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Applied and Environmental Microbiology, December 2001, p. 5750-5760, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.21.5750-5760.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Archaeal Diversity in Waters from Deep South African Gold Mines

Ken Takai,1,2,dagger Duane P. Moser,3 Mary DeFlaun,4 Tullis C. Onstott,3 and James K. Fredrickson1,*

Pacific Northwest National Laboratory, Richland, Washington 993521; Subground Animalculae Retrieval (SUGAR) Project, Frontier Research Program for Deep-Sea Environments, Japan Marine Science and Technology Center, Yokosuka 237-0061, Japan2; Department of Geosciences, Princeton University, Princeton, New Jersey 085443; and Envirogen Inc., Lawrenceville, New Jersey 086484

Received 12 June 2001/Accepted 10 September 2001

A culture-independent molecular analysis of archaeal communities in waters collected from deep South African gold mines was performed by performing a PCR-mediated terminal restriction fragment length polymorphism (T-RFLP) analysis of rRNA genes (rDNA) in conjunction with a sequencing analysis of archaeal rDNA clone libraries. The water samples used represented various environments, including deep fissure water, mine service water, and water from an overlying dolomite aquifer. T-RFLP analysis revealed that the ribotype distribution of archaea varied with the source of water. The archaeal communities in the deep gold mine environments exhibited great phylogenetic diversity; the majority of the members were most closely related to uncultivated species. Some archaeal rDNA clones obtained from mine service water and dolomite aquifer water samples were most closely related to environmental rDNA clones from surface soil (soil clones) and marine environments (marine group I [MGI]). Other clones exhibited intermediate phylogenetic affiliation between soil clones and MGI in the Crenarchaeota. Fissure water samples, derived from active or dormant geothermal environments, yielded archaeal sequences that exhibited novel phylogeny, including a novel lineage of Euryarchaeota. These results suggest that deep South African gold mines harbor novel archaeal communities distinct from those observed in other environments. Based on the phylogenetic analysis of archaeal strains and rDNA clones, including the newly discovered archaeal rDNA clones, the evolutionary relationship and the phylogenetic organization of the domain Archaea are reevaluated.

* Corresponding author. Mailing address: MS P7-50, P.O. 999, Richland, WA 99352. Phone: (509) 376-7063. Fax: (509) 376-9650. E-mail: jim.fredrickson{at}pnl.gov.

dagger Present address: Deep-Sea Microorganisms Research Group (DEEP-STAR), Japan Marine Science and Technology Center (JAMSTEC), Yokosuka 237-0061, Japan.

Applied and Environmental Microbiology, December 2001, p. 5750-5760, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.21.5750-5760.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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