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Phylogenetic and physiological diversity of Arthrobacter strains isolated from unconsolidated subsurface sediments

Department of Biological Science, Florida State University, 312 Nuclear Research Building, Tallahassee, FL 32306-4470, USA¹
Pacific Northwest National Laboratory, Richland, WA 99352, USA²
Center for Environmental Biotechnology, University of Tennessee, Knoxville, TN 37932, USA³

Author for correspondence: D. L. Balkwill. Tel: +1 850 644 5719. Fax: +1 850 644 4865. e-mail: balkwill{at}bio.fsu.edu

Forty strains of Gram-positive, aerobic, heterotrophic bacteria isolated from saturated subsurface lacustrine, paleosol and fluvial sediments at the US Department of Energy’s Hanford Site in south central Washington State were characterized by phylogenetic analysis of 16S rRNA gene sequences and by determination of selected morphological, physiological and biochemical traits. Phylogenetic analyses of 16S rDNA sequences from subsurface isolates in the context of similar sequences from previously described bacterial species indicated that 38 of the subsurface strains were most closely related to Arthrobacter. The other two strains appeared to be most closely related to Kocuria. The subsurface isolates fell into seven phylogenetically coherent and distinct clusters, indicating that there was a significant degree of diversity among them. Additional diversity was detected by analysis of cellular fatty acids and physiological traits. The general morphological, physiological and biochemical traits of the subsurface strains were consistent with those of Arthrobacter, Micrococcus and genera recently separated from Micrococcus, such as Kocuria. Some of the subsurface strains were phylogenetically closely related to certain species of Arthrobacter (16S rDNA sequence similarities >99%). However, most of the subsurface isolates did not cluster with previously established species in phylogenetic analyses of 16S rRNA gene sequences or with hierarchical cluster analysis of cellular fatty acid profiles. Moreover, many of the subsurface isolates that were most closely related to Arthrobacter also differed from all established species of that genus in several of their specific physiological characteristics. Most of the subsurface isolates, then, are likely to be novel strains or species of Arthrobacter.

Keywords: Arthrobacter, subsurface microbiology, microbial phylogeny

The GenBank accession numbers for the sequences determined in this study are given in Methods.

^a Present address: Center for Microbial Ecology, Michigan State University, East Lansing, MI 48824, USA.

^b Present address: Waterways Experiment Station, Vicksburg, MS 39180, USA.

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