Suchergebnisse

Explosives manufacturing and military activities generated large quantities of waste products, contaminating soil and groundwater at many military training ranges and manufacturing sites in Belgium. Consequently there is a need to develop strategies to efficiently remediate these sites. While many microorganisms are known to naturally metabolise certain energetic compounds in contaminated sites (bioremediation) and in the root-zone of plants, a process known as rhizoremediation, removal of 2,4,6-trinitrotoluene (TNT) is challenging due to its toxicity and recalcitrance to biological break-down. Treatments directed at stimulating microbial TNT-transformation activity, e.g. by 'one-microorganism' introduction or biostimulation have varied in effectiveness. The objective of this study was to explore the TNT-denitrating and TNT-detoxifying abilities of natural, plant-associated bacteria from Acer pseudoplatanus (sycamore) trees in TNT-contaminated soil, and to understand how TNT affects bacterial community composition in the rhizosphere of young sycamore trees and surrounding bulk soil. From this we determined whether the use of tailored bacterial consortia can improve TNT remediation. ; De productie van explosieven en militaire activiteiten genereerde grote hoeveelheden afvalstromen van explosieven, die terechtkwamen in het milieu en leidden tot een ernstige vervuiling van bodem en grondwater op verschillende militaire schietvelden en explosieven productie faciliteiten, in België. Er is dus nood aan technieken om deze vervuilde bodems op een doeltreffende manier te saneren. Ook al zijn er veel bacteriën gekend die van nature explosieven kunnen afbreken en ontgiftigen in vervuilde bodems, hetgeen gekend is als rhizoremediatie als de bacteriën zich in de wortel-zone van planten bevinden, toch blijft de eliminatie van 2,4,6-trinitrotolueen uit de bodem een uitdaging, omwille van de hoge toxiciteit en de recalcitrantie voor biologische afbraak. Ingrepen die gericht zijn op het stimuleren van de microbiële activiteit voor de afbraak van TNT, zoals de introductie van bacteriën of bio-stimulatie (bv. het toevoegen van nutriënten), heeft tot wisselende resultaten geleid. Het doel van deze studie was enerzijds om diepere inzichten te verwerven in bacteriën geassocieerd met Acer pseudoplatanus (gewone esdoorn) op een TNT-verontreinigde bodem, en hun capaciteiten om de bodem te ontgiftigen door TNT te reduceren. Anderzijds was het doel om beter te begrijpen hoe TNT de samenstelling van de bacteriële gemeenschap beïnvloedt in de rhizosfeer van jonge esdoorns en de rest van de bodem. Met deze verworven kennis kon dieper ingegaan worden op de vraag of TNT-remediatie bevorderd kan worden door bacteriële consortia in te schakelen.

Background: A diverse community of microbes naturally exists on the phylloplane, the surface of leaves. It is one of the most prevalent microbial habitats on earth and bacteria are the most abundant members, living in communities that are highly dynamic. Today, one of the key challenges for microbiologists is to develop strategies to culture the vast diversity of microorganisms that have been detected in metagenomic surveys. Results: We isolated bacteria from the phylloplane of Hedera helix (common ivy), a widespread evergreen, using five growth media: Luria–Bertani (LB), LB01, yeast extract–mannitol (YMA), yeast extract–flour (YFlour), and YEx. We also included a comparison with the uncultured phylloplane, which we showed to be dominated by Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes. Inter-sample (beta) diversity shifted from LB and LB01 containing the highest amount of resources to YEx, YMA, and YFlour which are more selective. All growth media equally favoured Actinobacteria and Gammaproteobacteria, whereas Bacteroidetes could only be found on LB01, YEx, and YMA. LB and LB01 favoured Firmicutes and YFlour was most selective for Betaproteobacteria. At the genus level, LB favoured the growth of Bacillus and Stenotrophomonas, while YFlour was most selective for Burkholderia and Curtobacterium. The in vitro plant growth promotion (PGP) profile of 200 isolates obtained in this study indicates that previously uncultured bacteria from the phylloplane may have potential applications in phytoremediation and other plant-based biotechnologies. Conclusions: This study gives first insights into the total bacterial community of the H. helix phylloplane, including an evaluation of its culturability using five different growth media. We further provide a collection of 200 bacterial isolates underrepresented in current databases, including the characterization of PGP profiles. Here we highlight the potential of simple strategies to obtain higher microbial diversity from environmental samples and the use of high-throughput sequencing to guide isolate selection from a variety of growth media. ; Funding This work was supported by the Hasselt University Methusalem project 08M03VGRJ, by the project G0D0916N ("Plant–microbe associations to reduce particulate matter concentration and toxicity in urban areas: a multidisciplinary approach") financed by the Research Foundation – Flanders (FWO) and by a personal grant of VS provided by the Research Foundation – Flanders (FWO). The funding bodies had no role in the design of the study and collection, analysis, interpretation of data, or manuscript preparation. Acknowledgements We thank Prof. Jonathan Van Hamme for proofreading the language in this manuscript. We thank the VSC (Flemish Supercomputer Center), funded by the Research Foundation – Flanders (FWO) and the Flemish Government – department EWI for providing the computational resources and services used in this work.

The presence of explosives in soils and the interaction with drought stress and nutrient limitation are among the environmental factors that severely affect plant growth on military soils. In this study, we seek to isolate and identify the cultivable bacteria of a 2,4-dinitrotoluene (DNT) contaminated soil (DS) and an adjacent grassland soil (GS) of a military training area aiming to isolate new plant growth-promoting (PGP) and 2,4-DNT-degrading strains. Metabolic profiling revealed disturbances in Ecocarbon use in the bare DS; isolation of cultivable strains revealed a lower colony-forming-unit count and a less diverse community associated with DS in comparison with GS. New 2,4-DNT-tolerant strains were identified by selective enrichments, which were further characterized by auxanography for 2,4-DNT use, resistance to drought stress, cold, nutrient starvation and PGP features. By selecting multiple beneficial PGP and abiotic stress-resistant strains, efficient 2,4-DNT-degrading consortia were composed. After inoculation, consortium UHasselt Sofie 3 with seven members belonging to Burkholderia, Variovorax, Bacillus, Pseudomonas and Ralstonia species was capable to successfully enhance root length of Arabidopsis under 2,4-DNT stress. After 9 days, doubling of main root length was observed. Our results indicate that beneficial bacteria inhabiting a disturbed environment have the potential to improve plant growth and alleviate 2,4-DNT stress. ; This study was supported by a PhD grant to ST from the Fund for Scientific Research-Flanders, Belgium (FWO-Vlaanderen) and the UHasselt Methusalem project 08M03VGRJ.

The presence of explosives in soils and the interaction with drought stress and nutrient limitation are among the environmental factors that severely affect plant growth on military soils. In this study, we seek to isolate and identify the cultivable bacteria of a 2,4-dinitrotoluene (DNT) contaminated soil (DS) and an adjacent grassland soil (GS) of a military training area aiming to isolate new plant growth-promoting (PGP) and 2,4-DNT-degrading strains. Metabolic profiling revealed disturbances in Ecocarbon use in the bare DS; isolation of cultivable strains revealed a lower colony-forming-unit count and a less diverse community associated with DS in comparison with GS. New 2,4-DNT-tolerant strains were identified by selective enrichments, which were further characterized by auxanography for 2,4-DNT use, resistance to drought stress, cold, nutrient starvation and PGP features. By selecting multiple beneficial PGP and abiotic stress-resistant strains, efficient 2,4-DNT-degrading consortia were composed. After inoculation, consortium UHasselt Sofie 3 with seven members belonging to Burkholderia, Variovorax, Bacillus, Pseudomonas and Ralstonia species was capable to successfully enhance root length of Arabidopsis under 2,4-DNT stress. After 9 days, doubling of main root length was observed. Our results indicate that beneficial bacteria inhabiting a disturbed environment have the potential to improve plant growth and alleviate 2,4-DNT stress.

We report the draft genome of Raoultella ornithinolytica TNT, a Gram-negative bacterium of the Enterobacteriaceae isolated from military soil in Belgium. Strain TNT uses nitrite released from trinitrotoluene (TNT) for growth and is a potent plant growth promoter. An analysis of its 5.6-Mb draft genome will bring insights into TNT degradation-reinforcing bioremediation applications.

We report the draft genome of Raoultella ornithinolytica TNT, a Gram-negative bacterium of the Enterobacteriaceae isolated from military soil in Belgium. Strain TNT uses nitrite released from trinitrotoluene (TNT) for growth and is a potent plant growth promoter. An analysis of its 5.6-Mb draft genome will bring insights into TNT degradation-reinforcing bioremediation applications.

Lead (Pb) and cadmium (Cd) are major environmental pollutants, and the accumulation of these elements in soils and plants is of great concern in agricultural production due to their toxic effects on crop growth. Also, these elements can enter into the food chain and severely affect human and animal health. Bioaugmentation with plant growth-promoting bacteria (PGPB) can contribute to an environmentally friendly and effective remediation approach by improving plant survival and promoting element phytostabilization or extraction under such harsh conditions. We isolated and characterised Pb and Cd-tolerant root-associated bacteria from Helianthus petiolaris growing on a Pb/Cd polluted soil in order to compose inoculants that can promote plant growth and also ameliorate the phytostabilization or phytoextraction efficiency. One hundred and five trace element-tolerant rhizospheric and endophytic bacterial strains belonging to eight different genera were isolated from the aromatic plant species Helianthus petiolaris. Most of the strains showed multiple PGP-capabilities, ability to immobilise trace elements on their cell wall, and promotion of seed germination. Bacillus paramycoides ST9, Bacillus wiedmannii ST29, Bacillus proteolyticus ST89, Brevibacterium frigoritolerans ST30, Cellulosimicrobium cellulans ST54 and Methylobacterium sp. ST85 were selected to perform bioaugmentation assays in greenhouse microcosms. After 2 months, seedlings of sunflower (H. annuus) grown on polluted soil and inoculated with B. proteolyticus ST89 produced 40% more biomass compared to the non-inoculated control plants and accumulated 20 % less Pb and 40% less Cd in the aboveground plant parts. In contrast, B. paramycoides ST9 increased the bioaccumulation factor (BAF) of Pb three times and of Cd six times without inhibiting plant growth. Our results indicate that, depending on the strain, bioaugmentation with specific beneficial bacteria can improve plant growth and either reduce trace element mobility or enhance plant trace element uptake. ; This research was funded by the UHasselt-Methusalem project 08M03VGRJ, as got an UHasselt BOF-BILA grant in the frame of collaboration between Hasselt University, Belgium and the University of Buenos Aires, and Ibero-American Development Bank under FONTAGRO project ATN/RF-16110-RG. ; Thijs, S (reprint author), Hasselt Univ, Ctr Environm Sci, Environm Biol, Agoralaan Bldg D, B-3590 Diepenbeek, Belgium. saran.anabel@inta.gob.ar; valeria.imperato@uhasselt.be; fernandez.lucia@inta.gob.ar; panos.gkorezis@uhasselt.be; jan.d'haen@uhasselt.be; merini.luciano@inta.gob.ar; jaco.vangronsveld@uhasselt.be; sofie.thijs@uhasselt.be

Military activities have worldwide introduced toxic explosives into the environment with considerable effects on soil and plant-associated microbiota. Fortunately, these microorganisms, and their collective metabolic activities, can be harnessed for site restoration via in situ phytoremediation. We characterized the bacterial communities inhabiting the bulk soil and rhizosphere of sycamore maple (Acer pseudoplatanus) in two chronically 2,4,6-trinitrotoluene (TNT) polluted soils. Three hundred strains were isolated, purified and characterized, a majority of which showed multiple plant growth promoting (PGP) traits. Several isolates showed high nitroreductase enzyme activity and concurrent TNT-transformation. A 12-member bacterial consortium, comprising selected TNT-detoxifying and rhizobacterial strains, significantly enhanced TNT removal from soil compared to non-inoculated plants, increased root and shoot weight, and the plants were less stressed than the un-inoculated plants as estimated by the responses of antioxidative enzymes. The sycamore maple tree (SYCAM) culture collection is a significant resource of plant-associated strains with multiple PGP and catalytic properties, available for further genetic and phenotypic discovery and use in field applications.

Despite great advances, experiments concerning the response of ecosystems to climate change still face considerable challenges, including the high complexity of climate change in terms of environmental variables, constraints in the number and amplitude of climate treatment levels, and the limited scope of responses and interactions covered. Drawing on the expertise of researchers from a variety of disciplines, this Perspective outlines how computational and technological advances can help in designing experiments that can contribute to overcoming these challenges, and also outlines a first application of such an experimental design. ; We thank the Flemish government (through Hercules Stichting big infrastructure and the Fund for Scientific Research Flanders project G0H4117N) and LSM (Limburg Sterk Merk, project 271) for providing funds to build the UHasselt Ecotron; Hasselt University for both funding and policy support (project BOF12BR01 and Methusalem project 08M03VGRJ); and the ecotron research committee for comments on the experimental design. We also thank Regional Landscape Kempen and Maasland for its collaboration and support. N.W., S.L., A.N. and I.V. are funded by Research Foundation-Flanders (FWO).