DETECTION OF GENETICALLY MODIFIED RICE LOCALLY AVAILABLE IN PUNJAB, PAKISTAN

Abstract

Rice (Oryza sativa L.) is an important cereal crop that provides food to half of the world's population. Pakistan's traditional and premium quality rice is mostly exported to Europe. Transgenic or genetically modified rice (GM RICE) has been developed in the USA, China, and other countries, like herbicide-tolerant Liberty-Link rice (LLRICE06, LLRICE62, LLRICE601) and insect-resistant Bt-63. International and national biosafety regulations allow countries to stop the import of any genetically modified organism (GMO) due to its possible safety concerns. PCR-based methods are recommended for the accurate detection of GM rice. Qualitative PCR is used to detect the presence or absence of GMOs in a rice sample. The objective of this study was to check the presence of GMOs in the rice samples collected from rice-growing areas in Punjab. Eighty-two rice samples were collected randomly from various markets/locations in rice-growing areas of Punjab for this study. Twelve bulks were prepared to represent eighty-two rice samples. The CTAB method was used to extract the DNA from rice samples to analyze it. PCR of rice bulk samples was performed using primers specific for SPS, CP, p35S, tNOS, nptII, Hpt and Bt 63 rice. Positive and negative controls were included in each PCR. Primers specific for chloroplast (CP) and Sucrose phosphate synthase (SPS) genes were used to confirm DNA quality for amplification. Then, primers specific for the cauliflower mosaic virus (CaMV) 35S promoter and the Agrobacterium nopaline synthase (NOS) terminator were used for the detection of GM rice through PCR, neomycin phosphotransferase II (nptII) and Hygromycin phosphotransferase (Hpt) were used as a selectable marker and Bt 63 rice was used in insect resistance. The gel electrophoresis of PCR was conducted using a DNA ladder and then taking of the image was through gel documentation system for visualization and analysis. According to the results, all the rice bulks representing the eighty-two rice samples were negative for p35S, tNOS, nptII, Hygro and Bt 63 rice event. These findings are encouraging and demonstrate that rice grown in kalar track of Punjab, Pakistan, is non-GMO. Such surveillance and monitoring would be largely beneficial to maintain the purity of our traditional non-GM rice and to avoid any possible mixing of GM rice seeds, especially at the time of import. All these efforts will ensure and protect our national share in the international trade of rice.