Establishment of Accurate Quantitative Detection Method for Herbicide-tolerant Soybean (Glycine max) 'DBN9004'
ZHAO Xin1, LIU Shuang1, LIU Na1, LI Rui-Huan1, GAO Fang-Rui1, LAN Qing-Kuo1, LIANG Jin-Gang2,*, WANG Yong1,*
1 Tianjin Academy of Agricultural Sciences, Tianjin 300384, China; 2 Development Center of Science and Technology, Ministry of Agriculture and Rural Affairs, Beijing 100176, China
Abstract：Herbicide-tolerant soybean (Glycine) 'DBN9004' is a new genetically modified soybean event with tolerance to both glyphosate and glufosinate. It has obtained safety certificate and has important industrial application prospect in China. In order to meet the technical requirements of safety supervision for genetically modified crops, it is urgent to develop quantitative detection methods for 'DBN9004'. In this study, the 3' flanking sequence of herbicide-tolerant soybean 'DBN9004' was used as the target, primers and probes were designed, and a qPCR assay was established. The specificity, accuracy, precision, detection limit, quantitative limit and other indicators were tested. The results showed that the method was specific and reproducible, with a quantitative limit of 0.1% (40 copies) and a detection limit of 0.05% (20 copies) , the relative standard deviation (RSD) was 6.00%~10.95%, and the quantitative results of qPCR and droplet digital PCR method were consistent. This method provides a new technical means for the accurate quantitative detection of new herbicide-tolerant soybean event 'DBN9004', and provides technical support for the regulation of agricultural transgene crops.
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