Fig. S1. The probes, Ant4D3 and Arctic96B-16–196, detected overlapping taxa in the Balneatrix genus, as analysed in the ARB SILVA release 114, non-redundant database. The members of this group detected only by Ant4D3 are in green, Arctic96B-16–196 in red, and those detected by both probes in blue. No colour indicates this taxon is not detected by either probe. Scale bar shows 10% estimated sequence divergence using the distance matrix algorithm in ARB SILVA.


Fig. S2. Distribution of silver grain areas of all cells active for leucine uptake. Frequency distribution of silver grain areas binned by 0.5 μm2. For example, symbols at 0 include all cells with silver grain area from 0.00 to 0.49 μm2.


Fig. S3. DNA and FISH volume for single-cells labelled with the EUB338 probe in summer (open circles) and fall (filled circles). Lowest cut-off for FISH volume was 0.03 μm3, as any smaller objects were found to be artefacts. The relationship was not significant in either season. Number of cells used for the analysis was 2377 cells in summer (r = 0.009) and 942 cells in fall (r = 0.100).


Fig. S4. Single-cell DNA area and FISH area (A) in summer for EUB338-labelled cells were not strongly correlated (r = 0.17, n = 113). There was no correlation between DNA area and silver grain area (r = 0.002, n = 113) (B).


Table S1. Oligonucleotide probes used for FISH and MAR FISH. All probe sequences were tested for number of hits in the ARB SILVA online database, release 114, partial ( Single probes were used. Target site of the probe on Escherichia coli 16S rRNA gene as well as the family and genera recognized by the probes are also given. The order for the organisms recognized by the Ant4D3, Arctic 96B16-196 and SAR86-1249 probes is Oceanospirillales.


Table S2. Surface water temperature, total prokaryotes and leucine incorporation for sites B and E during the three field seasons. Standard deviation is given for temperature and total prokaryotes, and standard error for leucine incorporation. Total number of samples was seven in summer 2011, six in fall 2011 and seven in summer 2012. Dates with an asterisk are samples used for FISH and MAR FISH analysis. n/d = not done.

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