DOI: 10.5061/dryad.hf33q77
title: Data from: Diel patterning in the bacterial community associated with the sea anemone Nematostella vectensis

description: Microbes can play an important role in the physiology of animals by
 providing essential nutrients, inducing immune pathways, and influencing
 the specific species that compose the microbiome through competitive or
 facilitatory interactions. The community of microbes associated with
 animals can be dynamic depending on the local environment, and factors
 that influence the composition of the microbiome are essential to our
 understanding of how microbes may influence the biology of their animal
 hosts. Regularly repeated changes in the environment, such as diel
 lighting, can result in two different organismal responses: a direct
 response to the presence and absence of exogenous light and endogenous
 rhythms resulting from a molecular circadian clock, both of which can
 influence the associated microbiota. Here, we report how diel lighting and
 a potential circadian clock impacts the diversity and relative abundance
 of bacteria in the model cnidarian Nematostella vectensis using an
 amplicon-based sequencing approach. Comparisons of bacterial communities
 associated with anemones cultured in constant darkness and in light:dark
 conditions revealed that individuals entrained in the dark had a more
 diverse microbiota. Overall community composition showed little variation
 over a 24-hour period in either treatment; however, abundances of
 individual bacterial OTUs showed significant cycling in each treatment. A
 comparative analysis of genes involved in the innate immune system of
 cnidarians showed differential expression between lighting conditions in
 N. vectensis, with significant up-regulation during long-term darkness for
 a subset of genes. Together, our studies support a hypothesis that the
 bacterial community associated with this species is relatively stable
 under diel light conditions when compared with static conditions and that
 particular bacterial members may have time-dependent abundance that
 coincides with the diel photoperiod in an otherwise stable community.
description: Supplemental
 FiguresSupplemental Figure 1.
 Alpha rarefaction curves for LD and DD entrained NV. Alpha rarefaction
 curves for the associated microbiota for NV that were entrained to either
 a 12-hour light: 12-hour dark light cycle (red squares) or 24 hours of
 darkness (blue circles) based on the rarefaction depth (1,080 sequences)
 used for all PCoA plots. Supplemental Figure 2. Number of OTUs for the
 ‘core’ bacterial community. Enumeration of the bacterial phylotypes of the
 ‘core’ bacterial community associated with NV for LD and DD entrained
 individuals for 60%, 70%, 80%, 90%, and 100% of sampled anemones.
 Supplemental Figure 3. Beta-diversity statistics for ‘core’ diversity
 analyses. Representation of statistical comparisons (ANOSIM) between the
 ‘core’ (at 60%, 70%, 80%, 90%, and 100%) bacterial communities associated
 with LD and DD entrained NV. Supplemental Figure 4. ‘Core’ bacterial taxa
 associated with adult NV across treatments. Classes of bacteria associated
 with LD and DD entrained NV for the ‘core’ community at the 60%, 70%, 80%,
 90%, and 100% level.Leach_etal_SuppFigures.docxSupplemental NoteThis Supplementary Note outlines the bioinformatic pipeline used to analyze the 16S rRNA data in this study via QIIME.Leach_etal_SuppNote.txtSupplemental TablesSupplemental Table 1. PCR recipe and thermal conditions for 16S rRNA amplification. Supplemental Table 2. Differentially abundant OTUs between LD and DD entrained NV. Supplemental Table 3. Alpha diversity estimates of LD and DD entrained NV and corresponding statistics.Leach_etal_SuppTables.xlsxRaw Sequence FilesAdult Nematostella vectensis were used for these experiments. Individuals were reared at room temperature and ambient lighting conditions. In preparation for the experiment, adults were split into glass dishes and transferred to an incubator at 25C. Full spectrum LED lights were used to simulate diel light (LD). One group of animals were exposed to LD conditions and the second group was maintained in complete darkness (DD) for 30 days. Four (4) replicates from each condition were sampled every 4-hour for a total of 11 time points. Using high throughput sequencing of 16S amplicons representing the bacterial communities associated with N. vectensis, we assayed community composition and membership as well as cycles of differential abundance.Dataset.zip