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A bioluminescent whole-cell reporter for detection of 2,4-dichlorophenoxyacetic acid and 2,4-dichlorophenol in soil

Hay, A.G., Rice, J.F., Applegate, B.M., Bright, N.G., Sayler, G.S.
Applied and environmental microbiology 2000 v.66 no.10 pp. 4589-4594
luciferase, recombinant DNA, transcription factors, gene expression, bioluminescence, gene transfer, genetic transformation, detection, herbicide residues, 2,4-D, 2,4-dichlorophenol, polluted soils, slurries, Ralstonia eutropha
A bioreporter was made containing of tfdRP(DII)-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 micromolar to 5.0 mM. This response was linear (R2 = 0.9825) in the range of 2.0 micromolar to 1.1 X 10(2) micromolar. 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 X 10(2) micromolar; 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.