蓖麻子叶节遗传转化研究

摘要

利用前期构建的P450基因的RNAi植物表达载体侵染蓖麻受体材料,旨在获得转P450基因的阳性株.以蓖麻品种通蓖5号为试验材料,利用根癌农杆菌介导法转化蓖麻子叶节,研究影响子叶节遗传转化的若干因素.结果表明:子叶节对Kan较为敏感,适宜筛选质量浓度为250 mg/L；外植体预培养2～3d,侵染用农杆菌菌液OD600=0.8,侵染时间10 min,共培养3d,可获得较高的遗传转化效率.再生植株经PCR和Southern Blot检测,证明为转基因植株.%In order to gain positive strains of transgenic using pre-built RNAi expression vector infection castor receptor material. In this study, several factors affecting transformation rate were studied by Agrobacterium mediated in Tong bi No. 5 castor. The results showed that:cotyledonary node of castor were rather sensitive to kanamycin,and corresponding concentration was 250 mg/L . The pre-culture time of 2 - 3 days,the bacterial concentration of OD600 = 0. 8 was the best inoculum density and 10 minutes was the best inoculation time,co-culture for 3 days were beneficial to improve transformation frequency. The PCR and Southern blot analysis proved that the target gene was suc-essfully integrated into the genome of castor.