Abstract
Bt cotton expressing Cry1Ac is being cultivated in Pakistan. It has been observed that pink bollworm may have developed resistance against single Bt gene (Cry1Ac). For durable resistance, insect resistant NIBGE-1601 cotton harboring double gene Cry1Ac-Cry2Ab construct was developed. There was a need to characterize NIBGE-1601 event for intellectual property rights protection. The Presence of NIBGE Cry1Ac and NIBGE Cry2Ab genes was checked in NIBGE-1601 cotton plants through PCR, while there was no amplification using primers specific for Monsanto events (MON531, MON15985, MON1445). Using genome walking technology, NIBGE-601 event has been characterized. Event-specific primers of NIBGE-1601 were designed and evaluated to differentiate it from other cotton events mentioned above. NIBGE-1601 event detection primers are highly specific, therefore, can detect NIBGE 1601 event at different conditions using single or multiplex PCR. In the qualitative PCR, using NIBGE-1601 event specific primers, 0.05 ng was the limit of detection for NIBGE-1601double gene cotton genomic DNA. Thus event characterization and development of event-specific diagnostics will help in breeding new cotton varieties resistant to cotton bollworms.
Highlights
Modified (GM) crops have been developed in many countries of the world for tolerance to biotic and abiotic stresses
NIBGE-1601 cotton event is different from Monsantro cotton events because Monsanto events (MON531, MON15985, MON1445) specific PCR products were not detected in the NIBGE-1601 cotton line (Table 2, Figs. 1, 2, 3, 4, Supplementary Fig. S1–S4)
Positive plants of T1 and T2 progenies of NIBGE-1601 were tested using single and multiplexing PCRs; the results were similar to that of its T0 parent (Supplementary Fig. S6A and S6B). These results indicate that NIBGE-1601 event specific primers are highly specific and can detect NIBGE-1601 event at different PCR conditions using single or multiplexing method
Summary
Modified (GM) crops have been developed in many countries of the world for tolerance to biotic and abiotic stresses. The detection of GMOs has become necessary to comply with labeling regulations because several countries have imposed threshold levels for labeling GM products to meet the demand of traceability and consumers right to know about the product[3]. International regulations make it imperative for the governments, food or feed producers and diagnostic labs to develop reliable methods for detection of GMOs. Genetically modified crops (GMCs) can be commercialized if found safe after thorough biosafety studies. To meet regulatory requirements and properly address the IPRs, NIBGE-1601 cotton event has been successfully characterized using genome walking technology. NIBGE-1601 event specific PCR primers have been evaluated to differentiate from Monsanto and other cotton lines including non-transgenic coker-312 and triple gene NIBGE cotton event
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