Monitoring gene expression in mixed microbial communities by using DNA microarrays.
A DNA microarray to monitor the expression of bacterial metabolic genes within mixed microbial communities was designed and tested. Total RNA was extracted from pure and mixed cultures containing the 2,4-dichlorophenoxyacetic acid (2,4-D)-degrading bacterium Ralstonia eutropha JMP134, and the inducing agent 2,4-D. Induction of the 2,4-D catabolic genes present in this organism was readily detected 4, 7, and 24 h after the addition of 2,4-D. This strain was diluted into a constructed mixed microbial community derived from a laboratory scale sequencing batch reactor. Induction of two of five 2,4-D catabolic genes (tfdA and tfdC) from populations of JMP134 as low as 10(5) cells/ml was clearly detected against a background of 10(8) cells/ml. Induction of two others (tfdB and tfdE) was detected from populations of 10(6) cells/ml in the same background; however, the last gene, tfdF, showed no significant induction due to high variability. In another experiment, the induction of resin acid degradative genes was statistically detectable in sludge-fed pulp mill effluent exposed to dehydroabietic acid in batch experiments. We conclude that microarrays will be useful tools for the detection of bacterial gene expression in wastewaters and other complex systems.
Dennis
Philip
P
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Ontario, Canada.
Edwards
Elizabeth A
EA
Liss
Steven N
SN
Fulthorpe
Roberta
R
D015084:2,4-Dichlorophenoxyacetic Acid / Q000378:metabolism; D001419:Bacteria / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001673:Biodegradation, Environmental; D019149:Bioreactors; D020118:Cupriavidus necator / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D015342:DNA Probes; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D017753:Ecosystem; D020869:Gene Expression Profiling; D007220:Industrial Waste; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods / Q000592:standards; D010209:Paper; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D014865:Waste Disposal, Fluid / Q000379:methods*
D001426:Bacterial Proteins; D015342:DNA Probes; D018076:DNA, Complementary; D007220:Industrial Waste; D015084:2,4-Dichlorophenoxyacetic Acid