关键词: 转基因大豆, g10-epsps基因, ZUTS-33, 实时荧光定量PCR, 定量检测

Abstract: ZUTS-33 is the transgenic soybean line developed by Zhejiang University with the modified agronomic trait of herbicide tolerance, which has entered the stage of safety assessment before commercial release. So far, there is no literature report on the detection method of this new transgenic variety, so it is urgent to establish an accurate quantitative detection method to provide technical support for the safety management of agricultural genetically modified organisms. Primers and TaqMan probes were designed according to the specific sequences of  the transgene and the insertion site of the T-DNA in herbicide-resistant soybean ZUTS-33 genome. The specificity, accuracy, precision and repeatability of the primer pairs and probes were evaluated by the optimized real-time fluorescence quantitative PCR detection method, and the detection limit of detection (LOD) and quantitative limit (LOQ) of this detection method were determined. The experimental results showed that the established real-time fluorescence quantitative PCR detection method for transformant specificity of transgenic soybean ZUTS-33 had high specificity of line identification, and accuracy and precision met the requirements, and repeatability was good. LOD and LOQ of the detection method reached 20 copies and 40 copies respectively. The results provided an effective method for identification, detection and monitoring of transgenic soybean ZUTS-33 with the g10-epsps gene.

Key words: transgenic soybean, g10-epsps gene, ZUTS-33, real-time fluorescence quantitative PCR, quantitative detection