Qualitative and Quantitative Molecular Testing Methodologies and Traceability Systems for Commercialised Bt Cotton Events and Other Bt Crops Under Field Trials in India

Abstract

Qualitative and quantitative PCR assays were developed for detection of commercialised Bt cotton events, i.e. MON531, MON15985 and other Bt crops, which are under different stages of field trials in India, i.e. Bt brinjal, Bt rice, Bt cauliflower, Bt potato and Bt okra. Multiplex PCR assays simultaneously detecting specific cry1Ac, cry1Ab, cry2Ab genes, Cauliflower Mosaic Virus 35S promoter, nptII marker gene along with species-or taxonspecific endogenous gene in these Bt crops have been developed. The quantitative real-time PCR assays were also reported for cry1Ac gene using designed primers and TaqMan probe. The sensitivity of developed assays for detection of specific transgene was established up to 0.01%. The analytical methods developed in the present report will be of immense use for qualitative screening and detection of Bt crops along with the quantitative analysis of inserted cry1Ac gene to meet the threshold level for regulatory compliance.

Detection and identification of genetically modified EE‐1 brinjal (Solanum melongena) by single, multiplex and SYBR® real‐time PCR

“…Soya, maize and cotton being the principal GM crops, various PCR‐based methods are available for their detection . The analytical methods reported for the detection of insect‐resistant crops target common genetic elements such as cry1Ac, nptII, CaMV 35S, etc . Although useful for screening, these assays are limited by the fact that several genetically modified organisms (GMOs) have the same detectable elements.…”

“…Although useful for screening, these assays are limited by the fact that several genetically modified organisms (GMOs) have the same detectable elements. The method does not differentiate between Bt crops such as cotton, brinjal, rice, cauliflower, potato and okra . The focus of the present study was to establish a very specific detection method for insect‐resistant EE‐1 brinjal harbouring cry1Ac.…”

“…9 -11 The analytical methods reported for the detection of insect-resistant crops target common genetic elements such as cry1Ac, nptII, CaMV 35S, etc. 12 Although useful for screening, these assays are limited by the fact that several genetically modified organisms (GMOs) have the same detectable elements. The method does not differentiate between Bt crops such as cotton, brinjal, rice, cauliflower, potato and okra.…”

“…The method does not differentiate between Bt crops such as cotton, brinjal, rice, cauliflower, potato and okra. 12 The focus of the present study was to establish a very specific detection method for insect-resistant EE-1 brinjal harbouring cry1Ac. An event-specific PCR based on the 3 flanking region for the unequivocal identification and quantification of insect-resistant brinjal EE-1, employing both a conventional and a real-time PCR format, is described.…”

Detection of Unlabeled Genetically Modified Soybean in the Omani Market

“…In order to support legislation, reliable, accurate methods for identification of GMOs in raw materials and food products are required [5,7,8]. Many analytical methods are available, including PCR methods for screening and for qualitative and quantitative detection of GMOs [5,9,10,11]. A review of methods has been published [12].…”