Skip to main content
SpringerLink
Log in

A Molecular Approach to Identify Active Microbes in Environmental Eukaryote Clone Libraries

  • Published:
Microbial Ecology Aims and scope Submit manuscript

Abstract

A rapid method for the simultaneous extraction of RNA and DNA from eukaryote plankton samples was developed in order to discriminate between indigenous active cells and signals from inactive or even dead organisms. The method was tested using samples from below the chemocline of an anoxic Danish fjord. The simple protocol yielded RNA and DNA of a purity suitable for amplification by reverse transcription-polymerase chain reaction (RT-PCR) and PCR, respectively. We constructed an rRNA-derived and an rDNA-derived clone library to assess the composition of the microeukaryote assemblage under study and to identify physiologically active constituents of the community. We retrieved nearly 600 protistan target clones, which grouped into 84 different phylotypes (98% sequence similarity). Of these phylotypes, 27% occurred in both libraries, 25% exclusively in the rRNA library, and 48% exclusively in the rDNA library. Both libraries revealed good correspondence of the general community composition in terms of higher taxonomic ranks. They were dominated by anaerobic ciliates and heterotrophic stramenopile flagellates thriving below the fjord’s chemocline. The high abundance of these bacterivore organisms points out their role as a major trophic link in anoxic marine systems. A comparison of the two libraries identified phototrophic dinoflagellates, “uncultured marine alveolates group I,” and different parasites, which were exclusively detected with the rDNA-derived library, as nonindigenous members of the anoxic microeukaryote community under study.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  1. Acinas, SG, Klepac-Ceraj, V, Hunt, DE, Pharino, C, Ceraj, I, Distel, DL, Polz, MF (2004) Fine-scale phylogenetic architecture of a complex bacterial community. Nature 430: 551–554

    Article  PubMed  CAS  Google Scholar 

  2. Altschul, SF, Madde, TL, Schaffer, AA, Zhang, JH, Zhang, Z, Miller, W, Lipman, DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25: 3389–3402

    Article  PubMed  CAS  Google Scholar 

  3. Amaral-Zettler, LA, Gómez, F, Zettler, E, Keenan, BG, Amils, R, Sogin, ML (2002) Eukaryotic diversity in Spain’s River of Fire. Nature 417: 137

    Article  PubMed  CAS  Google Scholar 

  4. Baldauf, SL, Bhattacharya, D, Cockrill, J, Hugenholtzt, P, Pawlowski, J, Simpson, GB (2004) The tree of life: an overview. In: Cracraft, J, Donoghue, MJ (Eds.) Assembling the Tree of Life. Oxford University Press, Oxford, pp 43–75

    Google Scholar 

  5. Baumgartner, M, Stetter, KO, Foissner, W (2002) Morphological, small subunit rRNA, and physiological characterization of Trimyema minutum (Kahl 1931), an anaerobic ciliate from submarine hydrothermal vents growing from 28°C to 52°C. J Eukaryot Microbiol 49: 227–238

    Article  PubMed  CAS  Google Scholar 

  6. Behnke, A, Bunge, J, Barger, K, Breiner, H-W, Alla, V, Stoeck, T (2006) Microeukaryote community patterns along an O2/H2S gradient in a super-sulfidic anoxic fjord (Framvaren, Norway). Appl Environ Microbiol 72: 3626–3636

    Article  PubMed  CAS  Google Scholar 

  7. Bernard, C, Fenchel, T (1994) Chemosensory behaviour of Strombidium purpureum, an anaerobic oligotrich with endosymbiotic purple non-sulphur bacteria. J Eukaryot Microbiol 41: 391–396

    Google Scholar 

  8. Blumberg, DD (1987) Creating a ribonuclease-free environment. Methods Enzymol 152: 20–24

    Article  PubMed  CAS  Google Scholar 

  9. Buckley, BA, Szmant AM (2004) RNA/DNA ratios as indicators of metabolic activity in four species of Caribbean reef-building corals. Mar Ecol Prog Ser 282: 143–149

    CAS  Google Scholar 

  10. Calijuri, MC, Dos Santos, ACA, Jati, S (2002) Temporal changes in the phytoplankton community structure in a tropical and eutrophic reservoir Barra Bonita, S.P., Brazil. J Plankton Res 24: 617–634

    Article  CAS  Google Scholar 

  11. Caron, DA, Countway, P, Brown, MV (2004) The growing contributions of molecular biology and immunology to protistan ecology: molecular signatures as ecological tools. J Eukaryot Microbiol 51: 38–48

    Article  PubMed  CAS  Google Scholar 

  12. Clarke, KJ, Finlay, BJ, Esteban, G, Guhl, BE, Embley, TM (1993) Cyclidium porcatum n. sp.: a free-living anaerobic scuticociliate containing a stable complex of hydrogenosomes, eubacteria and archaeobacteria. Eur J Protistol 29: 262–270

    Google Scholar 

  13. Countway, PD, Gast, RJ, Savai, P, Caron, DA (2005) Protistan diversity estimates based on 18S rDNA from seawater incubations in the western North Atlantic. J Eukaryot Microbiol 52: 1–12

    Article  Google Scholar 

  14. Dickerson, HW, Dawe, DL (1995) Ichthyophthirius multifiliis and Cryptocaryon irritans. In: Woo, PTK (Eds.) Fish Diseases and Disorders, Vol 1, Protozoan and Metazoan Infections. CAB International, Cambridge, pp 181–227

    Google Scholar 

  15. Edgcomb, VP, Kysela, DT, Teske, A, de Vera Gomez, A, Sogin ML (2002) Benthic eukaryotic diversity in the Guaymas Basin hydrothermal vent environment. Proc Natl Acad Sci 99: 7658–7662

    Article  PubMed  CAS  Google Scholar 

  16. Ehrenberg, CC (1838) Die Infusionstierchen als Vollkommene Organismen. Leopold Voss, Leipzig, Germany

    Google Scholar 

  17. Epstein, SS (1997) Microbial food webs in marine sediments. II. Seasonal changes in trophic interactions in a sandy tidal flat community. Microb Ecol 34: 199–209

    Article  PubMed  Google Scholar 

  18. Falkowski, PG, Schofield, O, Katz, ME, Van De Schootbrugge, B, Knoll, AH (2004) Why is the land green and the ocean red? In: Therstein, H, Young, JR (Eds.) Coccolithophores: from Molecular Processes to Global Impact. Springer, Berlin, pp 429–453

    Google Scholar 

  19. Fenchel, T, Kristensen, LD, Rasmussen, L (1990) Water column anoxia: vertical zonation of planktonic protozoa. Mar Ecol Prog Ser 62: 1–10

    Google Scholar 

  20. Fenchel, T, Finlay, BJ (1995) Ecology and Evolution in Anoxic Worlds. Oxford University Press, Oxford

    Google Scholar 

  21. Fenchel, T, Bernard, C, Esteban, G, Finlay, BJ, Hansen, PJ, Iversen, N (1995) Microbial diversity and activity in a Danish fjord with anoxic deep water. Ophelia 43: 45–100

    Google Scholar 

  22. Foissner, W, Blatterer, H, Berger, H, Kohmann, F (1991) Taxonomische und ökologische Revision der Ciliaten des Saprobiensystems—Band I: Cyrtophorida, Oligotrichida, Hypotrichia, Colpodea. Informationsberichte des Bayer. Landesamtes für Wasserwirtschaft, 1/91. Bartels und Wernitz Druck, München

  23. Foissner W, Blatterer H, Berger H, Kohmann F (1992) Taxonomische und ökologische Revision der Ciliaten des Saprobiensystems—Band II: Peritrichia, Heterotrichida, Odontomastida. Informationsberichte des Bayer. Landesamtes für Wasserwirtschaft, 5/92. Bartels und Wernitz Druck, München

  24. Foissner, W, Berger, H, Kohmann, F (1994) Taxonomische und ökologische Revision der Ciliaten des Saprobiensystems—Band III: Hymenostomata, Protomatida, Nassulida. Informationsberichte des Bayer. Landesamtes für Wasserwirtschaft, 1/94. Bartels und Wernitz Druck, München

  25. Foissner, W, Berger, H, Blatterer, H, Kohmann, F (1995) Taxonomische und ökologische Revision der Ciliaten des Saprobiensystems—Band IV: Gymnostomatea, Loxodes, Suctoria. Informationsberichte des Bayer. Landesamtes für Wasserwirtschaft, 1/95. Bartels und Wernitz Druck, München

  26. Fried, J, Ludwig, W, Psenner, R, Schleifer, KH (2002) Improvement of ciliate identification and quantification: a new protocol for fluorescence in situ hybridization (FISH) in combination with silver stain techniques. Syst Appl Microbiol 25: 555–571

    Article  PubMed  CAS  Google Scholar 

  27. Griffiths, RI, Whiteley, AS, O’Donnell, AG, Bailey, MJ (2000) Rapid method for coextraction of DNA and RNA from natural environments for analysis of ribosomal DNA- and rRNA-based microbial community composition. Appl Environ Microbiol 66: 5488–5491

    Article  PubMed  CAS  Google Scholar 

  28. Hällförs, G (2004) Helsinki commission and baltic marine environment protection commission checklist of Baltic Sea phytoplankton species. Baltic Sea Environ Proc 95

  29. Hugenholtz, P, Huber, T (2003) Chimeric 16S rDNA sequences of diverse origin are accumulating in the public databases. Int J Syst Evol Microbiol 53: 289–293

    Article  CAS  Google Scholar 

  30. Iwama, GK, Pickering, AD, Sumpter, JP, Schreck, CB (1997) Fish Stress and Health in Aquaculture. Cambridge University Press, New York, NY

    Google Scholar 

  31. Kahl, DM (1930–1935) Urtiere oder Protozoa. I: Wimpertiere oder Ciliata (Infusoria), eine Bearbeitung der freilebenden und ectocomensalen Infusorien der Erde, unter Ausschluss der marinen Tintinnidae. Gustav Fischer, Jena, Germany

    Google Scholar 

  32. Kerkhof, L, Ward, BB (1993) Comparison of nucleic acid hybridization for measurement of the relationship between RNA/DNA ratio and growth rate in a marine bacterium. Appl Environ Microbiol 59: 1303–1309

    PubMed  CAS  Google Scholar 

  33. Klaveness, D, Shalchian-Tabrizi, K, Thomsen, HA, Eikrem, W Jakobsen, KS (2005) Telonema antarcticum sp. nov., a common marine phagotrophic flagellate. Int J Syst Evol Microbiol 55: 2595–2604

    Article  PubMed  CAS  Google Scholar 

  34. Krsek, M, Wellington, EMH (1999) Comparison of different methods for the isolation of total community DNA from soil. J Microbiol Methods 39: 1–16

    Article  PubMed  CAS  Google Scholar 

  35. Lane, DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt, E, Goodfellow, M (Eds.) Nucleic Acid Techniques in Bacterial Systematics. Wiley, New York, pp 115–148

    Google Scholar 

  36. Lemieux, C, Otis, C, Turmel, M (2000) Ancestral chloroplast genome in Mesostigma viride reveals an early branch of green plant evolution. Nature 403: 649–652

    Article  PubMed  CAS  Google Scholar 

  37. López-García, P, Rodriguez-Valera, F, Pedros-Alio, C, Moreira, D (2001) Unexpected diversity of small eukaryotes in deep-sea Antarctic plankton. Nature 409: 603–607

    Article  PubMed  Google Scholar 

  38. López-García, P, Philippe, H, Gail F, Moreira, D (2003) Autochthonous eukaryotic diversity in hydrothermal sediment and experimental microcolonizers at the Mid-Atlantic Ridge. Proc Natl Acad Sci 100: 697–702

    Article  PubMed  CAS  Google Scholar 

  39. Lorenz, MG, Wackernagel, G (1987) Adsorption of DNA to sand and variable degradation rates of adsorbed DNA. Appl Environ Microbiol 67: 2354–2359

    Google Scholar 

  40. Lynn, DH, Small, EB (2000) Phylum Ciliophora. In: Lee, JJ, Leedale, GF, Bradbury, P (Eds.) An Illustrated Guide to the Protozoa, vol. 1. Allen Press, Lawrence, KS, pp 371–656

    Google Scholar 

  41. Ludwig, W, Strunk, O, Westram, R and 29 others (2004) ARB: a software environment for sequence data. Nucleic Acids Res 32: 1363–1371

    Google Scholar 

  42. MacGregor, BJ, Moser, DP, Baker, BJ, Alm, EW, Maurer, M, Nealson, KH, Stahl, DA (2001) Seasonal and spatial variability in Lake Michigan sediment small-subunit rRNA concentrations. Appl Environ Microbiol 67: 3908–3922

    Article  PubMed  CAS  Google Scholar 

  43. Maidak, BL, Cole, JR, Lilburn, TG, Parker, CT Jr, Saxman, PR, Farris, RJ, Garrity, GM, Olsen, GJ, Schmidt, TM, Tiedje, JM (2001) The RDP-II (Ribosomal Database Project). Nucleic Acids Res 29: 173–174

    Article  CAS  Google Scholar 

  44. Massana, R, Balagué, V, Guillou, L, Pedrós-Alió, C (2004) Picoeukaryotic diversity in an oligotrophic coastal site studied by molecular and culturing approaches. FEMS Microbiol Ecol 50: 231–243

    Article  CAS  PubMed  Google Scholar 

  45. Medlin, L, Elwood, HJ, Stickel, S, Sogin, ML (1988) The characterization of enzymatically amplified eukaryotic 16S-like rRNA-coding regions. Gene 71: 491–499

    Article  PubMed  CAS  Google Scholar 

  46. Mills, HJ, Martinez, RJ, Story, S, Sobecky, PA (2004) Identification of members of the metabolically active microbial populations associated with Beggiatoa species mat communities form Gulf of Mexico cold-seep sediments. Appl Environ Microbiol 70: 5447–5458

    Article  PubMed  CAS  Google Scholar 

  47. Mills, HJ, Martinez, RJ, Story, S, Sobecky, PA (2005) Characterization of microbial community structure in Gulf of Mexico gas hydrates: comparative analysis of DNA- and RNA-derived libraries. Appl Environ Microbiol 71: 3235–3247

    Article  PubMed  CAS  Google Scholar 

  48. Milner, MG, Saunders, JR, McCarthy, AJ (2001) Relationship between nucleic acid ratios and growth in Listeria monocytogenes. Microbiology-SGM 147: 2689–2696

    CAS  Google Scholar 

  49. Miskin, IP, Farrimond, P, Head, M (1999) Identification of novel bacterial lineages as active members of microbial populations in a freshwater sediment using a rapid RNA extraction procedure and RT-PCR. Microbiology 145: 1977–1987

    Article  PubMed  CAS  Google Scholar 

  50. Moon-van der Staay, SY, De Wachter, R, Vaulot, D (2001) Oceanic 18S rDNA sequences from picoplankton reveal unsuspected eukaryotic diversity. Nature 409: 607–610

    Article  PubMed  CAS  Google Scholar 

  51. Moreira, D, López-García, P (2002) The molecular ecology of microbial eukaryotes unveils a hidden world. Trends Microbiol 10: 31–38

    Article  PubMed  CAS  Google Scholar 

  52. Nikolausz, M, Marialigeti, K, Kovacs, G (2004) Comparison of RNA- and DNA-based species diversity investigations in rhizoplane bacteriology with respect to chloroplast sequence exclusion. J Microbiol Methods 56: 365–373

    Article  PubMed  CAS  Google Scholar 

  53. Nogales, B, Moore, ERB, Llobet-Brossa, E, Rossello-Mora, R, Amann, R, Timmis, KN (2001) Combined use of 16S ribosomal DNA and 16S rRNA to study the bacterial community of polychlorinated biphenyl-polluted soil. Appl Environ Microbiol 67: 1874–1884

    Article  PubMed  CAS  Google Scholar 

  54. Norris, TB, Wraith, JM, Castenholz, RW, McDermott, TR (2002) Soil microbial community structure across a thermal gradient following a geothermal heating event. Appl Environ Microbiol 68: 6300–6309

    Article  PubMed  CAS  Google Scholar 

  55. Nuyts, S, Van Mellaert, L, Lambin, P, Anne, J (2001) Efficient isolation of total RNA from Clostridium without DNA contamination. J Microbiol Methods 44: 235–238

    Article  PubMed  CAS  Google Scholar 

  56. Purdy, KJ, Embley, TM, Takii, S, Nedwell, DB (1996) Rapid extraction of DNA and RNA from sediments by a novel hydroxyapatite spin-column method. Appl Environ Microbiol 62: 3905–3907

    PubMed  CAS  Google Scholar 

  57. Ramsing, NB, Fossing, H, Ferdelman, TG, Andersen, F, Thamdrup, B (1996) Distribution of bacterial populations in a stratified fjord (Mariager Fjord, Denmark) quantified by in situ hybridization and related to chemical gradients in the water column. Appl Environ Microbiol 62: 1391–1404

    PubMed  CAS  Google Scholar 

  58. Rappé, MS, Giovannoni, SJ (2003) The uncultured microbial majority. Annu Rev Microbiol 57: 369–394

    Article  PubMed  CAS  Google Scholar 

  59. Robison-Cox, JF, Bateson, MM, Ward, DM (1995) Evaluation of nearest-neighbor methods for detection of chimeric small-subunit rRNA sequences. Appl Environ Microbiol 61: 1240–1245

    PubMed  CAS  Google Scholar 

  60. Rosati, G, Modeo, L, Melai, M, Petroni, G, Verni, F (2004) A multidisciplinary approach to describe protists: a morphological, ultrastructural, and molecular study on Peritromus kahli Villeneuve-Brachon, 1940 (Ciliophora, Heterotrichea). J Eukaryot Microbiol 51: 49–59

    Article  PubMed  Google Scholar 

  61. Sambrook, J, Fritsch, EF, Maniatis, T (Eds.) (1989) Molecular Cloning—A Laboratory Manual, 2nd edn. CSHL Press, New York

  62. Seguritan, V, Rohwer, F (2001) FastGroup: a program to dereplicate libraries of 16S rDNA sequences. BMC Bioinformatics 2: 9–16

    Article  PubMed  CAS  Google Scholar 

  63. Sessitsch, A, Gyamfi, S, Stralis-Pavese, N, Weilharter, A, Pfeifer U (2002) RNA isolation from soil for bacterial community and functional analysis: evaluation of different extraction and soil conservation protocols. J Microbiol Methods 51: 171–179

    Article  PubMed  CAS  Google Scholar 

  64. Sharkey, FH, Banat, IM, Marchant, R (2004) A rapid and effective method of extracting fully intact RNA from thermophilic geobacilli that is suitable for gene expression analysis. Extremophiles 8: 73–77

    Article  PubMed  CAS  Google Scholar 

  65. Shopsin, B, Gomez, M, Montgomery, SO, Smith, DH, Waddington, M, Dodge, DE, Bost, DA, Riehman, M, Naidich, S, Kreiswirth, BN (1999) Evaluation of protein A gene polymorphic region DNA sequencing for typing of Staphylococcus aureus strains. J Clin Microbiol 37: 3556–3563

    PubMed  CAS  Google Scholar 

  66. Stackebrandt, E, Liesack, W (1993) Nucleic acids and classification. In: Goodfellow, M, O’Donnel, A (Eds.) Modern Approaches in Bacterial Systematics, Academic Press, London, pp 151–194

  67. Stoeck, T, Epstein, SS (2003) Novel eukaryotic lineages inferred from SSU rRNA analyses in oxygen-depleted marine environments. Appl Environ Microbiol 69: 2657–2663

    Article  PubMed  CAS  Google Scholar 

  68. Stoeck, T, Taylor, G, Epstein, SS (2003) Novel eukaryotes from a permanently anoxic Cariaco Basin (Caribbean Sea). Appl Environ Microbiol 69: 5656–5663

    Article  PubMed  CAS  Google Scholar 

  69. Stoeck, T, Hayward, B, Taylor, GT, Varela, R, Epstein, SS (2006) A multiple PCR-primer approach to access the microeukaryotic diversity in environmental samples. Protist 157: 31–43

    Article  PubMed  CAS  Google Scholar 

  70. Taylor, GT, Scranton, MI, Iabichella, M, Ho, T-Y, Thunell, RC, Muller-Karger, F, Varela, R (2001) Chemoautotrophy in the redox transition zone of the Cariaco Basin: a significant midwater source of organic carbon production. Limnol Oceanogr 46: 148–163

    Article  CAS  Google Scholar 

  71. Teske, A, Wawer, C, Muyzer, G, Ramsing, NB (1996) Distribution of sulfate-reducing bacteria in a stratified fjord (Mariager Fjord, Denmark) as evaluated by most-probable-number counts and denaturing gradient gel electrophoresis of PCR-amplified ribosomal DNA fragments. Appl Environ Microbiol 62: 1405–1415

    PubMed  CAS  Google Scholar 

  72. Throndsen, J (1988) Cymbomonas Schiller (Prasinophyceae) reinvestigated by light and electron microscopy. Arch Protistenkd 136: 327–335

    Google Scholar 

  73. Tong, SM (1997) Heterotrophic flagellates and other protists from Southampton water, UK. Ophelia 47: 71–131

    Google Scholar 

  74. Turmel, M, Otis, C, Lemieux, C (2002) The complete mitochondrial DNA sequence of Mesostigma viride identifies this green alga as the earliest green plant divergence and predicts a highly compact mitochondrial genome in the ancestor of all green plants. Mol Biol Evol 19(1): 24–38

    CAS  Google Scholar 

  75. Wagner, R (1994) The regulation of ribosomal RNA synthesis and bacterial cell growth. Arch Microbiol 63: 3741–3751

    Google Scholar 

  76. Weinbauer, MG, Fritz, I, Wenderoth, DF, Höfle, MG (2002) Simultaneous extraction from bacterioplankton of total RNA and DNA suitable for quantitative structure and function analyses. Appl Environ Microbiol 68: 1082–1087

    Article  PubMed  CAS  Google Scholar 

  77. Weller, R, Ward, DM (1989) Selective recovery of 16S rRNA sequences from natural microbial communities in the form of cDNA. Appl Environ Microbiol 55: 1818–1822

    PubMed  CAS  Google Scholar 

  78. Weller, R, Weller, JW, Ward, DM (1991) 16S rRNA sequences of uncultivated hot spring cyanobacterial mat inhabitants retrieved as randomly primed cDNA. Appl Environ Microbiol 57: 1146–1151

    PubMed  CAS  Google Scholar 

  79. Wuyts, J, Perriere, G, Van de Peer, Y (2004) The European ribosomal RNA database. Nucleic Acids Res 32: D101–D103

    Article  PubMed  CAS  Google Scholar 

  80. Zohary, T, Pais-Madeira, AM, Robarts, R, Hambright, KD (1996) Interannual phytoplankton dynamics of a hypertrophic African lake. Arch Hydrobiol 136: 105–126

    Google Scholar 

Download references

Acknowledgments

We thank U. Frisenette, Captain of the RV Maria for all his support during sampling in Mariager and providing his expertise and all facilities that enabled sampling of the Mariager Fjord. We are grateful to Anika Knapp for initial RNA work. K. Jürgens provided valuable comments on an earlier draft of the manuscript and E. Jarroll helped to improve the language. We greatly appreciate the comments of two anonymous reviewers, who helped to significantly improve our manuscript. This study was funded by grants from the Deutsche Forschungsgemeinschaft (DFG) to TS (STO414/2-2) and the Stiftung Rheinland-Pfalz für Innovation (project 736).

Author information

Authors and Affiliations

  1. School of Biology, University of Kaiserslautern, Erwin-Schroedinger-Str. 14, D-67663, Kaiserslautern, Germany

    Thorsten Stoeck, Alexandra Zuendorf, Hans-Werner Breiner & Anke Behnke

Authors
  1. Thorsten Stoeck
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alexandra Zuendorf
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hans-Werner Breiner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Anke Behnke
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Thorsten Stoeck.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stoeck, T., Zuendorf, A., Breiner, HW. et al. A Molecular Approach to Identify Active Microbes in Environmental Eukaryote Clone Libraries. Microb Ecol 53, 328–339 (2007). https://doi.org/10.1007/s00248-006-9166-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00248-006-9166-1

Keywords

Search

Navigation