS terminator” elements in food and feed products. Eur. Food Res. Technol. 230:383-393. doi:10.1007/s00217-009-1170-5
Barbau-Piednoir, E., A. Lievens, E. Vandermassen, E.G. Mbongolo-Mbella, A. Leunda-Casi, N. Roosens, M. Sneyers, and M. Van-den-Bulcke. 2012. Four new SYBR®Green qPCR screening methods for the detection of Roundup Ready®, LibertyLink®, and Cry1Ab traits in genetically modified products. Eur. Food Res. Technol. 234:13-23. doi:10.1007/s00217-011-1605-7
BCH (Biosafety Clearing-House). 2018. Country profile. Costa Rica. Risk assessment. Conservation on Biological Diversity, BEL. https://bch.cbd.int/about/countryprofile.shtml?country=cr (accessed Jan 6, 2018).
Bonfini, L., P. Heinze, S. Kay, and E.G. Van-den. 2001. Review of GMO detection and quantification techniques. Publications Office of the European Union. EU. https://publications.europa.eu/en/publication-detail/-/publication/47b0a030-b501-4a33-ab9a-28baf6eade60/language-en (accessed Dec. 10, 2018).
Bravo, E. 2015. Transgénicos en Panamá. GRAIN, WA, USA. https://www.grain.org/article/entries/5167-transgenicos-en-panama (consultado Sept 30. 2018).
Carvajal, P., H. Ureña, J. Umaña, C. Sancho, F. Solano, M. Arleo, C. Martínez-Debat, y R. Umaña. 2017. Detección molecular de secuencias de ADN transgénico en alimentos de consumo humano y animal en Costa Rica. Agron. Costarricense 41(1):53-68. doi:10.15517/rac.v41i1.29751
Coello, R.P., J.P. Justo, A.F. Mendoza, and E.S. Ordoñez. 2017. Comparison of three DNA extraction methods for the detection and quantification of GMO in Ecuadorian manufactured food. BMC Res. Notes 10:758. doi:10.1186/s13104-017-3083-x
Conner, A.J., T.R. Glare, and J.P. Nap. 2003. The release of genetically modified crops into the environment. Part II. Overview of ecological risk assessment. Plant J. Cell Mol. Biol. 33:19-46. doi:10.1046/j.0960-7412.2002.001607.x
Datukishvili, N., T. Kutateladze, I. Gabriadze, K. Bitskinashvili, and B. Vishnepolsky. 2015. New multiplex PCR methods for rapid screening of genetically modified organisms in foods. Front. Microbiol. 6:757. doi:10.3389/fmicb.2015.00757
Debode, F., A. Marien, É. Janssen, C. Bragard, and G. Berben. 2017. The influence of amplicon length on real-time PCR results. Biotechnol. Agron. Soc. Environ. 21(1):3-11. doi.10.25518/1780-4507.13461
De-Faria, F. 2005. Granos y semillas transgénicos en cadena alimentaria: Costa Rica. Ambientico 137:19-21.
De-Jesús-Martínez, J. 2015. Evaluación económica del uso de maíz transgénico en el departamento de Olancho, Honduras. Tesis Lic., Zamorano, Francisco Morazan, HND.
Doyle, J.J., and J.L. Doyle. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 19:11-15.
Eckerstorfer, MF., M. Engelhard, A. Heissenberger, S. Simon, and H. Teichmann. 2019. Plants developed by new genetic modification techniques-comparison of existing regulatory frameworks in the EU and non-EU countries. Front. Bioeng. Biotechnol. 19:7-26. doi:10.3389/fbioe.2019.00026
Fraiture, M.A., P. Herman, I. Taverniers, M. De-Loose, D. Deforce, and N.H. Roosens. 2015. Current and new approaches in GMO detection: challenges and solutions. BioMed. Res. Int. 2015:392872. doi:10.1155/2015/392872
Freitas, M., W. Correr, J. Cancino-Bernardi, M.F. Barroso, C. Delerue-Matos, and V. Zucolotto. 2016. Impedimetric immunosensors for the detection of Cry1Ab protein from genetically modified maize seeds. Sens. Actuat. B: Chem. 237:702-709. doi:10.1016/J.SNB.2016.06.149
Galeano, P., C. Martínez-Debat, F. Ruibal, L. Franco-Fraguas, and G.A. Galván. 2010. Cross-fertilization between genetically modified and non-genetically modified maize crops in Uruguay. Environ. Biosaf. Res. 9:147-154. doi:10.1051/ebr/2011100
García, J.E. 2007. Cultivos genéticamente modificados: las promesas y las buenas intenciones no bastan. Rev. Biol. Trop. 55:347-364. doi:10.15517/RBT.V55I2.6015
García-González, J. 2010. La contaminación silenciosa. Biocenosis 23(1):38-49.
González-Ortega, E., A. Piñeyro-Nelson, E. Monterrubio-Vázquez, E. Gómez-Hernández, M. Arleo, J. Dávila-Velderrain, C. Martínez-Debat, and E.R. Álvarez-Buylla. 2017. Pervasive presence of transgenes and glyphosate in industrialized maize- derived food in Mexico. Agroecol. Sustain. Food Syst. 41:1146-1161. doi:10.1080/21683565.2017.1372841
ISAAA (International Service for the Acquisition of Agri-biotech Applications). 2018. Global status of commercialized biotech/GM crops: 2018. Brief Nº 54. ISAAA, NY, USA. https://www.isaaa.org/resources/publications/briefs/54/default.asp (accessed Oct. 15, 2019).
Kamle, S., and S. Ali. 2013. Genetically modified crops: detection strategies and biosafety issues. Gene 522:123-132. doi:10.1016/j.gene.2013.03.107
Leão-Buchir, J., G.V.M. Pereira, A.L.L. Silva, S. Alban, M.C. Rocha, J. Polettini, V. Thomaz-Soccol, and C.R. Soccol. 2018. Real-time PCR for traceability and quantification of genetically modified seeds in lots of non-transgenic soybean. Biosci. J. 34:34-41. doi:10.14393/BJ-v34n1a2018-37236
Lu, B.R. 2008. Transgene escape from GM crops and potential biosafety consequences: an environmental perspective. Collect Biosaf. Rev. 4(4):66-141.
Manzur, M.I., y M.I. Cárcamo. 2014. América Latina: La transgénesis de un continente. Visión crítica de una expansión descontrolada. 2da ed. Ediciones Boll, Santiago de Chile, CHL.
Mathur, R. 2018. Genetic engineering and biosafety in the use of genetically modified foods. IJASRM 2018(Special I):76-82.
Pacheco-Rodríguez, F., y J.E. García-González. 2014. Situación de los cultivos transgénicos en Costa Rica. Act. Acad. 54:29-60.
Paull, J. 2018. Genetically modified organism (GMOs) as invasive species. J. Environ. Protec. Sustain. Dev. 4(3):31-37.
Peng, C., P. Wang, X. Xu, X. Wang, W. Wei, X. Chen, and J. Xu. 2016. Development of a qualitative real-time PCR method to detect 19 targets for identification of genetically modified organisms. SpringerPlus 5:889. doi:10.1186/s40064-016-2395-y
Pierboni, E., L. Curcio, G.R. Tovo, M. Torricelli, and C. Rondini. 2016. Evaluation of systems for nopaline synthase terminator in fast and standard real-time PCR to screen genetically modified organisms. Food Anal. Methods 9:1009-1019. doi:10.1007/s12161-015-0283-7
Randhawa, G., M. Singh, and P. Sood. 2016. DNA-based methods for detection of genetically modified events in food and supply chain. Curr. Sci. 110:1000-1008. doi:10.18520/cs/v110/i6/1000-1009
Rocha-Munive, M.G., M. Soberón, S. Castañeda, E. Niaves, E. Scheinvar, L.E. Eguiarte, D. Mota-Sánchez, E. Rosales-Robles, U. Nava-Camberos, J.L. Martínez-Carrillo, C.A. Blanco, A. Bravo, and V. Souza. 2018. Evaluation of the impact of genetically modified cotton after 20 years of cultivation in Mexico. Front. Bioeng. Biotechnol. 6:82. doi:10.3389/fbioe.2018.00082
Rostoks, N., L. Grantiņa-Ieviņa, B. Ieviņa, V. Evelone, O. Valciņa, and I. Aleksejeva. 2019. Genetically modified seeds and plant propagating material in Europe: potential routes of entrance and current status. Heliyon 5(2):e01242. doi:10.1016/j.heliyon.2019.e01242.
Sprenger, U. 2008. La contaminación oculta: semilla transgénica, bioseguridad e intervenciones de la sociedad civil en Costa Rica. AGRECOL e.V., Guggenhausen, DEU. http://www.agrecol.de/la-contaminación-oculta-semillas-transgénicas-bioseguridad-e-intervenciones-de-la-sociedad-civil-en (consultado 30 oct. 2018).
Takabatake, R., M. Onishi, S. Futo, Y. Minegishi, A. Noguchi, K. Nakamura, and K. Kitta. 2015. Comparison of the specificity, stability, and PCR efficiency of six rice endogenous sequences for detection analyses of genetically modified rice. Food Control 50:949-955. doi:10.1016/j.foodcont.2014.05.043
Van-den-Eede, G., L. Bonfini, L. Cengia, C. Iannini, L. Kluga, and M. Mazzara. 2011. Compendium of reference methods for GMO analyses. Publications Office of the European Union, Luxembourg, LUX. doi:10.2788/16745
Wei, S., C. Wang, P. Zhu, G. Zhou, W. Fu, and X. Wu. 2018. A high-throughput multiplex tandem PCR assay for the screening of genetically modified maize. LWT 87:169-176. doi:10.1016/j.lwt.2017.08.061
Yang, L., A. Pan, K. Zhang, C. Yin, B. Qian, J. Chen, and D. Zhang. 2005. Qualitative and quantitative PCR methods for event-specific detection of genetically modified cotton Mon1445 and Mon531. Transgenic Res. 14:817-831. doi:10.1007/s11248-005-0010-z
Yeaman, G.R., S. Paul, I. Nahirna, Y. Wang, A.E. Deffenbaugh, Z.L. Liu, and K.C. Glenn. 2016. Development and validation of a fluorescent multiplexed immunoassay for measurement of transgenic proteins in cotton (Gossypium hirsutum). J. Agric. Food Chem. 64:5117-5127. doi:10.1021/acs.jafc.6b01441
Yılmaz, R., C. Bayraç, A. Başman, and H. Köksel. 2019. Development of SYBR green-based real time PCR assays for detection and quantification of adulteration in wheat-based composite breads and their in-house validation. J. Cereal Sci. 85:91-97. doi:10.1016/j.jcs.2018.11.020
Downloads
Additional Files
Published
How to Cite
Issue
Section
License
1. Proposed policy for open access journals
Authors who publish in this journal accept the following conditions:
a. Authors retain the copyright and assign to the journal the right to the first publication, with the work registered under the attribution, non-commercial and no-derivative license from Creative Commons, which allows third parties to use what has been published as long as they mention the authorship of the work and upon first publication in this journal, the work may not be used for commercial purposes and the publications may not be used to remix, transform or create another work.
b. Authors may enter into additional independent contractual arrangements for the non-exclusive distribution of the version of the article published in this journal (e.g., including it in an institutional repository or publishing it in a book) provided that they clearly indicate that the work was first published in this journal.
c. Authors are permitted and encouraged to publish their work on the Internet (e.g. on institutional or personal pages) before and during the review and publication process, as it may lead to productive exchanges and faster and wider dissemination of published work (see The Effect of Open Access).
