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See detailDiscovering novel enzymes y functional screening of plurigenomic libraries from alga-associated Flavobacteriia and Gammaproteobacteria
Martin, Marjolaine ULg; Vandermies, Marie ULg; Joyeux, Coline et al

in Microbiological Research (2016), 186

Alga-associated microorganisms, in the context of their numerous interactions with the host and thecomplexity of the marine environment, are known to produce diverse hydrolytic enzymes with ... [more ▼]

Alga-associated microorganisms, in the context of their numerous interactions with the host and thecomplexity of the marine environment, are known to produce diverse hydrolytic enzymes with originalbiochemistry. We recently isolated several macroalgal-polysaccharide-degrading bacteria from the sur-face of the brown alga Ascophyllum nodosum. These active isolates belong to two classes: the Flavobacteriiaand the Gammaproteobacteria. In the present study, we constructed two “plurigenomic” (with multi-ple bacterial genomes) libraries with the 5 most interesting isolates (regarding their phylogeny andtheir enzymatic activities) of each class (Fv and Gm libraries). Both libraries were screened for diversehydrolytic activities. Five activities, out of the 48 previously identified in the natural polysaccharolyticisolates, were recovered by functional screening: a xylanase (GmXyl7), a beta-glucosidase (GmBg1), anesterase (GmEst7) and two iota-carrageenases (Fvi2.5 and Gmi1.3). We discuss here the potential role ofthe used host-cell, the average DNA insert-sizes and the used restriction enzymes on the divergent screen-ing yields obtained for both libraries and get deeper inside the “great screen anomaly”. Interestingly, thediscovered esterase probably stands for a novel family of homoserine o-acetyltransferase-like-esterases,while the two iota-carrageenases represent new members of the poorly known GH82 family (contain-ing only 19 proteins since its description in 2000). These original results demonstrate the efficiencyof our uncommon “plurigenomic” library approach and the underexplored potential of alga-associatedcultivable microbiota for the identification of novel and algal-specific enzymes. [less ▲]

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See detailNo favorable effect of reduced tillage on microbial community diversity in a silty loam soil (Belgium)
Degrune, Florine ULg; Theodorakopoulos, Nicolas ULg; Dufrêne, Marc ULg et al

in Agriculture, Ecosystems & Environment (2016)

Among the soil management practices used to promote sustainable agriculture, reduced tillage and retention of residues from the previous crop are reported to enhance significantly both soil fertility and ... [more ▼]

Among the soil management practices used to promote sustainable agriculture, reduced tillage and retention of residues from the previous crop are reported to enhance significantly both soil fertility and crop productivity. Here, high-throughput sequencing (454 technology) was used to see how the tillage regime (conventional vs. reduced tillage) and the fate of crop residues (retention or removal) affect microbial communities at two sampling depths (top soil: 0–5 cm and deeper soil: 15–20 cm) in a fertile silty loam soil in Belgium. All combinations of these three factors were studied. After 6 years of conversion from conventional to reduced tillage, depth emerged as the main factor responsible for variation in microbial diversity, tillage regime ranked second, and finally, crop residue fate had no influence on microbial diversity. For both bacteria and fungi, the diversity appeared higher in the top soil than in the deeper soil, and surprisingly, higher under conventional than under reduced tillage. These differences are explained by changes in community composition due to taxon loss rather than taxon replacement. The specific local set of environmental conditions (a loess-derived soil and an oceanic temperate climate) may explain these results. These observations raise the question: does impoverishment in indicator taxa influence soil processes, and thus crop production? To answer this question, we discuss how the presence of certain indicator taxa liable to play an ecological role might relate to crop productivity. [less ▲]

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See detailNo favorable effect of reduced tillage on microbial communities in a silty loam soil (Belgium)
Degrune, Florine ULg; Theodorakopoulos, Nicolas ULg; Dufrêne, Marc ULg et al

Poster (2015, December 01)

To date, only a few studies have applied metagenomics to investigate the influence of different tillage regimes and types of crop residue management on soil microbial communities. These studies were ... [more ▼]

To date, only a few studies have applied metagenomics to investigate the influence of different tillage regimes and types of crop residue management on soil microbial communities. These studies were conducted under specific climates on soils characterized by particular land-use histories. A very different ecological context is to be found in certain areas of Western Europe, such as central Belgium, whose loess-derived soils are among the most fertile in the world and have long been used for intensive agriculture. Specific objectives were to determine diversity levels and changes in microbial community composition under different combinations of tillage regime (conventional vs. reduced) and crop residue fate (residue removal R- vs. residues left R+ on the field). As reduced tillage results in two contrasting zones (the first centimeters of soil are mixed each year, while the soil below remains unperturbed), we chose to perform the analysis at two depths: 0 to 5 cm and 15 to 20 cm. [less ▲]

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See detailThe cultivable surface microbiota of the brown alga Ascophyllum nodosum is enriched in macroalgal-polysaccharide-degrading bacteria
Martin, Marjolaine ULg; Barbeyron, Tristan; Martin, Renée ULg et al

in Frontiers in Microbiology (2015), 6(december),

Bacteria degrading algal polysaccharides are key players in the global carbon cycle and in algal biomass recycling. Yet the water column, which has been studied largely by metagenomic approaches, is poor ... [more ▼]

Bacteria degrading algal polysaccharides are key players in the global carbon cycle and in algal biomass recycling. Yet the water column, which has been studied largely by metagenomic approaches, is poor in such bacteria and their algal-polysaccharide-degrading enzymes. Even more surprisingly, the few published studies on seaweed-associated microbiomes have revealed low abundances of such bacteria and their specific enzymes. However, as macroalgal cell-wall polysaccharides do not accumulate in nature, these bacteria and their unique polysaccharidases must not be that uncommon. We, therefore, looked at the polysaccharide-degrading activity of the cultivable bacterial subpopulation associated with Ascophyllum nodosum. From A. nodosum triplicates, 324 bacteria were isolated and taxonomically identified. Out of these isolates, 78 (∼25%) were found to act on at least one tested algal polysaccharide (agar, ι- or κ-carrageenan, or alginate). The isolates “active” on algal-polysaccharides belong to 11 genera: Cellulophaga, Maribacter, Algibacter, and Zobellia in the class Flavobacteriia (41) and Pseudoalteromonas, Vibrio, Cobetia, Shewanella, Colwellia, Marinomonas, and Paraglaceciola in the class Gammaproteobacteria (37). A major part represents likely novel species. Different proportions of bacterial phyla and classes were observed between the isolated cultivable subpopulation and the total microbial community previously identified on other brown algae. Here, Bacteroidetes and Gammaproteobacteria were found to be the most abundant and some phyla (as Planctomycetes and Cyanobacteria) frequently encountered on brown algae weren’t identified. At a lower taxonomic level, twelve genera, well-known to be associated with algae (with the exception for Colwellia), were consistently found on all three A. nosodum samples. Even more interesting, 9 of the 11 above mentioned genera containing polysaccharolytic isolates were predominant in this common core. The cultivable fraction of the bacterial community associated with A. nodosum is, thus, significantly enriched in macroalgal-polysaccharide-degrading bacteria and these bacteria seem important for the seaweed holobiont even though they are under-represented in alga-associated microbiome studies. [less ▲]

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See detailEffects of Plant Growth-Promoting Rhizobacteria on Wheat Growth under Greenhouse and Field Conditions in combination with different nitrogen fertilizer levels
Nguyen, Minh ULg; Ongena, Marc ULg; Colinet, Gilles ULg et al

Poster (2015, November 23)

Many Plant Growth-Promoting Rhizobacteria (PGPR) are able to enhance root growth, mineral availability, and nutrient use efficiency of crops. The aim of this project is to screen commercially available ... [more ▼]

Many Plant Growth-Promoting Rhizobacteria (PGPR) are able to enhance root growth, mineral availability, and nutrient use efficiency of crops. The aim of this project is to screen commercially available PGPR formulations and lab strains to increase wheat growth and yield in combination with an optimized nitrogen (N) fertilizer application scheme. This could lead to a significant reduction of N fertilizer application without affecting the subsequent grain yields. The screened products collection includes (1) Mix1 (a mix of Azospirillum sp., Azorhizobium sp., and Azoarcus sp.), (2) Mix2 (a mix of Mix1 complemented with two strains of phosphorus-solubilizing Bacillus sp.), (3) Bacillus amyloliquefaciens a, (4) B. subtilis, and (5) B. amyloliquefaciens b. These products were screened under greenhouse and field conditions in 2014 by using spring and winter wheat varieties, respectively. Under greenhouse conditions, there was a significant increase in root dry weight and in root per shoot ratio of plants inoculated with Mix1. Under field conditions, the interaction between PGPR inoculation and different N fertilizer doses was assessed. The grain yield was negatively impacted by low N fertilizer applications. Under such conditions, the inoculation of the wheat rhizosphere with B. subtilis increased the grain yield by 15% relative to the water control. However, in the field trial, the variability between plot replicates was high and lead to non-significant results. Based on these results, modified screening strategies for PGPR selection were set up for the next trials. [less ▲]

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See detailImpacts of Plant Growth-Promoting Rhizobacteria-based Biostimulants on Wheat Growth under Greenhouse and Field Conditions
Nguyen, Minh ULg; Ongena, Marc ULg; Colinet, Gilles ULg et al

Poster (2015, November 16)

Plant Growth-Promoting Rhizobacteria (PGPR) are one of the main biostimulant classes due to their capacity of stimulating root growth and enhancing soil mineral availability, hence increasing nutrient use ... [more ▼]

Plant Growth-Promoting Rhizobacteria (PGPR) are one of the main biostimulant classes due to their capacity of stimulating root growth and enhancing soil mineral availability, hence increasing nutrient use efficiency in crops. The aim of this study is to screen commercially PGPR-containing products to enhance wheat growth and yield in combination with an optimized nitrogen (N) fertilizer application scheme. This could lead to a significant reduction of N fertilizer application without affecting the subsequent grain yields. The screened products collection includes (1) Mix1 (a mix of Azospirillum sp., Azorhizobium sp., and Azoarcus sp.), (2) Mix2 (a mix of Mix1 complemented with two strains of phosphorus-solubilizing Bacillus sp.), (3) Bacillus amyloliquefaciens a, (4) B. subtilis, and (5) B. amyloliquefaciens b. These biostimulants were screened under greenhouse and field conditions in 2014 by using spring and winter wheat varieties respectively. There was a significant increase in root dry weight and in root per shoot ratio of plants inoculated with Mix1. Under field conditions, the interaction between PGPR inoculation and N fertilizer application was assessed. The grain yield was negatively impacted by low N fertilizer applications. Under such conditions, the inoculation of the wheat rhizosphere with Bacillus subtilis increased the grain yield by 15% relative to the water control. However, in the field trial, the variability between plot replicates was high and lead to non-significant results. Based on those results, modified screening strategies for PGPR selection were set up for the 2015 trials to reduce field variability and possibly achieve higher yield increases. [less ▲]

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See detailBiotechnological potential of the microflora associated with the brown alga Ascophyllum nodosum
Martin, Marjolaine ULg; Martin, Renée ULg; Barbeyron, Tristan et al

in Journal of Microbial and Biochemical Technology (2015, August 18), 7(4), 179

Bacteria associated with algae are underexplored despite their huge biodiversity and the fact that they differ markedly from those living freely in seawater. These bacterial communities are known to ... [more ▼]

Bacteria associated with algae are underexplored despite their huge biodiversity and the fact that they differ markedly from those living freely in seawater. These bacterial communities are known to represent great potential for the production of diverse bioactive compounds, such as specific glycoside hydrolases, as they interact in multiple complex ways with their host. Furthermore, enzymes from marine bacteria have original properties, like cold-adapted, halotolerant and highly stable, which are constantly searched out by bio-industries. The aim of our study was to identify bacteria, associated with the brown alga Ascophyllum nodosum, showing diverse polysaccharolytic activities. To isolate cultivable microorganisms, algal thalli of Ascophyllum nodosum were swabbed with sterile cotton tips and marine agar plates were inoculated. Three-hundred isolated bacteria were screened for agarase, kappa- and iota-carrageenase, and sulfatase activities on specific marine media. Thirty-two bacteria with polysaccharolytic activities were isolated and a part of their 16S rDNA (8F-1492R) were amplified and sequenced. Twenty-seven were classified as Flavobacteriia and five as Gammaproteobacteria. Putative new strains and species of Zobellia, Maribacter, Cellulophaga, Shewanella, Glaciecola, Pseudoalteromonas and Colwellia were identified by phylogenetic analysis. All those genera are well-known to colonize algal surface but only some of them are famous to degrade algal polysaccharides (Zobellia, Maribacter, Cellulophaga, and Pseudoalteromonas). However, all those novel bacterial strains/species showed multiple and diverse enzymatic activities (agarase, iota-and kappa-carrageenase, cellulase, beta-glucosidase, sulfatase and/or amylase activities). Genomics libraries with their DNA were constructed in Escherichia coli and Bacillus subtilis and are screened to identify the genes coding for the observed enzymatic activities. Those novel glycoside hydrolases from unknown marine bacteria should have original and innovative properties with great biotechnological potential. [less ▲]

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See detailImpacts of Plant Growth-Promoting Rhizobacteria on Wheat Growth under Greenhouse and Field Conditions
Nguyen, Minh ULg; du Jardin, Patrick ULg; Jijakli, Haissam ULg et al

Poster (2015, June 16)

Plant Growth-Promoting Rhizobacteria (PGPR) are well-known on stimulating root growth, enhancing mineral availability, and nutrient use efficiency in crops, and therefore become promising tool for ... [more ▼]

Plant Growth-Promoting Rhizobacteria (PGPR) are well-known on stimulating root growth, enhancing mineral availability, and nutrient use efficiency in crops, and therefore become promising tool for sustainable agriculture. The aim of this project is to screen PGPR strains to enhance wheat growth and yield in combination with an optimised nitrogen (N) fertilizer dose, and thus finally reduce the use of N fertilizer with equivalent yield as the recommended N dose. A list of PGPR has been collected, including (1) Mix1 (a mix of Azospirillum sp., Azorhizobium sp., and Azoarcus sp.), (2) Mix2 (a mix of Mix1 plus with two strains phosphorus-solubilizing Bacillus sp.), (3) Bacillus amyloliquefaciens a, (4) Bacillus subtilis, and (5) Bacillus amyloliquefaciens b. The PGPR were screened in both greenhouse and field condition 2014. There was significant increase in root dry weight and in root per shoot ratio of plants inoculated with Mix1 in the greenhouse. Under field condition, besides the first factor PGPR, an additional factor, i.e. four N fertilizer doses, was applied in the combination with PGPR. Without or at low N fertilizer doses, the results showed that the grain yield declined significantly. The highest grain yield increase was fifteen per cent above the control and achieved by inoculating Bacillus subtilis without application of N fertilizer. However, there was statistically insignificant in all treatments due to variability between plot replicates. Based on these results, a modified protocol plus new strategies for PGPR selection has been built up for 2015 trial to reduce the influence of variability on field and possibly achieve the higher yield increase. [less ▲]

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See detailStudying the great potential of cultivable bacteria associated with the brown alga Ascophyllum nodosum
Martin, Marjolaine ULg; Martin, Renée ULg; Barbeyron, Tristan et al

Poster (2015, June 09)

Bacteria associated with algae differ markedly from those living freely in seawater and represent great potential for the production of diverse bioactive compounds as they interact in multiple complex ... [more ▼]

Bacteria associated with algae differ markedly from those living freely in seawater and represent great potential for the production of diverse bioactive compounds as they interact in multiple complex ways with their host. Here we identified new bacterial species, and their polysaccharolytic activities, associated with the brown alga Ascophyllum nodosum. To isolate cultivable microorganisms, algal thalli of Ascophyllum nodosum were swabbed with sterile cotton tips and marine agar plates were inoculated. Three-hundred isolated bacteria were screened for agarase, kappa- or iota-carrageenase activities on specific marine media. Thirty-two bacteria with polysaccharolytic activities were isolated and a part of their 16S rDNA (8F-1492R) were amplified and sequenced. Twenty-seven were classified as Flavobacteriia and five as Gammaproteobacteria. Putative new strains and species of Zobellia, Maribacter, Cellulophaga, Shewanella, Glaciecola, Pseudoalteromonas and Colwellia were identified by phylogenetic analysis. Genomics libraries with their DNA were constructed in Escherichia coli and Bacillus subtilis and are currently screened for diverse enzymatic activities (agarases, iota-and kappa-carrageenases, cellulases, beta-glucosidases, sulfatases and amylases). In an era where high throughput sequencing is mostly used to study bacterial communities, cultivation methods are underestimated. Here, we revealed that only ten percent of the cultivable bacteria on this brown alga could degrade algal polysaccharides, which lead to asking us; who and what are the 90 other percents doing there? Furthermore, by this cultivation method we could also identify putative new bacterial strains/species, which are screened for polysaccharidases. Novel glycoside hydrolases from unknown marine bacteria represent great biotechnological potential as they should have original industrial properties. [less ▲]

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See detailMultiple analyses of microbial communities applied to the gut of the wood-feeding termite Reticulitermes flavipes fed on artificial diets
Tarayre, Cédric ULg; Bauwens, Julien ULg; Mattéotti, Christel et al

in Symbiosis (2015)

The purpose of this work was the observation of the differences between the microbial communities living in the gut of the termite Reticulitermes flavipes fed on different diets. The termites were fed on ... [more ▼]

The purpose of this work was the observation of the differences between the microbial communities living in the gut of the termite Reticulitermes flavipes fed on different diets. The termites were fed on poplar wood (original diet) and artificial diets consisting of crystalline cellulose (with and without lignin), α-cellulose (with and without lignin) and xylan. The termites were then dissected and the protist communities were analyzed through microscopy, leading to the conclusion that protist species are strongly influenced by diets. BIOLOG ECO Microplates® were used to assess the metabolic properties of the different types of consortia, highlighting strong differences on the basis of principal component analysis and calculation of similarity rates. The microorganisms were cultivated in liquid media corresponding to the artificial diets before being characterized through a metagenetic analysis of gut microbiota (16S ribosomal DNA). This analysis identified several phyla: Acidobacteria, Actinobacteria, Bacteroidetes, Cyanobacteria, Fibrobacteres, Firmicutes, Nitrospirae, OP9, Planctomycetes, Proteobacteria, Spirochaetes, TM6, Tenericutes, Verrucomicrobia and WS3. The OTUs were also determined and confirmed the abundance of Proteobacteria, Bacteroidetes, Firmicutes and Verrucomicrobia. It was possible to isolate several strains from the liquid media, and one bacterium and several fungi were found to produce interesting enzymatic activities. The bacterium Chryseobacterium sp. XAvLW produced α-amylase, β-glucosidase, endo-1,4-β-D-glucanase, endo-1,4-β-D-xylanase and filter paper-cellulase, while the fungi Sarocladium kiliense CTGxxyl and Trichoderma virens CTGxAviL generated the same activities added with endo-1,3-β-D-glucanase. [less ▲]

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See detailSoil microbial community composition changes according to the tillage practice and plant development stage
Degrune, Florine ULg; Dufrêne, Marc ULg; Taminiau, Bernard ULg et al

Poster (2015, April)

Agricultural practices have a strong impact on soil bacterial and fungal community composition. Furthermore, microbial community composition can change with the stage of plant development. We are ... [more ▼]

Agricultural practices have a strong impact on soil bacterial and fungal community composition. Furthermore, microbial community composition can change with the stage of plant development. We are interested in exploring these effects in relation to changes induced by agriculture (conventional and reduced tillage) and plant stage (germination and flowering) in soil conditions. Here, instead of examining this impact at a high taxonomic level such as phylum and/or class, thus missing potentially relevant information from lower levels, we propose an original method: exploiting the available sequence information at the lowest taxonomic level attainable for each operational taxonomic unit. Results show that some microbial communities were impacted only by the tillage practice , while others were impacted only by the stage of plant. Changes in microbial community composition could be due to the soil conditions induced by the soil practice and the stage of plant. [less ▲]

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See detailA novel sub-phylum method discriminates better the impact of crop management on soil microbial community
Degrune, Florine ULg; Dufrêne, Marc ULg; Colinet, Gilles ULg et al

in Agronomy for Sustainable Development (2015)

Soil microorganisms such as mycorrhizae and plant-growth-promoting rhizobacteria have beneficial effects on crop productivity. Agricultural practices are known to impact soil microbial communities, but ... [more ▼]

Soil microorganisms such as mycorrhizae and plant-growth-promoting rhizobacteria have beneficial effects on crop productivity. Agricultural practices are known to impact soil microbial communities, but past studies examining this impact have focused mostly on one or two taxonomic levels, such as phylum and class, thus missing potentially relevant information from lower levels. Therefore we propose here an original, sub-phylum method for studying how agricultural practices modify microbial communities. This method involves exploiting the available sequence information at the lowest taxonomic level attainable for each operational taxonomic unit. In order to validate this novel method we assessed microbial community composition using 454 pyrosequencing of 16S and 28S rRNA genes, then we compared the results with results of a phylum-level analysis. Agricultural practices included conventional tillage, reduced tillage, residue removal and residue retention. Results show that, at the lowest taxonomic level attainable, tillage is the main factor influencing both bacterial community composition, accounting for 13% of the variation, and fungal community composition, accounting for 18% of the variation. Whereas phylum-level analysis failed to reveal any effect of soil practice on bacterial community composition, and missed the fact that different members of the same phylum responded differently to tillage practice. For instance, the fungal phylum Chytridiomycota showed no impact of soil treatment, while sub-phylum-level analysis revealed an impact of tillage practice on the Chytridiomycota sub-groups Gibberella, which includes a notorious wheat pathogen, and Trichocomaceae. This clearly demonstrates the necessity of exploiting the information obtainable at sub-phylum level when assessing the effects of agricultural practice on microbial communities. [less ▲]

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See detailNew lipolitic enzymes identified by screening two metagenomic libraries derived from the soil of a winter wheat field
Stroobants, Aurore ULg; Martin, Renée ULg; Roosens, Lyse et al

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (2015), 19(2), 125-131

Lipolytic enzymes are widely distributed and fulfil important physiological functions in the microorganisms inhabiting diverse environments. Soils are rich, diversified environments containing microbial ... [more ▼]

Lipolytic enzymes are widely distributed and fulfil important physiological functions in the microorganisms inhabiting diverse environments. Soils are rich, diversified environments containing microbial communities that remain largely unknown. [less ▲]

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See detailDetecting microbial patterns in relation to soil agricultural practices and the plant development stage
Degrune, Florine ULg; Dufrêne, Marc ULg; Taminiau, Bernard ULg et al

Poster (2014, December 02)

Agricultural practices have a strong impact on soil bacteria and fungi. Furthermore, microbial community composition can change with the stage of plant development. We are interested in exploring these ... [more ▼]

Agricultural practices have a strong impact on soil bacteria and fungi. Furthermore, microbial community composition can change with the stage of plant development. We are interested in exploring these effects in relation to changes induced by agriculture and plant stage in soil conditions. Some bacteria are influenced only by the plant stage, which induces changes in soil humidity, pH, nitrates, and carbon. We would thus expect these bacteria to be highly sensitive to these parameters. Other bacteria are affected only by the tillage practice applied. Further study is needed to identify the soil parameters responsible for this effect. The plant stage also has a great impact on fungal community composition. [less ▲]

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See detailWood digestion in lower termites: multidisciplinary approaches based on differential feeding
Bauwens, Julien ULg; Brasseur, Catherine ULg; Tarayre, Cédric ULg et al

Poster (2014, December)

Termites digestive tract and hindgut especially still holds many secrets despites hundreds of years of research. The complexity of the symbiotic microbial community and the contrast of physio-chemical ... [more ▼]

Termites digestive tract and hindgut especially still holds many secrets despites hundreds of years of research. The complexity of the symbiotic microbial community and the contrast of physio-chemical environments found in lower termites paunch are potentially the key point to explain the efficiency of ligno-cellulose digestion. Contribution of advancing technologies accelerates the progress of our knowledge in this field. Here, we present multiple approaches combining old and recent techniques used to highlight the effect of ligno-cellulosic compounds on termite gut and the role of populations from the symbiotic microbial community. Termites Reticulitermes flavipes (Kollar) submitted to various artificial diets showed variations in flagellates populations profile and enzymatic activities. Differential protein expression was investigated using 2D-DIGE MALDI-TOF-TOF and 2D-LC-MS/MS using high resolution orbitrap analyzer. Results from both proteomic experiments tend to support each-other and bring complementary points of view. The gel-free analysis resulted in highly contrasted identification of enzymes involved in ligno-cellulose digestion and metabolism. Finally, differential feeding experiments leaded to in vivo selection of different symbiotic communities. These communities were characterized following some metabolism assays and allowed the cultivation of diverse microbial consortia using media closely related to the respective artificial diets. This work provides relevant data on termite and associated microbial community response to alimentary diets. [less ▲]

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See detailIsolation and cultivation of xylanolytic and cellulolytic Sarocladium kiliense and Trichoderma virens from the gut of the termite Reticulitermes santonensis
Tarayre, Cédric ULg; Bauwens, Julien ULg; Brasseur, Catherine ULg et al

in Environmental Science and Pollution Research (2014)

The purpose of this work was the isolation and cultivation of cellulolytic and xylanolytic microorganisms extracted from the gut of the lower termite Reticulitermes santonensis. Microcrystalline cellulose ... [more ▼]

The purpose of this work was the isolation and cultivation of cellulolytic and xylanolytic microorganisms extracted from the gut of the lower termite Reticulitermes santonensis. Microcrystalline cellulose (with and without lignin) and beech wood xylan were used as diets instead of poplar wood in order to select cellulose and hemicellulose-degrading fungi. The strain Sarocladium kiliense (Acremonium kiliense) CTGxxyl was isolated from the termites fed on xylan, while the strain Trichoderma virens CTGxAviL was isolated from the termites fed on cellulose (with and without lignin). Both molds were cultivated in liquid media containing different substrates: agro-residues or purified polymers. S. kiliense produced maximal β-glucosidase, endo-1,4-β-D-glucanase, exo-1,4-β-D-glucanase and endo-1,4-β-D-xylanase activities of 0.103, 3.99, 0.53, and 40.8 IU/ml, respectively. T. virens produced maximal β-xylosidase, endo-1,4-β-D-glucanase, exo-1,4-β-D-glucanase, and endo-1,4-β-D-xylanase activities of 0.38, 1.48, 0.69, and 426 IU/ml. The cellulase and the xylanase of S. kiliense, less common than T. virens, were further investigated. The optimal activity of the xylanase was observed at pH 9–10 at 60 °C. The cellulase showed its maximal activity at pH 10, 70 °C. Zymography identified different xylanases produced by both molds, and some fragment sizes were highlighted: 35, 100, and 170 kDa for S. kiliense and 20, 40, 80, and 170 kDa for T. virens. In both cases, endo-1,4-β-D-xylanase activitieswere confirmed through mass spectrometry. [less ▲]

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See detailImpacts of biostimulant products on the growth of wheat and the microbial communities of its rhizosphere under contrasted production systems
Nguyen, Minh ULg; Bodson, Bernard ULg; Colinet, Gilles ULg et al

Poster (2014, August 24)

Plant growth-promoting rhizobacteria (PGPR) are one of the major biostimulant classes due to their ability to stimulate root growth, enhance mineral availability, and nutrient use efficiency in crops ... [more ▼]

Plant growth-promoting rhizobacteria (PGPR) are one of the major biostimulant classes due to their ability to stimulate root growth, enhance mineral availability, and nutrient use efficiency in crops. PGPR-containing biostimulant products could therefore make agriculture more sustainable by reducing demand for chemical fertilizer and lessen their negative environmental impacts. The aim of this project is to screen PGPR strains to (1) enhance wheat fitness level (growth, photosynthesis efficiency, stress tolerance, and yield) in combination with an optimised fertilizer level, (2) stimulate the increase in beneficial microorganism communities and suppress pathogenic ones in the wheat rhizosphere, (3) link wheat productivity to the composition of the microbial communities found in its rhizosphere, and (4) measure the impacts of such changes on soil fertility. A list of PGPR-containing biostimulants have been collected from screening, including several commercially available products (e.g. TwinN and NitroGuard, Mabiotec; Rhizocell GC, Ithec; B. subtilis FZB24 fl and Rhizo Vital 42, Abitep) as well as newly discovered PGPR strains. The biostimulants from that list have been screened in greenhouse and we expect to obtain results within next month. In parallel, several levels of nitrogen supply have been tested in combination with biostimulants to optimize agricultural practices and achieve the highest yield on field condition. A soil analysis protocols will also be built up to measure the influence of those PGPR strains on soil fertility changes and root uptake efficiency. In order to assess changes in the rhizomicrobial communities including fungi and bacteria (either pathogenic, neutral, or beneficial) under controlled or field conditions, metagenomic approaches will be set up. Finally, a maximum of three promising PGPR strains will be selected for practical agronomical application in larger field trials. [less ▲]

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See detailMultidisciplinary approaches and fractionations to study lower termite symbiotic system and ligno-cellulose digestion
Bauwens, Julien ULg; Brasseur, Catherine ULg; Tarayre, Cédric ULg et al

Poster (2014, August)

Wood-feeding termites are a considerable source of enzymes active on ligno-cellulosic compounds. These enzymes are produced by the termite host and some representatives of its symbiotic microbial ... [more ▼]

Wood-feeding termites are a considerable source of enzymes active on ligno-cellulosic compounds. These enzymes are produced by the termite host and some representatives of its symbiotic microbial community, and are of particular interest in regard second generation biofuel. However, the complexity of microbial interactions renders micro-organisms isolation very difficult. Culture-independent methods permitted to gather a large amount of data and to understand a little more the role of each microbial population, particularly the prokaryotes. Proteomics allows working on the final product of gene expression, and corresponds more to the real operation of the digestive system. In order to investigate such a complex system, it is necessary to use multidisciplinary approaches and to fractionate this system. Zymography or affinity chromatography are used in parallel of routine proteomics techniques such as two-dimensional gel electrophoresis associated to MALDI-TOF mass spectrometry and nano-LC ESI-MS/MS. We used an artificial-diet based rearing to induce changes in microbial population balance. We performed preliminary assay to investigate the glycosylated proteome in the hindgut of a lower termite, using Multi-Lectin Affinity Chromatography (M-LAC) and enzymatic activity of harvested fractions was assessed on cellulosic substrates. [less ▲]

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See detailIdentification and Characterization of a Halotolerant, Cold-Active Marine Endo-β-1,4-Glucanase by Using Functional Metagenomics of Seaweed-Associated Microbiota
Martin, Marjolaine ULg; Biver, Sophie ULg; Steels, Sébastien ULg et al

in Applied and Environmental Microbiology (2014), 80(16), 4958-4967

A metagenomic library was constructed from microorganisms associated with the brown alga Ascophyllum nodosum. Functional screening of this library revealed 13 novel putative esterase loci and two ... [more ▼]

A metagenomic library was constructed from microorganisms associated with the brown alga Ascophyllum nodosum. Functional screening of this library revealed 13 novel putative esterase loci and two glycoside hydrolase loci. Sequence and gene cluster analysis showed the wide diversity of the identified enzymes and gave an idea of the microbial populations present during the sample collection period. Lastly, an endo-β-1,4-glucanase having less than 50% identity to sequences of known cellulases was purified and partially characterized, showing activity at low temperature and after prolonged incubation in concentrated salt solutions. [less ▲]

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