‘南林895杨’遗传转化体系的优化

The optimization on a genetic transformation system for Populus deltoides × P. euramericana ‘Nanlin 895'

以‘南林895杨’为研究材料,采用正交设计方法,对以农杆菌介导法转基因过程中的几个关键因素进行了筛选比较。并利用GUS染色对转化效果进行统计分析,发现‘南林895杨’合适的遗传转化体系为:预培养3 d,菌液浓度OD600值为0.8,侵染8 min,共培养3 d。通过抗性筛选确定卡那霉素在分化阶段的筛选浓度为20 mg/L,在生根阶段的筛选浓度为10 mg/L,头孢霉素浓度为100 mg/L。优化了‘南林895杨’遗传转化体系,为利用转基因技术开展杨树的遗传改良研究奠定了基础。

Taken Populus deltoides × P. euramericana ‘ Nanlin 895＇ as research material, several key factors of poplar ge-netic transformation mediated by Agrobacterium were secreened by using the orthogonal design method. The statistical analy- sis on transformation effeciency was carried out by using GUS dyeing method. The rsults showed that the genetic transforma-tion system of Populus deltoides × P. euramericana ‘ Nanlin 895＇ was developed including 3 days pre-cuhure, 8 minutes Agrobacterium （OD600 = 0.8） infection and 3 days co-culture. The mediea added with 20 mg/L kanamycin was better in differentiation stage and 10 mg/L kanamyein was better in rooting stage, and the concentration of cefotaxime 100 mg/L was better also.