关键词: 大豆, 根瘤农杆菌, 外植体, 转化

Abstract: To develop a rapid and high efficient technique for soybean transformation, this report concentrated on resolving intractable problems, such as explants tissue culture, Agrobacterium tumeficiences infection and plant regeneration. Mature soybean seeds were germinated on MSB5-medium supplemented with 1 mg/L N6-benzylaminopurine (BAP) at 25~28℃ for 24 h and the explants were pre-cultured at the same condition as above. Explants were inoculated with Agrobacterium tumeficiences strain GV3101 harboring the vector pBI121 and cultured at 28 ℃ for 21 h in MSB5-medium suspension supplemented with 1mg/L N6-benzylaminopurine (BAP) and 100 μM acetosyringone. After co-cultivation for 3 days in the dark at 28 ℃, transformants were transferred to sterilized soil directly and incubated with suitable light and temperature conditions. The results of molecular test indicated that the regeneration ratio of this system was more than 90% (95% for Zhongpin661, 93% for Kennong18 and 90% for Lv75, respectively) compared to cotyledonary nodes no more than 50% and embryonic tips no more than 70%, and the transformation ratio was as high as 10% (Kennong18). Furthermore, as no complicated and time-consuming tissue culture operations were used in the latter cultivation for transformants, the whole regeneration period were shortened evidently. Therefore, this technique system for soybean transformation mediated by Agrobacterium tumeficiences was more fast and high efficient than others, and a powerful technical platform for Agrobacterium tumeficiences-mediated genetic transformation and mutant construction of genomic research in soybean.

Key words: Soybean (Glycine max L. Merrill), Agrobacterium tumefaciens, Explant, Transformation