Xin Yanjuan, Huang Jianyu, Deng Maicun, Zhang Wei
Marine Bioproducts Engineering Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China.
Antonie Van Leeuwenhoek. 2008 Nov;94(4):533-42. doi: 10.1007/s10482-008-9270-y. Epub 2008 Aug 1.
A culture-independent nested polymerase chain reaction (PCR) technique was used to investigate the diversity of actinobacteria communities associated with the sponges Hymeniacidon perleve and Sponge sp. The phylogenetic affiliation of sponge-derived actinobacteria was then assessed by 16S rRNA sequencing of cloned DNA fragments. A total of 196 positive clones were screened by restriction fragment length polymorphism (RFLP) analysis; 48 unique operational taxonomic units (OTUs) were selected for sequencing. Rarefaction analysis indicated that the clone libraries represented 93% and 94% of the total estimated diversity for the two species, respectively. Phylogenetic analysis of sequence data revealed representatives of various phylogenetic divisions, which were related to the following ten actinobacterial genera: Acidimicrobium, Corynebacterium, Propionibacterium, Actinomyces, Micrococcus, Microbacterium, Streptomyces, Mycobacterium, Cellulosimicrobium, Sporichthya, and unidentified actinobacterial clones. A sponge-specific, previously uncultured actinobacteria community grouped within the subclass Acidimicrobidae was discovered from both H. perleve and Sponge sp. Sequences belonging to Acidimicrobium in the H. perleve and the Sponge sp. clone libraries represented 33% and 24% of the clones, respectively. In the Sponge sp. clone library Mycobacterium dominated, accounting for 70% of all clones. The presence of Acidimicrobium and mycobacteria within two sponges can lay the groundwork for attempts to culture these interesting bacteria for industrial applications.
采用一种非培养的巢式聚合酶链反应（PCR）技术，研究与海绵 Hymeniacidon perleve 和海绵物种 Sponge sp. 相关的放线菌群落的多样性。然后通过对克隆的 DNA 片段进行 16S rRNA 测序，评估源自海绵的放线菌的系统发育归属。通过限制性片段长度多态性（RFLP）分析筛选了总共 196 个阳性克隆；选择了 48 个独特的操作分类单元（OTU）进行测序。稀疏分析表明，克隆文库分别代表了这两个物种估计总多样性的 93%和 94%。序列数据的系统发育分析揭示了各种系统发育分支的代表，它们与以下十个放线菌属相关：嗜酸微菌属、棒状杆菌属、丙酸杆菌属、放线菌属、微球菌属、微杆菌属、链霉菌属、分枝杆菌属、纤维微菌属、芽孢鱼菌属以及未鉴定的放线菌克隆。从 Hymeniacidon perleve 和海绵物种 Sponge sp. 中均发现了一个海绵特异性的、以前未培养的放线菌群落，它们聚集在嗜酸微菌亚科内。Hymeniacidon perleve 和海绵物种 Sponge sp. 克隆文库中属于嗜酸微菌属的序列分别占克隆的 33%和 24%。在海绵物种 Sponge sp. 的克隆文库中，分枝杆菌占主导，占所有克隆的 70%。两种海绵中嗜酸微菌属和分枝杆菌的存在可为尝试培养这些有趣的细菌用于工业应用奠定基础。
