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Nitrogen-Cycling Genes in Epilithic Biofilms of Oligotrophic High-Altitude Lakes (Central Pyrenees, Spain)

  • Microbiology of Aquatic Systems
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Abstract

Microbial biofilms in oligotrophic environments are the most reactive component of the ecosystem. In high-altitude lakes, exposed bedrock, boulders, gravel, and sand in contact with highly oxygenated water and where a very thin epilithic biofilm develops usually dominate the littoral zone. Traditionally, these surfaces have been considered unsuitable for denitrification, but recent investigations have shown higher biological diversity than expected, including diverse anaerobic microorganisms. In this study, we explored the presence of microbial N-cycling nirS and nirK (denitrification through the conversion of NO2 − to NO), nifH (N2 fixation), anammox (anaerobic ammonium oxidation), and amoA (aerobic ammonia oxidation, both bacterial and archaeal) genes in epilithic biofilms of a set of high-altitude oligotrophic lakes in the Pyrenees. The concentrations of denitrifying genes determined by quantitative PCR were two orders of magnitude higher than those of ammonia-oxidizing genes. Both types of genes were significantly correlated, suggesting a potential tight coupling nitrification-denitrification in these biofilms that deserves further confirmation. The nifH gene was detected after nested PCR, and no signal was detected for the anammox-specific genes used. The taxonomic composition of denitrifying and nitrogen-fixing genes was further explored by cloning and sequencing. Interestingly, both microbial functional groups were richer and more genetically diverse than expected. The nirK gene, mostly related to Alphaproteobacteria (Bradyrhizobiaceae), dominated the denitrifying gene pool as expected for oxygen-exposed habitats, whereas Deltaproteobacteria (Geobacter like) and Cyanobacteria were the most abundant among nitrogen fixers. Overall, these results suggest an epilithic community more metabolically diverse than previously thought and with the potential to carry out an active role in the biogeochemical nitrogen cycling of high-altitude ecosystems. Measurements of activity rates should be however carried out to substantiate and further explore these findings.

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Acknowledgments

We are thankful to the Centre de Recerca d’Alta Muntanya (CRAM), Universitat de Barcelona, Vielha, for laboratory facilities. X Triadó, L Banyeras, A Calviño, and A Fernández-Guerra are acknowledged for their technical and bioinformatics assistance, and S Merbft and J-C Auguet for qPCR compiled data. We thank two anonymous reviewers for the helpful comments on the manuscript. MVC was benefited by the Program Juan de la Cierva, Spanish Office for Science (MINECO) and currently by Beatriu de Pinós (Generalitat de Catalunya) postdoctoral fellowships. This study was supported by MINECO grants DARKNESS (CGL2012-32747) to EOC and NITROPIR (CGL2010-19373) to JC.

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Authors and Affiliations

  1. Biogeodynamics&Biodiversity Group (B&B), Centre for Advanced Studies of Blanes, CEAB-CSIC, Blanes, Spain

    Maria Vila-Costa, Mireia Bartrons, Jordi Catalan & Emilio O. Casamayor

  2. Department of Ecology, Department of Environmental Chemistry, University of Barcelona, IDAEA-CSIC, Barcelona, Spain

    Maria Vila-Costa

  3. Department of Environmental Sciences and Food, University of Vic, Barcelona, Spain

    Mireia Bartrons

  4. B&B-CREAF, Campus UAB, Cerdanyola del Vallès, Catalunya, Spain

    Jordi Catalan

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  1. Maria Vila-Costa
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  2. Mireia Bartrons
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  4. Emilio O. Casamayor
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Correspondence to Emilio O. Casamayor.

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Vila-Costa, M., Bartrons, M., Catalan, J. et al. Nitrogen-Cycling Genes in Epilithic Biofilms of Oligotrophic High-Altitude Lakes (Central Pyrenees, Spain). Microb Ecol 68, 60–69 (2014). https://doi.org/10.1007/s00248-014-0417-2

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  • DOI: https://doi.org/10.1007/s00248-014-0417-2

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