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Applied and Environmental Microbiology, November 1998, p. 4185-4193, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Evolution of a Pathway for Chlorobenzene Metabolism Leads to
Natural Attenuation in Contaminated Groundwater
Jan Roelof
van der
Meer,1
Christoph
Werlen,1
Shirley F.
Nishino,2 and
Jim C.
Spain2,*
Swiss Federal Institute for Environmental
Science and Technology (EAWAG), CH 8600 Dübendorf,
Switzerland,1 and
Air Force
Research Laboratory/MLQR, Tyndall Air Force Base, Florida
32403-53232
Received 1 May 1998/Accepted 18 August 1998
Complete metabolism of chlorinated benzenes is not a feature that
is generally found in aerobic bacteria but is thought to be due to a
novel recombination of two separate gene clusters. Such a recombination
could be responsible for adaptation of a natural
microbial community in response to contamination with synthetic
chemicals. This hypothesis was tested in a chlorobenzene (CB)-contaminated aquifer. CB-degrading bacteria from a contaminated site were characterized for a number of years by examining a
combination of growth characteristics and DNA-DNA hybridization, PCR,
and DNA sequence data. The genetic information obtained for the CB pathway of the predominant microorganism, Ralstonia sp.
strain JS705, revealed a unique combination of (partially duplicated) genes for chlorocatechol degradation and genes for a benzene-toluene type of aromatic ring dioxygenase. The organism was detected in CB-polluted groundwater by hybridizing colonies cultivated on low-strength heterotrophic media with probes for the CB pathway. Southern hybridizations performed to determine the organization of the
CB pathway genes and the 16S ribosomal DNA indicated that CB-degrading
organisms isolated from different wells at the site were identical to
JS705. Physiological characterization by the Biolog test system
revealed some differences. The genes for the aromatic ring dioxygenase
and dihydrodiol dehydrogenase of JS705 were detected in toluene and
benzene degraders from the same site. Our results suggest that recent
horizontal gene transfer and genetic recombination of existing genes
between indigenous microorganisms were the mechanisms for
evolution of the catabolic pathway. Evolution of the CB pathway seems
to have created the capacity for natural attenuation of CB at the
contaminated site.
*
Corresponding author. Mailing address: Air Force
Research Laboratory/MLQR, 139 Barnes Dr., Ste. 2, Tyndall AFB, FL
32403-5323. Phone: (850) 283-6058. Fax: (850) 283-6090. E-mail:
JSpain{at}ccmail.aleq.tyndall.af.mil.
Applied and Environmental Microbiology, November 1998, p. 4185-4193, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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