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Écosystèmes forestiers : diversité et fonction des champignons

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Abstract

Les champignons, par leur abondance et la diversité de formes, sont une composante importante du paysage de nos sous-bois en automne. Mais sait-on que pour la moitié d'entre elles, ces espèces vivent en symbiose avec les racines et qu'elles sont indispensables à la nutrition des arbres ? Chercheurs et forestiers s'intéressent de plus en plus à ce réseau trophique souterrain, à sa diversité fonctionnelle et à sa réponse aux perturbations auxquelles les forêts sont soumises.

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... We designed a total of 95 oligonucleotides, from which 89 were species-specific for ECM fungal species. According to regular fruiting body surveys, these 89 ECM species are the most common species to be found in the long-term observatory of the Breuil-Chenue forest over the last ten years [32]. The ease with which high-quality species-specific oligonucleotides could be selected (mismatch in the middle of the designed oligonucleotide, without forming secondary structures), depended on the fungal genera. ...
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In forest ecosystems, communities of ectomycorrhizal fungi (ECM) are influenced by several biotic and abiotic factors. To understand their underlying dynamics, ECM communities have been surveyed with ribosomal DNA-based sequencing methods. However, most identification methods are both time-consuming and limited by the number of samples that can be treated in a realistic time frame. As a result of ongoing implementation, the array technique has gained throughput capacity in terms of the number of samples and the capacity for parallel identification of several species. Thus far, although phylochips (microarrays that are used to detect species) have been mostly developed to trace bacterial communities or groups of specific fungi, no phylochip has been developed to carry oligonucleotides for several ectomycorrhizal species that belong to different genera. We have constructed a custom ribosomal DNA phylochip to identify ECM fungi. Specific oligonucleotide probes were targeted to the nuclear internal transcribed spacer (ITS) regions from 95 fungal species belonging to 21 ECM fungal genera. The phylochip was first validated using PCR amplicons of reference species. Ninety-nine percent of the tested oligonucleotides generated positive hybridisation signals with their corresponding amplicons. Cross-hybridisation was mainly restricted at the genus level, particularly for Cortinarius and Lactarius species. The phylochip was subsequently tested with environmental samples that were composed of ECM fungal DNA from spruce and beech plantation fungal communities. The results were in concordance with the ITS sequencing of morphotypes and the ITS clone library sequencing results that were obtained using the same PCR products. For the first time, we developed a custom phylochip that is specific for several ectomycorrhizal fungi. To overcome cross-hybridisation problems, specific filter and evaluation strategies that used spot signal intensity were applied. Evaluation of the phylochip by hybridising environmental samples confirmed the possible application of this technology for detecting and monitoring ectomycorrhizal fungi at specific sites in a routine and reproducible manner.
... La moitié des espèces sont des champignons ectomycorhiziens (ECM) et l'autre moitié comprend des champignons saprophytes vivant sur branches et tronc morts (SBM), ou s ur arbre vivant (SBV) ou sur brindilles (SB), des saprophytes décomposeurs de litière (SDL) et d'autres saprophytes (AS). D'aprèsBuée et al. (2006). ...
Thesis
Les sols forestiers sont des habitats hétérogènes, véritables réservoirs de microorganismes. Parmi ces microorganismes, les eucaryotes filamenteux (champignons et oomycètes) sont très divers et jouent un rôle important dans le fonctionnement et la durabilité des écosystèmes forestiers. Leur diversité et leur répartition spatiale à différentes échelles sont encore peu connues et les facteurs qui sous-tendent cette dispersion sont encore peu étudiés. Aussi, les objectifs étaient (i) d'exploiter le séquençage haut-débit pour des études d'écologie microbienne à large échelle et valider son application aux communautés d'oomycètes pathogènes en milieu forestier, (ii) de décrire ces communautés microbiennes, en termes de diversité et de structure, à différentes échelles spatiales (locale, régionale et continentale), (iii) de caractériser les variables biotiques et abiotiques structurant ces communautés et (iv) d'évaluer la réponse éventuelle des communautés aux variations climatiques. Une première étude pilote à l'échelle de la parcelle a été suivi de deux études à grande échelle spatiale le long de gradients environnementaux. Des gradients d'altitude et un gradient latitudinal, à l'échelle continentale, ont été utilisés comme gradient climatique. L'étude préliminaire a donc validé l'utilisation du pyroséquençage pour les communautés fongiques, et en particulier pour les espèces ectomycorhiziennes, et apporté des éléments pour établir une méthodologie d'échantillonnage couplée à cette technique. L'application de ces outils moléculaires à l'étude des communautés oomycètes pathogènes reste à optimiser. Les résultats obtenus sur les communautés fongiques telluriques suggèrent que dans l'hypothèse d'un réchauffement climatique, la richesse fongique ne serait pas directement affectée mais la composition des communautés le serait. La composition des communautés fongiques est également fortement liée au pH du sol. Ces résultats sont à affiner en étudiant plus en détail divers groupes taxonomiques et écologiques en lien avec des variables climatiques plus précises. Par ailleurs, de nombreuses perspectives sont envisageables pour améliorer la détection des oomycètes dans les sols forestiers, qui reste un challenge en écologie microbienne
... Moreover, its mycorrhizae can be unambiguously identified in situ because of its color, morphology, and typical smell. In the experimental forest site, an inventory of the epigeous sporocarps, performed three times each autumn during 5 years, revealed 138 ectomycorrhizal species [6]. Among them, S. citrinum represented the ninth most abundant. ...
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The ectomycorrhizal symbiosis alters the physicochemical and biological conditions in the surrounding soil, thus creating a particular environment called ectomycorrhizosphere, which selects microbial communities suspected to play a role in gross production and nutrient cycling. To assess the ectomycorrhizosphere effect on the structure of microbial communities potentially involved in the mobilization of nutrients from the soil minerals in a poor-nutrient environment, we compared the functional diversity of soil and ectomycorrhizosphere bacterial communities in a forest stand. Two hundred and sixty-four bacterial strains and 107 fungal strains were isolated from the bulk soil of an oak (Quercus petraea) stand and from oak-Scleroderma citrinum ectomycorrhizosphere and ectomycorrhizae, in two soil organo-mineral horizons (0 to 3 cm and 5 to 10 cm). They were characterized using two in vitro tests related to their capacities to mobilize iron and phosphorus. We demonstrated that the oak-S. citrinum ectomycorrhizosphere significantly structures the culturable bacterial communities in the two soil horizons by selecting very efficient strains for phosphorus and iron mobilization. This effect was also observed on the diversity of the phosphate-solubilizing fungal communities in the lower soil horizon. A previous study already demonstrated that Laccaria bicolor-Douglas fir ectomycorrhizosphere structures the functional diversity of Pseudomonas fluorescens population in a forest nursery soil. Comparing to it, our work highlights the consistency of the mycorrhizosphere effect on the functional diversity of bacterial and fungal communities in relation to the mineral weathering process, no matter the fungal symbiont, the age and species of the host tree, or the environment (nursery vs forest). We also demonstrated that the intensity of phosphorus and iron mobilization by the ectomycorrhizosphere bacteria isolated from the lower soil horizon was significantly higher compared to that which was isolated from the upper horizon. This reveals for the first time a stratification of the functional diversity of the culturable soil bacterial communities as related to phosphorus and iron mobilization.
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