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为提高农杆菌介导转化大豆子叶节再生体系的遗传转化效率,优化了激素水平、基因型、抗生素及筛选压力等影响植株再生的多个因素,并用EPSPS基因转化大豆子叶节。结果表明,不定芽诱导培养基中6-苄氨基嘌呤(6-BA)浓度为1.6mg/L时,不定芽诱导率最高,黑农37和合丰35的不定芽诱导率较吉育91高,吲哚丁酸(IBA)诱导生根的适宜浓度为0.5～1.0mg/L。头孢霉素的最适抑菌浓度为500mg/L,草甘膦有效筛选压力为8mg/L,并获得转EPSPS基因的抗性植株。
In order to improve the transformation efficiency of soybean cotyledon regeneration system mediated by Agrobacterium tumefaciens, several factors influencing plant regeneration such as hormone level, genotype, antibiotic and screening pressure were optimized and EPSPS gene was used to transform soybean cotyledonary node. The results showed that the induction rate of adventitious buds was highest when the concentration of 6-BA was 1.6mg / L in adventitious buds induction medium, and the induction rate of adventitious buds of Heinong 37 and Hefeng 35 was higher than that of Jiyu 91, Indole butyrate (IBA) suitable for inducing rooting is 0.5 ~ 1.0mg / L. The optimum inhibitory concentration of cefotaxime was 500 mg / L, the effective screening pressure of glyphosate was 8 mg / L, and the plants resistant to EPSPS were obtained.