Supplementary Information

Time series analysis of two hydrothermal plumes at 9˚50'N East Pacific Rise reveals distinct, heterogeneous bacterial populations

Jason B Sylvan1*, Benjamin C Pyenson2, Olivier Rouxel3,4, Christopher R German3 and Katrina J Edwards1

1University of Southern California, Department of Biological Sciences, Marine

Environmental Biology Section, 3616 Trousdale Parkway, Los Angeles, CA 90089

2Haverford College, 794 College Avenue, Haverford, PA 19041

3Woods Hole Oceanographic Institution, 266 Woods Hole Rd, Woods Hole, MA 02543

4 Université Européenne de Bretagne, Université de Brest, Institut Universaire Européen de la

Mer (IUEM), UMR 6538, Insititut Français de Recherche pour l’Exploitation de la Mer

(IFREMER), BP 80 F- 29280 Plouzané, France

*email: [email protected]

Supplementary Information

Sylvan et. al. Supplementary Information "!

Table S1- Distribution of OTUs in clone libraries from this study. OTUs with an asterisk (*) next to their name are from the group "other (non-plume)" in Fig. 2.

Number of clones from Phylogeny Isolated From OTU Accession Number 1-8 1-13 1-21 2-8 2-13 2-21 Aquificae Desulfurobacterium sp. str. SL19 AY268937 113A07 1 Hydrogenobacter subterraneus str. AB026268 HGP1 28E03 1 Aquifex pyrophilus str. Kol5a M83548 28G03 1

GNO2 Helicobacter anseris str. MIT 04-9362 DQ415545 213F05 1

Firmicutes Spiroplasma culicicola str. AES-1 NR_025701 113F05 1 Umboniibacter marinipuniceus str. KMM 3891 AB467279 113H05 1 4 Streptococcus australis str. ATCC 700641 AY485604 121H11 1 Fusibacter paucivorans str. SEBR oil well NR_024886 4211 213H09* 1 Geobacillus pallidus str. B1 oil resevoir HM030740 213G05* 1

Actinobacteria Rothia mucilaginosa str. J04 tumulus DQ409140 18B07 1 Propionibacterium acnes str. SK137 skin CP001977 121G01 1 Sylvan et. al. Supplementary Information #!

Cyanobacteria chloroplast 213E10* 1 Synechococcus 213H01 4 4 9 28H06 1 28F08 2 28F07 1 Synechocystis 213C03* 1 Prochlorococcus 18E05 1 1 28A04 2 213E04* 1 213H02* 1 213A04* 1

Acidobacteria Anaeromyxobacter dehalogenans str. 2CP-1 marine sediment NC_011891 121D12 1 Geobacter sp. str. FRC-32 marine sediment NZ_AASH01000049 113B09 2

Verrucomicrobia Anaeromyxobacter dehalogenans str. 2CP-1 sediment NC_011891 28E02 1 Rubritalea sp. str. MN1-1042 marine sediment AB543685 18E09 2 18A06 1 18A05 1 213D04* 1 Rubromonas marinus str. Pol012 marine sponge DQ302104 18D11 1 18G03 1 unclassified Verrucomicrobia 213E03* 1

Planctomycetes Phycisphaera mikrensis str. NBRC102666 marine alga AB447464 28G11 1 Sylvan et. al. Supplementary Information $!

Planctomyces maris str. DSM 8797 marine ABCE01000043 28F10 1 28C09 1 Piruella sp. str. 302 chalk mine X81942 213C10* 1 Piruella marina deep marine X62912 sediments 28C05 1 Piruella sp. str. 139 X81945 28B04 1 28F03 1 Rhodopiruella sp. str. SM49 marine FJ624355 113F02 1 1 18E10 1 4

Deferribacteres Calditerrivibrio nitroreducens str. Yu37-1 hot spring AB364234 18E11 1

Bacteroidetes Owenweeksia hongkongensis str. UST20020801 marine AB125062 18D04 1 113D02 2 28A06 1 Brumimicrobium mesophilum str. YH207 marine sediment DQ660382 113H12 1 Polaribacter dokdonensis str. MED152 marine DQ481463 18E07 1 28E09 1 Flexibacter echinicida str. F11 sea urchin AY006470 18F01 1 213E07 1 1 Algibacter mikhailovii str. LMG 23988 marine AM491809 113G12 1 Sylvan et. al. Supplementary Information %!

Winogradskyella sp. str. NF1-23 marine sediment FJ196009 28D10 1 deep-sea sediment clone BD2-17 deep-sea sediment AB015545 28D07 1 28G06 1 Winogradskyella poriferia str. UST030701-295 microbial mat AY848823 28C02 1 28A09 2 18C10 1 Flexibacter tractuosus str. IFO 15981 AB078071 18G08 1 Alvinella pompejana Cytophaga sp. str. Dex80-37 tubes AJ431253 18B01 1 deep-sea sediment clone BD2-2 deep-sea sediment AB015532 113D09 1 Fluviicola taffensis DSM 16823 freshwater CP002542 213B04* 1 Cellulophaga sp. str. MED134 marine AAMZ01000009 18C04 1

Deltaproteobacteria unclassified Deltaproteobacteria 213F03* 1 Desulfatibacillum alkenivorans str. sediment CP001322 AK-01 18D09 1 Geobacter metallireducens str. GS-15 freshwater sediment AAAS03000001 28B07 1 Geobacter bemidjiensis str. Bem freshwater sediment CP001124 113D05 1 Desulfotalea psychrophila str. LSv54 marine sediment NC_006138 18E01 1 113E11 1 Desulfobulbus mediterraneus str. 86FS1 marine sediment AF354663 113B11 1 1

Epsilonproteobacteria Sylvan et. al. Supplementary Information &!

Caminibacter mediatlanticus str. TB-2 deep-sea vent NZ_ABCJ01000002 121G08 1 113E08 1 18C07 1 28D01 1 121E07 1 113C03 1 113C10 3 1 Caminibacter profundus str. CR deep-sea vent AJ535664 113E01 1 Caminibacter hydrogeniphilus str. deep-sea vent AJ309655 AM1116 113E07 1 Nautilia profundicola str. AmH deep-sea vent CP001279 113F07 1 Nitratifractor salsuginis str. E9I37-1 deep-sea vent AB175500 28G01 1 1 1 121B08 2 113C07 1 Hydrogenimonas thermophila str. EP1-55-1 deep-sea vent AB105048 28F12 1 Arcobacter sp. str. MA5 abalone AB542077 213D07 1 28H10 1 18F04 1 1 Sulfurimonas autotrophica str. OK10 hydrothermal AB088431 sediments 18G01 3 2 1 1 18C03 1 1 1 28G04 1 18E03 3 1 18C06 1 1 28B05 1 121F08 1 Sulfurimonas denitrificans str. ATCC marine CP000153 33889 113A03 1 18A09 1 2 Sylvan et. al. Supplementary Information '!

213C08 1 18H12 4 2 18F05 1 3 Sulfurimonas autotrophica str. OK5 hydrothermal AB088432 sediments 113B04 1 18B08 2 18D03 3 3 28F05 1 113A02 1 Sulfurovum sp. str. NBC37-1 deep-sea vent AP009179 113A08 3 18B04 1 3 1 18G06 1 1 18A02 2 3 121A12 1 18H07 1 2 113F09 2 113C11 4 1 1 18E08 1 1 113H03 1 113D10 1 18D12 1 28B06 1 28E04 1 113C12 1 28B11 1 18G09 1 7 28F06 1 18D01 1 1 28C04 1 28D12 1 18B11 9 Sulfurovum lithotrophicum str. 42BKT hydrothermal NR_024802 sediments 28D09 1 28C12 1 113G06 2 28C03 1 Sylvan et. al. Supplementary Information !"

221B05 1 18G12 1 18B10 1 1 28B10 1 18G04 1 4 18H01 1 2 1 1

Zetaproteobacteria Mariprofundus ferrooxydans str. PV-1 Loihi Seamount AATS01000005 221C01 1

Alphaproteobacteria Kopriimonas byunsanensis str. KOPRI marine biofilm DQ167245 28B01 1 Erythrobacter longus str. EG6 hypersaline spring AM691106 18D02 1 2 5 1 213A06* 2 213D06* 1 Erythrobacter citreus str. RE25F/1 marine AF118020 221D02* 1 Stella humosa str. DSM 5900 marine NR_025582 213B08* 1 Croceitomaculum antarcticum str. marine EF495229 IMCC3195 113F11 1 Silicibacter sp. str. LS103 marine sponge FJ937914 18C02 2 2 121G02 1 Sulfitobacter dubius str. KMM 3554 sea grass NR_025691 221A01* 22 221F07* 1 221G04* 1 221H02* 1 221A06* 1 221F01* 1 Octadecabacter sp. str. NF3-22 marine FJ196058 213G09* 1 Thalassobius sp. str. ZS6-23 marine FJ196059 18B02 3 3 Sylvan et. al. Supplementary Information !"

18B06 1 Ruegeria atlantica str. SS-05 marine sediments EU624444 28H02 1 28G08 1 28G07 1 1 213B10* 1 213C01* 3 Pseudomonas sp. str. MK12 rhizosphere AY690720 213F02* 2 Maricaulis maris str. ATCC 15269 marine AJ227803 221C05* 1 marine Dinoroseobacter shibae str. DFL 12 dinoflagellates AAVE01000004 213E09* 3 Roseobacter sp. str. P81 marine sediments EU195951 113F03 1

Betaproteobacteria Lautropia sp. str. TeTO human oral cavity FJ823153 121G03 1

Gammaproteobacteria Codakia orbicularis gill symbiont clam symbiont X84979 28F11 1 Calyptogena phaseoliformis symbiont sulfur oxidizing str. KT-2 symbiont AB479082 113G05 1 Legionella pneumophila str. Lens CR628337 113E02 1 121D05 1 Haliea sp. str. SY02 marine sediment GU124598 113D11 1 113B10 1 2 213D03* 1 Vibrio splendidus str. ATCC33789 marine AB038030 213B02* 2 Photobacterium leiognathi str. RM1 squid symbiont AY292947 18A04 1 1 8 213B09* 1 Sylvan et. al. Supplementary Information !"#

Pseudoalteromonas sp. str. JL-S1 marine AY745839 18F06 1 113B08 1 2 Colwellia psychrerythraea str. 34H marine CP000083 28C07 1 28C08 1 213C07* 4 213E11* 1 Alcanivorax sp. str. Oha-1 marine AB053130 121E02 4 Marinobacter flavimaris str. SW-145 marine AY517632 121A02 25 15 121B06 10 121E08 2 2 121A01 4 121C06 1 121B03 1 121H05 1 Marinobacter flavimaris str. 2PR54-4 marine EU440995 121H01 1 121B07 1 Marinobacter sp. str. EZ44 Prochlorococcus co- EU591709 culture 121D04 1 1 121D10 2 121B12 2 121H02 1

Marinobacter sp. str. NT N31 marine AB166980 121C12 1 121G09 1 121H07 1 121D01 1 121B05 1 221G02* 1 221G07* 1 221G10* 1 Marinobacter exellens str. KMM 3809 marine sediments AY180101 221E01* 1 Sylvan et. al. Supplementary Information !!"

Marinobacter lutaoensis str. T5054 shallow marine vent NR_025116 18D05 2 Halomonas sulfidaeris str. Esulfide1 deep-sea vent AF212204 221A12* 2 Halomonas meridiana str. PR51-13 marine EU441001 221A10* 3 221F06* 1 Umboniibacter marinipuniceus str. KMM 3892 mollusc AB467280 113H01 1 1 Saccharophagus degradans str. 2-40 marine NC_007912 18A03 1 1 113E10 1 213E02* 2 Kangiella koreensis str. DSM 16069 marine tidal flats CP001707 113G04 1 Cellvibrio japonicus str. Ueda107 soil CP000934 113H06 1 Cycloclasticus pugetii str. PS-1 marine sediments NR_025955 113C01 1 Alteromonas genoviensis str. LMG marine biofilm AM887686 24079 221E03* 1 Psychromonas profunda str. 2825 marine sediments AJ416756 213F04* 1 Thioalkalivibrio sp. str. HL-EbGR7 soda lake sediment CP001339 113E12 2 Sylvan et. al. Supplementary Information !"#

Table S2 - Diversity indices as calculated by MOTHUR. Ranges given are 95% confidence intervals for each index. # Sample n Chao1 Ace Inverse Simpson 1-8 83 120-446 320-651 31-156 1-13 89 94-249 92-208 60-156 1-21 79 54-207 63-269 5.0-14 2-8 70 144-623 611-1276 38-589 2-13 78 57-145 113-230 22-64 2-21 64 66-440 49-338 4.3-11

Sylvan et. al. Supplementary Information !"#

Table S3- Relative hydrothermal component of the particulates collected (calculated as the ratio of Fe to (Fe+Mn+Al)). Samples were not collected from Trap R2 beyond sample R2-11 for Fe/(Fe+Mn+Al).

|------Fe/(Fe+Mn+Al) ------| Trap R1, Trap R1, Trap R2, Trap R2, Sample Bottle Sampling Day >1mm <1mm >1mm <1mm 1 6 0.92 0.93 0.63 0.67 2 12 0.86 0.92 0.56 0.69 3 18 0.85 0.90 0.77 0.71 4 24 0.79 0.91 0.63 0.57 5 30 0.84 0.88 0.71 0.61 6 36 0.90 0.96 0.88 0.68 7 42 0.93 0.97 0.78 0.66 8 48 0.91 0.97 1.02 0.78 9 54 0.94 0.92 0.54 0.62 10 60 0.78 0.90 0.55 0.43 11 66 0.77 0.80 0.54 0.43 12 72 0.93 0.89 13 78 0.96 0.93 14 84 0.91 0.92 15 90 0.92 0.93 16 96 0.95 0.88 17 102 0.83 0.94 18 108 0.90 0.93 19 114 0.95 0.92 20 120 0.97 0.95 21 126 0.88 0.91 mean 0.89 0.92 0.69 0.62

Sylvan et. al. Supplementary Information !"#

Table S4 - Ecophysiology of Zetaproteobacteria, Alphaproteobacteria and Gammaproteobacteria clones recovered from time points 13 and 21. Included are only clones that were present during at least time point 13 or 21. Ecophysiology was deduced from papers referenced in the fourth column. For strains referenced as "unknown (NCBI)", these strains have been deposited into the NCBI database but not yet published in the primary literature.

Number of clones from Phylogeny Ecophysiology OTU Reference 1-8 1-13 1-21 2-8 2-13 2-21 Alphaproteobacteria Erythrobacter longus str. EG6 aerobic anogygenic Csotonyi et al., 2008 phototroph 18D02 1 2 5 1 213A06* 2 213D06* 1 Erythrobacter citreus str. RE25F/1 heterotroph Denner et al., 2002 221D02* 1 Stella humosa str. DSM 5900 heterotroph Vasilyeva, 1985 213B08* 1 Croceitomaculum antarcticum str. heterotroph Lee et al., 2007 IMCC3195 113F11 1 Silicibacter sp. str. LS103 unknown unknown (NCBI) 18C02 2 2 121G02 1 Sulfitobacter dubius str. KMM 3554 sulfite oxidizer Ivanova et al., 2004

221A01* 22 221F07* 1 221G04* 1 221H02* 1 221A06* 1 221F01* 1 Octadecabacter sp. str. NF3-22 unknown unknown (NCBI) 213G09* 1 Thalassobius sp. str. ZS6-23 unknown unknown (NCBI) 18B02 3 3 Ruegeria atlantica str. SS-05 likely S-oxidizer Newton et al., 2010 28G07 1 1 213B10* 1 Sylvan et. al. Supplementary Information !"#

213C01* 3 Pseudomonas sp. str. MK12 unknown unknown (NCBI) 213F02* 2 Maricaulis maris str. ATCC 15269 unknown unknown (NCBI) 221C05* 1 Dinoroseobacter shibae str. DFL 12 S-oxidizer Newton et al., 2010 213E09* 3 Roseobacter sp. str. P81 likely S-oxidizer Newton et al., 2010 113F03 1

Gammaproteobacteria Calyptogena phaseoliformis symbiont sulfur oxidizing Fujikura et al., 1999 str. KT-2 symbiont 113G05 1 Legionella pneumophila str. Lens human pathogen Cazalet et al., 2004 113E02 1 121D05 1 Haliea sp. str. SY02 unknown unknown (NCBI) 113D11 1 113B10 1 2 213D03* 1 Vibrio splendidus str. ATCC33789 mollusc pathogen Urakawa et al., 1999

213B02* 2 Photobacterium leiognathi str. RM1 luminescent unknown (NCBI) symbiont 18A04 1 1 8 213B09* 1 Pseudoalteromonas sp. str. JL-S1 heterotroph Du et al., 2006 113B08 1 2 Colwellia psychrerythraea str. 34H psychrophilic Huston et al., 2000 heterotroph 213C07* 4 213E11* 1 Alcanivorax sp. str. Oha-1 petroleum degrader unknown (NCBI) 121E02 4 Marinobacter flavimaris str. SW-145 heterotroph Yoon et al., 2004b 121A02 25 15 121B06 10 Sylvan et. al. Supplementary Information !"#

121E08 2 2 121A01 4 121C06 1 121B03 1 121H05 1 Marinobacter flavimaris str. 2PR54-4 petroleum degrader unknown (NCBI) 121H01 1 121B07 1 Marinobacter sp. str. EZ44 heterotroph Morris et al., 2008 121D04 1 1 121D10 2 121B12 2 121H02 1

Marinobacter sp. str. NT N31 unknown unknown (NCBI) 121C12 1 121G09 1 121H07 1 121D01 1 121B05 1 221G02* 1 221G07* 1 221G10* 1 Marinobacter exellens str. KMM 3809 heterotroph Gorshkova et al., 2003 221E01* 1 Halomonas sulfidaeris str. Esulfide1 heterotroph Kaye et al., 2004 221A12* 2 Halomonas meridiana str. PR51-13 petroleum degrader unknown (NCBI) 221A10* 3 221F06* 1 Umboniibacter marinipuniceus str. heterotroph Romanenko et al., KMM 3892 2010 113H01 1 1 Saccharophagus degradans str. 2-40 heterotroph Ekborg et al., 2005 18A03 1 1 113E10 1 213E02* 2 Kangiella koreensis str. DSM 16069 heterotroph Yoon et al., 2004a Sylvan et. al. Supplementary Information !"#

113G04 1 Cellvibrio japonicus str. Ueda107 heterotroph Humphry et al., 2003

113H06 1 Cycloclasticus pugetii str. PS-1 hydrocarbon Dyksterhouse et al., degrader 1995 113C01 1 Alteromonas genoviensis str. LMG heterotroph Vandecandelaere et 24079 al., 2008 221E03* 1 Psychromonas profunda str. 2825 heterotroph Xu et al., 2003 213F04* 1 Thioalkalivibrio sulfidophilus str. HL- autotrophic S- Muyzer et al., 2011 EbGR7 oxidizer 113E12 2 Sylvan et. al. Supplementary Information !"#

1-8 1-21 1-8 1-21 29 31 30 31

0 0 16 0 16 0 1-13 2-8 1-13 2-13 37 44 36 31 1 0 1 0

1 0 7 1 0 6 0 3 0 3

1 2

1-8 1-21 2-13 2-21 34 28 38 0 23

0 18 3 1-13 2-21 38 18 1 0 1 3 1

0 0 2 1 1 2-8 50 0

Figure S1 - Venn diagrams of number of shared OTUs between each pair of samples from this study. Four different Venn diagrams are necessary to show the relationships between all 6 clone libraries. OTUs are described using a 97% cutoff value and shared membership was calculated using MOTHUR.

Sylvan et. al. Supplementary Information !$#

99 RESET 121C12 99 RESET 121B07 96 RESET 121D01 99 RESET 121H07 RESET 121G09 RESET 121C06 RESET 121D10 31 96 RESET 121B12 77 98 RESET 121H05 RESET 121A01 RESET 121E08 Marinobacter RESET 121A02 RESET 121B03 Marinobacter flavimaris str. SW-145 AY517632.1 99 Marinobacter sp. str. NT N31 AB166980.1 RESET_121H01 RESET_121H02

99 RESET 121B06 Marinobacter sp. str. EZ44 EU591709.1 RESET_121D04 RESET_121B05 Marinobacter lutaoensis str. T5054 NR 025116 RESET 18D05 RESET 121E02 99 Alcanivorax sp. str. Oha-1 AB053130

0.05

Figure S2- Expansion of the Marinobacter branch of the Gammaproteobacteria tree (Fig. 4). One representative clone per OTU (defined by a 97% cutoff) was used to construct the tree. Bootstrap values >50% are reported. NCBI accession numbers for sequences from cultured representatives are given following the species name. Tree construction methods are reported in the manuscript.

Sylvan et. al. Supplementary Information %&#

60 1-13 2-8

50 1-8 2-13 40 1-21 30 2-21 20 OTUs (3% Cutoff)

0 0 20 40 60 80 # Clones Sequenced

Figure S3- Rarefaction curve for clone libraries from this study, calculated with MOTHUR.

OTUs are defined at the 97% similarity level. Samples from TrapR1 are represented by solid symbols, samples from Trap R2 are represented by open symbols of the same shape from the same time points as those in Trap R1.

Sylvan et. al. Supplementary Information %!#

Marinobacter

RESET_113E12 71 RESET_113F01 98 RESET_113E02 97 RESET_121D05 Legionella pneumophila str. Lens, CR628337 66 RESET_113G04 Kangiella koreensis str. DSM_16069, CP001707 RESET_113G05 68 Calyptogena phaseoliformis symbiont str. KT-2, AB479082 99 RESET_221E06* 55 RESET_221E09* 59 RESET_221F06* Halomonas Group 2A 91 RESET_221A10* 99 Halomonas meridiana str. PR51-13, EU441001 Halomonas sulfidaeris str. Esulfide1, AF212204 Halomonas Group 2C 99 RESET_221A12* 95 RESET_221E07* Alcanivorax sp. str. Oha-1, AB053130 RESET_121F12 99 RESET_121E04 65 91 RESET_121E02 99 RESET_121E05 RESET_18D05 99 RESET_18D07 99 RESET_18A03 99 RESET_113H11 89 RESET_113H01 Uncultured bacterium clone P7X3b1BG11, EU491090 99 99 Uncultured organism clone ctg_CGOF124, DQ395870 64 RESET_113D11 Haliea sp. str. SY02, GU124598 99 RESET_213D03* 91 RESET_113B10 99 RESET_213D05 RESET_113E10 Cellvibrio japonicus str. Ueda107, CP000934 70 Saccharophagus degradans str. 2-40, NC_007912 Umboniibacter marinipuniceus str. KMM 3892, AB467280 99 RESET_213E02* 99 RESET_213F10* 99 RESET_28C07 51 RESET_28C08 64 Colwellia psychrerythraea str. 34H, CP000083 93 RESET_213E01* 7496 RESET_213E05* 99 RESET_213C07* 99 RESET_213F07* RESET_213E11* RESET_221E03* RESET_213F04* 99 95 RESET_213F08 99 RESET_213G03 99 RESET_113B08 71 RESET_18F06 Pseudoalteromonas sp. str. JL-S1, AY745839 RESET_213H03* 99 Photobacterium leiognathi str. RM1, AY292947 99 Photobacterium sp. str. HAR72, AB038032 75 RESET_18A04 RESET_213B05 98 RESET_113B03 RESET_213H11 RESET_213C04 RESET_213B03 RESET_213D08 Uncultured bacterium clone S-DCM-106, GU061990 RESET_213G07 87 RESET_213G11 Thioalkalivibrio sp. str. HL-EbGR7, CP001339 Codakia orbicularis gill symbiont, X84979 RESET_113H06 RESET_113C01 58 Cycloclasticus pugetii str. PS-1, NR_025955 Kopriimonas byunsanensis str. KOPRI 13522, DQ167245 Erythrobacter longus str. EG6, AM691106 99 Thalassobius sp. str. ZS6-23, FJ196059 α-proteobacteria 70 Silicibacter sp. str. LS103, FJ937914 99 62 Oceanibulbus indolifex str. Hel 45, AJ550939 64 Roseobacter sp. str. P81, EU195951 Mariprofundus ferrooxydans str. PV-1, AATS01000005 ζ-proteobacteria Uncultured anammox sediment clone Kazan-3B-30/BC19-3B-30 AY592196 Loihi Seamount basalt clone P7X3b1D06 EU491086 75 89 Calditerrivibrio nitroreducens, AB364234 Deferribacteres 0.05 Sylvan et. al. Supplementary Information %%#

Figure S4 - 16S rRNA phylogenetic tree of Gammaproteobacteria clones recovered during this study, with representative Alphaproteobacteria, Zetaproteobacteria and Deferribacteres included to root the tree. All clones classified as Gammaproteobacteria are included here, versus Fig. 4, where a single representative for each OTU (defined at the 97% similarity level). Clones that were designated as "Other (non-plume)" in Fig. 2 and that were not included in Fig. 4 are designated here with an asterisk (*). Alignments of sequences from this study and their nearest cultured organisms in the Greengenes database as well as some close uncultured clones from the ARB database were constructed using MEGA5. The tree was constructed using Maximum Likelihood algorithm based on the Tamura-Nei model with 500 bootstraps. Bootstrap values >50% are reported. The Marinobacter branch is expanded in Fig. S5. NCBI accession numbers for sequences from cultured representatives and environmental clones are given following the species/clone name. #

# Sylvan et. al. Supplementary Information %'#

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