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Applied and Environmental Microbiology, July 1999, p. 3064-3070, Vol. 65, No. 7
Department of Biological Sciences, University
of Calgary, Calgary, Alberta, Canada T2N 1N4
Received 25 February 1999/Accepted 30 April 1999
Toluene-degrading bacteria were isolated from
hydrocarbon-contaminated soil by incubating liquid enrichment cultures
and agar plate cultures in desiccators in which the vapor pressure of
toluene was controlled by dilution with vacuum pump oil. Incubation in desiccators equilibrated with either 100, 10, or 1% (wt/wt) toluene in
vacuum pump oil and testing for genomic cross-hybridization resulted in
four genomically distinct strains (standards) capable of growth on
toluene (strains Cstd1, Cstd2, Cstd5, and Cstd7). The optimal toluene
concentrations for growth of these standards on plating media differed
considerably. Cstd1 grew best in an atmosphere equilibrated with 0.1%
(wt/wt) toluene, but Cstd5 failed to grow in this atmosphere.
Conversely, Cstd5 grew well in the presence of 10% (wt/wt) toluene,
which inhibited growth of Cstd1. 16S ribosomal DNA sequencing and
cross-hybridization analysis indicated that both Cstd1 and Cstd5 are
members of the genus Pseudomonas. An analysis of the
microbial communities in soil samples that were incubated with 10%
(wt/wt) toluene with reverse sample genome probing indicated that
Pseudomonas strain Cstd5 was the dominant community member.
However, incubation of soil samples with 0.1% (wt/wt) toluene resulted
in a community that was dominated by Pseudomonas strain Q7,
a toluene degrader that has been described previously (Y. Shen, L. G. Stehmeier, and G. Voordouw, Appl. Environ. Microbiol. 64:637-645,
1998). Q7 was not able to grow by itself in an atmosphere equilibrated
with 0.1% (wt/wt) toluene but grew efficiently in coculture with
Cstd1, suggesting that toluene or metabolic derivatives of toluene were
transferred from Cstd1 to Q7.
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Composition of Toluene-Degrading Microbial
Communities from Soil at Different Concentrations of Toluene
*
Corresponding author. Mailing address: Department of
Biological Sciences, University of Calgary, 2500 University Dr. NW,
Calgary, Alberta T2N 1N4, Canada. Phone: (403) 220-6388. Fax: (403)
289-9311. E-mail: voordouw{at}ucalgary.ca.
Applied and Environmental Microbiology, July 1999, p. 3064-3070, Vol. 65, No. 7
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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