Abstract
A metagenomic study of the Kolyma lowland permafrost samples, 20–35 thousand years, performed using a Geneclean for Ancient DNA kit (Bio101, United States), revealed 8 phylotypes which belonged to the phyla Actinobacteria and Proteobacteria. Analysis of the 16S rRNA gene clone library showed that most of the clones (48% and 29%) were represented by the genera Arthrobacter and Bradyrhizobium, respectively. For the first time microorganisms of the genera Williamsia, Bradyrhizobium, Filomicrobium and Hansschlegelia were observed in the ancient microbial communities of these ecosystems. Analysis of the isolates 16S rRNA genes revealed the presence of the microorganisms—the representatives of the phyla Firmicutes and Actinobacteria phylogenetically related to known species and being obvious representatives of novel taxa. In situ electron-microscope analysis of total preparations of the studied samples showed the presence of intact bacterial cells of different morphotypes.
Similar content being viewed by others
References
Duchkov, A.D., Characteristics of permafrost in Siberia, Advances in the Geological Storage of Carbon Dioxide. NATO Science Ser. IV, 2006, vol. 65, pp. 81–92.
Gilichinsky, D.A., Permafrost, in Encyclopedia of Environmental Microbiology, Bitton, G., Ed., New York: Wiley, 2002, pp. 2367–2385.
Soina, V.S., Vorobyova, E.A., Zvyagintsev, D., and Gilichinsky, D., Preservation of cells in permafrost: a model for exobiology, Adv. Space Res., 1995, vol. 15, no. 3, pp. 237–242.
Vorobyova, E., Soina, V., Gorlenko, M., Minkovskaya, N., Zalinova, N., Mamukelashvili, A., Gilichinsky, D., Rivkina, E., and Vishnivetskaya, T., The deep cold biosphere: facts and hypothesis, FEMS Microbiol. Rev., 1997, vol. 20, pp. 277–290.
Tiedje, J.M., Petrova, M.A., and Moyer, C., Phylogenetic diversity of archaea from ancient siberian permafrost, Abstr. 8th Int. Symp. Microb. Ecol. (ISME-8), Halifax: Atlantic Canada Soc. Microb. Ecol., 1998, p. 323.
Vishnivetskaya, T.A., Petrova, M.A., Urbance, J., Ponder, M., Moyer, C.L., Gilichinsky, D.A., and Tiedje, J.M., Bacterial community in ancient siberian permafrost as characterized by culture and cultureindependent methods, Astrobiology, 2006, vol. 6, no. 3, pp. 400–414.
Khlebnikova, G.M., Gilichinsky, D.A., FedorovDavydov, D.G., and Vorobyeva, E.A., Quantitative evaluation of microorganisms in permafrost deposits and buried soils, Microbiology, 1990, vol. 59, no. 1, pp. 106–111.
Bej, A.K., Mahbubani, M.H., Dicesare, J.L., and Atlas, R.M., Polymerase chain reaction-gene probe detection of microorganisms by using filter-concentrated samples, Appl. Environ. Microbiol., 1991, vol. 57, pp. 3529–3534.
Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A., and Struhl, K., Current Protocols in Molecular Biology, New York: Wiley, 1994.
Alexandrov, A., Kaushik, D., and Pascal, S.M., Streamline method to Analyze 16S rRNA gene clone libraries, BioTechniques, 2001, vol. 30, pp. 938–944.
Cole, J.R., Wang, Q., Cardenas, E., Fish, J., Chai, B., Farris, R.J., Kulam-Syed-Mohideen, A.S., McGarrell, D.M., Marsh, T., Garrity, G.M., and Tiedje, J.M., The Ribosomal database project: improved alignments and new tools for RRNA analysis, Nucleic Acid Res., 2008, vol. 37(Database issue) D141–145. doi: 10.1093/nar/gkn879
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., and Kumar, S., MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods, Mol. Biol. Evol., 2011, vol. 28, no. 10, pp. 2731–2739. doi: 10.1093/molbev/msr121
Giandomenico, A.R., Cerniglia, G.E., Biaglow, J.E., Stevens, C.W., and Koch, C.J., The importance of sodium pyruvate in assessing damage produced by hydrogen peroxide, Free Radic. Biol. Med., 1997, vol. 23, no. 3, pp. 426–434.
Ameyama, M., Shinagawa, E., Matsushita, K., and Adachi, O., Growth stimulating activity for microorganisms in naturally occurring substances and partial characterization of the substance for the activity as pyrroloquinoline quinine, Agric. Biol. Chem., 1985, vol. 49, no. 3, pp. 699–709.
Krivan, H.C., Franklin, D.P., Wang, W., Laux, D.C., and Cohen, P.S., Phosphatidylserine found in intestinal mucus serves as a sole source of carbon and nitrogen for salmonellae and Escherichia coli, Infect. Immun., 1992, vol. 60, no. 9, pp. 3943–3946.
Bruns, A., Cypionka, H., and Overmann, J., Cyclic AMP and acyl homoserine lactones increase the cultivation efficiency of heterotrophic bacteria from the central Baltic sea, Appl. Environ. Microbiol., 2002, vol. 68, no. 8, pp. 3978–3987.
Suzina, N.E., Esikova, T.Z., Akimov, V.N., Abashina, T.N., Dmitriev, V.V., Polivtseva, V.N., Duda, V.I., and Boronin, A.M., Study of ectoparasitism of ultramicrobacteria of the genus Kaistia, strains NF1 and NF3 by electron and fluorescece microscopy, Microbiology, 2008, vol. 77, no. 1, pp. 47–54.
Vainshtein, M., Suzina, N., Kudryashova, E., and Ariskina, E., New magnet-sensitive structures in bacterial and archaeal cells, Biol. Cell, 2002, vol. 94, pp. 29–35.
Tindall, B.J., Rossello-Móra, R., Busse, H.-J., Ludwig, W., and KäMpfer, P., Notes on the characterization of prokaryote strains for taxonomic purposes, Int. J. Syst. Evol. Microbiol., 2010, vol. 60, pp. 249–266.
Dmitriev, V.V., Suzina, N.E., Rusakova, T.G., Gilichinskii, D.A., and Duda, V.I., Ultrastructural characteristics of natural forms of microorganisms isolated from permafrost grounds of Eastern Siberia by the method of low-temperature fractionation, Doklady Biol. Sci., 2001, vol. 378, pp. 304–307.
Steven, B., Briggs, G., McKay, C.P., Pollard, W.H., Greer, C.W., and Whyte, L.G., Characterization of the microbial diversity in a permafrost sample from the canadian high arctic using culture-dependent and culture-independent methods, FEMS Microbiol. Ecol., 2007, vol. 59, pp. 513–523.
Katayama, T., Tanaka, M., Moriizumi, J., Nakamura, T., Brouchkov, A., Douglas, T.A., Fukuda, M., Tomita, F., and Asano, K., Phylogenetic analysis of bacteria preserved in a permafrost ice wedge for 25.000 years, Appl. Environ. Microbiol., 2007, vol. 73, pp. 2360–2363.
Spirina, E.V., Cole, J.R., Chai, B., Gilichinsky, D.A., and Tiedje, J.M., Phylogenetic diversity of tundra cryosol microbial community in Arctic ecosystems, Abstr. 2nd Eur. Conf. Permafrost. EUCOP 2005, Potsdam, 2005, p. 51.
Karasev, S.G., Gurina, L.V., Gavrish, E.Yu., Adanin, V.M., Gilichinskii, D.A., and Evtushenko, L.I., Viable actinobacteria form Siberian ancient permafrost deposits, Kriosfera Zemli, 1998, vol. 2, no. 2, pp. 69–75.
Kochkina, G.A., Ivanushkina, N.E., Karasev, S.G., Gavrish, E.Yu., Gurina, L.V., Evtushenko, L.I., Spirina, E.V., Vorob’Eva, E.A., Gilichinskii, D.A., and Ozerskaya, S.M., Survival of micromycetes and actinobacteria under conditions of long-term natural cryopreservation, Microbiology, 2001, vol. 70, no. 3, pp. 356–364.
Gilichinsky, D.A., Wilson, G.S., Friedmann, E.I., McKay, C.P., Sletten, R.S., Rivkina, E.M., Vishnivetskaya, T.A., Erokhina, L.G., Ivanushkina, N.E., Kochkina, G.A., Shcherbakova, V.A., Soina, V.S., Spirina, E.V., Vorobyova, E.A., Fyodorov-Davydov, D.G., Hallet, B., Ozerskaya, S.M., Sorokovikov, V.A., Laurinavichyus, K.S., Shatilovich, A.V., Chanton, P., Ostroumov, V.E., and Tiedje, J.M., Microbial populations in Antarctic permafrost: biodiversity, state, age and implication for astrobiology, Astrobiology, 2007, vol. 7, pp. 275–311.
Spirina, E., Cole, J., Chai, B., Gilichinsky, D., and Tiedje, J., High throughput approach to study ancient microbial phylogenetic diversity in permafrost as a terrestrial model of Mars, Astrobiology, 2003, vol. 2, pp. 542–543.
Marteinsson, V.T., Kristjansson, J.K., Kristmannsdottir, H., Dahlkvist, M., Saemundsson, K., Hannington, M., Petursdottir, S.K., Geptner, A., and Stoffers, P., Discovery and description of giant submarine smectite cones on the seafloor in Eyjafjordur, Northern Iceland, and a novel thermal microbial habitat, Appl. Environ. Microbiol., 2001, vol. 67, pp. 827–833.
Takeuchi, M., Komai, T., Hanada, S., Tamaki, H., Tanabe, S., Miyachi, Y., Uchiyama, M., Nakazawa, T., Kimura, K., and Kamagata, Y., Bacterial and archaeal 16S rRNA genes in Late Pleistocene to Holocene muddy sediments from the Kanto Plain of Japan, Geomicrobiol. J., 2009, vol. 26, pp. 104–118.
Mulyukin, A.L., Demkina, E.V., Kozlova, A.N., Soina, V.S., and El’Registan, G.I., Synthesis of Anabiosis autoinducers by non-spore-forming bacteria as a mechanism regulating their activity in soil and subsoil sedimentary rocks, Microbiology, 2001, vol. 70, no. 5, pp. 535–541.
Johnson, S.S., Hebsgaard, M.B., Christensen, T.R., Mastepanov, M., Neilson, R., Munch, K., Brand, T., Gilbert, M., Zuber, M., Bunce, M., Ronn, R., Gilichinsky, D., Froese, D., and Willerslev, E., Ancient bacteria show evidence of DNA repair, Proc. Natl. Acad. Sci. USA, 2007, vol. 104, no. 36, pp. 14401–14405.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © E.B. Kudryashova, E.Yu. Chernousova, N.E. Suzina, E.V. Ariskina, D.A. Gilichinsky, 2013, published in Mikrobiologiya, 2013, Vol. 82, No. 3, pp. 351–361.
The article was translated by the authors.
Rights and permissions
About this article
Cite this article
Kudryashova, E.B., Chernousova, E.Y., Suzina, N.E. et al. Microbial diversity of Late Pleistocene Siberian permafrost samples. Microbiology 82, 341–351 (2013). https://doi.org/10.1134/S0026261713020082
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S0026261713020082