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桑树遗传转化技术中抗生素的浓度优化研究
引用本文:钟名其,楼程富,谈建中.桑树遗传转化技术中抗生素的浓度优化研究[J].汕头大学学报(自然科学版),2001,16(2):1-6.
作者姓名:钟名其  楼程富  谈建中
作者单位:1. 汕头大学生物学系,汕头,515063
2. 浙江大学蚕学系,杭州,310029
3. 苏州大学蚕桑学院,苏州,215151
基金项目:国家自然科学基金资助项目(编号:39570556)
摘    要:调查了抗生素对根癌农杆菌介导的桑品种“新一之濑”遗传转化不同培养阶段外植体生长、分化或生根的影响及其对农杆菌的抑制效果 ,并确定 :在叶盘 (或茎尖 )转化筛选培养阶段 ,Kan (卡那霉素 )和 Cab (羧苄青霉素 )适宜浓度分别为 3 0~ 40 mg/ L和 2 0 0~40 0 mg/ L (或 6 0~ 80 mg/ L和 2 0 0~ 6 0 0 mg/ L ) ,在抗性芽继代 (或生根 )培养阶段 ,Kan和Cab分别为 40~ 5 0 mg/ L和 1 0 0~ 2 0 0 mg/ L (或 5~ 1 0 mg/ L和 5 0~ 1 0 0 mg/ L) .Cef(头孢霉素 )对桑树各种外植体的影响效果与 Cab相似 ,但两者的抑菌效果因农杆菌菌株、外植体类型以及培养阶段的不同而存在差异 ,用含 Kan、Rif(利福平 )和 Str(链霉素 )的 LB培养基浸渍叶盘则出现分化率下降、褐化死亡较严重的现象 .

关 键 词:抗生素  根癌农杆菌  桑树  遗传转化
文章编号:1001-4217(2001)02-0001-06
修稿时间:2001年2月14日

Optimization of antibiotic concentration in genetic transformation technique for mulberry
Abstract:In this paper were investigated the effects of several antibiotics upon the growth and differentiation or rooting of various explants excised from the mulberry cultivar shinichinose at different culture stages of genetic transformation mediated by Agrobacterium tumefaciens.Also investigated were the inhibitive effects of antibiotics upon Agrobacterium tumefaciens.It has been found that the optimum concentrations of kanamycin and carbenicillin are respectively 30~40mg/L and 200~400mg/L(60~80mg/L and 200~600mg/L)at the stage of screening culture where leaf discs or shoot apices were used as explants,and are respectively 40~50mg/L and 100~200mg/L(or 5~10mg/L and 50~100mg/L)at the stage of subculture or rooting of resistance buds;and that the effects of cefotaxime upon the same items under investigation are similar to those of carbenicillin,but its inhibitive effects upon Agrobacterium tumefaciens vary with the strains of Agrobacterium,the types of explant of mulberry,or the stages of culture.It has also been found that leaves,when immersed in LB medium containing kanamycin,rifampicin or streptomycin,have a lower differentiation rate,and in serious cases browning occurs or death ensues.
Keywords:antibiotic  Agrobacterium tumefaciens  mulberry  genetic transformation
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