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以13种不同基因型的大豆为材料,研究了大豆外植体再生过程中不同大豆基因型对丛生芽数目的影响,选取优势基因型为受体材料,比较了在草丁膦和潮霉素筛选压力下外植体的再生情况,并确定其合适的筛选浓度,在对农杆菌介导的子叶节转化体系优化的基础上进行了大豆转化效率的研究。结果显示:在相同的芽诱导条件下,山宁14产生的丛生芽最多,更适合于大豆子叶节转化;与其它筛选剂相比,采用潮霉素的梯度筛选更有利于抗性苗的成活,其最适筛选浓度为8 mg.L-1;通过检测GUS基因的表达和分子鉴定证明外源的GUS基因已插入到转基因植株的基因组中,转化效率达到3.2%。
Thirteen different soybean genotypes were used to study the effect of different soybean genotypes on the number of clusters of buds during regeneration of soybean explants. The dominant genotypes were selected as acceptor materials. The effects of glufosinate and hygromycin The regeneration of explants under screening pressure was determined and the suitable screening concentration was determined. The conversion efficiency of soybean was studied based on the optimization of Agrobacterium-mediated cotyledonary node transformation system. The results showed that under the same shoot induction conditions, the shoots produced by Shanning 14 were the most suitable for the transformation of soybean cotyledons. Compared with other screening agents, the gradient screening with hygromycin was more conducive to the survival of resistant seedlings , The optimal screening concentration was 8 mg.L-1. The expression of GUS gene and molecular identification proved that exogenous GUS gene was inserted into the genome of transgenic plants, and the transformation efficiency reached 3.2%.