银白杨遗传转化中抗生素浓度优化的研究.pdfVIP

第 27 卷 　第 5 期 北 　京 　林 　业 　大 　学 　学 　报 Vol . 27 , No . 5 2005 年 9 月 JOURNAL OF BEIJ IN G FORESTRY UNIVERSITY Sep . , 2005 银白杨遗传转化中抗生素浓度优化的研究 李 　慧 　陈晓阳　李 　云 　李 　伟 　丁 　霞 北京林业大学林木 、花卉遗传育种教育部重点实验室 摘要 :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 10 mgL ,羧苄青霉素和头孢霉素的适宜浓度为 200～600 mg L 和 200～400 mgL ;在抗性芽生根培养时 ,卡那霉素和 G418 浓度分别为 20～25 mgL 和 15～20 mgL ,羧苄青霉素和 头孢霉素浓度为 200～800 mgL 和 200～600 mgL . 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. 关键词 : 银白杨 , 抗生素 , 遗传转化 中图分类号 :S792119 　　文献标识码 :A 　　文章编号 :1000 1522 2005 05 0118 04 L I Hui ;CHEN Xiaoyang ;L I Yun ;L I Wei ;DIN G Xia . Optimization of antibiotic concentration in genetic transformation of Populus alba . J ournal of B eij ing Forestry University 2005 27 5 118 12 1[ Ch , 9 ref . ] Key Laboratory for Genetics and Breeding in Forest Trees and Ornamental Plants , Ministry of Education , Beij ing Forestry University ,100083 ,P. R . China . Effects of antibiotics ,including kanamycin Kan , G4 18 ,carbenicillin Cab and cefotaxime Cef , on the growth and differentiation or rooting of various explants of Pop ulus alba were studied . The optimum concentrations of bacterium inhibition and genetically modified organism selection of Pop ulus alba were determined in A grobacteriummediated transformation . The optimum concentrations of Kan , G4 18 , Cab and Cef are 15 mgL ,10 mgL , 200 - 600 mgL and 200 - 400 mgL at the stage of screening culture where leaves were used as explants , and are 20 - 25 mgL ,15 - 20 mgL , 200 - 800 mgL and 200 - 600 mgL at the stage of subculture or rooting of resistant buds. The inhibitive effects of Cef and Cab upon A grobacterium tumef aciens vary with the strains of A grobacterium . Cab is a better choice to inhibit A grobacterium LBA4404 and Cef is more propitious to inhibit A grobacterium C58 Key word s 　Pop ulu