花生子叶遗传转化再生体系影响因素的研究

Research of Influence Factors on Peanut Cotyledon Regeneration in Genetic Transformation

本试验选用子叶为外植体研究了影响植株再生和遗传转化的因素。结果表明,细胞分裂素(6-BA)5.00 mg/L+(2,4-D)1.50 mg/L的诱导培养基子叶外植体丛生芽诱导率最高,分别达89.67%和83.33%。遗传转化影响因素结果表明,菌液OD600为0.5-0.7、侵染时间10min、共培养时间3d、每50mL菌液添加烟草提取物1.0mL可获得较好的转化效果。抗生素对芽分化率影响的研究结果表明,当头孢唑林钠(Cef)250 mg/L+羧苄青霉素(Carb)250-500 mg/L时效果最好,不但可以完全抑制农杆菌生长,而且有较高的芽诱导率。

Two peanut cultivars were used as source of cotyledon explants in this experiment. The percentage of shoots induced reached 89.67% and 83.33% respectively in MSC1 with 5.00 mg/L 6-BA and 1.50 mg/L 2,4-D. Transformation efficiency could be affected by many factors in the system of cotyledon explants. The better transformation efficiency was obtained by using Agrobacterium tumefaciens strain LBA4404 with OD600 as 0.5-0.7 and with 10 min infection, 3d co-cultivation and the proper concentration of wounded tobacco leaf extract （1.0 mL/50 mL）. Media with 250 mg/L Cef and 250-500 mg/L Carb could not only lower pollution ratio but also get higher transformation efficiency.