A new method for qualitative multi-scale analysis of bacterial biofilms on filamentous fungal colonies using confocal and electron microscopy

Bacterial biofilms frequently form on fungal surfaces and can be involved in numerous bacterial-fungal interaction processes, such as metabolic cooperation, competition, or predation. The study of biofilms is important in many biological fields, including environmental science, food production, and medicine. However, few studies have focused on such bacterial biofilms, partially due to the difficulty of investigating them. Most of the methods for qualitative and quantitative biofilm analyses described in the literature are only suitable for biofilms forming on abiotic surfaces or on homogeneous and thin biotic surfaces, such as a monolayer of epithelial cells.While laser scanning confocal microscopy (LSCM) is often used to analyze in situ and in vivo biofilms, this technology becomes very challenging when applied to bacterial biofilms on fungal hyphae, due to the thickness and the three dimensions of the hyphal networks. To overcome this shortcoming, we developed a protocol combining microscopy with a method to limit the accumulation of hyphal layers in fungal colonies. Using this method, we were able to investigate the development of bacterial biofilms on fungal hyphae at multiple scales using both LSCM and scanning electron microscopy (SEM). This report describes the protocol, including microorganism cultures, bacterial biofilm formation conditions, biofilm staining, and LSCM and SEM visualizations

Fungal Endophytes of Populus trichocarpa Alter Host Phenotype, Gene Expression, and Rhizobiome Composition.

Mortierella and Ilyonectria genera include common species of soil fungi that are frequently detected as root endophytes in many plants, including Populus spp. However, the ecological roles of these and other endophytic fungi with respect to plant growth and function are still not well understood. The functional ecology of two key taxa from the P. trichocarpa rhizobiome, M. elongata PMI93 and I. europaea PMI82, was studied by coupling forest soil bioassays with environmental metatranscriptomics. Using soil bioassay experiments amended with fungal inoculants, M. elongata was observed to promote the growth of P. trichocarpa. This response was cultivar independent. In contrast, I. europaea had no visible effect on P. trichocarpa growth. Metatranscriptomic studies revealed that these fungi impacted rhizophytic and endophytic activities in P. trichocarpa and induced shifts in soil and root microbial communities. Differential expression of core genes in P. trichocarpa roots was observed in response to both fungal species. Expression of P. trichocarpa genes for lipid signaling and nutrient uptake were upregulated, and expression of genes associated with gibberellin signaling were altered in plants inoculated with M. elongata, but not I. europaea. Upregulation of genes for growth promotion, downregulation of genes for several leucine-rich repeat receptor kinases, and alteration of expression of genes associated with plant defense responses (e.g., jasmonic acid, salicylic acid, and ethylene signal pathways) also suggest that M. elongata manipulates plant defenses while promoting plant growth

Genome-wide analysis of lectin receptor-like kinases in Populus

Transcript level of C-type PtLecRLK gene in 24 different datasets from the Populus Gene Atlas Study. RNA-seq data were collected from the Populus Gene Atlas Study in Phytozome v11.0 ( http://phytozome.jgi.doe.gov/pz/portal.html ). The transcript level was expressed as FPKM. The sheet labeled as “whole_set” contains the original FPKM values from Gene Atlas. The data of four different tissues under standard condition are sorted in the data sheet labeled as “standard”. (XLSX 10 kb

The obscure events contributing to the evolution of an incipient sex chromosome in Populus: a retrospective working hypothesis

Genetic determination of gender is a fundamental developmental and evolutionary process in plants. Although it appears that dioecy in [i]Populus[/i] is genetically controlled, the precise gender-determining systems remain unclear. The recently released second draft assembly and annotated gene set of the [i]Populus[/i] genome provided an opportunity to revisit this topic. We hypothesized that over evolutionary time, selective pressure has reformed the genome structure and gene composition in the peritelomeric region of the chromosome XIX, which has resulted in a distinctive genome structure and cluster of genes contributing to gender determination in [i]Populus trichocarpa[/i]. Multiple lines of evidence support this working hypothesis. First, the peritelomeric region of the chromosome XIX contains significantly fewer single nucleotide polymorphisms than the rest of [i]Populus[/i] genome and has a distinct evolutionary history. Second, the peritelomeric end of chromosome XIX contains the largest cluster of the nucleotide-binding site–leucine-rich repeat (NBS–LRR) class of disease resistance genes in the entire [i]Populus[/i] genome. Third, there is a high occurrence of small microRNAs on chromosome XIX, which is coincident to the region containing the putative gender-determining locus and the major cluster of NBS–LRR genes. Further, by analyzing the metabolomic profiles of floral bud in male and female [i]Populus[/i] trees using a gas chromatography-mass spectrometry, we found that there are gender-specific accumulations of phenolic glycosides. Taken together, these findings led to the hypothesis that resistance to and regulation of a floral pathogen and gender determination coevolved, and that these events triggered the emergence of a nascent sex chromosome. Further studies of chromosome XIX will provide new insights into the genetic control of gender determination in [i]Populus[/i]

Contribution to the study of the structure and the polymorphism of the genome of the ectomycorrhizal basidiomycete Laccaria bicolor (Maire) Orton and identification of QTLs of mycorrhization in two Poplars, Populus trichocarpa Torr. & A. Gray ex Hook. and Populus deltoides (Bartr.) Marsh

Les symbioses mycorhiziennes, entre champignons et racines de plantes, concernent 95 % des espèces végétales. Les arbres sociaux des forêts boréales et tempérées forment avec les champignons un type particulier d’association : la symbiose ectomycorrhizienne. Les ectomycorrhizes jouent un rôle essentiel dans la nutrition hydrominérale des arbres, le cycle des éléments minéraux et la production primaire. Cependant, leur complexité n’a pas permis à ce jour de déchiffrer leurs rôles et leurs fonctions précises. La récente disponibilité du génome du champignon ectomycorhizien Laccaria bicolor et de celui de l'arbre hôte Populus trichocarpa fournit une occasion inégalée d’approfondir nos connaissances du développement et du fonctionnement de cette symbiose. Les objectifs de cette étude ont donc été de participer à la caractérisation et au décryptage du génome de L. bicolor puis à la recherche des gènes impliqués dans la formation des ectomycorhizes chez les deux partenaires. Dans un premier temps et afin d’aider à l’assemblage de la séquence génomique de L. bicolor, nous avons identifié les séquences répétées et construit une carte génétique. Sur les 60 Mb de ce génome, nous avons mis en évidence 8 % de séquences microsatellitaires et 24 % d’éléments transposables. Une carte génétique a été construite à partir de 111 monocaryons issus de L. bicolor S238N. Cette carte comprend 326 marqueurs (8 RAPD, 243 AFLP, 59 SSR et 14 SNP) répartis sur 10 groupes de liaison ancrés à la séquence génomique de L. bicolor. Dans un second temps, nous avons tenté d’identifier les gènes impliqués dans l’établissement des ectomycorhizes chez le peuplier en combinant une approche de détection par QTLs et par puces à ADN. Nous avons ciblé 81 gènes potentiellement impliqués dans l’établissement et/ou le fonctionnement de la symbiose.The mycorrhizal symbioses between fungi and roots concern 95 % of the plant species. Social trees of boreal and temperate forests form a particular type of root association with fungi: the ectomycorrhizal symbiosis. Ectomycorrhizas play a major role in tree hydromineral nutrition, nutrient cycles and primary production. However, their complexity have so far prevented from deciphering their precise function and role. The recent availability of the genome of the ectomycorrhizal fungus Laccaria bicolor and that of the host-tree Populus trichocarpa provides an unprecedented opportunity to decipher the key components of development and functioning of this symbiosis. The aims of this study were to participate to the characterization and deciphering of the genome of L. bicolor, and to determine the genes involved in the formation of ectomycorrhizas in both partners. Firstly, in order to facilitate the assembly of the genomic sequence of L. bicolor, we have identified the repeated sequences and generated a genetic map. On the 60 Mb of this genome, 8 % are microsatellite sequences and 24 % transposable elements. A genetic map was built from 111 monokaryons issued from L. bicolor S238N. This map includes 326 markers (8 RAPD, 243 AFLP, 59 SSR and 14 SNP) distributed on 10 linkage groups anchored onto the genomic sequence of L. bicolor. Secondly, we have identified the genes involved in the establishment of ectomycorrhizas in poplar by combining QTL detection and DNA microarrays. We targeted 81 genes which can be involved in the establishment and/or the functioning of the symbiosis

Contribution à l'étude de la structure et du polymorphisme du génome du basidiomycète ectomycorhizien "Laccaria bicolor" (Maire) Orton et identification de QTLs de mycorhization chez les peupliers, "Populus trichocarpa Torr. & A. Gray ex Hook. et "Populus deltoides (Bartr.) Marsh

The mycorrhizal symbioses between fungi and roots concern 95 % of the plant species. Social trees of boreal and temperate forests form a particular type of root association with fungi: the ectomycorrhizal symbiosis. Ectomycorrhizas play a major role in tree hydromineral nutrition, nutrient cycles and primary production. However, their complexity have so far prevented from deciphering their precise function and role. The recent availability of the genome of the ectomycorrhizal fungus Laccaria bicolor and that of the host-tree Populus trichocarpa provides an unprecedented opportunity to decipher the key components of development and functioning of this symbiosis. The aims of this study were to participate to the characterization and deciphering of the genome of L. bicolor, and to determine the genes involved in the formation of ectomycorrhizas in both partners. Firstly, in order to facilitate the assembly of the genomic sequence of L. bicolor, we have identified the repeated sequences and generated a genetic map. On the 60 Mb of this genome, 8 % are microsatellite sequences and 24 % transposable elements. A genetic map was built from 111 monokaryons issued from L. bicolor S238N. This map includes 326 markers (8 RAPD, 243 AFLP, 59 SSR and 14 SNP) distributed on 10 linkage groups anchored onto the genomic sequence of L. bicolor. Secondly, we have identified the genes involved in the establishment of ectomycorrhizas in poplar by combining QTL detection and DNA microarrays. We targeted 81 genes which can be involved in the establishment and/or the functioning of the symbiosis.Les symbioses mycorhiziennes, entre champignons et racines de plantes, concernent 95 % des espèces végétales. Les arbres sociaux des forêts boréales et tempérées forment avec les champignons un type particulier d'association : la symbiose ectomycorrhizienne. Les ectomycorrhizes jouent un rôle essentiel dans la nutrition hydrominérale des arbres, le cycle des éléments minéraux et la production primaire. Cependant, leur complexité n'a pas permis à ce jour de déchiffrer leurs rôles et leurs fonctions précises. La récente disponibilité du génome du champignon ectomycorhizien Laccaria bicolor et de celui de l'arbre hôte Populus trichocarpa fournit une occasion inégalée d'approfondir nos connaissances du développement et du fonctionnement de cette symbiose. Les objectifs de cette étude ont donc été de participer à la caractérisation et au décryptage du génome de L. bicolor puis à la recherche des gènes impliqués dans la formation des ectomycorhizes chez les deux partenaires. Dans un premier temps et afin d'aider à l'assemblage de la séquence génomique de L. bicolor, nous avons identifié les séquences répétées et construit une carte génétique. Sur les 60 Mb de ce génome, nous avons mis en évidence 8 % de séquences microsatellitaires et 24 % d'éléments transposables. Une carte génétique a été construite à partir de 111 monocaryons issus de L. bicolor S238N. Cette carte comprend 326 marqueurs (8 RAPD, 243 AFLP, 59 SSR et 14 SNP) répartis sur 10 groupes de liaison ancrés à la séquence génomique de L. bicolor. Dans un second temps, nous avons tenté d'identifier les gènes impliqués dans l'établissement des ectomycorhizes chez le peuplier en combinant une approche de détection par QTLs et par puces à ADN. Nous avons ciblé 81 gènes potentiellement impliqués dans l'établissement et/ou le fonctionnement de la symbiose

Mycorrhiza helper bacteria

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