Quantification of bacterial and archaeal symbionts in high and low microbial abundance sponges using real-time PCR.

In spite of considerable insights into the microbial diversity of marine sponges, quantitative information on microbial abundances and community composition remains scarce. Here, we established qPCR assays for the specific quantification of four bacterial phyla of representative sponge symbionts as well as the kingdoms Eubacteria and Archaea. We could show that the 16S rRNA gene numbers of Archaea, Chloroflexi, and the candidate phylum Poribacteria were 4-6 orders of magnitude higher in high microbial abundance (HMA) than in low microbial abundance (LMA) sponges and that actinobacterial 16S rRNA gene numbers were 1-2 orders higher in HMA over LMA sponges, while those for Cyanobacteria were stable between HMA and LMA sponges. Fluorescence in situ hybridization of Aplysina aerophoba tissue sections confirmed the numerical dominance of Chloroflexi, which was followed by Poribacteria. Archaeal and actinobacterial cells were detected in much lower numbers. By use of fluorescence-activated cell sorting as a primer- and probe-independent approach, the dominance of Chloroflexi, Proteobacteria, and Poribacteria in A. aerophoba was confirmed. Our study provides new quantitative insights into the microbiology of sponges and contributes to a better understanding of the HMA/LMA dichotomy.

Bayer K ，Kamke J ，Hentschel U 《-》

Chloroflexi bacteria are more diverse, abundant, and similar in high than in low microbial abundance sponges.

Some marine sponges harbor dense and phylogenetically complex microbial communities [high microbial abundance (HMA) sponges] whereas others contain only few and less diverse microorganisms [low microbial abundance (LMA) sponges]. We focused on the phylum Chloroflexi that frequently occurs in sponges to investigate the different associations with three HMA and three LMA sponges from New Zealand. By applying a range of microscopical and molecular techniques a clear dichotomy between HMA and LMA sponges was observed: Chloroflexi bacteria were more abundant and diverse in HMA than in LMA sponges. Moreover, different HMA sponges contain similar Chloroflexi communities whereas LMA sponges harbor different and more variable communities which partly resemble Chloroflexi seawater communities. A comprehensive phylogenetic analysis of our own and publicly available sponge-derived Chloroflexi 16S rRNA gene sequences (> 780 sequences) revealed the enormous diversity of this phylum within sponges including 29 sponge-specific and sponge-coral clusters (SSC/SCC) as well as a 'supercluster' consisting of > 250 sponge-derived and a single nonsponge-derived 16S rRNA gene sequence. Interestingly, the majority of sequences obtained from HMA sponges, but only a few from LMA sponges, fell into SSC/SCC clusters. This indicates a much more specific association of Chloroflexi bacteria with HMA sponges and suggests an ecologically important role for these prominent bacteria.

Schmitt S ，Deines P ，Behnam F ，Wagner M ，Taylor MW ... -  《-》

Removing environmental sources of variation to gain insight on symbionts vs. transient microbes in high and low microbial abundance sponges.

In this study, we pursue unravelling the bacterial communities of 26 sponges, belonging to several taxonomical orders, and comprising low microbial abundance (LMA) and high microbial abundance (HMA) representatives. Particularly, we searched for species-specific bacteria, which could be considered as symbionts. To reduce temporal and spatial environmentally caused differences between host species, we sampled all the sponge species present in an isolated small rocky area in a single dive. The bacterial communities identified by pyrosequencing the 16S rRNA gene showed that all HMA species clustered separated from LMA sponges and seawater. HMA sponges often had highest diversity, but some LMA sponges had also very diverse bacterial communities. Network analyses indicated that no core bacterial community seemed to exist for the studied sponges, not even for such a space and time-restricted sampling. Most sequences, particularly the most abundant ones in each species, were species-specific for both HMA and LMA sponges. The bacterial sequences retrieved from LMA sponges, despite being phylogenetically more similar to seawater, did not represent transient seawater bacteria. We conclude that sponge bacterial communities depend more on the host affiliation to the HMA or LMA groups than on host phylogeny.

Blanquer A ，Uriz MJ ，Galand PE 《-》

Molecular microbial diversity survey of sponge reproductive stages and mechanistic insights into vertical transmission of microbial symbionts.

Schmitt S ，Angermeier H ，Schiller R ，Lindquist N ，Hentschel U ... -  《-》

Molecular diversity of fungal and bacterial communities in the marine sponge Dragmacidon reticulatum.

The present work aimed to investigate the diversity of bacteria and filamentous fungi of southern Atlantic Ocean marine sponge Dragmacidon reticulatum using cultivation-independent approaches. Fungal ITS rDNA and 18S gene analyses (DGGE and direct sequencing approaches) showed the presence of representatives of three order (Polyporales, Malasseziales, and Agaricales) from the phylum Basidiomycota and seven orders belonging to the phylum Ascomycota (Arthoniales, Capnodiales, Dothideales, Eurotiales, Hypocreales, Pleosporales, and Saccharomycetales). On the other hand, bacterial 16S rDNA gene analyses by direct sequencing approach revealed the presence of representatives of seven bacterial phyla (Cyanobacteria, Proteobacteria, Actinobacteria, Bacteroidetes, Lentisphaerae, Chloroflexi, and Planctomycetes). Results from statistical analyses (rarefaction curves) suggested that the sampled clones covered the fungal diversity in the sponge samples studied, while for the bacterial community additional sampling would be necessary for saturation. This is the first report related to the molecular analyses of fungal and bacterial communities by cultivation-independent approaches in the marine sponges D. reticulatum. Additionally, the present work broadening the knowledge of microbial diversity associated to marine sponges and reports innovative data on the presence of some fungal genera in marine samples.

Passarini MR ，Miqueletto PB ，de Oliveira VM ，Sette LD ... -  《-》