, Volume 12, Issue 4, pp 481–490 | Cite as

Metabolic activity of Siberian permafrost isolates, Psychrobacter arcticus and Exiguobacterium sibiricum, at low water activities

  • Monica A. Ponder
  • Michael F. Thomashow
  • James M. Tiedje
Original Paper


The Siberian permafrost is an extreme, yet stable environment due to its continuously frozen state. Microbes maintain membrane potential and respiratory activity at average temperatures of −10 to −12°C that concentrate solutes to an a w = 0.90 (5 osm), The isolation of viable Psychrobacter arcticus sp. 273-4 and Exiguobacterium sibiricum sp. 255-15 from ancient permafrost suggests that these bacteria have maintained some level of metabolic activity for thousands of years. Permafrost water activity was simulated using ½ TSB + 2.79 m NaCl (5 osm) at and cells were held at 22 and 4°C. Many cells reduced cyano-tetrazolium chloride (CTC) indicating functioning electron transport systems. Increased membrane permeability was not responsible for this lack of electron transport, as more cells were determined to be intact by LIVE/DEAD staining than were reducing CTC. Low rates of aerobic respiration were determined by the slope of the reduced resazurin line for P. arcticus, and E. sibiricum. Tritiated leucine was incorporated into new proteins at rates indicating basal level metabolism. The continued membrane potential, electron transport and aerobic respiration, coupled with incorporation of radio-labeled leucine into cell material when incubated in high osmolarity media, show that some of the population is metabolically active under simulated in situ conditions.


Psychrobacter Siberian permafrost Exiguobacterium Salt tolerance Low temperature Low water activity 



This research was funded by the National Astrobiology Institute of NASA. We thank Tatiana Vishnivetskaya for input based on preliminary physiological data which allowed us to focus on a narrower number of interesting permafrost isolated strains for further studies within our laboratory. We acknowledge the assistance of Chia-Kai Chang, Gisel Rodriguez, and Matt Campbell. We thank Richard Lenski and Corien Bakermans for strains E.coli 606 and P. cryohalolentis, respectively.


  1. Albers SV, Van de Vossenberg JL, Driessen AJ, Konings WN (2001) Bioenergetics and solute uptake under extreme conditions. Extremophiles 5:285–294PubMedGoogle Scholar
  2. Bakermans C, Ayala-del-Rio HL, Ponder MA, Vishnivetskaya T, Gilichinsky D, Thomashow MF, Tiedje JM (2006) Psychrobacter cryohalolentis sp. nov. and Psychrobacter arcticus sp. nov., isolated from Siberian permafrost. Int J Syst Evol Microbiol 56:1285–1291PubMedCrossRefGoogle Scholar
  3. Bakermans C, Tsapin AI, Souza-Egipsy V, Gilichinsky DA, Nealson KH (2003) Reproduction and metabolism at −10°C of bacteria isolated from Siberian permafrost. Environ Microbiol 5:321–326PubMedCrossRefGoogle Scholar
  4. Brinton KLF, Tsapin AI, Gilichinsky D, McDonald GD (2002) Aspartic acid racemization and age-depth relationships for organic carbon in Siberian permafrost. Astrobiology 2:77–82PubMedCrossRefGoogle Scholar
  5. Buesing N, Gessner MO (2003) Incorporation of radiolabeled leucine into protein to estimate bacterial production in plant litter, sediment, epiphytic biofilms, and water samples. Microb Ecol 45:291–301PubMedCrossRefGoogle Scholar
  6. Carpenter EJ, Lin S, Capone DG (2000) Bacterial activity in South Pole snow. Appl Environ Microbiol 66:4514–4517PubMedCrossRefGoogle Scholar
  7. Christner BC (2002) Incorporation of DNA and protein precursors into macromolecules by bacteria at −15°C. Appl Environ Microbiol 68:6435–6438PubMedCrossRefGoogle Scholar
  8. Friedmann EI (1994) Permafrost as microbial habitat. Viable microorganisms in permafrost. D. Gilichinsky. Puschinio, Russian Academy of Sciences, pp 21–26Google Scholar
  9. Gilichinsky D (1993) Viable microorganisms in permafrost: the spectrum of possible applications to investigations in science for cold regions. Fourth International symposium on thermal engineering and science for cold regions, Hanover, NH, US. Army Corp of EngineersGoogle Scholar
  10. Gilichinsky D, Rivkina E, Shcherbakova V, Laurinavichuis K, Tiedje J (2003) Supercooled water brines within permafrost—an unknown ecological niche for microorganisms: a model for astrobiology. Astrobiology 3:331–341PubMedCrossRefGoogle Scholar
  11. Gilichinsky DA, Wagener S, Vishnevetskaya T (1995) Permafrost microbiology. Permafrost Periglacial Process 6:281–291CrossRefGoogle Scholar
  12. Gilichinsky DA, Wilson GS, Friedmann EI, McKay CP, Sletten RS, Rivkina EM, Vishnivetskaya TA, Erokhina LG, Ivanushkina NE, Kochkina GA, Shcherbakova VA, Soina VS, Spirina EV, Vorobyova EA, Fyodorov-Davydov DG, Hallet B, Ozerskaya SM, Sorokovikov VA, Laurinavichyus KS, Shatilovich AV, Chanton JP, Ostroumov VE, Tiedje JM (2007) Microbial populations in Antarctic permafrost: biodiversity, state, age, and implication for astrobiology. Astrobiology 7:275–311PubMedCrossRefGoogle Scholar
  13. Junge K, Eicken H, Deming JW (2004) Bacterial activity at −2 to −20 degrees C in Arctic wintertime sea ice. Appl Environ Microbiol 70:550–557PubMedCrossRefGoogle Scholar
  14. Karl DM, Bird DF, Bjorkman K, Houlihan T, Shackelford R, Tupas L (1999) Microorganisms in the accreted ice of Lake Vostok, Antarctica. Science 286:2144–2147PubMedCrossRefGoogle Scholar
  15. Krumbein WE, Gorbushina AA, Holtkamp-Tacken E (2004) Hypersaline microbial systems of sabkhas: examples of life’s survival in “extreme” conditions. Astrobiology 4:450–459PubMedCrossRefGoogle Scholar
  16. Litchfield CD (1998) Survival strategies for microorganisms in hypersaline environments and their relevance to life on early Mars. Meteorit Planet Sci 33:813–819PubMedCrossRefGoogle Scholar
  17. Ostroumov VE, Siegert C (1996) Exobiological aspects of mass transfer in microzones of permafrost deposits. Life Sci Space Mars Recent Results 18:79–86Google Scholar
  18. Pewe T (1995) Permafrost. Encylopedia Britannica, pp 752–759Google Scholar
  19. Ponder M (2005) Characterization of physiological and transcriptome changes in the ancient Siberian permafrost bacterium Psychrobacter arcticus 273-4 with low temperature and increased osmotica. Microbiology and molecular genetics. Michigan State University, East Lansing, p 215Google Scholar
  20. Ponder MA, Gilmour SJ, Bergholz PW, Mindock CA, Hollingsworth R, Thomashow MF, Tiedje JM (2005) Characterization of potential stress responses in ancient Siberian permafrost psychroactive bacteria. FEMS Microbiol Ecol 53:103–115PubMedCrossRefGoogle Scholar
  21. Price PB, Sowers T (2004) Temperature dependence of metabolic rates for microbial growth, maintenance, and survival. Proc Natl Acad Sci USA 101:4631–6PubMedCrossRefGoogle Scholar
  22. Rand RP (2004) “Osmotic Stress Pressure Measurements.” Retrieved 11-6-07, 2007, from http://www.brocku.ca/researchers/peter_rand/osmotic/osfile.html
  23. Rivkina EM, Friedmann EI, McKay CP, Gilichinsky DA (2000) Metabolic activity of permafrost bacteria below the freezing point. Appl Environ Microbiol 66:3230–3233PubMedCrossRefGoogle Scholar
  24. Rodrigues D, Goris J, Vishnivetskaya T, Gilichinsky D, Thomashow M, Tiedje J (2006) Characterization of exiguobacterium isolates from the Siberian permafrost. Description of Exiguobacterium sibiricum sp. nov. Extremophiles 10:285–294PubMedCrossRefGoogle Scholar
  25. Soina VS, Mulyukin AL, Demkina EV, Vorobyova EA, El-Registan GI (2004) The structure of resting bacterial populations in soil and subsoil permafrost. Astrobiology 4:345–358PubMedCrossRefGoogle Scholar
  26. Vishnivetskaya T, Kathariou S, McGrath J, Gilichinsky D, Tiedje JM (2000) Low-temperature recovery strategies for the isolation of bacteria from ancient permafrost sediments. Extremophiles 4:165–173PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2008

Authors and Affiliations

  • Monica A. Ponder
    • 1
    • 4
    • 5
  • Michael F. Thomashow
    • 1
    • 2
    • 3
    • 4
  • James M. Tiedje
    • 1
    • 2
    • 4
  1. 1.Department of Microbiology and Molecular GeneticsMichigan State UniversityEast LansingUSA
  2. 2.Department of Crop and Soil SciencesMichigan State UniversityEast LansingUSA
  3. 3.MSU-DOE Plant Research LabMichigan State UniversityEast LansingUSA
  4. 4.NASA Astrobiology Institute’s Center for Genomic and Evolutionary Studies on Microbial Life at Low TemperaturesMichigan State UniversityEast LansingUSA
  5. 5.101 Food Science and Technology (0418), Department of Food Science and TechnologyVirginia Polytechnic Institute and State UniversityBlacksburgUSA

Personalised recommendations