A survey of indigenous microbial hydrocarbon degradation genes in soils from Antarctica and Brazil.
Total community DNA from 29 noncontaminated soils and soils impacted by petroleum hydrocarbons and chloro-organics from Antarctica and Brazil were screened for the presence of nine catabolic genes, encoding alkane monooxygenase or aromatic dioxygenases, from known bacterial biodegradation pathways. Specific primers and probes targeting alkane monooxygenase genes were derived from Pseudomonas putida ATCC 29347 (Pp alkB), Rhodococcus sp. strain Q15 (Rh alkB1, Rh alkB2), and Acinetobacter sp. ADP-1 (Ac alkM). In addition, primers and probes detecting aromatic dioxygenase genes were derived from P. putida ATCC 17484 (ndoB), P. putida F1 (todC1), P. putida ATCC 33015 (xylE and cat23), and P. pseudoalcaligenes KF707 (bphA). The primers and probes were used to analyze total community DNA extracts by using PCR and hybridization analysis. All the catabolic genes, except the Ac alkM, were detected in contaminated and control soils from both geographic regions, with a higher frequency in the Antarctic soils. The alkane monooxygenase genes, Rh alkB1 and Rh alkB2, were the most frequently detected alk genes in both regions, while Pp alkB was not detected in Brazil soils. Genes encoding the aromatic dioxygenases toluene dioxygenase (todC1) and biphenyl dioxygenase (bphA) were the most frequently detected in Antarctica, and todC1 and catechol-2,3-dioxygenase (cat23) were the most frequent in Brazil soils. Hybridization analysis confirmed the PCR results, indicating that the probes used had a high degree of homology to the genes detected in the soil extracts and were effective in detecting biodegradative potential in the indigenous microbial population.
Luz
A P
AP
Biomedical Sciences Institute, Department of Microbiology, University of São Paulo, Brazil. apluz@usp.br
Pellizari
V H
VH
Whyte
L G
LG
Greer
C W
CW
D000150:Acinetobacter / Q000235:genetics; D000864:Antarctic Regions; D001419:Bacteria / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics; D001938:Brazil; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D004734:Energy Metabolism; D006838:Hydrocarbons / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000235:genetics; D009693:Nucleic Acid Hybridization; D010105:Oxygenases / Q000235:genetics; D016133:Polymerase Chain Reaction; D044205:Pseudomonas pseudoalcaligenes / Q000235:genetics; D016958:Pseudomonas putida / Q000235:genetics; D012240:Rhodococcus / Q000235:genetics; D012988:Soil Microbiology
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D006838:Hydrocarbons; D007506:Iron-Sulfur Proteins; D010105:Oxygenases; C074161:biphenyl-2,3-dioxygenase