[en] Sea ice is a biome of immense size and provides a range of habitats for diverse microbial communities many of which are adapted to living at low temperatures and high salinities in brines. We measured simultaneous incorporation of thymidine (TdR) and leucine (Leu), bacterial cell abundance and cell population properties (by flow cytometry) in subarctic sea ice in SW Greenland. Short-term temporal variability was moderate, and steep environmental gradients, typical for sea ice, were the main drivers of the variability in bacterial cell properties and activity. Low nucleic acid (LNA) bacteria, previously linked to oligotrophic ecotypes in marine habitats, were more abundant in the upper ice layers, whereas High nucleic acid (HNA) bacteria dominated in lower ice, where organic carbon was in high concentrations. Leucine incorporation was saturated at micro molar concentrations, as known from freshwater and marine biofilm systems. Leu:TdR ratios were high (up to >300) in lowermost ice layers and when they are compared to published respiration measurements these results suggest non-specific leucine incorporation. There was evidence of polyhydroxyalkanoate (PHA) containing bacteria in the sea ice, shown by brightly fluorescing intracellular inclusions after Nile Blue A staining. High leucine saturating concentrations coupled with the occurrence of PHA producing organisms further highlight the similarity of sea ice internal habitats to biofilm-like systems rather than to open-water systems.
Disciplines :
Earth sciences & physical geography
Author, co-author :
Kaartokallio, H.
Søgaard, D.H.
Norman, L.
Rysgaard, S.
Tison, Jean-Louis
Delille, Bruno ; Université de Liège - ULiège > Département d'astrophys., géophysique et océanographie (AGO) > Unité d'Océanographie chimique (UOC)
Thomas, D.
Language :
English
Title :
Short-term variability in bacterial abundance, cell properties, and incorporation of leucine and thymidine in subarctic sea ice
Alonso-Sáez L, Galand PE, Casamayor EO, Pedrós-Alió C, Bertilsson S (2010) High bicarbonate assimilation in the dark by Arctic bacteria. ISME J 4:1581-1590
Amon RMW, Fitznar HP, Benner R (2001) Linkages among the bioreactivity, chemical composition, and diagenetic state of marine dissolved organic matter. Limnol Oceanogr 46:287-297
Anderson AJ, Dawes EA (1990) Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoate. Microbiol Rev 54:450-472
Ayo B, Unanue M, Azua I, Gorsky G, Turley C, Iriberri J (2001) Kinetics of glucose and amino acid uptake by attached and free-living marine bacteria in oligotrophic waters. Mar. Biol 138:1071-1076
Ayub ND, Tribelli PM, López NI (2009) Polyhydroxyalkanoates are essential for maintenance of redox state in the Antarctic bacterium Pseudomonas sp. 14-3 during low temperature adaptation. Extremophiles 13:59-66
Azam F, Hodson RE (1981) Multiphasic kinetics for D-glucose uptake by assemblages of natural marine bacteria. Mar. Ecol Prog Ser 6:213-222
Bao H, Duong F (2012) Discovery of an auto-regulation mechanism for the maltose ABC transporter MalFGK2. PLoS ONE 7:e34836
Becquevort S, Dumont I, Tison J-L, Lannuzel D, Sauvee M-L, Chou L, Schoemann V (2009) Biogeochemistry and microbial community composition in sea ice and underlying seawater off East Antarctica during early spring. Polar Biol 32:879-895
Belzile C, Brugel S, Nozais C, Gratton Y, Demers S (2008) Variations of the abundance and nucleic acid content of heterotrophic bacteria in Beaufort Shelf waters during winter and spring. J Mar. Syst 74:946-956
Bjørnsen PK, Kuparinen J (1991) Determination of bacterioplankton biomass, net production and growth efficiency in the Southern Ocean. Mar. Ecol Prog Ser 71:185-194
Buesing N, Gessner MO (2003) Incorporation of radiolabeled leucine into protein to estimate bacterial production in plant litter, sediment, epiphytic biofilms, and water samples. Microb Ecol 45:291-301
Calvo JM, Matthews RG (1994) The leucine-responsive regulatory protein, a global regulator of metabolism in Escherichia coli. Microbiol Rev 58:466-490
Campisano A, Overhage J, Rehm BHA (2008) The polyhydroxyalkanoate biosynthesis genes are differentially regulated in planktonic- and biofilm-grown Pseudomo nas aeruginosa. J Biotechnol 133:442-452
Chin-Leo G, Kirchman DL (1990) Unbalanced growth in natural assemblages of marine bacterioplankton. Mar. Ecol Prog Ser 63:1-8
Cho JC, Giovannoni SJ (2004) Oceanicola granulosus gen. nov., sp. nov. and Oceanicola batsensis sp. nov., poly-phydroxybutyrate-producing marine bacteria in the order 'Rhodobacterales'. Int J Syst Evol Microbiol 54:1129-1136
Collins RE, Rocap G, Deming JW (2010) Persistence of bacterial and archaeal communities in sea ice through an Arctic winter. Environ Microbiol 12:1828-1841
Cox GFN, Weeks WF (1983) Equations for determining the gas and brine volumes in sea ice samples. J Glaciol 29:306-316
Deming JW (2010) Sea ice bacteria and viruses. In: Thomas DN, Dieckmann GS (eds) Sea ice, 2nd edn. Wiley-Blackwell, Oxford, p 247-282
Ducklow HW (2003) Seasonal production and bacterial utilization of DOC in the Ross Sea, Antarctica. Antarctic Res Ser 78:143-158
Ducklow HW, Yager PL (2007) Pelagic bacterial processes in Polynyas. In: Smith WO Jr., Barber DG (eds) Polynyas: Windows into polar oceans. Elsevier Oceanography Series 74, Elsevier BV, p 323-361
Ducklow H, Dickson M, Kirchman D, Steward G, Orchardo J, Marra J, Azam F (2000) Constraining bacterial production, conversion efficiency and respiration in the Ross Sea, Antarctica, January-February, 1997. Deep-Sea Res II 47:3227-3247
Fischer H, Pusch M (1999) Use of the [14C] leucine incorporation technique to measure bacterial production in river sediments and the epiphyton. Appl Environ Microbiol 65:4411-4418
Fransson A, Chierici M, Yager PL, Smith WO Jr. (2011) Antarctic sea ice carbon dioxide system and controls. J Geophys Res 116(C12), C12035, doi.org/:10. 1029/2010 JC00 6844
Fuhrman JA, Azam F (1980) Bacterioplankton secondary production estimates for coastal waters of British Columbia, Antarctica, and California. Appl Environ Microbiol 39:1085-1095
Fuhrman JA, Azam F (1982) Thymidine incorporation as a measure of heterotrophic bacterioplankton production in marine surface waters: evaluation and field results. Mar. Biol 66:109-120
Fuhrman JA, Ferguson RL (1986) Nanomolar concentrations and rapid turnover of dissolved free amino acids in sea - water: agreement between chemical and microbiological measurements. Mar. Ecol Prog Ser 33:237-242
Garneau MÈ, Roy S, Lovejoy C, Gratton Y, Vincent WF (2008) Seasonal dynamics of bacterial biomass and production in a coastal arctic ecosystem: Franklin Bay, western Canadian Arctic. J Geophys Res 113(C7), C07S91, doi.org/:10. 1029/2007 JC004281
Gasol JM, del Giorgio PA (2000) Using flow cytometry for counting natural planktonic bacteria and understanding the structure of planktonic bacterial communities. Sci Mar. 64:197-224
Gasol JM, Zweifel UL, Peters F, Fuhrman JA, Hagström Å (1999) Significance of size and nucleic acid content hetero geneity as measured by flow cytometry in natural planktonic bacteria. Appl Environ Microbiol 65:4475-4483
Gillies JE, Kuehn KA, Francoeur SN, Neely RK (2006) Application of the [3H]leucine incorporation technique for quantification of bacterial secondary production associated with decaying wetland plant litter. Appl Environ Microbiol 72:5948-5956
Gosselin M, Legendre L, Therriault J-C, Demers S, Rochet M (1986) Physical control of the horizontal patchiness of seaice microalgae. Mar. Ecol Prog Ser 29:289-298
Grasshoff K, Ehrhardt M, Kremling K (1983) Methods of seawater analysis, 2nd edn. Verlag Chemie, Wienheim
Grossmann S, Dieckmann GS (1994) Bacterial standing stock, activity, and carbon production during formation and growth of sea ice in the Weddell Sea, Antarctica. Appl Environ Microbiol 60:2746-2753
Guglielmo L, Carrada G, Catalano G, Dell'Anno A and others (2000) Structural and functional properties of sympagic communities in the annual sea ice at Terra Nova Bay (Ross Sea, Antarctica). Polar Biol 23:137-146
Hales B, van Greer A, Takahashi T (2004) High-frequency measurements of seawater chemistry: Flow-injection analysis of macronutrients. Limnol Oceanogr Methods 2:91-101
Helmke E, Weyland H (1995) Bacteria in sea ice and underlying water of the eastern Weddell Sea in midwinter. Mar. Ecol Prog Ser 117:269-287
Holmes RM, Aminot A, Kerouel R, Hocker BA, Peterson BJ (1999) A simple and precise method for measuring ammonium in marine and fresh water. Can J Fish Aquat Sci 56:1801-1808
Ishige T, Tani A, Sakai Y, Kato N (2003) Wax ester production by bacteria. Curr Opin Microbiol 6:244-250
Jørgensen NOG (1992) Incorporation of [3H] leucine and [3H] valine into protein of freshwater bacteria: Uptake kinetics and intracellular isotope dilution. Appl Environ Microbiol 58:3638-3646
Junge K, Imhoff F, Staley T, Deming JW (2002) Phylogenetic diversity of numerically important Arctic sea-ice bacteria cultured at subzero temperature. Microb Ecol 43:315-328
Kaartokallio H (2004) Food web components, and physical and chemical properties of Baltic Sea ice. Mar. Ecol Prog Ser 273:49-63
Kaartokallio H, Kuosa H, Thomas DN, Granskog MA, Kivi K (2006) Biomass, composition and activity of organism assemblages along a salinity gradient in sea ice subjected to river discharge in the Baltic Sea. Polar Biol 30:183-197
Kaartokallio H, Tuomainen J, Kuosa H, Kuparinen J, Martikainen PJ, Servomaa K (2008) Succession of sea-ice bacterial communities in the Baltic Sea fast ice. Polar Biol 31:783-793
Kazakov AE, Rodionov DA, Alm E, Arkin AP, Dubchak I, Gelfand MS (2009) Comparative genomics of regulation of fatty acid and branched-chain amino acid utilization in Proteobacteria. J Bacteriol 191:52-64
Kirchman DL (1992) Incorporation of thymidine and leucine in the subarctic Pacific: application to estimating bacterial production. Mar. Ecol Prog Ser 82:301-309
Kirchman D, K'nees E, Hodson R (1985) Leucine incorporation and its potential as a measure of protein synthesis by bacteria in natural aquatic systems. Appl Environ Microbiol 49:599-607
Kirchman DL, Hill V, Cottrell MT, Gradinger R, Malmstrom RR, Parker A (2009) Standing stocks, production, and respiration of phytoplankton and heterotrophic bacteria in the western Arctic Ocean. Deep-Sea Res II 56:1237-1248
Koller M, Gasser I, Schmid F, Berg G (2011) Linking ecology with economy: Insights into polyhydroxyalkanoate-producing microorganisms. Eng Life Sci 11:222-237
Kottmeier S, Grossi SM, Sullivan CW (1987) Sea ice microbial communities. VIII. Bacterial production in annual sea ice of McMurdo Sound, Antarctica. Mar. Ecol Prog Ser 35:175-186
Krembs C, Deming JW (2008) The role of exopolymers in microbial adaptation to sea ice. In: Margesin R, Schinner F, Marx J-C, Gerday C (eds) Psychrophiles: from biodiversity to biotechnology. Springer-Verlag, Heidelberg, p 247-264
Krembs C, Eicken H, Deming JW (2011) Exopolymer alteration of physical properties of sea ice and implications for ice habitability and biogeochemistry in a warmer Arctic. Proc Natl Acad Sci USA 108:3653-3658
Kroon H (1993) Determination of nitrogen in water: comparison of continuous flow method with on-line UV digestion with the original Kjeldahl method. Anal Chim Acta 276:287-293
Kuosa H, Kaartokallio H (2006) Experimental evidence on nutrient and substrate limitation of Baltic Sea sea-ice algae and bacteria. Hydrobiologia 554:1-10
Kuparinen J, Autio R, Kaartokallio H (2011) Sea ice bacterial growth rate, growth efficiency and preference for inorganic nitrogen sources in the Baltic Sea. Polar Biol 34:1361-1373
Lebaron P, Servais P, Agogué H, Courties C, Joux F (2001) Does the high nucleic-acid content of individual bacterial cells allow to discriminate active cells in aquatic systems? Appl Environ Microbiol 67:1775-1782
Logan BE, Fleury RC (1993) Multiphasic kinetics can be an artifact of the assumption of saturable kinetics for microorganisms. Mar. Ecol Prog Ser 102:115-124
Long MH, Koopmans D, Berg P, Rysgaard S, Glud RN, Søgaard DH (2012) Oxygen exchange and ice melt measured at the ice-water interface by eddy correlation. Biogeosciences 9:1957-1967
Madison LL, Huisman GW (1999) Metabolic engineering of poly (3-hydroxyalkanoates): from DNA to plastic. Microbiol Mol Biol Rev 63:21-53
Maranger R, Bird DF, Juniper SK (1994) Viral and bacterial dynamics in Arctic sea ice during the spring algal bloom near Resolute, N. W. T., Canada. Mar. Ecol Prog Ser 111:121-127
Martin A, Anderson MJ, Thorn C, Davy SK, Ryan KG (2011) Response of sea-ice microbial communities to environmental disturbance: an in situ transplant experiment in the Antarctic. Mar. Ecol Prog Ser 424:25-37
Mary I, Heywood JL, Fuchs BM, Amann R, Tarran GA, Burkill PH, Zubkov MV (2006) SAR11 dominance among metabolically active low nucleic acid bacterioplankton in surface waters along an Atlantic meridional transect. Aquat Microb Ecol 45:107-113
Methé BA, Nelson KE, Deming JW, Momen B and others (2005). The psychrophilic lifestyle as revealed by the genome sequence of Colwellia psychrerythraea 34H through genomic and proteomic analyses. Proc Natl Acad Sci USA 102:10913-10918
Middelboe M, Glud RN, Sejr MK (2012) Bacterial carbon cycling in a subarctic fjord: A seasonal study on microbial activity, growth efficiency, and virus-induced mortality in Kobbefjord, Greenland. Limnol Oceanogr 57:1732-1742
Mikkelsen DM, Rysgaard S, Glud RN (2008) Microalgal composition and primary production in Arctic sea ice: a seasonal study from Kobbefjord (Kangerluarsunnguaq), West Greenland. Mar. Ecol Prog Ser 368:65-74
Miranda MR, Guimarães JRD, Coelho-Souza AS (2007) [3H]Leucine incorporation method as a tool to measure secondary production by periphytic bacteria associated to the roots of floating aquatic macrophyte. J Microbiol Methods 71:23-31
Mizushima T, Yokoyama K, Mima S, Tsuchiya T, Sekimizu K (1997) Inhibition of thymidine transport in dnaA mutants of Escherichia coli. J Biol Chem 272:21195-21200
Mock T, Meiners KM, Giesenhagen HC (1997) Bacteria in sea ice and underlying brackish water at 54° 26' 50" N (Baltic Sea, Kiel Bight). Mar. Ecol Prog Ser 158:23-40
Morán XAG, Bode A, Ángel Suárez L, Nogueira E (2007) Assessing the relevance of nucleic acid content as an indicator of marine bacterial activity. Aquat Microb Ecol 46:141-152
Mundy CJ, Gosselin M, Ehn JK, Belzile C and others (2011) Characteristics of two distinct high-light acclimated algal communities during advanced stages of sea ice melt. Polar Biol 34:1869-1896
Nguyen D, Maranger R (2011) Respiration and bacterial carbon dynamics in Arctic sea ice. Polar Biol 34:1843-1855
Nissen H, Nissen P, Azam F (1984) Multiphasic uptake of D-glucose by an oligotrophic marine bacterium. Mar. Ecol Prog Ser 16:155-160
Oksanen JF, Blanchet G, Kindt R, Legendre P and others (2012) Vegan: Community Ecology Package. R package version 2.0-5. CRAN. R-project.org/package= vegan
Ostle AG, Holt JG (1982) Nile blue A as a fluorescent stain for poly-beta-hydroxybutyrate. Appl Environ Microbiol 44:238-241
Petrich C, Eicken H (2010) Growth, structure and properties of sea ice. In: Thomas DN, Dieckmann GS (eds) Sea ice, 2nd edn. Wiley-Blackwell, Oxford, p 23-78
Piiparinen J, Kuosa H (2011) Impact of UVA radiation on algae and bacteria in Baltic Sea ice. Aquat Microb Ecol 63:75-87
Pollard PC (2010) Bacterial activity in plant (Schoenoplectus validus) biofilms of constructed wetlands. Water Res 44:5939-5948
Pomeroy LR, Wiebe WJ (2001) Temperature and substrates as interactive limiting factors for marine heterotrophic bacteria. Aquat Microb Ecol 23:187-204
Poretsky RS, Sun S, Mou X, Moran MA (2010) Transporter genes expressed by coastal bacterioplankton in response to dissolved organic carbon. Environ Microbiol 12:616-627
Pusceddu A, Dell Anno A, Vezzulli L, Fabiano M and others (2008) Microbial loop malfunctioning in the annual sea ice at Terra Nova Bay (Antarctica). Polar Biol 32:337-346
Qian J, Mopper K (1996) High performance high temperature combustion total organic carbon analyzer. Anal Chem 68:3090-3097
R Development Core Team (2011) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna
Rappé MS, Connon SA, Vergin KL, Giovannoni SJ (2002) Cultivation of the ubiquitous SAR11 marine bacterioplankton clade. Nature 418:630-633
Riedel A, Michel C, Gosselin M (2007) Grazing of large-sized bacteria by sea-ice heterotrophic protists on the Mackenzie Shelf during the winter-spring transition. Aquat Microb Ecol 50:25-38
Rivkin RB, Legendre L (2001) Biogenic carbon cycling in the upper ocean: effects of microbial respiration. Science 291:2398-2400
Robinson C (2008) Heterotrophic bacterial respiration. In: Kirchman DL (ed) Microbial ecology of the oceans. Wiley-Blackwell, Oxford, p 299-334
Saier MH (2000) Families of transmembrane transporters selective for amino acids and their derivatives. Microbiology 146:1775-1795
Shiah FK, Ducklow HW (1997) Bacterioplankton growth responses to temperature and chlorophyll variations in estuaries measured by thymidine: leucine incorporation ratio. Aquat Microb Ecol 13:151-159
Simon M, Azam F (1989) Protein content and protein synthesis rates of planktonic marine bacteria. Mar. Ecol Prog Ser 51:201-213
Smith REH, Clement P (1990) Heterotrophic activity and bacterial productivity in assemblages of microbes from sea ice in the high Arctic. Polar Biol 10:351-357
Søgaard DH, Kristensen M, Rysgaard S, Glud RN, Hansen PJ, Hilligsøe KM (2010) Autotrophic and heterotrophic activity in Arctic first-year sea ice: seasonal study from Malene Bight, SW Greenland. Mar. Ecol Prog Ser 419:31-45
Søgaard DH, Thomas DN, Rysgaard S, Glud RN and others (2013) The relative contributions of biological and abiotic processes to the carbon dynamics in subarctic sea ice. Polar Biol. doi.org:/10.1007/s00300-013-1396-3
Thomas DN, Dieckmann GS (2002) Antarctic sea ice - a habitat for extremophiles. Science 295:641-644
Thomas DN, Lara RJ, Haas C, Schnack-Schiel SB and others (1998) Biological soup within decaying summer sea ice in the Amundsen Sea, Antarctica. In: Lizotte M, Arrigo K (eds) Antarctic sea ice biological processes, interactions and variability. Antarctic Res Ser 73:161-171
Thomas DN, Papadimitriou S, Michel C (2010) The biogeochemistry of sea ice. In: Thomas DN, Dieckmann GS (eds) Sea ice, 2nd edn. Wiley-Blackwell, Oxford, p 425-467
Törnblom E, Søndergaard M (1999) Seasonal dynamics of bacterial biomass and production on eelgrass Zostera marina leaves. Mar. Ecol Prog Ser 179:231-240
Tribelli PM, Di Martino C, López NI, Raiger Iustman LJ (2012) Biofilm lifestyle enhances diesel bioremediation and biosurfactant production in the Antarctic polyhydroxyalkanoate producer Pseudomonas extremaustralis. Biodegradation 23:645-651
Tuominen L (1995) Comparison of leucine uptake methods and a thymidine incorporation method for measuring bacterial activity in sediment. J Microbiol Methods 24:125-134
Unanue M, Ayo B, Agis M, Slezak D, Herndl G, Iriberri J (1999) Ectoenzymatic activity and uptake of monomers in marine bacterioplankton described by a biphasic kinetic model. Microb Ecol 37:36-48
Underwood GJC, Fietz S, Papadimitriou S, Thomas DN, Dieckmann GS (2010) Distribution and composition of dissolved extracellular polymeric substances (EPS) in Antarctic sea ice. Mar. Ecol Prog Ser 404:1-19
Underwood GJC, Aslam SN, Michel C, Niemi A and others (2013). Broad-scale predictability of carbohydrates and exopolymers in Antarctic and Arctic sea ice. Proc Natl Acad Sci USA 110:15734-15739
van Looij A, Riemann B (1993) Measurements of bacterial production in coastal marine environments using leucine: application of a kinetic approach to correct for isotope dilution. Mar. Ecol Prog Ser 102:97-104
Wältermann M, Steinbüchel A (2005) Neutral lipid bodies in prokaryotes: recent insights into structure, formation, and relationship to eukaryotic lipid deposits. J Bacteriol 187:3607-3619
Wang Y, Hammes F, Boon N, Chami M, Egli T (2009) Isolation and characterization of low nucleic acid (LNA)-content bacteria. ISME J 3:889-902
Zhou J, Delille B, Eicken H, Vancoppenolle M and others (2013) Physical and biogeochemical properties in landfast sea ice (Barrow, Alaska): Insights on brine and gas dynamics across seasons. J Geophys Res Oceans 118:3172-3189
