银白杨遗传转化中抗生素浓度优化的研究

Optimization of antibiotic concentration in genetic transformation of Populus alba.

该文探讨了卡那霉素、G418、羧苄青霉素和头孢霉素4种抗生素对银白杨不同培养阶段外植体生长、分化或生根的影响,确立了由农杆菌介导的银白杨遗传转化研究中抗生素种类和转化体的筛选浓度.结果表明:在叶片转化筛选阶段,卡那霉素和G418的适宜浓度分别为15和10mgL,羧苄青霉素和头孢霉素的适宜浓度为200～600mgL和200～400mgL;在抗性芽生根培养时,卡那霉素和G418浓度分别为20～25mgL和15～20mgL,羧苄青霉素和头孢霉素浓度为200～800mgL和200～600mgL.头孢霉素或羧苄青霉素的抑菌效果因农杆菌菌株的不同而存在差异,羧苄青霉素对农杆菌LBA4404抑制效果好,头孢霉素对农杆菌C58抑制效果理想.

Effects of antibiotics,including kanamycin （Kan）, G418, carbenicillin （Cab） and cefotaxime （Cef）, on the growth and differentiation or rooting of various explants of Populus alba were studied. The optimum concentrations of bacterium inhibition and genetically modified organism selection of Populus alba were determined in Agrobacterium-mediated transformation. The optimum concentrations of Kan, G418, Cab and Cef are 15mg/L, 10 mg/L, 200 - 600 mg/L and 200 - 400 mg/L at the stage of screening culture where leaves were used as explants, and are 20 - 25 mg/L, 15 - 20 mg/L, 200 - 800 mg/L and 200 - 600 mg/L at the stage of subculture or rooting of resistant buds. The inhibitive effects of Cef and Cab upon Agrobacterium tumefaciens vary with the strains of Agrobacterium. Cab is a better choice to inhibit Agrobacterium LBA4404 and Cef is more propitious to inhibit Agrobacterium C58.