Search results for "soil DNA"
showing 9 items of 9 documents
Estimation of the degrading genetic potential of soil bacterial communities: a new tool for evaluating and predicting soil contamination by organic m…
2007
During the last two centuries, industrialisation has led to increasing contamination of the environment by xenobiotics, notably aromatic compounds (i.e. PAHs, pesticides). These widespread pollutants are a major threat affecting soil quality and human health. In this context, numerous policies have been drawn up notably in Europe, to monitor the threats of pollution to ecosystems and to detect and implement measures to counterbalance the damage. Nevertheless, the European Commission and scientific community are still confronted with a lack of appropriate tools to monitor biodiversity in relation to environmental processes affecting global warming and soil protection. This study proposes too…
Estimation of the density of the protocatechuate-degrading bacterial community in soil by real-time PCR
2008
Summary The β-ketoadipate pathway is the major route for degradation of aromatic compounds by various soil microorganisms. Protocatechuate 3,4-dioxygenase, a key enzyme of this pathway and which is encoded by pcaGH genes, catalyses the ring cleavage of protocatechuate. Microorganisms harbouring pcaGH genes are widely distributed in the environment but little is known about their relative abundance within the total microflora. Hence, this paper reports the development of a real-time PCR assay to quantify the bacterial pcaH sequence that encodes the β sub-unit of the protocatechuate 3,4-dioxygenase. This real-time PCR assay was linear over seven orders of magnitude with a calculated efficienc…
Biogeographical patterns of soil molecular microbial biomass
2011
Affiche - résumé. Session GC: Microbes in the Changing Environmenl: Global Climate Change and Soif under Human Impact; International audience; The spatial organization of soil microbial communities over large areas and the identification of environmental factors structuring their distribution have been liUle investigated. The overall objective of this study was to determine the spatial paUerning of microbial biomass in soils on wide extent and to rank the environmental fiUers most influencing this distribution, by using the French Soil Quality Monitoring Network. This network covers ail the French terrilory and soils were sampled at2,150 sites along a systematic grid sampling. The soil DNA …
Inter-laboratory evaluation of the ISO standard 11063 "Soil quality -- Method to directly extract DNA from soil samples"
2011
International audience; Extracting DNA directly from micro-organisms living in soil is a crucial step for the molecular analysis of soil microbial communities. However, the use of a plethora of different soil DNA extraction protocols, each with its own bias, makes accurate data comparison difficult. To overcome this problem, a method for soil DNA extraction was proposed to the International Organization for Standardization (ISO) in 2006. This method was evaluated by 13 independent European laboratories actively participating in national and international ring tests. The reproducibility of the standardized method for molecular analyses was evaluated by comparing the amount of DNA extracted, …
Molecular analysis of the catechol-degrading bacterial community in a coal wasteland heavily contaminated with PAHs
2010
International audience; A PCR-based molecular tool was developed to estimate the diversity of the catechol-degrading bacterial community in a coal wasteland heavily contaminated with PAHS. A degenerate primer pair specific to catA sequences was designed by multiple alignment of known sequences coding a key intermediate of the β-ketoadiapate pathway degrading catechol, namely catechol 1,2-dioxygenase. The specificity of this primer pair was assessed in 21 pure strains by PCR and sequencing. Comparison of the 16S rDNA and catA phylogenies revealed an absence of congruence between these two genes. The primer set was able to amplify catA sequences in DNA extracts from an industrial soil highly …
Occurrence of CTX-M Producing Escherichia coli in Soils, Cattle, and Farm Environment in France (Burgundy Region).
2012
Article en open access; International audience; CTX-M [a major type of extended-spectrum beta-lactamase (ESBU] producing Escherichia coli are increasingly involved in human infections worldwide. The aim of this study was to investigate potential reservoirs for such strains: soils, cattle, and farm environment. The prevalence of bla(CTX-M) genes was determined directly from soil DNA extracts obtained from 120 sites in Burgundy (France) using real-time PCR. bla(CTX-M) targets were found in 20% of the DNA extracts tested. Samples of cattle feces (n = 271) were collected from 182 farms in Burgundy. Thirteen ESBL-producing isolates were obtained from 12 farms and further characterized for the pr…
Inter-laboratory evaluation of the ISO standard 11063 "Soil quality - Method to directly extract DNA from soil samples"
2011
International audience; Extracting DNA directly from micro-organisms living in soil is a crucial step for the molecular analysis of soil microbial communities. However, the use of a plethora of different soil DNA extraction protocols, each with its own bias, makes accurate data comparison difficult. To overcome this problem, a method for soil DNA extraction was proposed to the International Organization for Standardization (ISO) in 2006. This method was evaluated by 13 independent European laboratories actively participating in national and international ring tests. The reproducibility of the standardized method for molecular analyses was evaluated by comparing the amount of DNA extracted, …
pcaH, a molecular marker for estimating the diversity of the protocatechuate-degrading bacterial community in the soil environment
2007
Microorganisms degrading phenolic compounds play an important role in soil carbon cycling as well as in pesticide degradation. The pcaH gene encoding a key ring-cleaving enzyme of the -ketoadipate pathway was selected as a functional marker. Using a degenerate primer pair, pcaH fragments were cloned from two agricultural soils. Restriction fragment length polymorphism (RFLP) screening of 150 pcaH clones yielded 68 RFLP families. Comparison of 86 deduced amino acid sequences displayed 70% identity to known PcaH sequences. Phylogenetic analysis results in two major groups mainly related to PcaH sequences from Actinobacteria and Proteobacteria phyla. This confirms that the developed primer pai…
Long-term impact of 19 years' farmyard manure or sewage sludge application on the structure, diversity and density of the protocatechuate-degrading b…
2012
International audience; Impact of long-term biosolids application on soil-living micro-organisms key players of ecosystemic services is scarcely reported. Here, the impact of the 19 year-long application of farmyard manure (FM) and sewage sludge (SS) organic fertilisation regimes on the protocatechuate-degrading bacterial (pca) community was estimated by comparison to a mineral fertilisation regime (U). The structure, diversity and density of the pca community were determined using pcaH, a gene encoding the protocatechuate 3,4-dioxygenase. Ten years after the last application, the structure of the pca community in soils amended with 55100 (100 t/ha/2 years) and to a lesser extent with FM (1…