MPMS诱变结合抗生素抗性选育多杀菌素高产菌株  

Breeding of spinosad high-producing strains through MPMS mutagenesis combined with antibiotics

作　　者：王海霞[1,2] 陈园[2] 王超[2] 黎琪[2] 陈新[1] 张晓琳[2] 

机构地区：[1]武汉轻工大学生物与制药工程学院,湖北武汉430023 [2]国家粮食局科学研究院,北京100037

出　　处：《粮油食品科技》2017年第3期82-86,共5页Science and Technology of Cereals,Oils and Foods

基　　金：粮食公益性行业科研专项(201313002);农业科技成果转化资金项目(SQ2013EC4490009)

摘　　要：采用多功能等离子体诱变系统(MPMS)对菌株ASAGF 13-8-1进行诱变,并通过链霉素、庆大霉素、利福平、氯霉素四种抗生素抗性筛选多杀菌素高产菌株。利用96孔板发酵培养结合生物检测的快速方法进行高产菌株高通量初筛,摇瓶发酵进行复筛。通过5轮复筛验证,获得1株产量较出发菌株提高28.68%且遗传稳定的突变菌14-2。比较MPMS诱变和MPMS诱变结合0.9μg/mL链霉素、14μg/mL庆大霉素、400μg/mL利福平、2μg/mL氯霉素的四重抗性筛选对出发菌株ASAGF 13-8-1的作用效果,结果显示MPMS诱变结合抗生素抗性筛选更有利于多杀菌素高产菌株的选育。Spinosad high - producing strains were breeded from strains ASAGF 13 - 8 - 1 mutated by multi-functional plasma mutagenesis system （MPMS） , combined with streptomycin, gentamicin, chloramphenicol and rifampicin resistance screening. A fast and high throughput screening method of 96 - well plate and bi-oassay as well as the shake flask culture were employed to primary and secondary screen respectively. One mutant 14-2 have been obtained from isolates of MPMS mutagenesis combined with antibiotic of 0.9μg/ mL streptomycin, 14 μg / m L gentamicin, 400μ g / m L rifampicin, 2 μ g / m L chloramphenicol through five rounds rescreening, and showed spinosad production enhanced by 28.68% , compared to original strain. By comparing the impact of MPMS mutagenesis and MPMS mutagenesis combined with antibiotic combination of Str^0.9Gen^14Rif^400Chl^2 on ASAGF 13 -8 -1, the results showed MPMS mutagenesis combined with antibi-otics help breed of spinosad high - producing strains.

关 键 词：刺糖多孢菌 多杀菌素 诱变 抗生素 抗性筛选 

分 类 号：TS201.3[轻工技术与工程—食品科学]