BackgroundStony corals provide the structural foundation of coral reef ecosystems and are termed holobionts given they engage in symbioses, in particular with photosynthetic dinoflagellates of the genus Symbiodinium. Besides Symbiodinium, corals also engage with bacteria affecting metabolism, immunity, and resilience of the coral holobiont, but the role of associated viruses is largely unknown. In this regard, the increase of studies using RNA sequencing (RNA-Seq) to assess gene expression provides an opportunity to elucidate viral signatures encompassed within the data via careful delineation of sequence reads and their source of origin.ResultsHere, we re-analyzed an RNA-Seq dataset from a cultured coral symbiont (Symbiodinium microadriaticum, Clade A1) across four experimental treatments (control, cold shock, heat shock, dark shock) to characterize associated viral diversity, abundance, and gene expression. Our approach comprised the filtering and removal of host sequence reads, subsequent phylogenetic assignment of sequence reads of putative viral origin, and the assembly and analysis of differentially expressed viral genes. About 15.46% (123 million) of all sequence reads were non-host-related, of which <1% could be classified as archaea, bacteria, or virus. Of these, 18.78% were annotated as virus and comprised a diverse community consistent across experimental treatments. Further, non-host related sequence reads assembled into 56,064 contigs, including 4856 contigs of putative viral origin that featured 43 differentially expressed genes during heat shock. The differentially expressed genes included viral kinases, ubiquitin, and ankyrin repeat proteins (amongst others), which are suggested to help the virus proliferate and inhibit the algal host’s antiviral response.ConclusionOur results suggest that a diverse viral community is associated with coral algal endosymbionts of the genus Symbiodinium, which prompts further research on their ecological role in coral health and resilience.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-017-1084-5) contains supplementary material, which is available to authorized users.

Current research posits that all multicellular organisms live in symbioses with associated microorganisms and form so-called metaorganisms or holobionts. Cnidarian metaorganisms are of specific interest given that stony corals provide the foundation of the globally threatened coral reef ecosystems. To gain first insight into viruses associated with the coral model system Aiptasia (sensu Exaiptasia pallida), we analyzed an existing RNA-Seq dataset of aposymbiotic, partially populated, and fully symbiotic Aiptasia CC7 anemones with Symbiodinium. Our approach included the selective removal of anemone host and algal endosymbiont sequences and subsequent microbial sequence annotation. Of a total of 297 million raw sequence reads, 8.6 million (∼3%) remained after host and endosymbiont sequence removal. Of these, 3,293 sequences could be assigned as of viral origin. Taxonomic annotation of these sequences suggests that Aiptasia is associated with a diverse viral community, comprising 116 viral taxa covering 40 families. The viral assemblage was dominated by viruses from the families Herpesviridae (12.00%), Partitiviridae (9.93%), and Picornaviridae (9.87%). Despite an overall stable viral assemblage, we found that some viral taxa exhibited significant changes in their relative abundance when Aiptasia engaged in a symbiotic relationship with Symbiodinium. Elucidation of viral taxa consistently present across all conditions revealed a core virome of 15 viral taxa from 11 viral families, encompassing many viruses previously reported as members of coral viromes. Despite the non-random selection of viral genetic material due to the nature of the sequencing data analyzed, our study provides a first insight into the viral community associated with Aiptasia. Similarities of the Aiptasia viral community with those of corals corroborate the application of Aiptasia as a model system to study coral holobionts. Further, the change in abundance of certain viral taxa across different symbiotic states suggests a role of viruses in the algal endosymbiosis, but the functional significance of this remains to be determined.

Current research posits that all multicellular organisms live in symbioses with associated microorganisms and form so-called metaorganisms or holobionts. Cnidarian metaorganisms are of specific interest given that stony corals provide the foundation of the globally threatened coral reef ecosystems and their well-being strongly relies on forming mutualistic relationships with endosymbiotic algae of the genus Symbiodinium. So far, only few studies characterized viral diversity and the potential underlying functional importance to coral holobionts. Here we analyzed an existing RNA-Seq dataset of the coral model metaorganism Aiptasia CC7 (sensu Exaiptasia pallida) associated with aposymbiotic, partially populated, and fully symbiotic anemones with Symbiodinium to gain further insight into viral community composition and the relation to the algal endosymbiosis. Our approach included the selective removal of anemone host and algal endosymbiont sequences and subsequent microbial sequence annotation. Of a total of 297 million raw sequence reads, 8.6 million (~ 3%) remained after host and endosymbiont sequence removal. Of these, 3,293 sequences (paired-end read pairs) could be assigned as of viral origin. Taxonomic annotation shows that Aiptasia is associated with a diverse viral community consisting of 116 viral taxa covering 40 families. The viral community was dominated by viruses from the families Herpesviridae (12.00%), Partitiviridae (9.93%), and Picornaviridae (9.87%). Despite an overall stable viral community, we found that some viral taxa significantly changed in relative abundance when Aiptasia engage in a symbiotic relationship with Symbiodinium. Elucidation of viral taxa consistently present in all samples revealed an Aiptasia core virome of 15 viral taxa from 11 viral families that was comprised of many viruses previously reported in coral viromes. Our study provides a first insight into the viral community of Aiptasia. Aiptasia seem to harbor a diverse and overall stable viral community, although certain members change in abundance when the anemone host associates with its algal endosymbiont. However, the functional significance of this remains to be determined. , 2014;Weynberg et al., 2015 Weynberg et al., , 2017 Correa et al., 2016; Levin et al., 2016; 67 Brüwer et al., 2017; Vega Thurber et al., 2017). 6869 Unfortunately, corals are under increasing threat from anthropogenic influences, in particular 70 climate change (Hoegh-Guldberg, 1999; Hughes et al., 2003 Hughes et al., , 2017 IPCC, 2014), and 71 understanding coral metaorganisms is critical in order to mitigate strategies to conserve coral 72 reef ecosystems. To this end, the sea anemone Aiptasia (sensu Exaiptasia pallida) is becoming a 73 popular model system to investigate the coral-dinoflagellate symbiosis (Weis et al., 2008; 74 Voolstra, 2013;Baumgarten et al., 2015). While some studies looked into the association of 109 fully symbiotic anemones, animals were kept in autoclaved and sterile-filtered artificial 110 seawater (AFSW; other c...

Net growth of microbial populations, that is, changes in abundances over time, can be studied using 16S rRNA fluorescence in situ hybridization (FISH). However, this approach does not differentiate between mortality and cell division rates. We used FISH-based image cytometry in combination with dilution culture experiments to study net growth, cell division, and mortality rates of four bacterial taxa over two distinct phytoplankton blooms: the oligotrophs SAR11 and SAR86, and the copiotrophic phylum Bacteroidetes , and its genus Aurantivirga . Cell volumes, ribosome content, and frequency of dividing cells (FDC) co-varied over time. Among the three, FDC was the most suitable predictor to calculate cell division rates for the selected taxa. The FDC-derived cell division rates for SAR86 of up to 0.8/day and Aurantivirga of up to 1.9/day differed, as expected for oligotrophs and copiotrophs. Surprisingly, SAR11 also reached high cell division rates of up to 1.9/day, even before the onset of phytoplankton blooms. For all four taxonomic groups, the abundance-derived net growth (−0.6 to 0.5/day) was about an order of magnitude lower than the cell division rates. Consequently, mortality rates were comparably high to cell division rates, indicating that about 90% of bacterial production is recycled without apparent time lag within 1 day. Our study shows that determining taxon-specific cell division rates complements omics-based tools and provides unprecedented clues on individual bacterial growth strategies including bottom–up and top–down controls. IMPORTANCE The growth of a microbial population is often calculated from their numerical abundance over time. However, this does not take cell division and mortality rates into account, which are important for deriving ecological processes like bottom–up and top–down control. In this study, we determined growth by numerical abundance and calibrated microscopy-based methods to determine the frequency of dividing cells and subsequently calculate taxon-specific cell division rates in situ . The cell division and mortality rates of two oligotrophic (SAR11 and SAR86) and two copiotrophic ( Bacteroidetes and Aurantivirga ) taxa during two spring phytoplankton blooms showed a tight coupling for all four taxa throughout the blooms without any temporal offset. Unexpectedly, SAR11 showed high cell division rates days before the bloom while cell abundances remained constant, which is indicative of strong top–down control. Microscopy remains the method of choice to understand ecological processes like top–down and bottom–up control on a cellular level.