Open AccessDetection and quantification of degradative genes in soils contaminated by toluene
Author(s) -
HallierSoulier Sylvie,
Ducrocq Véronique,
Mazure Nathalie,
Truffaut Nicole
Publication year - 1996
Publication title -
fems microbiology ecology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.377
H-Index - 155
eISSN - 1574-6941
pISSN - 0168-6496
DOI - 10.1111/j.1574-6941.1996.tb00311.x
Subject(s) - biology , pseudomonas putida , polymerase chain reaction , microcosm , bacteria , soil microbiology , soil contamination , gene , pseudomonas , bioremediation , microbiology and biotechnology , soil water , contamination , toluene , dna , dna–dna hybridization , biodegradation , biochemistry , genetics , chemistry , ecology , organic chemistry
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 10 2 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.