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Applied and Environmental Microbiology, August 2008, p. 4910-4922, Vol. 74, No. 15
0099-2240/08/$08.00+0     doi:10.1128/AEM.00233-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of the Diversity and Distribution of a Thermal Spring Microbial Community by Using rRNA and Metabolic Genes{triangledown} ,{dagger}

Justine R. Hall,1 Kendra R. Mitchell,1 Olan Jackson-Weaver,1,{ddagger} Ara S. Kooser,2 Brandi R. Cron,1 Laura J. Crossey,2 and Cristina D. Takacs-Vesbach1*

Department of Biology, University of New Mexico, 167 Castetter Hall, MSC03-2020 1, University of New Mexico, Albuquerque, New Mexico 87131,1 Department of Earth and Planetary Sciences, University of New Mexico, Northrop Hall, MSC03-2040 1, University of New Mexico, Albuquerque, New Mexico 871312

Received 25 January 2008/ Accepted 21 May 2008

The diversity and distribution of a bacterial community from Coffee Pots Hot Spring, a thermal spring in Yellowstone National Park with a temperature range of 39.3 to 74.1°C and pH range of 5.75 to 6.91, were investigated by sequencing cloned PCR products and quantitative PCR (qPCR) of 16S rRNA and metabolic genes. The spring was inhabited by three Aquificae genera—Thermocrinis, Hydrogenobaculum, and Sulfurihydrogenibium—and members of the Alpha-, Beta-, and Gammaproteobacteria, Firmicutes, Acidobacteria, Deinococcus-Thermus, and candidate division OP5. The in situ chemical affinities were calculated for 41 potential metabolic reactions using measured environmental parameters and a range of hydrogen and oxygen concentrations. Reactions that use oxygen, ferric iron, sulfur, and nitrate as electron acceptors were predicted to be the most energetically favorable, while reactions using sulfate were expected to be less favorable. Samples were screened for genes used in ammonia oxidation (amoA, bacterial gene only), the reductive tricarboxylic acid (rTCA) cycle (aclB), the Calvin cycle (cbbM), sulfate reduction (dsrAB), nitrogen fixation (nifH), nitrite reduction (nirK), and sulfide oxidation (soxEF1) by PCR. Genes for carbon fixation by the rTCA cycle and nitrogen fixation were detected. All aclB sequences were phylogenetically related and spatially correlated to Sulfurihydrogenibium 16S rRNA gene sequences using qPCR (R2 = 0.99). This result supports the recent finding of citrate cleavage by enzymes other than ATP citrate lyase in the rTCA cycle of the Aquificaceae family. We briefly consider potential biochemical mechanisms that may allow Sulfurihydrogenibium and Thermocrinis to codominate some hydrothermal environments.

* Corresponding author. Mailing address: Department of Biology, University of New Mexico, 167 Castetter Hall, MSC03-2020 1, University of New Mexico, Albuquerque, NM 87131. Phone: (505) 277-3418. Fax: (505) 277-0304. E-mail: cvesbach{at}unm.edu

{triangledown} Published ahead of print on 6 June 2008.

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

{ddagger} Present address: Department of Cell Biology and Physiology, University of New Mexico, 232 Biomedical Research Facility, MSC08-4750 1, University of New Mexico, Albuquerque, NM 87131.

Applied and Environmental Microbiology, August 2008, p. 4910-4922, Vol. 74, No. 15
0099-2240/08/$08.00+0     doi:10.1128/AEM.00233-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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