Supplementary Figure Legends, Imachi Et Al

1Supplementary Figure legends, Imachi et al.

3Fig. S1. The procedure for inoculation of the sediment slurry into the sponge carriers. (a) The

4sponge carriers were submerged into the sediment slurry and then squeezed by hand to fill the

5sediment slurry. During the filling process, the sediment slurry was flushed by nitrogen gas.

6(b) The color of sponge materials changed from white to brown. (c) The sponge carriers were

7inserted into net-like cylindrical plastic covers and placed into the PVC box.

9Fig. S2. Detailed design of the distributor unit.

11Fig. S3. Sampling of the sponge carriers from the DHS reactor. (a) During the sampling of the

12sponge carriers, nitrogen gas was flushed into the sampling port. The sponge carriers were

13removed from the reactor with tweezers. (b) Sponge carriers after removal. (c) After removing

14the net-like cylindrical plastic covers, sediments were squeezed from the sponges in anaerobic

15synthetic seawater. (d) The sediment slurry was used for further analysis and cultivation.

17Fig. S4. Time course of changes in (a) ORP and (b) pH values of the effluent seawater.

19Fig. S5. Phylogenetic tree of archaeal phylotypes obtained in this study. (a) A tree for most of

20methanogenic and euryarchaeotic phylotypes and (b) an expanded bacterial phylogenetic tree

21for the phylum Crenarchaeota and deeply branching Archaea. Sequences obtained in this

22study are marked in blue (the inoculum sample) and red (the enrichment samples on the DHS

23reactor and batch-type enrichment cultures). The number in parentheses after the phylotypes

24obtained from the inoculum sample indicates the number of identical clones. The number in

25parentheses after the phylotypes obtained from the enrichment samples on the DHS reactor

1 1 1indicate the number of identical clones for each phylotype and the frequency of each clone

2library in the following order; DNA-based clone library from day 357, DNA-based clone

3library from day 560, RNA-based clone library from day 560, DNA-based clone library from

4day 761, and RNA-based clone library from day 761. The number in parentheses after the

5phylotypes obtained from the batch-type enrichment cultures indicates the numbers of

6identical clones obtained per number of clones analyzed for each phylotype. The accession

7numbers are also given after each phylotype name. The scale bar represents the estimated

8number of nucleotide changes per sequence position. The symbols at the nodes show the

9bootstrap values (>70% indicated only) obtained after 1,000 resampling.

11Fig. S6. Phylogenetic tree of deduced McrA amino acid sequences showing the phylogenetic

12positions of clones and isolates obtained in this study. The bar indicates 10% estimated

13sequence divergence. The meanings of the colored sequences and the numbers in parentheses

14are the same as in Fig. 3.

16Fig. S7. Phylogenetic tree of bacterial 16S rRNA gene phylotypes obtained in this study. (a) A

17large bacterial tree for diverse bacterial group and (b-e) expanded bacterial phylogenetic trees

18for the phyla (b) Proteobacteria, (c) Planctomycetes, (d) Firmicutes and (e) Chloroflexi. The

19scale bar represents the estimated number of nucleotide changes per sequence position. The

20symbols at the nodes show the bootstrap values obtained after 1,000 resampling. The

21meanings of the colored sequences and the numbers in parentheses are the same as in Fig. 3.

23Movie S1. The DHS bioreactor system used in this study.