Supplementary data. Dissolved extracellular polymeric substance (deps) dynamics and bacterial growth

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1 Polar Biology Supplementary data Dissolved extracellular polymeric substance (deps) dynamics and bacterial growth during sea ice formation in an ice tank study Shazia N Aslam 1*, Graham J.C. Underwood 1, Hermanni Kaartokallio, Louiza Norman, Riitta Autio, Michael Fischer, Harri Kuosa 5, Gerhard S. Dieckmann, and David N. Thomas, 1 Department of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex, CO SQ, U.K. Finnish Environment Institute (SYKE), Marine Research Centre, P.O. Box 1, FI-51 Helsinki. Finland. Ocean Sciences, College of Natural Science, Bangor University, Menai Bridge, Anglesey, LL5 5AB, U.K. Alfred Wegener Institute for Polar and Marine Research, Am Handelshafen 1, D-757 Bremerhaven, Germany. 5 Tvärminne Zoological Station, University of Helsinki, Finland Corresponding author; shaziaa@essex.ac.uk

2 ESM_1 Diatom extracellular polymeric substances (EPS), ethanol based deps fractionation, and percentage contribution of deps fractions to dissolved carbohydrates (dcho). TdCHO represent dissolved carbohydrate concentration before dialysis, while dcho represent carbohydrate concentration after dialysis. Adapted from Underwood et al. (1) Cell associated Carbohydrates Cell Structural complexity Ethanol based EPS fractionation %age contribution of EPS fraction to dcho Dissolved Carbohydrates (TdCHO) Highly complex polysaccharides Complex polysaccharides Less Complex polysaccharides Polysaccharides kda (dcho) EPS- EPS-5 EPS-7 %CHO - %CHO -5 %CHO 5-7 Non EPS carbohydrates non-precipitable CHO CHO < kda %CHO 7-1

3 ESM_ Vertical profile of a temperature ( o C) and b salinity in ice cores collected on each sampling day. Except for d (N = 1 and for NSW and ADOM respectively on d), ice cores were collected on each sampling day for both NSW and ADOM series and average of ice cores was used for all graphs. (a) -1-5 Temperature ( o C) Temperature ( o C) Depth (cm) (b) Salinity (PSU) Salinity (PSU) Depth (cm) 1 1 d d5 d d d5 d NSW ADOM

4 ESM_ Abundances (cells ml -1 ) of flagellates and ice algae (diatoms) over the experimental period. Diatoms were not counted after d, as after d all diatoms encountered were empty frustules. Except for samples collected on d (N > on d) N (number of samples) was for all sample types collected from both NSW and ADOM mesocosm on each day. Values are mean ± SE. Sampling day n.d.= Not detected Seawater Sea ice Brines Frost flowers NSW ADOM NSW ADOM NSW ADOM NSW ADOM Flagellates d 7.1 ±.. ± 1. d 17. ± ± 1. d ±.5 1. ± 1.. ± d5 7.5 ± ±. 7.1 ± ± 1.. ± 1..7 ±.7 d 5 ± ± 1..1 ± 5.. ± ± ± 1.7 Algae d. ± ± 1.1 d. ±. ± 11.5 d n.d. n.d. n.d. n.d. d5 n.d. n.d. n.d. n.d. n.d. n.d. d n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d.

5 ESM_ High-DNA/ low-dna bacterial ratios for all sample types from sea ice generated in a d mesocosm experiment using North Sea Water (NSW) and North Sea Water enriched with additional algal-derived DOM (ADOM). Seawater Sea ice Brines Frost flowers NSW ADOM NSW ADOM NSW ADOM NSW ADOM d 1.5. d d d d

6 ESM_5 Correlation between total bacterial numbers (TBN) and inorganic nutrients in seawater, sea ice and brines. Values in bold are significant at P <.5* or <.1** Seawater Sea Ice Brines NSW ADOM NSW ADOM NSW ADOM NH -.1 () PO -. () Si -.5 () NO -. () NO.7** ().1** (1) -.7 (1) -. (1) -.5** (1).71* (1).1 (1).5* (1).5* (1). (11). (1) -. (1). (1). (1).7 (1) -. (1) -.57.**

7 ESM_ Dissolved oxygen levels (µmol l -1 ) in the seawater on each sampling day in the NSW and ADOM mesocosms. Unfilled circles represent NSW and filled circles represent ADOM samples. N (sample numbers) for all sample types collected from both NSW and ADOM mesocosm on each sampling day. NSW ADOM O µ mol l d d d d5 d Sampling day

8 ESM_7: Concentration of salinity-normalised (normalised to salinity ) (a) organic carbon (POC and DOC), (b) dissolved carbohydrate (dcho and dua) and (c) EPS fractions (EPS-7 and EPS-) in all samples vs. salinity. Open symbols represent samples collected from NSW bags and filled symbols represent samples collected from ADOM bags. Lines (dotted line for NSW and solid line for ADOM samples) in the graphs represent average values in seawater on d; values above these lines represent production of new material after d till the end of exp. (a) POC DOC [µmol l -1 C]s [µmol l -1 C]s [µmol l -1 C]s (b) (c) dcho EPS-7 dua EPS- Salinity

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