Abstract
Culture conditions for the maintenance of previously uncultured members of the Archaea thriving in anoxic water layers of stratified freshwater lakes are described. The proposed enrichment conditions, based on the use of defined medium composition and the maintenance of anoxia, have been proven effective for the maintenance of the archaeal community with virtually no changes over time for periods up to 6 months as revealed by a PCR-DGGE analysis. Phylotypes belonging to groups poorly represented in culture collections such as the Deep-Sea Hydrothermal Vent Euryarchaeota (DHVE) and the Miscellaneous Crenarchaeotic Group (MCG) were maintained and selectively enriched when compared to the correspondent indigenous planktonic archaeal community.
References
Auguet JC, Casamayor EO (2008) A hotspot for cold Crenarchaeota in the neuston of high mountain lakes. Environ Microbiol 10:1080–1086
Balch WE, Fox GE, Magrum LJ, Woese CR, Wolfe RS (1979) Methanogens: reevaluation of a unique biological group. Microbiol Rev 43:260–296
Casamayor EO, Borrego CM (2009) Archaea. In: Likens GE (ed) Encyclopedia of inland waters, vol 3. Elsevier, Oxford, pp 167–181
Casamayor EO, Muyzer G, Pedrós-Alió C (2001) Composition and temporal dynamics of planktonic archaeal assemblages from anaerobic sulfurous environments studied by 16S rDNA denaturing gradient gel electrophoresis and sequencing. Aquat Microb Ecol 25:237–246
Chaban B, Ng SYM, Jarrell KF (2006) Archaeal habitats from the extreme to the ordinary. Can J Microbiol 52:73–116
de la Torre JR, Walker CB, Ingalls AE, Könneke M, Stahl DA (2008) Cultivation of a thermophilic ammonia oxidizing archaeon synthesizing crenarchaeol. Environ Microbiol 10:810–818
DeLong EF (1992) Archaea in coastal marine environments. Proc Natl Acad Sci USA 89:5685–5689
DeLong EF (1998) Everything in moderation: archaea as ‘non-extremophiles’. Curr Opin Genet Dev 8:649–654
Donachie SP, Foster JS, Brown MV (2007) Culture clash: challenging the dogma of microbial diversity. ISME J 1:97–99
Giovannoni S, Stingl U (2007) The importance of culturing bacterioplankton in the ‘omics’ age. Nat Rev Microbiol 5:820–826
Gray ND, Head IM (2007) Linking genetic identity and function in communities of uncultured bacteria. Environ Microbiol 3:481–492
Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98
Hatzenpichler R, Lebedeva EV, Spieck E, Stoecker K, Richter A, Daims H, Wagner M (2008) A moderately thermophilic ammonia-oxidizing crenarchaeote from a hot spring. Proc Natl Acad Sci USA 105(6):2134–2139
Huber T, Faulkner G, Hugenholtz P (2004) Bellerophon: a program to detect chimeric sequences in multiple sequence alignments. Bioinformatics 20:2317–2319
Inagaki F, Suzuki M, Takai K, Oida H, Sakamoto T, Aoki K et al (2003) Microbial communities associated with geological horizons in coastal subseafloor sediments from the Sea of Okhotsk. Appl Environ Microbiol 69:7224–7235
Könneke M, Bernhard AE, de la Torre JR, Walker CB, Waterbury JB, Stahl DA (2005) Isolation of an autotrophic ammonia-oxidizing marine archaeon. Nature 437:543–546
Kozubal M, Macur RE, Korf S, Taylor WP, Ackerman GG, Nagy A, Inskeep WP (2008) Isolation and distribution of a novel iron-oxidizing crenarchaeon from acidic geothermal springs in yellowstone National Park. Appl Environ Microbiol 74(4):942–949
Llirós M, Casamayor EO, Borrego CM (2008) High archaeal richness in the water column of a freshwater sulphurous karstic lake along an inter-annual study. FEMS Microbiol Ecol 66:331–342
Ludwig W, Strunk O, Westram R, Richter L, Meier H, Yadhukumar, Buchner A, Lai T, Steppi S, Jobb G, Forster W, Brettske I, Gerber S, Ginhart AW, Gross O, Grumann S, Hermann S, Jost R, Konig A, Liss T, Lussmann R, May M, Nonhoff B, Reichel B, Strehlow R, Stamatakis A, Stuckmann N, Vilbig A, Lenke M, Ludwig T, Bode A, Schleifer KH (2004) ARB: a software environment for sequence data. Nucleic Acids Res 32:1363–1371
Muyzer G, Brinkhoff T, Nübel U, Santegoeds C, Schäfer H, Wawer C (1998) Denaturing gradient gel electrophoresis (DGGE) in microbial ecology. In: Akkermans ADL, van Elsas JD, de Bruijn FJ (eds) Molecular microbial ecology manual. Kluwer Academic Publishers, Dordrecht, pp 1–27
Nichols D (2007) Cultivation gives context to the microbial ecologist. FEMS Microbiol Ecol 60:351–357
Prosser JI, Nicol GW (2008) Relative contributions of archaea and bacteria to aerobic ammonia oxidation in the environment. Environ Microbiol 10:2931–2941
Schleper C (2008) Metabolism of the deep. Nature 456(11):712–714
Takai K, Horikoshi K (1999) Genetic diversity of archaea in deep-sea hydrothermal vent environments. Genetics 152:1285–1297
Takai K, Moser DP, DeFlaun M, Onstott TC, Fredrickson JK (2001) Archaeal diversity in waters from deep south african gold mines. Appl Environ Microbiol 67:5750–5760
Teske A, Sorensen KB (2008) Uncultured archaea in deep marine subsurface sediments: have we caught them all? ISME J 2:3–18
Teske A, Hinrichs K-U, Edgcomb V, de Vera Gomez A, Kysela D, Sylva SP, Sogin ML, Jannasch HW (2002) Microbial diversity of hydrothermal sediments in the Guaymas basin: Evidence for anaerobic methanotrophic communities. Appl Environ Microbiol 68:1994–2007
Widdel F, Bak F (1992) Gram-negative mesophilic sulphate reducing bacteria. In: Balows THG, Dworkin AM, Harder W, Schleifer KH (eds) The prokaryotes. Springer, New York, pp 3353–3378
Acknowledgments
This work was supported by coordinated projects REN2003-08333 and CRENYC CGL2006-12058 from the Spanish Ministerio de Educación y Ciencia (MEC) to CMB and EOC. We thank A. Camacho and A. Picazo from the Institut Cavanilles de Biodiversitat i Biologia Evolutiva of the University of Valencia for their assistance and valuable support during the sampling campaign in Cuenca. A. Plasencia and M. Llirós benefit from a FI doctoral fellow from the Generalitat de Catalunya and from a FPI doctoral fellow from the Spanish MEC, respectively.
Author information
Authors and Affiliations
Corresponding author
Additional information
Sequences accession numbers
Archaeal 16S rRNA sequences obtained in this study have been deposited in the GenBank under accession numbers EU595421 to EU595447.
Rights and permissions
About this article
Cite this article
Plasencia, A., Bañeras, L., Llirós, M. et al. Maintenance of previously uncultured freshwater archaea from anoxic waters under laboratory conditions. Antonie van Leeuwenhoek 99, 403–408 (2011). https://doi.org/10.1007/s10482-010-9412-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10482-010-9412-x