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Applied and Environmental Microbiology
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Environmental Microbiology

Characterization of the Community Structure of a Dechlorinating Mixed Culture and Comparisons of Gene Expression in Planktonic and Biofloc-Associated “Dehalococcoides” and Methanospirillum Species

Annette R. Rowe, Brendan J. Lazar, Robert M. Morris, Ruth E. Richardson
Annette R. Rowe
1Field of Microbiology
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Brendan J. Lazar
2School of Civil and Environmental Engineering, Hollister Hall, Cornell University, Ithaca, New York 14853
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Robert M. Morris
2School of Civil and Environmental Engineering, Hollister Hall, Cornell University, Ithaca, New York 14853
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Ruth E. Richardson
2School of Civil and Environmental Engineering, Hollister Hall, Cornell University, Ithaca, New York 14853
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  • For correspondence: rer26@cornell.edu
DOI: 10.1128/AEM.00445-08
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ABSTRACT

This study sought to characterize bacterial and archaeal populations in a perchloroethene- and butyrate-fed enrichment culture containing hydrogen-consuming “Dehalococcoides ethenogenes” strain 195 and a Methanospirillum hungatei strain. Phylogenetic characterization of this microbial community was done via 16S rRNA gene clone library and gradient gel electrophoresis analyses. Fluorescence in situ hybridization was used to quantify populations of “Dehalococcoides” and Archaea and to examine the colocalization of these two groups within culture bioflocs. A technique for enrichment of planktonic and biofloc-associated biomass was developed and used to assess differences in population distribution and gene expression patterns following provision of substrate. On a per-milliliter-of-culture basis, most D. ethenogenes genes (the hydrogenase gene hupL; the highly expressed gene for an oxidoreductase of unknown function, fdhA; the RNA polymerase subunit gene rpoB; and the 16S rRNA gene) showed no statistical difference in expression between planktonic and biofloc enrichments at either time point studied (1 to 2 and 6 h postfeeding). Normalization of transcripts to ribosome (16S rRNA) levels supported that planktonic and biofloc-associated D. ethenogenes had similar gene expression profiles, with one notable exception; planktonic D. ethenogenes showed higher expression of tceA relative to biofloc-associated cells at 6 h postfeeding. These trends were compared to those for the hydrogen-consuming methanogen in the culture, M. hungatei. The vast majority of M. hungatei cells, ribosomes (16S rRNA), and transcripts of the hydrogenase gene mvrD and the housekeeping gene rpoE were observed in the biofloc enrichments. This suggests that, unlike the comparable activity of D. ethenogenes from both enrichments, planktonic M. hungatei is responsible for only a small fraction of the hydrogenotrophic methanogenesis in this culture.

FOOTNOTES

    • Received 23 February 2008.
    • Accepted 27 August 2008.
  • † Supplemental material for this article may be found at http://aem.asm.org/ .

  • Copyright © 2008 American Society for Microbiology
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Characterization of the Community Structure of a Dechlorinating Mixed Culture and Comparisons of Gene Expression in Planktonic and Biofloc-Associated “Dehalococcoides” and Methanospirillum Species
Annette R. Rowe, Brendan J. Lazar, Robert M. Morris, Ruth E. Richardson
Applied and Environmental Microbiology Oct 2008, 74 (21) 6709-6719; DOI: 10.1128/AEM.00445-08

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Characterization of the Community Structure of a Dechlorinating Mixed Culture and Comparisons of Gene Expression in Planktonic and Biofloc-Associated “Dehalococcoides” and Methanospirillum Species
Annette R. Rowe, Brendan J. Lazar, Robert M. Morris, Ruth E. Richardson
Applied and Environmental Microbiology Oct 2008, 74 (21) 6709-6719; DOI: 10.1128/AEM.00445-08
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KEYWORDS

biodiversity
chlorine
Chloroflexi
Gene Expression Profiling
Methanospirillum

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