Polar Biology

, Volume 29, Issue 1, pp 27–39 | Cite as

Antarctic marine bacterioplankton subpopulations discriminated by their apparent content of nucleic acids differ in their response to ecological factors

  • A. Corzo
  • S. Rodríguez-Gálvez
  • L. Lubian
  • C. Sobrino
  • P. Sangrá
  • A. Martínez
Original Paper


Bacterial abundances determined in Drake Passage and Bransfield and Gerlache Straits (Antarctica) in the Austral summer ranged from 0.78 to 9.4×105 cells ml−1, and were positively correlated with standing stocks of Chl a. Two bacterial subpopulations were discriminated based in their different levels of green fluorescence and wide angle light scatter (SSC) per cell after SYTO-13 staining for the first time in Antarctic waters. High nucleic acid (HNA) and low nucleic acid (LNA) subpopulations differed considerably in their response to changes in environmental variables. The apparent content of nucleic acids per cell for the HNA subpopulation (FL1-HNA) showed vertical profiles similar to those of Chl a, including the presence of a maximum at the subsurface chlorophyll maximum. FL1-HNA was positively correlated with Chl a. No similar trends were observed for the LNA fraction. HNA and LNA subpopulations differed in the response of the wide angle light scatter signal to environmental factors as well. SSC-HNA decreased strongly with depth and was positively correlated with Chl a. Again, no similar trends were observed for the LNA subpopulation. The percentage of HNA cells (%HNA) ranged between 35.0 and 76.7% and showed a general tendency to increase with depth. This increase seemed to be larger when the stratification of the water column was higher. Differences in grazing pressure could be responsible of the unexpected vertical distribution of HNA cells. Our results shows that in situ LNA and HNA bacterioplankton subpopulations are under different ecological controls and likely to play different trophodynamic roles in Antarctic waters.


Heterotrophic Bacterium Drake Passage Antarctic Water Total Bacterial Abundance High Nucleic Acid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was financially supported by grants MAT2000-0261-P4-04, REN2002-01281/MAR, and REN2001-2650/ANT from the Ministerio de Ciencia y Tecnología, Spain. We acknowledge the support and help of the Officers and Crew of BIO-Hespérides during cruise CIEMAR. Authors thanks Pep Gasol, Carles Pedrós-Alió and several anonymous referees for their time, comments and suggestions.


  1. Andersson A, Larsson U, Hagström A (1986) Size-selective grazing by a microfagellate on pelagic bacteria. Mar Ecol Progr Ser 33:51–57CrossRefGoogle Scholar
  2. Basterretxea G, Arístegui J (1999) Pytoplankton biomass and production during late austral spring (1991) and summer (1993) in the Bransfield Strait. Polar Biol 21:11–22CrossRefGoogle Scholar
  3. Bernard L, Courties C, Servais P, Trousellier M, Petit M, Lebaron P (2000). Relationships among bacterial cell size, productivity, and genetic diversity in aquatic environments using cell sorting and flow cytometry. Microb Ecol 40:148–158Google Scholar
  4. Bird DF, Kalff J (1984) Empirical relationship between bacterial abundance and chlorophyll concentration in fresh and marine waters. Can J Fish Aquat Sci 41:1015–1023Google Scholar
  5. Boyd P and others (2000) A mesoscale phytoplankton bloom in the polar Southern Ocean stimulated by iron fertilization. Nature 407:695–702CrossRefPubMedGoogle Scholar
  6. Button DK, Robertson BR, JüttnerF (1996) Microflora of a subalpine lake: bacterial populations, size and DNA distributions, and their dependence on phosphate. FEMS Microbiol Ecol 21:87–101CrossRefGoogle Scholar
  7. Choi JW, Sherr EB, Sherr BF (1996) Relation between presence-absence of a visible nucleoid and metabolic activity in bacterioplankton cells. Limnol Oceanogr 41:1161–1168Google Scholar
  8. Cole JJ, Findlay S, Pace ML (1988) Bacterial production in fresh and saltwater ecosystems: a cross-systems overview. Mar Ecol Prog Ser 43:1–10CrossRefGoogle Scholar
  9. Cole JJ, Caraco NF (1993) The pelagic microbial food web of oligotrophic lakes, pp. 101–112. In: T. Ford (ed) Aquatic microbiology, Blackwell Scientific PressGoogle Scholar
  10. Corzo A, Rodríguez-Gálvez S, Lubian L, Sangrá P, Martínez A, Morillo JA (2005) Spatial distribution of transparent exopolymer particles (TEP) in the Bransfield Strait (Antarctica). J Plank Res (in press)Google Scholar
  11. Davey HM, Kell DB (1996) Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analysis. Microbiol Rev 60:641–696PubMedGoogle Scholar
  12. DeLong EF, Wu KY, Prezelin BB, Jovine RVM (1994) High abundance of archaea in Antractic marine picoplankton. Nature 371:695–697CrossRefPubMedGoogle Scholar
  13. del Giorgio P, Gasol JM (1995) Biomass distribution in freshwater plankton communities. Am Nat 146:135–152CrossRefGoogle Scholar
  14. del Giorgio PA, Bird DF, Prairie YT, Planas D (1996a) Flow cytometric determination of bacterial abundance in lake plankton with the green nucleid acid stain SYTO 13. Limnol Oceanogr 41:783–789Google Scholar
  15. del Giorgio PA, Gasol JM, Vaqué D, Mura P, Agustí S, Duarte CM (1996b) Bacterioplankton community structure: protists control net production and the proportion of active bacteria in a coastal marine community. Limnol Oceanogr 41:1169–1179Google Scholar
  16. Dubelaar GBJ, Jonker BR (2000) Flow cytometry as a tool for the study of phytoplankton. Sci Mar 64:135–156Google Scholar
  17. Figueiras FG, Estrada M, López O, Arbones B (1998). Photosynthetic parameters and primary production in the Bransfield Strait: relationships with mesoscale hydrographic structures. J Mar Systems 17:129–141CrossRefGoogle Scholar
  18. García MA, Castro CG, Ríos AF, Doval MD, Rosón G, Gomis D, López O (2002) Water masses and the distribution of physico-chemical properties in the Western Bransfield Strait and Gerlache Strait during austral summer 1995/96. Deep-Sea Res II 49:585–602CrossRefGoogle Scholar
  19. Gasol JM, del Giorgio PA (2000) Using flow cytometry for counting natural planktonic bacteria and understanding the structure of planktonic bacterial communities. Sci Mar 64:197–224CrossRefGoogle Scholar
  20. Gasol JM, del Giorgio PA, Massana R, Duarte CM (1995) Active versus inactive bacteria: size-dependence in a coastal marine plankton community. Mar Ecol Prog Ser 128:91–97CrossRefGoogle Scholar
  21. Gasol JM, Zweifel UL, Peters F, Fuhrman JA, Hagström A (1999) Significance of size and nucleic acid content heterogeneity as measured by flow cytometry in natural plaktonic bacteria. Appl Environ Microbiol 65:4475–4483PubMedGoogle Scholar
  22. Grasshoff K, Ehrhardt M, Kremling K (1983) Methods of sea water analysis. Verlag ChemieGoogle Scholar
  23. Guindulain T, Comas J, Vives-Rego J (1997) Use of nucleic acid dyes SYTO-13, TOTO-1, and YOYO-1 in the study of Escherichia coli and marine prokaryotic populations by flow cytometry. Appl Environ Microbiol 63:4608–4611PubMedGoogle Scholar
  24. Guindulain T, Vives-Rego J (2002) Involvement of RNA and DNA in the stainig of Escherichia coli by SYTO 13. Lett Appl Microbiol 34:1–7CrossRefGoogle Scholar
  25. Guixa-Boixereu N, Vaqué D, Gasol JM, Sánchez-Camara J, Pedrós-Alió C. Viral distribution and activity in Antarctic waters. Deep-Sea Res II 49:827–846Google Scholar
  26. Heissenberger A, Leppard GG, Herndl GJ (1996) Relationship between the intracellular integrity and the morphology of the capsular envelope in attached and free-living marine bacteria. Appl Environ Microbiol 62:4521–4528PubMedGoogle Scholar
  27. Holm-Hansen O, Lorenzen C J, Holmes RW, Stickland JDH (1965) Fluorimetric determination of chlorophyll. J Cons Perm Int Explo Mer 30:3–15Google Scholar
  28. Holm-Hansen O, Mitchell BG (1991) Spatial and temporal distribution of phytoplankton and primary production in the western Bransfield strait region. Deep-Sea Res 38:961–980CrossRefGoogle Scholar
  29. Jacquet S, Lennon JF, Marie D, Vaulot D (1998) Picoplankton population dynamics in coastal waters of the northwestern Mediterranean Sea. Limnol Oceanogr 43:1916–1931Google Scholar
  30. Jellet JF, Li WKW, Dickie PM, Boraie A, Kepkay PE (1996) Metabolic activity of bacterioplankton communities assed by flow cytometry and single carbon substrate utilization. Mar Ecol Prog Ser 136:213–225CrossRefGoogle Scholar
  31. Jochem FJ (2000) Probing the physiological state of phytoplankton at the single-cell level. Sci Mar 64:117–268Google Scholar
  32. Jochem FJ (2001) Morphology and DNA content of bacterioplankton in the northern Gulf of Mexico: analysis by epifluorescence microscopy and flow cytometry. Aquat Microb Ecol 25:179–194CrossRefGoogle Scholar
  33. Jochem FJ, Lavrentyev PJ, First MR (2004) Growth and grazing rates of bacteria groups with different apparent DNA content in the Gulf of Mexico. Mar Biol 145:1213–1225CrossRefGoogle Scholar
  34. Jürgens K, Güde H (1994) The potential importance of grazing-resistant bacteria in planktonic systems. Mar Ecol Prog Ser 112:169–188CrossRefGoogle Scholar
  35. Karl DM, Holm-Hansen O, Taylor GT, Tien G, Bird DF (1991) Microbial biomass and productivity in the Western Bransfield Strait Antarctica during 1986–87 Austral winter. Deep-Sea Res II 38:1029–1055CrossRefGoogle Scholar
  36. Kemp PF, Lee S, LaRoche J (1993) Estimating the growth rate of slowly growing marine bacteria from RNA content. Appl Environ Microbiol 59:2594–2601PubMedGoogle Scholar
  37. Karner M, Fuhrman JA (1997) Determination of active marine bacterioplakton: a comparison of universal 16S rRNA probes, autoradiography, and nucleoid stainig. Appl Environ Microbiol 63:1208–1213PubMedGoogle Scholar
  38. Kramer J, Singleton FL (1993) Measurement of rRNA variations in natural communities of microorganisms in the southeastern U.S. continental shelf. Appl Environ Microbiol 59:2430–2436PubMedGoogle Scholar
  39. Lebaron P, Servais P, Agogué H, Courties C, Joux F (2001). Does the high nucleic acid content of individual bacterial cells allow us to discriminate between active cells and inactive cells in aquatic systems? Appl Environ Microbiol 67:1775–1782CrossRefPubMedGoogle Scholar
  40. Lebaron P, Servais P, Baudoux AC, Bourrain M, Curties C, Parthuisot N (2002). Variations of bacterial-specific activity with cell size and nucleic acid content assessed by flow cytometry. Aquat Microb Ecol 28:131–140CrossRefGoogle Scholar
  41. Li WKW, Jellet JF, Dikie PM (1995) DNA distributions in planktonic bacteria stained with TOTO or TO-PRO. Limnol Oceanogr 40:1485–1495Google Scholar
  42. Lipski M (1985) Chlorophyll a in the Bransfield Strait and the southern part of Drake Passage during BIOMASS_SIBEX (December 1983–January 1984). Pol Polar Res 6:21–30Google Scholar
  43. Longnecker K, Sherr BF, Sherr EB (2005) Activity and phylogenetic diversity of high and low nucleic acid content, ETS-active, bacterial cells in an upwelling ecosystem. Apply Envirom Microb (submitted)Google Scholar
  44. Maaløe O, Kjeldgaard NO (1966) Control of macromolecular synthesis. A study of DNA, RNA and protein synthesis in bacteria. W.A. Benjamin, Inc.Google Scholar
  45. Marie D, Partensky F, Jacquet S, Vaulot D (1997) Enumeration and cell cycle análisis of natural populations of marine picoplankton by flow cytometry using the nucleic acid stain SyberGreen I. Appl Environ Microbiol 63:186–193PubMedGoogle Scholar
  46. Massana R, Taylor LT, Murray AE, Wu KY, Jeffrey WH, DeLong EF (1998) Vertical distribution and temporal variation of marine planktonic archaea in the Gerlache Strait, Antarctica, during early spring. Limnol Oceanogr 43:607–617CrossRefGoogle Scholar
  47. Moran XAG, Estrada M (2002) Pytoplanktonic DOC and POC production in the Bransfield and Gerlache Straits as derived from kinetic experiments of 14C incorporation. Deep-Sea Res II 49:769–786CrossRefGoogle Scholar
  48. Monfort P, Baleux B (1992) Comparison of flow cytometry and epifluorescence microscopy for counting bacteria in aquatic ecosystems. Cytometry 13:188–192CrossRefPubMedGoogle Scholar
  49. Monger BC, Landry MR (1993) Flow cytometry analysis of some marine bacteria with Hoechst 33342. Mar Eol Prog Ser 59:905–911Google Scholar
  50. Murray AE, Preston CM, Massana R, Taylor LT, Blakis A, Wu K, DeLong EF (1998) Seasonal and spatial variability of bacterial and archaeal assemblages in the coastal waters near Anvers Island, Antarctica. Appl Environ Microbiol 64:2585–2595PubMedGoogle Scholar
  51. Pedrós-Alió C, Calderón-Paz JI, Gasol JM (2000) Comparative analysis shows that bacterivory, not viral lysis, controls the abundance of heterotrophic prokaryotic plankton. FEMS Microbiol Ecol 32:157–165CrossRefPubMedGoogle Scholar
  52. Pedrós-Alió C, Vaqué D, Guixa-Boixereu N, Gasol JM (2002) Prokaryotic plankton biomass and heterotrophic production in western Antartic waters during 1995–1996 Austral Summer. Deep-Sea Res II 49:805–825CrossRefGoogle Scholar
  53. Robertson BK, Button DK (1989) Characterizing aquatic bacteria according to population, cell size and apparent DNA content by flow cytometry. Cytometry 10:70–76CrossRefPubMedGoogle Scholar
  54. Rodríguez J, Jiménez-Gómez F, Blanco JM, Figueroa FL (2002) Physical gradients and spatial variability of the size structure and composition of phytoplankton in the Gerlache Strait (Antarctica). Deep-Sea Res II 49:693–706CrossRefGoogle Scholar
  55. Schloss I, Estrada M (1994) Phytoplankton composition in the Weddell-Scotia confluence area during austral spring in relation to hydrography. Polar Biol 14:77–90CrossRefGoogle Scholar
  56. Servais P, Courties C, Lebaron P, Troussellier M (1999) Coupling bacterial activity measurements with cell sorting by flow cytometry. Microb Ecol 38:180–189CrossRefPubMedGoogle Scholar
  57. Servais P, Casamayor EO, Courties C, Catala P, Partuisot N, Lebaron P (2003) Activity and diversity of bacterial cells with high and low nucleic acid content. Aquat Microb Ecol 33:41–51CrossRefGoogle Scholar
  58. Sherr, BF, Sherr, EB, McDaniel, J (1992). Effect of protistan grazing on the frequency of dividing cells (FDC) in bacterioplankton assemblages. Appl Environ Microbiol 58:2381–2385PubMedGoogle Scholar
  59. Sherr EB, Sherr BF (1994) Bacterivory and herbivory: Key roles of phagotrophic protists in pelagic food webs. Microb Ecol 28:127–133CrossRefGoogle Scholar
  60. Troussellier M, Courties C, Lebaron P, Servais P (1999) Flow cytometric discrimination of bacterial populations based on SYTO 13 staining of nucleic acids. FEMS Microbiol Ecol 29:319–330CrossRefGoogle Scholar
  61. Tyndall RL, Hand RE, Mann RC, Evans C, Jernigan R (1985) Application of flow cytometry to detection and characterization of Legionella spp. Appl Environ Microbiol 49:852–857PubMedGoogle Scholar
  62. Yentsch CS, Menzel DW (1963) A method for the determination of phytoplankton chlorophyll and phaeophytin by fluorescence. Deep-Sea Res 10:221–231Google Scholar
  63. UNESCO (1994) Protocols for the Joint Global Ocean Flux Study (JGOFS) core measurements. Manuals and Guides 29:1–170Google Scholar
  64. Vaqué D, Guixa-Boixerau N, Gasol JM, Pedros-Alío C (2002) Distribution of microbial biomass and importance of protists in regulating prokariotic assemblages in three areas close to the Antarctic Peninsula in spring and summer 1995–96. Deep-Sea Res II 49:847–867CrossRefGoogle Scholar
  65. Varela M, Fernández E, Serret P (2002) Size-fractionated phytoplankton biomass and primary production in the Gerlache and South Bransfield Straits (Antarctic Peninsula) in austral summer 1995–96. Deep-Sea Res II 49:749–768CrossRefGoogle Scholar
  66. Zubkov MV, Fuchs BM, Burkill PH, Amann R (2001) Comparison of cellular and biomass specific activities of dominant bacterioplankton groups in stratified waters of the Celtic Sea. Appl Environ Microbiol 67:5210–5218CrossRefPubMedGoogle Scholar
  67. Zweifel UL, Hagström A (1995) Total counts of marine bacteria include a large fraction of non-nucleoid-containing bacteria (ghosts). Appl Environ Microbiol 61:2180–2185PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • A. Corzo
    • 1
  • S. Rodríguez-Gálvez
    • 2
  • L. Lubian
    • 2
  • C. Sobrino
    • 2
  • P. Sangrá
    • 3
  • A. Martínez
    • 3
  1. 1.Departamento de Biología, Facultad de Ciencias del Mar y AmbientalesUniversidad de CádizPolígono Río San PedroSpain
  2. 2.Instituto de Ciencias Marinas de Andalucía (CSIC) Polígono Río San PedroSpain
  3. 3.Departamento de FísicaCampus Universitario de TafiraEdificio de Ciencias BásicasSpain

Personalised recommendations