生防放线菌原生质体融合条件的研究

Studies on protoplast fusion between strain of F46 and strain of SC1, SC11

利用重寄生链霉菌F46与生防放线菌SC1和SC11进行原生质体融合,以原生质体的形成率和再生率为指标,研究了Gly、蔗糖、酶及其质量浓度对原生质体融合的影响,并确定了最佳的酶解时间及灭活时间。结果表明,3个菌株培养液中加入Gly和蔗糖的质量浓度以0．5和30 g／mL为宜。对菌株SC1和SC11,分别用10 mg／mL和15 mg／mL,溶菌酶酶解60 min和100 min较合适;对菌株F46,以蜗牛酶(10 mg／mL)和溶菌酶(10 mg／mL)体积比为1:1的混合酶液酶解75 min为宜。菌株F46在55℃条件下热灭活45 min,菌株SC1和SC11则需在55℃条件下热灭活15 min;菌株F46,SC1和SC11的紫外灭活时间分别为16,2和3 min。

The key factors affecting the protoplasts formation and fusants regeneration were determined between hyperparasitic Streptomyces F46 and actinomycetes SC1 or/and SC11. The results showed that the suitable concentration of Gly and sucrose for strain F46, SC1 and SC11 were 0.5 and 30 g/mL . The high rate of protoplasts formation and fusants regeneration could be obtained for strain F46 lysing 75 min with macerozyme of 10 mg/mL snailase and 10 mg/mL lysozyme （1 ： 1） in 35 ℃ water bath, and for strains SC1 and SC11 lysing respectively 60 min and 100 min with 10 mg/mL Lysozyme and 15 mg/mL lysozyme in 35℃ water bath. The period of thermal inactivation in 55℃ was 45 min for strain F46, and was 15 min for stain SC1 and SC11. The suitable inactivation period with UV light was respectively 16, 2 and 3 min for stain F46, SC1 and SC11.