胡杨遗传转化体系的建立及抗生素浓度的优化(英文)

Establishment of genetic transformation system of Populus euphratica and optimization of antibiotic concentration

关键词:  胡杨  遗传转化体系  抗生素浓度  优化

Key words:  Populus euphratica  genetic transformation system  antibiotic concentration  optimization

摘要:

探讨了不同激素浓度对胡杨叶片分化以及卡那霉素、G418、羧苄青霉素和头孢霉素4种抗生素对胡杨不同培养阶段外植体生长、分化或生根的影响,确定了由农杆菌介导的胡杨遗传转化研究中抗生素种类和转化体的筛选浓度,建立了适于胡杨叶片转化的遗传转化体系。结果表明:胡杨叶片再生的最佳培养基为MS+BA 0.5 mg/L+NAA 0.1～0.2 mg/L+白砂糖25 g/L+琼脂5 g/L; 在叶片转化筛选阶段,卡那霉素和G418的适宜浓度分别为10 mg/L和7.5 mg/L,羧苄青霉素和头孢霉素的适宜浓度为200～600 mg/L和200～400 mg/L; 在抗性芽生根培养时,卡那霉素和G418分别为15～20 mg/L和10～15 mg/L,羧苄青霉素和头孢霉素为200～800 mg/L和200～600 mg/L。

Abstract:

Effects of hormones including NAA,IAA,6-BA on leaf differentiation of Populus euphratica and antibiotics including Kanamycin(Kan),G418,Carbenicillin(Cab),Cefotaxime(Cef)on the growth and differentiation or rooting of various explants of P.euphratica were studied. Genetic transformation system mediated by Agrobacterium and optimum antibiotic concentrations for modified organism selection of P.euphratica were founded. Results shown that:the optimal culture medium for leaf differentiation of P.euphratica was MS+BA 0.5 mg/L+NAA 0.1～0.2 mg/L+sugar 25 g/L+agar5 g/L; at co-culture stage which leaves were used as explants,the optimum concentrations of Kan,G418,Cab and Cef were 10 mg/L,7.5 mg/L,200～600 mg/L,200～400 mg/L respectively; at the stage of subculture or rooting of resistance buds,the optimum concentrations were 15～20 mg/L,10～15 mg/L,200～800 mg/L,200～600 mg/L respectively.