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Applied and Environmental Microbiology, September 2007, p. 5865-5874, Vol. 73, No. 18
0099-2240/07/$08.00+0     doi:10.1128/AEM.01207-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

"Candidatus Accumulibacter" Population Structure in Enhanced Biological Phosphorus Removal Sludges as Revealed by Polyphosphate Kinase Genes{triangledown}

Shaomei He, Daniel L. Gall, and Katherine D. McMahon*

Department of Civil and Environmental Engineering, University of Wisconsin—Madison, Madison, Wisconsin 53706

Received 30 May 2007/ Accepted 23 July 2007

We investigated the fine-scale population structure of the "Candidatus Accumulibacter" lineage in enhanced biological phosphorus removal (EBPR) systems using the polyphosphate kinase 1 gene (ppk1) as a genetic marker. We retrieved fragments of "Candidatus Accumulibacter" 16S rRNA and ppk1 genes from one laboratory-scale and several full-scale EBPR systems. Phylogenies reconstructed using 16S rRNA genes and ppk1 were largely congruent, with ppk1 granting higher phylogenetic resolution and clearer tree topology and thus serving as a better genetic marker than 16S rRNA for revealing population structure within the "Candidatus Accumulibacter" lineage. Sequences from at least five clades of "Candidatus Accumulibacter" were recovered by ppk1-targeted PCR, and subsequently, specific primer sets were designed to target the ppk1 gene for each clade. Quantitative real-time PCR (qPCR) assays using "Candidatus Accumulibacter"-specific 16S rRNA and "Candidatus Accumulibacter" clade-specific ppk1 primers were developed and conducted on three laboratory-scale and nine full-scale EBPR samples and two full-scale non-EBPR samples to determine the abundance of the total "Candidatus Accumulibacter" lineage and the relative distributions and abundances of the five "Candidatus Accumulibacter" clades. The qPCR-based estimation of the total "Candidatus Accumulibacter" fraction as a proportion of the bacterial community as measured using 16S rRNA genes was not significantly different from the estimation measured using ppk1, demonstrating the power of ppk1 as a genetic marker for detection of all currently defined "Candidatus Accumulibacter" clades. The relative distributions of "Candidatus Accumulibacter" clades varied among different EBPR systems and also temporally within a system. Our results suggest that the "Candidatus Accumulibacter" lineage is more diverse than previously realized and that different clades within the lineage are ecologically distinct.

* Corresponding author. Mailing address: Department of Civil and Environmental Engineering, University of Wisconsin—Madison, Madison, WI 53706. Phone: (608) 263-3137. Fax: (608) 262-5199. E-mail: tmcmahon{at}engr.wisc.edu

{triangledown} Published ahead of print on 3 August 2007.

Applied and Environmental Microbiology, September 2007, p. 5865-5874, Vol. 73, No. 18
0099-2240/07/$08.00+0     doi:10.1128/AEM.01207-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Gunther, S., Trutnau, M., Kleinsteuber, S., Hause, G., Bley, T., Roske, I., Harms, H., Muller, S. (2009). Dynamics of Polyphosphate-Accumulating Bacteria in Wastewater Treatment Plant Microbial Communities Detected via DAPI (4',6'-Diamidino-2-Phenylindole) and Tetracycline Labeling. Appl. Environ. Microbiol. 75: 2111-2121 [Abstract] [Full Text]  


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