繁茂膜海绵中可培养稀有放线菌的多样性

Phylogenetic diversity of the culturable rare actinomycetes in marine sponge Hymeniacidon perlevis by improved isolation media

摘要:

摘要：【目的】本文旨在尝试改进分离培养方法从大连海域繁茂膜海绵中筛选稀有放线菌，并对其多样性进行研究。【方法】根据繁茂膜海绵元素组成配制微量元素溶液，加入到放线菌分离培养基中，同时将部分培养基稀释成寡营养培养基，结合富集培养法，对繁茂膜海绵中放线菌进行分离培养。采用16S rDNA的限制性片断长度多态性(Restriction Fragment Length Polymorphism, RFLP)分析和序列分析，揭示其多样性。【结果】共获得可培养放线菌59株，通过形态、颜色观察，将其归为27个类群。RFLP分析表现为15种不同的图谱类型。16S rDNA序列分析表明：它们分别属于放线菌的10个属，其中布劳氏菌属（Prauseria）和糖单胞菌属（Saccharomonospora）是首次报道从海绵中分离培养。【结论】改进的分离培养基适合于繁茂膜海绵中稀有放线菌的分离培养，进一步揭示了该海绵中丰富的稀有放线菌，同样的方法有可能应用于其他海绵放线菌的分离培养。

Abstract:

Abstract: [Objective] Based on the molecular diversity information, seven actinomycete-selective culture media and isolation conditions were modified to isolate and cultivate diverse rare actinomycetes from Hymeniacidon perlevis. [Methods] Modified, selective cultivation and enrichment media were used, with the addition of an elemental solution of simulating the elemental composition of marine sponge H. perlevis. Restriction Fragment Length Polymorphism (RFLP) analysis of 16S rDNA sequence was used to reveal the diversity of culturable rare actinomycetes. [Results] A total of 59 actinomycete strains were isolated from the marine sponge H. perlevis. A total of 27 representative actinomycetes were selected according to their morphological feature, color and pigments. They gave 15 different RFLP patterns after digesting their PCR products of 16s rDNA with Hha I. The results showed that these isolates belonged to 10 genera: Streptomyces, Nocardiopsis, Micromonospora, Cellulosimicrobium, Gordonia, Nocardia, Prauseria, Pseudonocardia, Saccharomonospora and Microbacterium. [Conclusion] The modified isolation media and selective cultivation procedures are highly effective in the recovery of culturable actinomycetes from the marine sponge H. perlevis, resulting in the highest diversity of culturable rare actinomycetes from any sponges.