Detection and quantification of degradative genes in soils contaminated by toluene |
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Authors: | Sylvie Hallier-Soulier,Vé ronique Ducrocq,Nathalie Mazure,Nicole Truffaut |
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Affiliation: | Laboratoire de Génétique Microbienne, Universitéde Technologie de Compiègne, BP 649, 60206 Compiègne Cedex, France |
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Abstract: | Abstract: A method based on the polymerase chain reaction (PCR) was developed for a rapid and specific detection of toluene degradative genes in soil. The xylE gene coding for catechol 2,3-dioxygenase was chosen as a target gene. The detection threshold was evaluated in microcosms using a sterilized standard soil inoculated with various amounts of a degradative strain of Pseudomonas putida (mX). The extracted DNA was used as a template to amplify the xylE gene. PCR followed by hybridization with an internal probe allowed us to detect 102 bacteria per g of soil. In polluted soils, quantification of target DNA by competitive PCR was compared with enumeration of degradative microflora. This molecular method appeared to be rapid, sensitive and more suitable than the microbiological approach to estimate the biodegradative potential of a polluted soil. |
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Keywords: | Toluene biodegradation xylE gene Soil microcosm Polymerase chain reaction |
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