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Applied and Environmental Microbiology, October 2005, p. 6175-6184, Vol. 71, No. 10
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.10.6175-6184.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Diversity of the Microeukaryotic Community in Sulfide-Rich Zodletone Spring (Oklahoma){dagger}

Qingwei Luo,1 Lee R. Krumholz,1 Fares Z. Najar,2 Aaron D. Peacock,3 Bruce A. Roe,2 David C. White,3 and Mostafa S. Elshahed1*

Department of Botany and Microbiology and Institute for Energy and the Environment,1 Department of Chemistry and Biochemistry and Advanced Center for Genome Technology, University of Oklahoma, Norman, Oklahoma,2 Center for Biomarker Analysis, The University of Tennessee, Knoxville, Tennessee3

Received 25 October 2004/ Accepted 14 May 2005

The microeukaryotic community in Zodletone Spring, a predominantly anaerobic sulfide and sulfur-rich spring, was examined using an 18S rRNA gene cloning and sequencing approach. The majority of the 288 clones sequenced from three different locations at Zodletone Spring belonged to the Stramenopiles, Alveolata, and Fungi, with members of the phylum Cercozoa, order Diplomonadida, and family Jakobidae representing a minor fraction of the clone library. No sequences suggesting the presence of novel kingdom level diversity were detected in any of the three libraries. A large fraction of stramenopile clones encountered were monophyletic with either members of the genus Cafeteria (order Bicosoecida) or members of the order Labyrinthulida (slime nets), both of which have so far been encountered mainly in marine habitats. The majority of the observed fungal clone sequences belonged to the ascomycetous yeasts (order Saccharomycetales), were closely related to yeast genera within the Hymenobasidiomycetes (phylum Basidiomycetes), or formed a novel fungal lineage with several previously published or database-deposited clones. To determine whether the unexpected abundance of fungal sequences in Zodletone Spring clone libraries represents a general pattern in anaerobic habitats, we generated three clone libraries from three different anaerobic settings (anaerobic sewage digester, pond sediment, and hydrocarbon-exposed aquifer sediments) and partially sequenced 210 of these clones. Phylogenetic analysis indicated that clone sequences belonging to the kingdom Fungi represent a significant fraction of all three clone libraries, an observation confirmed by phospholipid fatty acid and ergosterol analysis. Overall, this work reveals an unexpected abundance of Fungi in anaerobic habitats, describes a novel, yet-uncultured group of Fungi that appears to be widespread in anaerobic habitats, and indicates that several of the previously considered marine protists could also occur in nonmarine habitats.

* Corresponding author. Mailing address: University of Oklahoma, Department of Botany and Microbiology, 770 Van Vleet Oval, Norman, OK 73019. Phone: (405) 325-5255. Fax: (405) 325-7619. E-mail: mostafa{at}ou.edu.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, October 2005, p. 6175-6184, Vol. 71, No. 10
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.10.6175-6184.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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