[Show abstract][Hide abstract] ABSTRACT: Very little is known about the influence of bacterial-fungal ecological interactions on polycyclic aromatic hydrocarbon (PAH) dissipation in soils. Fusarium solani MM1 and Arthrobacter oxydans MsHM11 can dissipate PAHs in vitro. We investigated their interactions and their effect on the dissipation of three PAHs-phenanthrene (PHE), pyrene (PYR) and dibenz(a,h)anthracene (DBA)-in planted microcosms, in sterile sand or non-sterile soil. In sterile sand microcosms planted with alfalfa, the two microbes survived and grew, without any significant effect of co-inoculation. Co-inoculation led to the dissipation of 46 % of PHE after 21 days. In soil microcosms, whether planted with alfalfa or not, both strains persisted throughout the 46 days of the experiment, without any effect of co-inoculation or of alfalfa, as assessed by real-time PCR targeting taxon-level indicators, i.e. Actinobacteria 16S rDNA and the intergenic transcribed spacer specific to the genus Fusarium. The microbial community was analyzed by temporal temperature gradient electrophoresis and real-time PCR targeting bacterial and fungal rDNA and PAH-ring hydroxylating dioxygenase genes. These communities were modified by PAH pollution, which selected PAH-degrading bacteria, by the presence of alfalfa and, concerning the bacterial community, by inoculation. PHE and PYR concentrations significantly decreased (91 and 46 %, respectively) whatever the treatment, but DBA concentration significantly decreased (30 %) in planted and co-inoculated microcosms only.
[Show abstract][Hide abstract] ABSTRACT: In contaminated soils, several natural processes (e.g. biodegradation, oxidation) can induce degradation of organic pollutants. To evaluate the effects of biodegradation on the organic matter and the main organic constituents (coal and coal tar) of a coking plant soil, we conducted a batch incubation experiment, whereby mineral nutrient solution and a microbial inoculum from the coking plant soil were added to the different samples. Microbial counts were monitored during the 9 months of the biodegradation experiment and the mineralization rate was quantified by measuring the CO2 produced and the content, molecular weight (MW) distribution and molecular composition of the solvent extractable organic matter. The microbial enumeration and mineralization rate demonstrated no biodegradation with the coal tar but biodegradation was observed for the coal and coking plant soil samples. The coking plant soil exhibited the highest biodegradation rate, probably due to complex interactions between the different organic and mineral phases and the presence of various nutrient and carbon sources. Biodegradation of the kerogen with subsequent production of lower MW compounds (n-alkanes and isoprenoids) was observed for both coal and coking plant soil samples. The results indicate that the kerogen could be a potential source of carbon for the microorganisms and showed the central role of coal in the changes observed for the coking plant soil.
[Show abstract][Hide abstract] ABSTRACT: Current soil quality evaluation does not include an assessment of metal bioavailability to organisms. However, sentinel soil-dwelling invertebrates can be used for such an assessment. This study aims to establish the modulating soil parameter of metal bioavailability to snails and a procedure for ranking field sites (n = 9; 43 plots) based on the evaluation of the transfer of metals to the land snails used as indicators of metal zooavailability. Multivariate regressions identify soil pH, organic carbon and iron oxides influence cadmium, chromium, copper, lead and zinc zooavailability to snails underlining the need to consider other parameter than total soil concentration during bioavailability assessment. However, for As, no influence of soil parameter on it bioavailability to snails was identified. Internal Concentrations of Reference (CIRef) of Cd, Pb, As, Cr, Cu and Zn were determined in Cantareus aspersus that were caged on unpolluted plots. CIRef allow for the identification of contaminated sites. CIRef have revealed unexpected metal transfer on some “unpolluted” sites and a lack of transfer on some contaminated sites, thus confirming the necessity for biological measures to evaluate metal mobility. The Sum of Excess of Transfers (SET) index ranked the industrially impacted sites as the top priorities for management. We recommend that the SET methodology be used for future environmental risk assessment. By highlighting real metal transfers and considering the numerous parameters influencing environmental bioavailability, the snails watch provides information on environmental quality.
[Show abstract][Hide abstract] ABSTRACT: The fungal communities of a multi-contaminated soil polluted by polycyclic aromatic hydrocarbons and heavy metals (NM) were studied within a long-term in situ experiment of natural attenuation assisted by plants. Three treatments were monitored: bare soil (NM-BS), soil planted with alfalfa and inoculated with mycorrhizal fungi (NM-Msm), and soil with spontaneous vegetation (NM-SV). The same soil after thermal desorption (TD) was planted with alfalfa and inoculated with mycorrhizal fungi (TD-Msm). Twice a year for 5 years, the fungal abundance and the community structure were evaluated by real-time PCR and temporal temperature gradient gel electrophoresis targeting 18S rRNA genes. The fungal abundance increased over time and was higher in planted than in bare NM soil and in TD than in NM soil. The Shannon diversity index (H') increased during the first 2 years with the emergence of more than 30 ribotypes, but decreased after 3 years with the selection of a few competitive species, mostly Ascomycetes. H' was higher under complex plant assemblage (NM-SV) than in the NM-BS plots but did not differ between NM and TD soils planted with alfalfa. These results indicated that even in a highly polluted soil, the plant cover was the main driver of the fungal community structure.
[Show abstract][Hide abstract] ABSTRACT: The polycyclic aromatic hydrocarbon (PAH) dissipation potential of Fusarium solani MM1 and Arthrobacter oxydans MsHM11, isolated from a PAH-polluted soil, was investigated in liquid cultures containing 500, 500, and 50 mg l−1 of phenanthrene (PHE), pyrene (PYR), and dibenz(a,h)anthracene (DBA), respectively. The contributions of adsorption, absorption and biotransformation were evaluated separately by extracting and analysing PAHs from the biomass and from the culture medium. In the pure culture, F. solani dissipated approximately 30% of each tested PAH in 28 days when glucose was supplied as the carbon source. PAH adsorption on F. solani hyphae was negligible, whereas absorption was found to account for one-third of the dissipation. Absorbed PAHs, reaching 3% of the fungal dry biomass, could be visualised in autofluorescent intracellular vesicles. In contrast, A. oxydans grew on PAHs as sole carbon source and led to 55% dissipation of PHE in 28 days without any significant absorption. This dissipation potential was inhibited by the presence of glucose, and the PYR and DBA contents were never dissipated in the bacterial pure culture. In the mixed co-culture, the PAH dissipation rates were lower, showing that antagonistic interactions, such as pH changes or competition for carbon source, indirectly inhibited both microbial potentials.
International Biodeterioration & Biodegradation 01/2012; 68(1):28-35. · 2.06 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Plant species (exotic invasive vs native non-invasive) colonization pattern and the relation with the soil nutrient availability and AM fungi abundance, was investigated. Soil samples were collected from two sites: one invaded by the exotic plant, Amaranthus viridis, and one uninvaded site for chemical and AM propagules density analyses. Additionally, we grew five Sahelian Acacia species in soil from the two sites, sterilized or not, to test the involvement of soil biota in the invasion process. While nutrient availability was significantly higher in soil samples from the invaded sites, a drastic reduction in AM fungal community density, was observed. Moreover, Acacia seedlings' growth was severely reduced in soils invaded by Amaranthus and this effect was similar to that of sterilized soil of both origins. The observed growth inhibition was accompanied by reduction of AM colonization and nodulation of the roots. Finally, the influence of soil chemistry and AM symbiosis on exotic plants' invasion processes is discussed.
[Show abstract][Hide abstract] ABSTRACT: Due to human activities, large volumes of soils are contaminated with organic pollutants such as polycyclic aromatic hydrocarbons, and very often by metallic pollutants as well. Multipolluted soils are therefore a key concern for remediation. This work presents a long-term evaluation of the fate and environmental impact of the organic and metallic contaminants of an industrially polluted soil under natural and plant-assisted conditions. A field trial was followed for four years according to six treatments in four replicates: unplanted, planted with alfalfa with or without mycorrhizal inoculation, planted with Noccaea caerulescens, naturally colonized by indigenous plants, and thermally treated soil planted with alfalfa. Leaching water volumes and composition, PAH concentrations in soil and solutions, soil fauna and microbial diversity, soil and solution toxicity using standardized bioassays, plant biomass, mycorrhizal colonization, were monitored. Results showed that plant cover alone did not affect total contaminant concentrations in soil. However, it was most efficient in improving the contamination impact on the environment and in increasing the biological diversity. Leaching water quality remained an issue because of its high toxicity shown by micro-algae testing. In this matter, prior treatment of the soil by thermal desorption proved to be the only effective treatment.
International Journal of Phytoremediation 01/2011; 13 Suppl 1:245-63. · 1.18 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Within the French National programme GESSOL and the GISFI consortium, the BIOTECHNOSOL project has been carried out. The purpose of the project aims to acquire information about the biodiversity of constructed Technosols that are used to restore brownfields. Results indicate an increase of biodiversity in the system within the two first study years.
[Show abstract][Hide abstract] ABSTRACT: Polycyclic aromatic hydrocarbon (PAH) dissipation efficiency can be increased in the plant rhizosphere, but may be affected by various environmental factors. We investigated the effects of the watering regime and phosphorus concentration on PAH dissipation in the rhizosphere of mycorrhizal plants in a pot experiment. Two plant species, alfalfa (Medicago sativa) and tall fescue (Festuca arundinacea), were co-cultured and inoculated with an arbuscular mycorrhizal (AM) fungus (Glomus intraradices) in PAH (phenanthrene (PHE)=500 mg kg(-1), pyrene (PYR)=500 mg kg(-1), dibenzo(a,h)anthracene (DBA)=65 mg kg(-1)) spiked agricultural soil for 6 weeks. Treatments with different phosphorus concentrations and watering regimes were compared. The PHE dissipation reached 90% in all treatments and was not affected by the treatments. The major finding was the significant positive impact of mycorrhizal plants on the dissipation of high molecular weight PAH (DBA) in high-water low-phosphorus treatment. Such an effect was not observed in high-water high-phosphorus and low-water low-phosphorus treatments, where AM colonization was very low. A positive linear relationship was detected between PYR dissipation and the percentage of Gram-positive PAH-ring hydroxylating dioxygenase genes in high-water high-phosphorus treatments, but not in the other two treatments with lower phosphorus concentrations and water contents. Such results indicated that the phosphorus and water regime were important parameters for the dissipation of HMW-PAH.
[Show abstract][Hide abstract] ABSTRACT: The objectives of this study were to determine whether the invasive plant Amaranthus viridis influenced soil microbial and chemical properties and to assess the consequences of these modifications on native plant growth. The experiment was conducted in Senegal at two sites: one invaded by A. viridis and the other covered by other plant species. Soil nutrient contents as well as microbial community density, diversity and functions were measured. Additionally, five sahelian Acacia species were grown in (1) soil disinfected or not collected from both sites, (2) uninvaded soil exposed to an A. viridis plant aqueous extract and (3) soil collected from invaded and uninvaded sites and inoculated or not with the arbuscular mycorrhizal (AM) fungus Glomus intraradices. The results showed that the invasion of A. viridis increased soil nutrient availability, bacterial abundance and microbial activities. In contrast, AM fungi and rhizobial development and the growth of Acacia species were severely reduced in A. viridis-invaded soil. Amaranthus viridis aqueous extract also exhibited an inhibitory effect on rhizobial growth, indicating an antibacterial activity of this plant extract. However, the inoculation of G. intraradices was highly beneficial to the growth and nodulation of Acacia species. These results highlight the role of AM symbiosis in the processes involved in plant coexistence and in ecosystem management programs that target preservation of native plant diversity.
[Show abstract][Hide abstract] ABSTRACT: The polycyclic aromatic hydrocarbon (PAH) contamination, bacterial community, and PAH-degrading bacteria were monitored in aged PAH-contaminated soil (Neuves-Maisons [NM] soil; with a mean of 1,915 mg of 16 PAHs.kg(-1) of soil dry weight) and in the same soil previously treated by thermal desorption (TD soil; with a mean of 106 mg of 16 PAHs.kg(-1) of soil dry weight). This study was conducted in situ for 2 years using experimental plots of the two soils. NM soil was colonized by spontaneous vegetation (NM-SV), planted with Medicago sativa (NM-Ms), or left as bare soil (NM-BS), and the TD soil was planted with Medicago sativa (TD-Ms). The bacterial community density, structure, and diversity were estimated by real-time PCR quantification of the 16S rRNA gene copy number, temporal thermal gradient gel electrophoresis fingerprinting, and band sequencing, respectively. The density of the bacterial community increased the first year during stabilization of the system and stayed constant in the NM soil, while it continued to increase in the TD soil during the second year. The bacterial community structure diverged among all the plot types after 2 years on site. In the NM-BS plots, the bacterial community was represented mainly by Betaproteobacteria and Gammaproteobacteria. The presence of vegetation (NM-SV and NM-Ms) in the NM soil favored the development of a wider range of bacterial phyla (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Verrucomicrobia, Actinobacteria, Firmicutes, and Chloroflexi) that, for the most part, were not closely related to known bacterial representatives. Moreover, under the influence of the same plant, the bacterial community that developed in the TD-Ms was represented by different bacterial species (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, and Actinobacteria) than that in the NM-Ms. During the 2 years of monitoring, the PAH concentration did not evolve significantly. The abundance of gram-negative (GN) and gram-positive (GP) PAH-degrading bacteria was estimated by real-time PCR quantification of specific functional genes encoding the alpha subunit of PAH-ring hydroxylating dioxygenase (PAH-RHD(alpha)). The percentage of the PAH-RHD(alpha) GN bacterial genes relative to 16S rRNA gene density decreased with time in all the plots. The GP PAH-RHD(alpha) bacterial gene proportion decreased in the NM-BS plots but stayed constant or increased under vegetation influence (NM-SV, NM-Ms, and TD-Ms).
Applied and Environmental Microbiology 08/2009; 75(19):6322-30. · 3.95 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Toxic metal accumulation in soils of agricultural interest is a serious problem needing more attention, and investigations on soil-plant metal transfer must be pursued to better understand the processes involved in metal uptake. Arbuscular mycorrhizal (AM) fungi are known to influence metal transfer in plants by increasing plant biomass and reducing metal toxicity to plants even if diverging results were reported. The effects of five AM fungi isolated from metal contaminated or non-contaminated soils on metal (Cd, Zn) uptake by plant and transfer to leachates was assessed with Medicago truncatula grown in a multimetallic contaminated agricultural soil. Fungi isolated from metal-contaminated soils were more effective to reduce shoot Cd concentration. Metal uptake capacity differed between AM fungi and depended on the origin of the isolate. Not only fungal tolerance and ability to reduce metal concentrations in plant but also interactions with rhizobacteria affected heavy metal transfer and plant growth. Indeed, thanks to association with nodulating rhizobacteria, one Glomus intraradices inoculum increased particularly plant biomass which allowed exporting twofold more Cd and Zn in shoots as compared to non-mycorrhizal treatment. Cd concentrations in leachates were variable among fungal treatments, but can be significantly influenced by AM inoculation. The differential strategies of AM fungal colonisation in metal stress conditions are also discussed.
[Show abstract][Hide abstract] ABSTRACT: Some plants are more mycorrhizal than others and mycorrhizal colonisation of plants in extreme environments is frequently
additionally reduced due to decreased spore density and/or diversity and therefore frequently overlooked. We analysed two
plant species from both metal polluted and saline enriched soils with differing mycorrhizal colonisation levels/status using
classical and molecular methods. The selected plant species were Sesleria caerulea (L.) Ard. and Thlaspi praecox Wulfen from a metal polluted site, and Limonium angustifolium (Tausch) Degen [Statice serotina Rchb., L. vulgare Mill. subsp. Serotinum (Rchb.) Gams] and Salicornia europaea L. from the Sečovlje salterns in Slovenia. Despite the high mycorrhizal frequencies (F%) observed, the presence of arbuscules (A%) was at best low in S. caerulea and T. praecox, and undetectable in L. angustifolium and S. europaea. Temporal temperature gradient gel electrophoresis (TTGE) was applied to field-collected samples from both burdened environments
and proved to be an effective technique for rapid profiling and identification of arbuscular mycorrhizal fungi (AMF). Sequencing
and phylogenetic analysis confirmed the association of AMF of the genus Glomus with roots of all four plant species. This is the first report on the identification and profiling of Glomeromycota in the
field-collected Cd/Zn metal hyperaccumulator T. praecox growing at a highly metal polluted site, as well as in L. angustifolium and S. europaea collected in a saline environment. The identification of AMF from both ecosystems only partially resembles previous identifications
on the basis of spores.
Plant and Soil 01/2009; 314(1):25-34. · 3.24 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study was designed to examine saprophytic fungi diversity under different tree species situated in the same ecological context. Further, the link between the diversity and decomposition rate of two broadleaved, two coniferous and two mixed broadleaved-coniferous litter types was targeted. Litter material was decomposed in litter bags for 4 and 24 months to target both early and late stages of the decomposition. Fungal diversity of L and F layers were also investigated as a parallel to the litter bag method. Temperature gradient gel electrophoresis fingerprinting was used to assess fungal diversity in the samples. Mass loss values and organic and nutrient composition of the litter were also measured. The results showed that the species richness was not strongly affected by the change of the tree species. Nevertheless, the community compositions differed within tree species and decomposition stages. The most important shift was found in the mixed litters from the litter bag treatment for both variables. Both mixed litters displayed the highest species richness (13.3 species both) and the most different community composition as compared to pure litters (6.3-10.7 species) after 24 months. The mass loss after 24 months was similar or greater in the mixed litter (70.5% beech-spruce, 76.2% oak-Douglas-fir litter) than in both original pure litter types. This was probably due to higher niche variability and to the synergistic effect of nutrient transfer between litter types. Concerning pure litter, mass loss values were the highest in oak and beech litter (72.8% and 69.8%) compared to spruce and D. fir (59.4% and 66.5%, respectively). That was probably caused by a more favourable microclimate and litter composition in broadleaved than in coniferous plantations. These variables also seemed to be more important to pure litter decomposition rates than were fungal species richness or community structure.
[Show abstract][Hide abstract] ABSTRACT: Arbuscular mycorrhizal (AM) fungi are known to have a beneficial effect on host plants growing in metal polluted soils. However, the potential contribution of AM fungi to cadmium (Cd) accumulation in plants is still uncertain and is rarely quantified in the context of phytoremediation. In a pot experiment, we quantified the partitioning of Cd in mycorrhizal (M) or non-mycorrhizal (NM) Medicago truncatula in four soils as a function of Cd availability. AM inoculation with Glomus intraradices increased plant growth and metal accumulation in above-ground biomass, even though Cd concentrations in M plants were lower than in NM plants. AM inoculation also increased the quantity of available Cd and reduced Cd in leachates. Results were consistent and significant in the four tested soils and were not related to Cd availability. Such AM fungi could be a good tool for phytoextraction.
Soil Biology and Biochemistry 08/2008; 40:2710-2712. · 4.41 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Real-Time PCR based assays were developed to quantify Gram positive (GP) and Gram negative (GN) bacterial populations that are capable of degrading the polycyclic aromatic hydrocarbons (PAH) in soil and sediment samples with contrasting contamination levels. These specific and sensitive Real-Time PCR assays were based on the quantification of the copy number of the gene that encodes the alpha subunit of the PAH-ring hydroxylating dioxygenases (PAH-RHDalpha), involved in the initial step of the aerobic metabolism of PAH. The PAH-RHDalpha-GP primer set was designed against the different allele types present in the data base (narAa, phdA/pdoA2, nidA/pdoA1, nidA3/fadA1) common to the Gram positive PAH degraders such as Rhodococcus, Mycobacterium, Nocardioides and Terrabacter strains. The PAH-RHDalpha-GN primer set was designed against the genes (nahAc, nahA3, nagAc, ndoB, ndoC2, pahAc, pahA3, phnAc, phnA1, bphAc, bphA1, dntAc and arhA1) common to the Gram negative PAH degraders such as Pseudomonas, Ralstonia, Commamonas, Burkholderia, Sphingomonas, Alcaligenes, Polaromonas strains. The PCR clones for DNA extracted from soil and sediment samples using the designed primers showed 100% relatedness to the PAH-RHDalpha genes targeted. Deduced from highly sensitive Real-Time PCR quantification, the ratio of PAH-RHDalpha gene relative to the 16S rRNA gene copy number showed that the PAH-bacterial degraders could represent up to 1% of the total bacterial community in the PAH-contaminated sites. This ratio highlighted a positive correlation between the PAH-bacterial biodegradation potential and the PAH-contamination level in the environmental samples studied.
Journal of Microbiological Methods 06/2008; 73(2):148-59. · 2.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We studied the effect of forest tree species on a community of decomposers that colonize cellulose strips. Both fungal and bacterial communities were targeted in a native forest dominated by beech and oak and 30-year-old beech and spruce plantations, growing in similar ecological conditions in the Breuil-Chenue experimental forest site in Morvan (France). Microbial ingrowths from the 3rd to 10th month of strip decomposition (May to December 2004) were studied. Community composition was assessed using temperature gradient gel electrophoresis with universal fungal (ITS1F, ITS2) and bacterial (1401r, 968f) primers. Soil temperature and moisture as well as fungal biomass were also measured to give additional information on decomposition processes. Changing the dominant tree species had no significant influence in the number of decomposer species. However, decomposer community composition was clearly different. If compared to the native forest, where community composition highly differed, young monocultures displayed similar species structure for fungi and bacteria. Both species numbers and community composition evolved during the decay process. Time effect was found to be more important than tree species. Nevertheless, the actual environmental conditions and seasonal effect seemed to be even more determining factors for the development of microbial communities. The course and correlations of the explored variables often differed between tree species, although certain general trends were identified. Fungal biomass was high in summer, despite that species richness (SR) decreased and conversely, that high SR did not necessarily mean high biomass values. It can be concluded that the growth and development of the microbiological communities that colonized a model material in situ depended on the combination of physical and biological factors acting collectively and interdependently at the forest soil microsite.
[Show abstract][Hide abstract] ABSTRACT: Bioremediation technologies of Polycyclic Aromatic Hydrocarbons (PAH) are often limited by the recalcitrance to biodegradation of high molecular weight (HMW) PAH. Rhizosphere is known to increase the biodegradation of PAH but little is known about the biodegradability of these HMW compounds by rhizosphere bacteria. This study compared the effects of a 3 and a 5-ring PAH, phenanthrene (PHE) and dibenzo[a,h]anthracene (dBA) respectively, on the composition of bacterial community, the bacterial density and the biodegradation activity. Compartmentalized devices were designed to harvest three consecutive sections of the rhizosphere. Rhizosphere and non-rhizosphere compartments were filled with PHE or dBA spiked or unspiked sand and inoculated with a soil bacterial inoculum. Different bacterial communities and degradation values were found 5 weeks after spiking with PHE (41-76% biodegradation) and dBA (12-51% biodegradation). In sections closer to the root surface, bacterial populations differed as a function of the distance to roots and the PAH added, whereas in further rhizosphere sections, communities were closer to those of the non-planted treatments. Biodegradation of PHE was also a function of the distance to roots, and decreased from 76 to 42% within 9 mm from the roots. However, biodegradation of dBA was significantly higher in the middle section (3-6 mm from roots) than the others. Rhizosphere degradation of PAH varies with the nature of the PAH, and C fluxes from roots could limit the degradation of dBA.