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Applied and Environmental Microbiology, August 1999, p. 3354-3359, Vol. 65, No. 8
Department of Civil Engineering, University
of Minnesota, Minneapolis, Minnesota 55455
Received 19 March 1999/Accepted 11 May 1999
Pseudomonas pseudoalcaligenes POB310(pPOB) and
Pseudomonas sp. strains B13-D5(pD30.9) and
B13-ST1(pPOB) were introduced into soil microcosms containing
3-phenoxybenzoic acid (3-POB) in order to evaluate and compare
bacterial survival, degradation of 3-POB, and transfer of plasmids to a
recipient bacterium. Strain POB310 was isolated for its ability
to use 3-POB as a growth substrate; degradation is initiated by
POB-dioxygenase, an enzyme encoded on pPOB. Strain B13-D5
contains pD30.9, a cloning vector harboring the genes
encoding POB-dioxygenase; strain B13-ST1 contains pPOB. Degradation of
3-POB in soil by strain POB310 was incomplete, and bacterial densities
decreased even under the most favorable conditions (100 ppm of 3-POB,
supplementation with P and N, and soil water-holding capacity of 90%).
Strains B13-D5 and B13-ST1 degraded 3-POB (10 to 100 ppm) to
concentrations of <50 ppb with concomitant increases in density from
106 to 108 CFU/g (dry weight) of soil. Thus, in
contrast to strain POB310, the modified strains had the following two
features that are important for in situ bioremediation: survival in
soil and growth concurrent with removal of an environmental
contaminant. Strains B13-D5 and B13-ST1 also completely degraded 3-POB
when the inoculum was only 30 CFU/g (dry weight) of soil. This suggests
that in situ bioremediation may be effected, in some cases, with low
densities of introduced bacteria. In pure culture, transfer of pPOB
from strains POB310 and B13-ST1 to Pseudomonas sp. strain
B13 occurred at frequencies of 5 × 10
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Copyright © 1999, American Society for Microbiology. All rights reserved.
Degradation of 3-Phenoxybenzoic Acid in Soil by
Pseudomonas pseudoalcaligenes POB310(pPOB) and Two Modified
Pseudomonas Strains


and
7 and
10
1 transconjugant per donor, respectively. Transfer of
pPOB from strain B13-ST1 to strain B13 was observed in autoclaved soil
but not in nonautoclaved soil; formation of transconjugant bacteria was
more rapid in soil containing clay and organic matter
than in sandy soil. Transfer of pPOB from strain POB310 to strain B13 in soil was never observed.
*
Corresponding author. Mailing address: Department of
Civil Engineering, University of Minnesota, Minneapolis, MN 55455. Phone: (612) 625-8582. Fax: (612) 626-7750. E-mail:
dwyer003{at}tc.umn.edu.
Present address: Lawrence Livermore National Laboratory,
Environmental Restoration Division, Livermore, CA 94551.
Present address: ThermoRetec Corp., St. Paul, MN 55101.
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