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Isolation of a 250 million-year-old halotolerant bacterium from a primary salt crystal

Nature volumeΒ 407,Β pages 897–900 (2000) Cite this article

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Abstract

Bacteria have been found associated with a variety of ancient samples1, however few studies are generally accepted due to questions about sample quality and contamination. When Cano and Borucki2 isolated a strain of Bacillus sphaericus from an extinct bee trapped in 25–30 million-year-old amber, careful sample selection and stringent sterilization techniques were the keys to acceptance. Here we report the isolation and growth of a previously unrecognized spore-forming bacterium (Bacillus species, designated 2-9-3) from a brine inclusion within a 250 million-year-old salt crystal from the Permian Salado Formation. Complete gene sequences of the 16S ribosomal DNA show that the organism is part of the lineage of Bacillus marismortui and Virgibacillus pantothenticus. Delicate crystal structures and sedimentary features indicate the salt has not recrystallized since formation. Samples were rejected if brine inclusions showed physical signs of possible contamination. Surfaces of salt crystal samples were sterilized with strong alkali and acid before extracting brines from inclusions. Sterilization procedures reduce the probability of contamination to less than 1 in 109.

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Figure 1: Halite crystals taken from the dissolution pipe at the 569 m (1850 ft) level of the Salado Formation (air intake shaft18 for the Waste Isolation Pilot Plant, Carlsbad, New Mexico) in October 1998.
Figure 2: Phylogenetic tree developed from the 16S rDNA sequence of unknown 2-9-3 compared with several other species of the family Bacillaceae.

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References

  1. Grant, W. D., Gemmell, R. T. & McGenity, T. J. Halobacteria: the evidence for longevity. Extremophiles 2, 279–287 (1998).

    ArticleΒ  CASΒ  PubMedΒ  Google ScholarΒ 

  2. Cano, R. J. & Borucki, M. Revival and identification of bacterial spores in 25 to 40 million year old Dominican amber. Science 268, 1060–1064 (1995).

    ADSΒ  CASΒ  PubMedΒ  Google ScholarΒ 

  3. Denner, E. B. M. et al. Halococcus salifodinae sp. nov., an archaeal isolate from an Austrian salt mine. Int. J. Syst. Bacteriol. 44, 774–780 (1994).

    ArticleΒ  Google ScholarΒ 

  4. Huval, J. H. & Vreeland, R. H. in General and Applied Aspects of Halophilic Bacteria. Vol. 201 (ed. Rodriguez-Valera, F.) 53–62 (Plenum, New York, 1991).

    Google ScholarΒ 

  5. Norton, C. F., McGenity, T. J. & Grant, W. D. Archaeal halophiles (halobacteria) from two British salt mines. J. Gen. Microbiol. 139, 1077 –1081 (1993).

    ArticleΒ  CASΒ  Google ScholarΒ 

  6. Greenblatt, C. L. et al. Diversity of microorganisms isolated from amber. Microb. Ecol. 38, 58–68 (1999).

    ArticleΒ  CASΒ  PubMedΒ  Google ScholarΒ 

  7. Lambert, L. H. et al. Staphylococcus succinus sp. nov., isolated from Dominican amber. Int. J. Syst. Bacteriol. 48, 511– 518 (1998).

    ArticleΒ  CASΒ  PubMedΒ  Google ScholarΒ 

  8. Vreeland, R. H. & Powers, D. W. in Microbiolology and Biogeochemistry of Hypersaline Environments (ed. Oren, A.) 53–74 (CRC, Boca Raton, Florida, 1999 ).

    Google ScholarΒ 

  9. Vreeland, R. H. & Rosenzweig, W. D. in Enigmatic and Extreme Microorganisms. (ed. Seckbach, J.) 387– 398 (Kluwer, Delft, 1998).

    Google ScholarΒ 

  10. Croft, J. S. Upper Permian conodonts and other microfossils from the Pinery and Lamar Limestone Members of the Bell Canyon Formation and from the Rustler Formation, west Texas. Thesis, Ohio State Univ. (1978).

  11. Walter, J. C. Paleontology of the Rustler Formation, Culberson County, Texas. J. of Paleontol. 27, 679–702 (1953).

    Google ScholarΒ 

  12. Renne, P. R., Steiner, M. B., Sharp, W. D., Ludwig, K. R. & Fanning, C. M. 40/39 Ar and U/Pb SHRIMP dating of latest Permian tephras in the Midland Basin Texas. EOS 77, 794 (1996).

    Google ScholarΒ 

  13. Renne, P. R., Sharp, W. D. & Becker, T. A. 40Ar/39Ar dating of langbeinite [K2Mg2(SO4)3] in late Permian evaporites of the Salado Formation, Southeastern New Mexico, USA. Mineral. Mag. 62A, 1253– 1254 (1998).

    ArticleΒ  ADSΒ  CASΒ  Google ScholarΒ 

  14. Hardie, L. A., Lowenstein, T. K. & Spencer, R. J. The problem of distinguishing between primary and secondary features in evaporites. Sixth Int. Symp. On Salt 1, 11–39 (1983).

    Google ScholarΒ 

  15. Roedder, E. The fluids in salt. Amer. Mineral. 69, 413 –439 (1984).

    CASΒ  MATHΒ  Google ScholarΒ 

  16. Lowenstein, T. K. & Hardie, L. A. Criteria for recognition of salt-pan evaporites. Sedimentol. 32, 627–644 (1985).

    ArticleΒ  ADSΒ  CASΒ  Google ScholarΒ 

  17. Lowenstein, T. K. Origin of depositional cycles in a Permian β€œsaline giant”: the Salado (McNutt zone) evaporites of New Mexico and Texas. Geol. Soc. Am. Bull. 100, 592–608 (1988).

    ArticleΒ  ADSΒ  CASΒ  Google ScholarΒ 

  18. Holt, R. M. & Powers, D. W. Geological mapping of the air intake shaft at the Waste Isolation Pilot Plant. Report no. DOE/WIPP 90-051, 1–90 (U. S. Department of Energy, Carlsbad NM, 1990).

  19. Lowenstein, T. K. & Spencer, R. J. Syndepositional origin of potash evaporites: petrographic and fluid inclusion evidence. Am. J. Science 290, 1–42 (1990).

    ArticleΒ  ADSΒ  CASΒ  Google ScholarΒ 

  20. Holt, R. M. & Powers, D. W. in Geological and Hydrological Studies of Evaporites in the Northern Delaware Basin for the Waste Isolation Pilot Plant (WIPP), New Mexico (eds Powers, D. W., Holt, R. M., Beauheim, R. L. & Rempe, N.). Geol. Soc. Am. Guidebook 14, 45–78 (1990).

    Google ScholarΒ 

  21. Powers, D. W. & Hassinger, B. W. Synsedimentary dissolution pits of halite of the Permian Salado Formation, southeastern New Mexico. J. Sed. Petrol. 55, 769–773 (1985).

    Google ScholarΒ 

  22. Onstott, T. C., Mueller, C., Mikulki, K., Vicenzi, E. & Powers, D. W. 40Ar/39Ar laser microprobe dating of polyhalite from bedded, late Permian evaporites. EOS 76, S285 (1995).

    Google ScholarΒ 

  23. Vreeland, R. H., Piselli, A. F., McDonnough, S. & Myer, S. S. Distribution and diversity of halophilic bacteria in a subsurface salt formation. Extremophiles 2, 321–331 (1998).

    ArticleΒ  CASΒ  PubMedΒ  Google ScholarΒ 

  24. Rosenzweig, W. D., Peterson, J., Woish, J. & Vreeland, R. H. Development of a protocol to retrieve microorganisms from ancient salt crystals. Geomicrobiol. (in the press).

  25. Vreeland, R. H., Anderson, R. & Murray, R. G. E. Cell wall and phospholipid composition and their contribution to the salt tolerance of Halomonas elongata. J. Bacteriol. 160, 879–883 (1984).

    CASΒ  PubMedΒ  PubMed CentralΒ  Google ScholarΒ 

  26. Arahal, D. R, Marquez, M. C., Volcani, B. E., Schleifer, K. H. & Ventosa, A. Bacillus marismortui sp. nov., a new moderately halophilic species from the Dead Sea. Int. J. Syst. Bacteriol. 49, 521–530 ( 1999).

    ArticleΒ  PubMedΒ  Google ScholarΒ 

  27. Heyndrickx, M. et al. Virgibacillus: a new genus to accommodate Bacillus pantothenticus (Proom and Knight 1950). Emended description of Virgibacillus pantothenticus. Int. J. Syst. Bacteriol. 48, 99–106 (1998).

    ArticleΒ  Google ScholarΒ 

  28. Garabito, M. J., Arahal, D. R., Mellado, E., Marquez, M. C. & Ventosa, A. Bacillus salexigens sp. nov., a new moderately halophilic Bacillus species. Int. J. Syst. Bacteriol. 47, 735–741 ( 1997).

    ArticleΒ  CASΒ  PubMedΒ  Google ScholarΒ 

  29. Waino, M., Tindall, B. J., Schumann, P. & Ingvorsen, K. Gracilibacillus gen nov., with description of Gracilibacillus halotolerans gen. nov. sp. nov.; transfer of Bacillus dipsosauri to Gracilibacillus dipsosauri comb. nov., and Bacillus salexigens to the genus Salibacillus gen. nov., as Salibacillus salexigens comb. nov. Int. J. Syst. Bacteriol. 49, 821–831 (1999).

    ArticleΒ  CASΒ  PubMedΒ  Google ScholarΒ 

  30. Cournoyer, B. & Lavire, C. Analysis of Frankia evolutionary radiation using glnII sequences. FEMS Microbiol. Lett. 177, 29–34 (1999).

    ArticleΒ  CASΒ  PubMedΒ  Google ScholarΒ 

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Acknowledgements

The authors acknowledge the following people who helped obtain the crystal samples for this research: D. Belski, N. Rempe, R. Carrasco, T. Garcia, D. Acevedo, S. Britain, E. Keyser, B. Kinsall, A. Morin and T. Padilla. This research was supported by the US National Science Foundation: Life in Extreme Environments Program (EAR Lexen).

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Authors and Affiliations

  1. Department of Biology, West Chester University, West Chester, 19383 , Pennsylvania, USA

    Russell H. VreelandΒ &Β William D. Rosenzweig

  2. Consulting Geologist, Box 87, Anthony, 79821, Texas, USA

    Dennis W. Powers

Authors
  1. Russell H. Vreeland
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  2. William D. Rosenzweig
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  3. Dennis W. Powers
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Correspondence to Russell H. Vreeland.

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Vreeland, R., Rosenzweig, W. & Powers, D. Isolation of a 250 million-year-old halotolerant bacterium from a primary salt crystal. Nature 407, 897–900 (2000). https://doi.org/10.1038/35038060

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Comments

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  1. AurΓ©lien laetitia

    Thank you, what about the bacillus marismortui close similarity with b. permians?

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