Abstract:The herbicide-tolerant genetically modified soybean DBN9004, which was independently developed in China, was granted the agricultural genetically modified organism safety certificate in 2021, demonstrating extensive application prospects. To provide prompt and reliable technical support for the transgenic biosafety supervision of this variety, loop-mediated isothermal amplification (LAMP) technology was employed in this study. By targeting the boundary between the 3' end of the insertion sequence and the soybean genome sequence in DBN9004, a rapid and convenient detection method for DBN9004 components was established. A specific primer set was designed, leading to the development of a rapid detection system characterized by an amplification time of approximately 45 minutes and high stability. This system was demonstrated to effectively distinguish DBN9004 from other transgenic soybean varieties, exhibiting strong specificity. Furthermore, the detection sensitivity for recombinant plasmids containing the DBN9004 sequence was determined to be 10 fg·μL-1, the detection sensitivity for genomic DNA was found to reach 1 ng·μL-1,and the detection sensitivity for positive mixed samples was found to reach 0.5%. The experimental results indicated that the system was characterized by advantages such as short processing time, high stability, good specificity, and high sensitivity. It was thus concluded that this methodology plays a critical role in advancing on-site nucleic acid detection technology for transgenic products of this specific soybean variety.Therefore, a practical and reliable technical foundation is provided by this system for the rapid screening and biosafety regulation of DBN9004.
