Figure S1. Mean alpha rarefaction curves across sediment samples from five sites in Belize. a) Culture-dependent results obtained using seawater agar (SWA) and marine agar (MA) media and b) Culture- independent results. Error bars represent standard error among replicates. a.
200
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Culture dependent MA Culture dependent SWA Culture independent Method
b.
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Culture dependent MA Culture dependent SWA Culture independent Method
Figure S2. Alpha diversity boxplots of marine sediment microbial communities from Carrie Bow Cay, Belize in culture-dependent and culture-independent samples determined using a) Faith’s Phylogenetic Diversity Index and b) Pielou’s Evenness. Culture-dependent methods include the use of marine agar medium (MA) and seawater agar medium (SWA. Data points are overlayed on the boxplot to show variation. a.
100% Proteobacteria Nitrospinae
Bacteroidetes Fibrobacteres
Planctomycetes Entotheonellaeota 90% Cyanobacteria Marinimicrobia (SAR406 clade)
Firmicutes Deinococcus-Thermus
Chloroflexi Margulisbacteria
[A] Thaumarchaeota Unassigned 80% Acidobacteria Archaea UA Verrucomicrobia Acetothermia
Actinobacteria Elusimicrobia 70% [A] Nanoarchaeaeota Tenericutes
Kiritimatiellaeota TA06
Latescibacteria WS2 60% Spirochaetes Dependentiae Patescibacteria LCP-89 Gemmatimonadetes FCPU426
Omnitrophicaeota WPS-2 50% Fusobacteria CK-2C2-2
Bacteria UA Armatimonadetes
[A] Euryarchaeota [A] Altiarchaeota
Relative Percent 40% Calditrichaeota Poribacteria Lentisphaerae Cloacimonetes
[A] Crenarchaeota AncK6 Halanaerobiaeota 30% Zixibacteria [A] Asgardaeota [A] Diapherotrites Dadabacteria GN01
Nitrospirae Eukaryota UA 20% Modulibacteria BHI80-139
Epsilonbacteraeota WOR-1
PAUC34f FBP 10% Chlamydiae Aegiribacteria BRC1 [A] Hadesarchaeaeota
Schekmanbacteria WS1 Hydrogenedentes Eukaryota SAR 0% Culture-Dependent MA Culture-Dependent SWA Culture-Independent Aerophobetes b. 100% Vibrio Shimia Ruegeria Erythrobacter Persicobacter Fulvivirga 90% Microbulbifer Rhodopirellula Alteromonas Urania-1B-19 marine sediment group Rhodobacteraceae UA OM190 UA 80% Pseudoalteromonas Blastopirellula
Woeseia Cobetia Ferrimonas Thiotrichaceae UA 70% Bacillus Winogradskyella Photobacterium Flavobacteriaceae UA
Celeribacter Desulfobacteraceae UA 60% Labrenzia Pelagibius
Halomonas Shewanella
Xenococcus PCC-7305 Tistlia 50% BD7-8 UA Thalassotalea Relative Percent Sva0081 sediment group Nitrosopumilaceae UA Zeaxanthinibacter Alphaproteobacteria UA 40% Kiloniellacea UA Idiomarina Gammaproteobacte UA alpha proteobacterium endosymbiont 2 of Inanidrilus makropetalos Cyclobacteriaceae UA 30% Propionigenium Anaerolineaceae UA Xenococcaceae uncultured Ascidiaceihabitans Flavobacteriales UA Candidatus Nitrosopumilus 20% Desulfobulbaceae UA Pleurocapsa PCC-7319 Endozoicomonas Pseudomonas Actibacter NB1-j UA 10% Desulfobacteraceae UA
Marinobacter Chroococcidiopsis PCC-6712 0% Culture-Dependent MA Culture-Dependent SWA Culture-Independent Figure S3. Relative abundance of microbial communities in marine sediments. a) Phylum level culture- dependent diversity on two growth media (MA and SWA, left) and culture-independent diversity (right). b) Genus level culture-dependent diversity on two growth media (MA and SWA, left) and culture-independent diversity (right). Legends lists a) all phyla and b) the top 50 genera in order from most to least abundant and six rare genera that had a notable percentage in at least one replicate. Note that bar colors repeat for some rare taxa. a.
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b.
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0.85 Pielou’s Evenness (CI Samples) Pielou’s Evenness
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12345 Site (Culture-Independent Samples n=24)
Figure S4. Culture-independent alpha diversity boxplots of marine sediment microbial communities from across five sites around Carrie Bow Cay, Belize determined using a) Faith’s Phylogenetic Diversity Index and b) Pielou’s Evenness. Culture Dependent ASVs (n=50) Culture Independent ASVs (n=24) 1500
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80 85 90 95 100 50 60 70 80 90 100 Percent Sequence Identity (%) Percent Sequence Identity (%)
Figure S5. Histograms of amplicon sequence variant (ASV) from culture-dependent and culture-independent samples and their similarity to previously cultured strains. Cultured representatives included both type and cultured strains extracted from SILVA v138.1. 1 km
Figure S6. Sediment sample collection site information. Domain Phylum Class
5 6 2 1
Culture-Dependent SWA
Culture-Dependent MA 63 243
Culture-Independent 2
Family Species ASV
Order 13 87 100 4 20 151 35 1 6 44 64 27 3 27 5 51 19 14 4 12 33 5
27151 631 1117 2544
Figure S7. Proportional Venn Diagrams denoting the number of taxa detected with culture-dependent and culture-independent methods (marine agar and seawater agar) across taxonomic levels other than genus.