Archives of Microbiology

, Volume 150, Issue 2, pp 178–183 | Cite as

An extremely thermophilic Methanococcus from a deep sea hydrothermal vent and its plasmid

  • Honxue Zhao
  • Alvin G. Wood
  • Friedrich Widdel
  • Marvin P. Bryant
Original Papers


An extremely thermophilic methanogen was isolated from a hydrothermal vent core sample from Guaymas Basin, Gulf of California, at a depth of 2003 m. The isolate, designated strain AG86, was a coccoid autotroph using H2-CO2 as energy and carbon source with a growth temperature range of 48 to 92°C, optimum, 85°C. AG86 required NaCl and Mg2+ and trace amounts of selenite and tungstate. Vitamins were not required. However, yeast extract, Casamino acids and Trypticase stimulated growth significantly. When grown in the presence of these stimulants and at the optimal growth temperature and pH 6.5, the minimum doubling time was 20 min. Cells were fragile and readily lysed by detergents. The mol% G+C was 33%. These results and partial 16S rRNA sequencing indicated that AG86 belonged to the genus Methanococcus and closely resembled Methanococcus jannaschii. Tests for extrachromosomal DNA revealed a plasmid in AG86 and two plasmids in M. jannaschii. Different patterns were obtained from restriction endonuclease digestion of the three plasmids, and no homology was observed with DNA-DNA hybridization.

Key words

Thermophilic Methanocoecus Deep sea hydrothermal vent methanogen Plasmids 



covalently close circular DNA


defined marine medium


Guanine plus cytosine


most probable number


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  1. Balch WE, Wolfe RS (1976) New approach to the cultivation of methanogenic bacteria: 2-mercaptoethanesulfonic acid (HS-CoM)-dependent growth of Methanobacterium ruminantium in a pressurized atmosphere. Appl Environ Microbiol 32:781–791Google Scholar
  2. Balch WE, Fox GE, Magrum LJ, Woese CR, Wolfe RS (1979) Methanogens: Reevaluation of a unique biological group. Microbiol Rev 43:260–296Google Scholar
  3. Bryant MP (1972) Commentary on the Hungate technique for culture of anaerobic bacteria. Am J Clin Nutr 25:1324–1328Google Scholar
  4. De Ley J (1970) Reexamination of the association between melting point, buoyant density and chemical base composition of deoxyribonucleic acid. J Bacteriol 101:738–754Google Scholar
  5. Fox GE, Peckman KJ, Woese CR (1977) Comparative cataloging of 16S ribosomal ribonucleic acid: molecular approach to prokaryotic systematics. Int J Syst Bacteriol 27:44–57Google Scholar
  6. Harris JE, Pinn RA, Davis RP (1984) Isolation and characterization of a novel thermophilic, freshwater methanogen. Appl Environ Microbiol 48:1123–1128Google Scholar
  7. Hermann M, Noll KM, Wolfe RS (1986) Improved agar bottle plate for isolation of methanogens or other anaerobes in a defined gas atmosphere. Appl Environ Microbiol 51:1124–1126Google Scholar
  8. Hungate RE (1950) The anaerobic mesophilic cellulolytic bacteria. J Bacteriol 149:316–319Google Scholar
  9. Jannasch HW (1988) Isolation of extremely thermophilic, fermentative archaebacteria from deep sea geothermal sediments. In: Wise DL (ed) Biotechnology applied to fossil fuels. CRC Press, Boca Raton (in press)Google Scholar
  10. Jones WJ, Leigh JA, Mayer F, Woese CR, Wolfe RS (1983) Methanococcus jannaschii sp. nov., an extremely thermophilic methanogen from a submarine hydrothermal vent. Arch Microbiol 136:254–261Google Scholar
  11. Lane DJ, Pace B, Olsen GJ, Stahl DA, Sogin ML, Pace NR (1985) Rapid determination of 16S ribosomal RNA sequences for phylogenetic analysis. Proc Natl Acad Sci USA 82:6955–6959Google Scholar
  12. Mandel M, Marmur J (1978) Use of ultraviolet absorbance temperature profile for determining the guanine pulse cytosine content of DNA. In: Grossman L, Moldave K (eds) Methods in enzymology, vol 12, part B. Academic Press, New York, pp 195–206Google Scholar
  13. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  14. Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3:208–218Google Scholar
  15. Paterek JR, Smith PH (1985) Isolation and characterization of a halophilic methanogen from Great Salt Lake. Appl Environ Microbiol 50:877–881Google Scholar
  16. Pfennig N, Wagener S (1986) An improved method of preparing wet mounts for photomicrographs of microorganisms. J Microbiol Meth 4:303–306Google Scholar
  17. Razin S, Harasawa R, Barile MF (1983) Cleavage patterns of the mycoplasma chromosome, obtained by using restriction endonucleases, as indicators of genetic relatedness among strains. Int J Syst Bacteriol 33:201–206Google Scholar
  18. Rodina AG (1972) Methods in aquatic microbiology. University Park Press, Baltimore, pp 177–180Google Scholar
  19. Saito H, Miura KH (1963) Preparation of transforming deoxyribonucleic acid by phenol treatment. Biochim Biophys Acta 72:619–629Google Scholar
  20. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517Google Scholar
  21. Whitman WB, Ankwand E, Wolfe RS (1982) Nutrition and carbon metabolism of Methanococcus voltae. J Bacteriol 149:852–862Google Scholar
  22. Widdel F (1986) Growth of methanogenic bacteria in pure culture with 2-propanol and other alcohols and hydrogen donors. Appl Environ Microbiol 51:1056–1062Google Scholar
  23. Widdel F, Pfennig N (1984) Dissimilatory sulfate-sulfur-reducing bacteria. In: Krieg NR, Holt JG (eds) Bergey's manual of systematic bacteriology, vol 1. The William and Wilkins Co., Baltimore, pp 663–679Google Scholar
  24. Wood AG, Whitman WB, Konisky J (1985) A newly-isolated marine methanogen harbors a small cryptic plasmid. Arch Microbiol 142:259–261Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Honxue Zhao
    • 1
  • Alvin G. Wood
    • 1
  • Friedrich Widdel
    • 1
  • Marvin P. Bryant
    • 1
  1. 1.Department of Animal Sciences and MicrobiologyUniversity of IllinoisUrbanaUSA

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