Development of technologies for detection of GMOs tolerant to herbicide glyphosate

MARIELA DEL GIUDICE; GABRIELA SOTO; JUAN UGALDE; CAROLINA LUCERO; PABLO SIPOWICZ; DIEGO REY SERANTES; ARIEL ODORIZI; DIEGO COMERCI

Mendoza

Congreso; LVIII Annual Meeting of the Argentine Society for Biochemistry and Molecular Biology Research; 2022

In the last decades, agrobiotechnology has been favored by the advance of genetic engineering and molecular biology techniques that allow the genetic modification of plant organisms. These modifications consist in the introduction of genes that confer specific improvements to plants, such as resistance to insect damage, tolerance to herbicides or to different environmental stress factors, and as a result, generate genetically modified organisms (GMOs). Argentina is one of the major producers of GMOs, and the largest production volume correspond to crops that carry herbicide tolerance genes, especially those that confer resistance to glyphosate. To regulate GMOs production and commercialization, it is necessary to determine special cultivation conditions and quality control systems to ensure the agro-ecological balance. Therefore, the use of appropriate detection methods is required. Despite the great demand generated by the massive production and commercialization of transgenic crops in Argentina, national diagnostic tools have not yet been developed and most of the ones currently available involve expensive and imported reagents. Our aim is to develop technologies to identify genetically modified plants tolerant to herbicides, based on “in house” produced biomolecules. These technologies will allow detecting the presence of GMOs in the main crops produced and commercialized by Argentina.In this study we report the production of biomolecules (antigens and antibodies) and the development of a capture ELISA (Enzyme-Linked ImmunoSorbent Assay) to detect the transgenic protein that confers tolerance to glyphosate: CP4 EPSPS. We produced and purified the recombinant protein from E. coli and obtained monoclonal and polyclonal antibodies against it. With these antibodies we developed a capture ELISA and optimized it using different combinations of them to capture the recombinant CP4 EPSPS. The results showed that the system is able to detect low protein concentrations. Moreover, we analyzed the ability to capture the native protein in two plant species: Alfalfa (Medicago sativa L.) and Soybean (Glycine max L.). Our results indicated that the developed technology allows to detect the presence of GMOs in different crop samples, with high sensitivity and specificity.