正交设计在杨树最佳遗传转化体系的建立  

Establishing Optimum Genetic Transformation System for Poplar with Orthogonal Design

作　　者：蔡诚[1] 吴大强[1] 纵方[1] 项艳[1] 

机构地区：[1]安徽农业大学林学与园林学院,合肥230036

出　　处：《安徽林业科技》2012年第2期40-44,共5页Anhui Forestry Science and Technology

基　　金：安徽省教育厅"十五"学术带头人基金项目(2007年度);国家"十一五"科技支撑项目(2006BAD03A15050-4)资助

摘　　要：本试验以建立的南林95杨高频再生体系为基础,利用正交试验设计,通过GUS组织染色分析法研究了预培养时间、菌液浓度、AS浓度、侵染时间和共培养时间5个因素在4个水平上对南林95杨遗传转化的影响,并探讨了南林95杨转化中添加卡那霉素的适宜浓度。运用DPS软件对试验结果进行分析,建立了杨树遗传转化体系。结果表明:外植体预培养7d,在含AS200μmol/L的菌液浓度为0.6的农杆菌液中侵染20min,共培养3d为最佳遗传转化体系,适宜的卡那霉素筛选浓度为50mg/L;本试验共获得6株转化植株,经GUS染色检测和PCR分析表明,GUS基因已初步整合进入南林95杨基因组中。Based on the established regeneration system of poplar Nanlin 95 （Populus deltoids Bartr. cv. Lux x P. euramericana （Dode）Guineir cv. 1-45 / 51）, the effects of the preculture period, concentrations of bacterium solution, AS concentrations, infection time and co-culture period on the genetic transformation of the poplar variety at 4 levels were studied with orthogonal experiment design and GUS tissue coloration analysis, and adding an optimal concentration of kanamycin in the genetic transformation of the poplar variety was also probed. The experimental data were analyzed with DPS software and a genetic transformation system for poplar was established. The results showed that the optimum genetic transformation system could be achieved with 7-day preculture, 20-minute infection of Agrobacterium （OD6oo = 0.6, 200 μmol/L AS） and 3-day co culture. The selected optimal concentration of kanamycin was 50 mg/L. 6 transgenetic plants were obtained in the experiment and preliminary integration of GUS genes in these poplar Nanhn 95 plants were confirmed by GUS coloration analysis and PCR tests.

关 键 词：杨树 正交设计 遗传转化 农杆菌 

分 类 号：S792.11[农业科学—林木遗传育种]