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以中黄20、周豆02005、周豆11和周豆18整体子叶节为材料,以MSB为基本培养基,通过L9(34)正交设计研究了6-BA、IBA和KT 9种浓度组合对大豆整体子叶节丛生芽再生的影响;并将大豆的整体子叶节应用于农杆菌介导的遗传转化研究。结果表明:9种激素组合中以MSB+3.0 mg.L-16-BA+0.1 mg.L-1IBA+0.5 mg.L-1KT最适合周豆02005丛生芽的诱导,而MSB+3.5 mg.L-16-BA+0.2 mg.L-1IBA+0.2 mg.L-1KT最适宜周豆11、周豆18和中黄20的丛生芽诱导;4个大豆基因型中,以中黄20的丛生芽数最多;丛生芽用于诱导生根时,IBA为0.8 mg.L-1、NAA为0.8～1.0 mg.L-1时较适合大豆生根,其中以中黄20生根数最多。以中黄20整体子叶节为材料,进行了农杆菌介导的遗传转化研究。PCR分析和GUS染色初步表明,外源基因转入了大豆的基因组中,转化率为7.8%,得到转基因植株的周期为35～42 d。
The total cotyledon nodes of Zhonghuang 20, Zhoudou 02005, Zhoudou 11 and Zhoudou 18 were used as materials, MSB was used as the basic medium, and 9 concentrations of 6-BA, IBA and KT were studied by L9 (34) On the whole cotyledon cotyledon regeneration of soybean; and the application of the whole cotyledon of soybean in Agrobacterium-mediated genetic transformation. The results showed that MSB + 3.0 mg.L-16-BA + 0.1 mg.L-1 IBA + 0.5 mg.L-1KT was most suitable for induction of cluster shoots of Zhou bean 02005, while MSB + 3.5 mg.L -16-BA + 0.2 mg.L-1IBA + 0.2 mg.L-1KT was most suitable for the induction of cluster shoots in Zhoudou 11, Zhoudou 18 and Zhonghuang 20. Among the four soybean genotypes, When the shoots were used to induce rooting, the rooting rate of IBA was 0.8 mg.L-1 and NAA was 0.8-1.0 mg.L-1, which was more suitable for soybean rooting. The whole cotyledon node of Zhonghuang 20 was used as material to study Agrobacterium-mediated genetic transformation. PCR analysis and GUS staining showed that exogenous genes were transferred into the soybean genome with a conversion rate of 7.8% and the cycle of the transgenic plants was 35-42 days.