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Fig. S1. Fluorescent in situ hybridization (FISH) images of ANME-1 archaea and Desulfosarcina-Desulfococcus sulfate reducing bacteria from three different sediment temperatures (20°C, 55°C and 90°C). Probe specificity and hybridization conditions are described in Girguis and colleagues (2005) and the supporting information.

Fig. S2. Maximum-likelihood phylogenetic tree illustrating the relationships of 16 ssu rRNA ANME-1 sequences recovered from Middle Valley sediments to archaeal sequences from NCBI non-redundant database. Phylogenetic tree was generated with FastTree 2.0.0 (Price et al., 2010) using minimum-evolution subtree-pruning-regrafting and maximum-likelihood nearest-neighbour interchanges. Local support values shown are based on the Shimodaira-Hasegawa (SH) test. The tree was rooted to Pyrodictium occultum (M21087). Scale = 0.09 substitutions per site.

Fig. S3. Fe K-edge XANES (left) and EXAFS (right) spectra for Middle Valley sediments. As revealed by a shift in the inflection point of the absorption edge to higher energies, XANES spectra reveal a dominance of Fe(II) in the upper sediments (0–3 cm) and mixed Fe(II)/Fe(III) in deeper sediments (below 6 cm). Spectra for the depths 0–3 and 3–6 cm, 6–9 and 9–12 cm, and 12–15 and 15–20 cm were nearly identical – one representative spectrum is shown for each interval. Standard spectra for iron sulfide (FeS) and lepidocrocite (γ-FeOOH) are included to illustrate the binding energies for Fe(II) and Fe(III) respectively. The Fe EXAFS spectra of sediments collected at 6–9 cm (black solid line) can be fit with a linear combination of standard spectra composed of green rust sulfate (32 mole %), siderite (24 mole %), and various Fe sulfides, including pyrrhotite, pyrite and mackinawite (44 mole %) (χ2red = 1.9; R-factor = 0.06).

Table S1. Total cell densities in sediments incubated on the thermal gradient, continuous flow bioreactor (cells ml sediment−1). Cells counts were obtained by filtering sediment cell suspensions, staining with SYBR I and counting at 100X on an epifluorescent scope with a gridded ocular.

FilenameFormatSizeDescription
emi2825_sm_FigS1-3-TabS1.docx98KSupporting info item
emi2825_sm_FigS1.eps965KSupporting info item
emi2825_sm_FigS2.eps3794KSupporting info item
emi2825_sm_FigS3.eps459KSupporting info item
emi2825_sm_TableS1.pdf25KSupporting info item

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