抗生素对细枝木麻黄愈伤组织再生及农杆菌生长的影响

Effects of Antibiotics on Callus Regeneration of Casuarina cunninghamiana and Inhibition of Agrobacterium tumefaciens

以细枝木麻黄3个无性系水培苗作为材料,研究了硫酸卡那霉素(kanamycin sulfate,Km)对细枝木麻黄愈伤组织诱导、增殖和芽分化的影响,以及头孢霉素(cefotaxime,Cef)、羧苄青霉素(carbenicillin,Cab)和氨苄青霉素(ampicillin,Amp)对农杆菌的抑制效果。结果表明,Km对细枝木麻黄愈伤组织诱导适宜的选择压力是20mg/L,Km选择性筛选愈伤组织增殖的适宜质量浓度为40mg/L,愈伤组织芽分化的选择压力是40～50mg/L。在抗生素抑菌效果方面,Cef质量浓度为200mg/L、Cab300mg/L和Amp500mg/L能完全抑制农杆菌的生长。综合比较,对细枝木麻黄遗传转化研究最理想的抑菌剂是Cef,使用质量浓度低且能有效抑制根癌农杆菌的生长。

An experiment was conducted to study the effects of kanamycin sulfate on callus induction,callus proliferation and bud differentiation of three clones of Casuarina cunninghamiana as well as the inhibitory effects of cefotaxime,carbenicillin and ampicillin on the bacterial growth of Agrobacterium tumefaciens C58C1.Results showed that the optimum concentration of kanamycin sulfate was 20 mg/L for callus induction,40 mg/L for callus proliferation,and 40-50 mg/L for bud differentiation.The growth of bacterium could be completely inhibited when concentrations of cefotaxime,carbenicillin and ampicillin were 200,300 and 500 mg/L respectively.Comprehensive comparison of antibacterial effects of the three antibiotics shows that cefotaxime is the best bacteriostat for the genetic transformation of C.cunninghamiana,and low concentration of cefotaxime can inhibit the bacterial growth of A.tumefaciens C58C1.