Fig. S1. Phylogenetic tree from the 16S rRNA gene sequence of the prokaryotic species to which we have produced antibodies and are represented in LDChip300.

Fig. S2. Sandwich microarray immunoassay analysis of the whole geo-biochemical extracts (with PBS-EDTA) with LDChip300. Several permafrost samples were extracted with PBS-EDTA buffer as indicated in Experimental procedures and then analysed with LDChip300 by sandwich microarray immunoassay. The images on the left show two positive reactions (IVE4C1, C2) corresponding to different antibodies against Acidithiobacillus thiooxidans cells (left panel) and the corresponding negative control (right panel). The charts (right) correspond to all the immunograms obtained for the different samples (0, 60, 201, etc) with LDChip300. Fluorescent intensity scale ranges from 0 to 30 000 relative units. Peaks numbers correspond to antibodies: 1, IA2S1; 2, IA3S1; 3, IC1C1; 4, IC2C3; 5, IC3S2; 6, IC4C1; 7, ID1S1; 8, 253; 9, IVE1S1; 10, IVE3S1; 11, IVE3S100; 12, IVE5S100; 13, IVF4S2; 14, IVI5S1 (Shewanella oneidensis, anaerobic growth in fumarate); 15, IVJ1C1 (Haloferax mediterranii); 16, IVJ8C1 (Halorubrum sp.); 17, FeReTs_983; 18, HscA2_1487; 19, ModA2_1494; 20, NifD2_1485; 21, AMP; 22, Dinitrofenol; 23, Humic acids; 24, Naphthalene; 25, Trp; 26, IVE4C1; 27, IVE4C2; 28, IVE4S100; 29, A185; 30, A1496 (see Table S2 for description).

Fig. S3. Viable aerobic cell counting in the Deception Island permafrost. A–E show the morphology after 9 days of incubation at 20°C of the colonies obtained from samples at different depths: A, 30; B, 51, C, 90; D, 111; E, 250. (Right) DAPI staining of the corresponding colonies (numbers) on the plates.

Fig. S4. Detection of prokaryotes in the Deception Island permafrost with prokaryotic acidophile microarray oligonucleotide microarray containing16S and 23S rRNA gene oligonucleotide probes for different phylogenetic groups of prokaryotes (Garrido et al., 2008). (Top) Scanned images obtained after hybridization of PCR amplified and fluorescently labeled environmental 16S and 23S rRNA genes from different depths (samples 0, 60, 70, 300, 340, 415) with prokaryotic acidophile microarray. Positive signals obtained with 16S rRNA gene (green rectangles) and with 23S rRNA one (red) are highlighted. (Bottom) Charts obtained after quantification of the fluorescent signals from the images. Probes showing most relevant positive results are indicated: Y2R and EU338, Bacteria; ALF968, Alphaproteobacteria; BET42a, Betaproteobacteria; GAM42a, Gammaproteobacteria; DELTA495a, Deltaproteobacteria and Gemmatimonadetes; DELTA385, Deltaproteobacteria; LGC354a,b,c, Low GC Gram-positive Firmicutes group of Bacteria; HGC236 and HGC1901, High GC Gram positive bacteria (mainly Actinobacteria); 1Af, Archaea. Green bars for 16S rRNA and red ones for 23S rRNA.

Fig. S5. Bacterial diversity of Deception Island permafrost obtained by 16S rRNA gene sequencing. Rarefaction curves obtained for the different clusters of samples, as a function of the depth intervals (top numbers), at genetic distances of 0, 0.03, 0.05 and 0.1.

Fig. S6. Phylogenetic tree with all OTU representative sequences from the 16S rRNA gene clones obtained from Deception Island permafrost. The tree contains representative 16S rRNA gene sequences from 117 OTUs defined at a genetic distance of 0.03, together with their best matching type strain and environmental sample sequences. The sequence of Sulfolobus acidocaldaricus was included as an out-group. OTU representative sequences are represented by an alphanumeric identifier, in bold type; numbers enclosed in square brackets indicate the number of sequences included in each OTU. Reference sequences are represented by accession number and name or condensed description. Branch labels correspond to bootstrap values. Main taxonomic groups are indicated on the right.

Table S1. List of the permafrost core samples taken in the drill.

Table S2. List of all the antibodies used in this work (also found in Parro et al., 2011a).

Table S3. List of the antibodies that showed positive reactions after analysing the surface samples with LDChip300.

Table S4. Viable aerobic cell count Aerobic viable cells as colony forming units per gram of sample along the permafrost column after 9 days at 20°C.

Table S5. List of the sequenced colonies.

EMI_2767_sm_GenBank.txt466KSupporting info item
EMI_2767_sm_Data.doc3018KSupporting info item

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