Hardoim C C P, Costa R, Araújo F V, Hajdu E, Peixoto R, Lins U, Rosado A S, van Elsas J D
Department of Microbial Ecology, Centre for Ecological and Evolutionary Studies, University of Groningen, Kerklaan 30, Groningen 9751 NN, The Netherlands.
Appl Environ Microbiol. 2009 May;75(10):3331-43. doi: 10.1128/AEM.02101-08. Epub 2009 Mar 20.
Microorganisms can account for up to 60% of the fresh weight of marine sponges. Marine sponges have been hypothesized to serve as accumulation spots of particular microbial communities, but it is unknown to what extent these communities are directed by the organism or the site or occur randomly. To address this question, we assessed the composition of specific bacterial communities associated with Aplysina fulva, one of the prevalent sponge species inhabiting Brazilian waters. Specimens of A. fulva and surrounding seawater were collected in triplicate in shallow water at two sites, Caboclo Island and Tartaruga beach, Búzios, Brazil. Total community DNA was extracted from the samples using "direct" and "indirect" approaches. 16S rRNA-based PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analyses of the total bacterial community and of specific bacterial groups--Pseudomonas and Actinobacteria--revealed that the structure of these assemblages in A. fulva differed drastically from that observed in seawater. The DNA extraction methodology and sampling site were determinative for the composition of actinobacterial communities in A. fulva. However, no such effects could be gleaned from total bacterial and Pseudomonas PCR-DGGE profiles. Bacterial 16S rRNA gene clone libraries constructed from directly and indirectly extracted DNA did not differ significantly with respect to diversity and composition. Altogether, the libraries encompassed 15 bacterial phyla and the candidate division TM7. Clone sequences affiliated with the Cyanobacteria, Chloroflexi, Gamma- and Alphaproteobacteria, Actinobacteria, Bacteroidetes, and Acidobacteria were, in this order, most abundant. The bacterial communities associated with the A. fulva specimens were distinct and differed from those described in studies of sponge-associated microbiota performed with other sponge species.
微生物在海洋海绵的鲜重中占比可达60%。据推测,海洋海绵是特定微生物群落的聚集点,但目前尚不清楚这些群落受生物体或环境的影响程度,还是随机形成的。为了解决这个问题,我们评估了与巴西海域常见海绵物种之一黄指海绵(Aplysina fulva)相关的特定细菌群落的组成。在巴西布齐奥斯的卡波克洛岛和塔尔图加海滩这两个地点的浅水区,对黄指海绵及其周围海水样本进行了三次重复采集。使用“直接”和“间接”方法从样本中提取总群落DNA。基于16S rRNA的聚合酶链反应-变性梯度凝胶电泳(PCR-DGGE)分析总细菌群落以及特定细菌类群——假单胞菌属和放线菌——结果显示,黄指海绵中这些菌群的结构与海水中观察到结构截然不同。DNA提取方法和采样地点对黄指海绵中放线菌群落的组成具有决定性作用。然而,从总细菌和假单胞菌的PCR-DGGE图谱中未发现此类影响。从直接和间接提取的DNA构建的细菌16S rRNA基因克隆文库在多样性和组成方面没有显著差异。这些文库总共包含15个细菌门和候选分类群TM7。与蓝细菌、绿弯菌门、γ-和α-变形菌纲、放线菌、拟杆菌门和酸杆菌门相关的克隆序列依次最为丰富。与黄指海绵样本相关的细菌群落是独特的,与其他海绵物种的海绵相关微生物群研究中所描述的不同。