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Applied and Environmental Microbiology, January 2001, p. 411-419, Vol. 67, No. 1
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.1.411-419.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Investigation of Candidate Division TM7, a Recently Recognized Major Lineage of the Domain Bacteria with No Known Pure-Culture Representatives

Philip Hugenholtz,1,* Gene W. Tyson,1 Richard I. Webb,2 Ankia M. Wagner,3 and Linda L. Blackall1

Advanced Wastewater Management Centre1 and Centre for Microscopy and Microanalysis,2 Department of Microbiology and Parasitology, The University of Queensland, Brisbane 4072, Australia, and Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria 0001, South Africa3

Received 31 May 2000/Accepted 29 September 2000

A molecular approach was used to investigate a recently described candidate division of the domain Bacteria, TM7, currently known only from environmental 16S ribosomal DNA sequence data. A number of TM7-specific primers and probes were designed and evaluated. Fluorescence in situ hybridization (FISH) of a laboratory scale bioreactor using two independent TM7-specific probes revealed a conspicuous sheathed-filament morphotype, fortuitously enriched in the reactor. Morphologically, the filament matched the description of the Eikelboom morphotype 0041-0675 widely associated with bulking problems in activated-sludge wastewater treatment systems. Transmission electron microscopy of the bioreactor sludge demonstrated that the sheathed-filament morphotype had a typical gram-positive cell envelope ultrastructure. Therefore, TM7 is only the third bacterial lineage recognized to have gram-positive representatives. TM7-specific FISH analysis of two full-scale wastewater treatment plant sludges, including the one used to seed the laboratory scale reactor, indicated the presence of a number of morphotypes, including sheathed filaments. TM7-specific PCR clone libraries prepared from the two full-scale sludges yielded 23 novel TM7 sequences. Three subdivisions could be defined based on these data and publicly available sequences. Environmental sequence data and TM7-specific FISH analysis indicate that members of the TM7 division are present in a variety of terrestrial, aquatic, and clinical habitats. A highly atypical base substitution (Escherichia coli position 912; C to U) for bacterial 16S rRNAs was present in almost all TM7 sequences, suggesting that TM7 bacteria, like Archaea, may be streptomycin resistant at the ribosome level.

* Corresponding author. Mailing address: Advanced Wastewater Management Centre, Department of Microbiology and Parasitology, The University of Queensland, Brisbane 4072, Australia. Phone: 617 3365 2485. Fax: 617 3365 4620. E-mail: philiph{at}biosci.uq.edu.au.

Applied and Environmental Microbiology, January 2001, p. 411-419, Vol. 67, No. 1
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.1.411-419.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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