印度洋深海沉积物可培养放线菌分离样品预处理条件的优化

Optimization of Cultivable Actinomyces Isolated Conditions From Indian Ocean Deep-Sea Sediment

海洋沉积物微生物资源丰富,放线菌是其中的优势类群之一,但是海洋放线菌纯培养难度较大。本文利用响应面法对采自印度洋的深海沉积物样品进行湿热处理条件优化,实验中选取了5个因素设计实验,生长的放线菌经纯化培养后,针对16S rRNA基因进行测序分析,以每种优化条件下培养出的放线菌种类数为响应值,优化结果最终为:湿热处理温度55℃,湿热处理时间1.2727 min,风干时间1.6970 d,抑制剂浓度0.9697倍,海盐添加量为2.1212%。最后,重新选择4种培养基,利用优化后的条件进行该样品纯培养分离,分离出大量放线菌及部分潜在分类单元。实验证明,多种分离方法的组合和优化可提高环境样品中纯培养微生物多样性及出菌率,有助于获得更多稀有微生物资源。本实验可为纯培养分离中不同培养基成分的组合设计提供方法参考。

Marine sediments are rich in microbial resources,in which the phylum Actinobacteria is the dominant group.However,it’s difficult to get pure culture of these marine Actinobacteria.In this study five experimental factors were selected to optimize bacteria separation conditions by response surface methodology for deep-sea sediment sample collected from Indian Ocean.The numbers of cultivable Actinobacteria were taken as the response values for optimization test after sequencing the 16S rRNA gene of isolated strains.The results of optimal conditions were bellow:wet and heat treatment temperature 55℃for 1.2727 min,air drying time 1.6970 days,inhibitor concentration 0.9697 times and sea salt 2.1212%.A large number of Actnobacteria strains and potential strains were isolated and cultivated using reselected four culture media based on optimized conditions.Accordingly,isolating methods and isolation media are the crucial factors to recover the microorganisms inhabiting in the sediment environments.This study provides a reference for the optimization of pure culture conditions of microorganisms.