为了改良多杀菌素生产菌种,提高多杀菌素产量,研究了甘氨酸添加浓度、溶菌酶作用时间、温度和浓度对多杀菌素生产菌刺糖多胞菌Saccharopolyspora spinosaSP06081菌株原生质体制备和再生的影响,并考察了不同再生培养基和渗透压稳定剂对其再生的影响,确定了该菌株原生质体制备和再生的最佳条件。同时,对原生质体再生菌株的形态与多杀菌素产量变化进行了比较研究。结果表明:菌体在添加0.2%的甘氨酸的TSB培养基中培养48h收集,0.1mg/mL溶菌酶,28oC作用20min制备原生质体,将原生质体涂布于以蔗糖为渗透压稳定剂的R2YE培养基中,原生质体再生数目最多,达108个/mL以上;原生质体再生菌株在形态和抗生素产量上产生分化,29.3%的再生菌株形态上保持与亲本菌株一致,具有菌丝松散,断裂分枝多的特点,其中53.2%的再生菌株多杀菌素产量变异向正方向移动,最高产量达到582.0mg/L,比亲本菌株提高85.6%。原生质体再生菌株的形态分化与多杀菌素产量具有重要相关性。 In order to improve spinosad production and spinosad production, the effects of glycine concentration, lysozyme activity time, temperature and concentration on the preparation and regeneration of protoplasts from sporopollen-producing strain Saccharopolyspora spinosaSP06081 were studied The effects of different regeneration media and osmotic pressure stabilizers on their regeneration were investigated. The optimum conditions for protoplast preparation and regeneration were determined. At the same time, the morphological changes of protoplast regeneration strains and spinosad production were compared. The results showed that protoplasts were cultured in TSB medium supplemented with 0.2% glycine for 48h and lysozyme (0.1mg / mL) for 20min at 28oC. Protoplasts were cultured on R2YE with sucrose as osmotic pressure stabilizer In the base, the number of protoplasts regenerated was up to 108 cells / mL. Protoplast regeneration strains were differentiated in morphology and antibiotic production. 29.3% of the regenerated strains remained the same morphologically as the parent strain, with hyphae loosens and rupture branches Among them, 53.2% of the yield of spinosad in regenerated strain moved in the positive direction with the highest yield reaching 582.0mg / L, which was 85.6% higher than that of the parental strain. The morphological differentiation of protoplast regeneration strains has an important correlation with spinosad production.