大豆子叶节植株再生体系的优化及转EPSPS基因的研究

Optimization of Plant Regeneration System via Cotyledonary Node and Transformation of EPSPS Gene in Soybean

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。

The EPSPS gene was transformed into soybean by Agrobactrium-mediated genetic transformation system with cotyledonary node as explants in soybean.Some major factors,which played important role in improving the regeneration efficiency of soybean,such as plant hormones,genotype,antibiotics and selective pressures were optimized.The result showed that,during the period of shoot induction,a higher shoot regenerated rate and lower deformity rate were obtained with 1.6mg/L 6-BA in this experiment.The rates of regeneration of Heinong37,Hefeng35 were higher than those in Jiyu91.For rooting,the optimal concentration of IBA was 0.5~1.0mg/L.And the optimal concentration of Cefotaxime Sodium in medium was 500mg/L.The effective glyphosate screening concentration was 8mg/L and some of the herbicide resistant soybean plants were obtained.