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Applied and Environmental Microbiology, October 2000, p. 4589-4594, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Bioluminescent Whole-Cell Reporter for Detection of 2,4-Dichlorophenoxyacetic Acid and 2,4-Dichlorophenol in Soil

Anthony G. Hay,dagger James F. Rice,Dagger Bruce M. Applegate, Nathan G. Bright, and Gary S. Sayler*

Center for Environmental Biotechnology, University of Tennessee, Knoxville, Knoxville, Tennessee 37996-1605

Received 1 May 2000/Accepted 25 June 2000

A bioreporter was made containing a tfdRPDII-luxCDABE fusion in a modified mini-Tn5 construct. When it was introduced into the chromosome of Ralstonia eutropha JMP134, the resulting strain, JMP134-32, produced a sensitive bioluminescent response to 2,4-dichlorophenoxyacetic acid (2,4-D) at concentrations of 2.0 µM to 5.0 mM. This response was linear (R2 = 0.9825) in the range of 2.0 µM to 1.1 × 102 µM. Saturation occurred at higher concentrations, with maximal bioluminescence occurring in the presence of approximately 1.2 mM 2,4-D. A sensitive response was also recorded in the presence of 2,4-dichlorophenol at concentrations below 1.1 × 102 µM; however, only a limited bioluminescent response was recorded in the presence of 3-chlorobenzoic acid at concentrations below 1.0 mM. A significant bioluminescent response was also recorded when strain JMP134-32 was incubated with soils containing aged 2,4-D residues.

* Corresponding author. Mailing address: Center for Environmental Biotechnology, 676 Dabney Hall, University of Tennessee, Knoxville, Knoxville, TN 37996-1605. Phone: (423) 974-8080. Fax: (423) 974-8086. E-mail: Sayler{at}utk.edu.

dagger Present address: Department of Microbiology, Cornell University, Ithaca, NY 14853.

Dagger Present address: Monsanto Company, St. Louis, MO 63167-0001.

Applied and Environmental Microbiology, October 2000, p. 4589-4594, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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