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Applied and Environmental Microbiology, November 2000, p. 4673-4678, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Rhizosphere Competitiveness of Trichloroethylene-Degrading, Poplar-Colonizing Recombinant Bacteria

Hojae Shim,1,dagger Sadhana Chauhan,1 Doohyun Ryoo,1,Dagger Kally Bowers,1 Stuart M. Thomas,2 Keith A. Canada,1 Joel G. Burken,3 and Thomas K. Wood1,*

Department of Chemical Engineering, University of Connecticut, Storrs, Connecticut 06269-32221; Experimental Station, E. I. DuPont de Nemours, Inc., Wilmington, Delaware 198802; and Department of Civil Engineering, University of Missouri, Rolla, Missouri 654093

Received 10 April 2000/Accepted 24 August 2000

Indigenous bacteria from poplar tree (Populus canadensis var. eugenei `Imperial Carolina') and southern California shrub rhizospheres, as well as two tree-colonizing Rhizobium strains (ATCC 10320 and ATCC 35645), were engineered to express constitutively and stably toluene o-monooxygenase (TOM) from Burkholderia cepacia G4 by integrating the tom locus into the chromosome. The poplar and Rhizobium recombinant bacteria degraded trichloroethylene at a rate of 0.8 to 2.1 nmol/min/mg of protein and were competitive against the unengineered hosts in wheat and barley rhizospheres for 1 month (colonization occurred at a level of 1.0 × 105 to 23 × 105 CFU/cm of root). In addition, six of these recombinants colonized poplar roots stably and competitively with populations as large as 79% ± 12% of all rhizosphere bacteria after 28 days (0.2 × 105 to 31 × 105 CFU/cm of root). Furthermore, five of the most competitive poplar recombinants (e.g., Pb3-1 and Pb5-1, which were identified as Pseudomonas sp. strain PsK recombinants) retained the ability to express TOM for 29 days as 100% ± 0% of the recombinants detected in the poplar rhizosphere expressed TOM constitutively.

* Corresponding author. Mailing address: Department of Chemical Engineering, University of Connecticut, Storrs, CT 06269-3222. Phone: (860) 486-2483. Fax: (860) 486-2959. E-mail: twood{at}engr.uconn.edu.

dagger Present address: Department of Civil and Environmental Engineering, Hanyang University, Kyungkido 425-791, Korea.

Dagger Present address: Department of Environmental Engineering, Jeonju University, Jeonju 560-759, Korea.

Applied and Environmental Microbiology, November 2000, p. 4673-4678, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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