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直接以农杆菌转化系统为平台 ,以栽培大豆 (GlycinemaxL )吉林 35、中黄 2 8、南农、铁丰 2 9、铁丰 30和开育 12的子叶节为外植体 ,用LBA4 4 0 4农杆菌 (含pPC KSA质粒 )研究了影响大豆子叶节转化的因素。结果表明 ,大豆品种之间转化存在着差异 ,吉林 35转化率最高 ,平均转化率为 2 16 % ,单次最高转化率达 6 12 % ;中黄 2 8次之 ,平均转化率为1 9% ,单次最高转化率达 3 33% ;南农转化率为 1 0 5 % ;铁丰 2 9居第 4位 ,转化率为 0 95 % ;铁丰 30和开育 12无转化植株。用 4d苗龄的子叶节 ,农杆菌浸染 30min ,共培养 3d比较合适。萌发培养基和不定芽诱导培养基对转化率的影响存在交互作用 ,对于吉林 35来说 ,大豆萌发培养基 (G)和不定芽诱导培养基 (Y)的最佳组配方式为G2 +Y1(转化率 6 12 % )和G1+Y2组合 (转化率 5 2 6 % )。农杆菌介导的转化系统中 ,转化植株的筛选方式对转化率影响很大 ,筛选培养初期 ,采用比较低的筛选浓度 ,在继代培养过程中逐次提高筛选压力 ,可以获得较好的转化效果。
Agrobacterium tumefaciens transformation system was used as the platform to cultivate soybean cotyledon (Glycine maxL) Jilin 35, Zhonghuang 28, Nannong, Tiefeng 29, Tiefeng 30 and C12 under the explants. LBA440 4 Agrobacterium (containing pPC KSA plasmid) The factors affecting the transformation of soybean cotyledons were studied. The results showed that there were differences in the transformation among soybean varieties. The conversion rate of Jilin 35 was the highest, with an average conversion rate of 2 16% and a maximum single conversion rate of 6 12%. The medium conversion rate was 28% with an average conversion rate of 19% , The highest single conversion rate was 333%; the conversion rate of southern farming was 105%; the second was Tiefeng 29, the conversion rate was 0 95%; With 4d seedling age of cotyledon section, Agrobacterium disseminated 30min, co-cultured 3d more appropriate. The effects of germination medium and adventitious bud induction medium on the transformation rate had an interaction. For Jilin 35, the optimal combination mode of soybean germination medium (G) and adventitious bud induction medium (Y) was G2 + Y1 (6 12% conversion) and G1 + Y2 (526% conversion). Agrobacterium-mediated transformation system, the transformation of plants screening methods have a great impact on the conversion rate, the initial selection of screening, the use of relatively low screening concentration in the subculturing process gradually increase the screening pressure, you can get better conversion results .