‘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'

作　　者：孙伟博[1] 于娟[1] 潘惠新[1] 诸葛强[1] 

机构地区：[1]南京林业大学林木遗传与生物技术省部共建教育部重点实验室,南京210037

出　　处：《林业科技开发》2013年第6期85-88,共4页China Forestry Science and Technology

基　　金：国家林业公益性行业科研重大专项(编号:201304102);国家"863"计划项目(编号:2013AA102703)

摘　　要：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.

关 键 词：'南林895杨’ 遗传转化 体系优化 农杆菌介导 

分 类 号：S513[农业科学—作物学]