郑州大学学报（理学版）

研究了抗生素浓度、菌液浓度、侵染时间以及共培养时间对转基因油菜中油821外植体分化的影响,并分别进行了分析,建立了高效的农杆菌介导的油菜遗传转化体系.实验结果表明:使用5 d苗龄的带柄子叶作为转化受体,转化率高;卡那霉素的筛选浓度对转化影响较大,最终确定11 mg/L为该实验筛选的临界浓度;侵染时间为5 min,菌液浓度OD600为0.4时,其外植体转化率最高;共培养时间为48 h时效果最好,不仅有利于外源基因的整合,也不会造成农杆菌的过度增殖;在培养基中分别加入200μmol/L乙酰丁香酮(AS)和5～10 mg/L的硝酸银,可明显提高转化率.

The influence of antibiotic concentration,bacteria liquid concentration,infection time,and cultivation time on the explant differentiation of transgenic napus Zhongyou 821 was researched,and separately analysed.The efficient transformation system of the agrobacterium mediated napus was established.The results showed that,the high transforming condition was achieved when 5-day cotyledons with petiole were used as transformation receptors;the concentration of kanamycin had great influence on transformation,and the critical concentration was 11 mg/L for this experiment selection;the explant had the highest conversion rate when the OD600 of the bacteria concentration was 0.4 and the time of immersion was 5 min;the best cultivation time was 48 h and beneficial to the integration of the exogenous gene and without excessive proliferation of the agrobacterium;and both 200 μmol/L AS and 5~10 mg/L AgNO3 could improve the rate of the conversion.