Establishment of the genetic transformation system of soybean and its optimum conditions

An exogenous gene transformation method used by germinating embryo co-culture with Agrobacterium tumefaciens GV3101 under vacuum permeation conditions was established in different soybean varieties.Several parameters of Agrobacterium-mediated gene transformation were compared to determine the optimum conditions.The results indicated that the optimal pre-training time,infection time,and co-culture time are respective 3 days,6 hours,and 3 days, the optimal concentration of Agrobacterium is OD600=0.6,the optimal concentration of acetosyringone is 200 μmol·L-1.Under optimized conditions,the pCAMBIA1304 vector was transferred by GV3101 into Soybean cv.Ludou 11 and Wei 6823,obtaining 510 Ludou 11 and 591 Wei 6823 regenerated plants,respectively.Among them,444 and 462 plants had the resistance on MB+0.6 mg·L-1 IAA medium with 25 mg·L-1 kanamycin.13 and 15 transgenic plants were identified from the resistance plants by GUS and PCR analysis.Their transformation efficiencies were 2.2% and 2.5% respectively.