Development And Characterization Of Transgenic Soybean With Insectresistance And Herbicide Tolerance

The rapid development of transgenic technology has revolutionarily altered the breeding methods. Soybeans are one of the "biotech food" crops that have been genetically modified. In 2014, about 82% of global soybeans cultivated for the commercial market States were genetically modified. Facing the great challenge of transgenic soybeans brings to soybean production in the country, development of genetically modified soybean with important agronomic traits is urgent.By using agrobacterium-mediated cotyledonary-node transformation system, we obtained dozens of transgenic soybean plants which contain glyphosate tolerance gene g10-epsps and insect resistant gene bt. In the study, seven major events of these transgenic lines were analyzed for the molecular patterns and agronomic traits. By herbicide spraying, PCR detection and semi-quantitative PCR analysis, we screened positive transgenic plants and determined the leaves of target gene expression. Using Western blot analysis of g10-epspsprotein, lines S8 and L57 were found to be high expressers of the g10-epsps protein. ELISA analysis showed that the expression of bt protein in the lines S8 and L57 is also relatively high in various tissues. Comparison of the fluorescence quantitative PCR (qPCR) and Southern hybridization for determining the transgene copy numbers showed that the qPCR is simple, efficient and applicable for detection of the copy number of exogenous gene.To rapidly determine the transgenic plants we tested the methods that germinated the putative transgenic seeds in filter paper or soil with different contents of the corresponding herbicide. Result showed that using filter paper for seeds germination with 100 mg/L glyphosate solution can rapidly distinguish the transgenic seedlings from the non-transformants. Similarly seeds germination in the soil with 500 mg/L glyphosate solution or 50 mg/L glufosinate solution can also rapidly identified transgenic plants containing the g10-epsps or bar gene, respectively. In addition, we conducted a field test for the herbicide resistance of three glyphosate tolerance lines. Line ZUTS-32, ZUTS-33 were selected for further environmental release test in the coming year.In summary, the study analyzed molecular characters, insect and herbicide resistance of transgenic soybean. The data obtained in the study is an important piece for development of new transgenic soybean varieties of our country.