Project description:Illumina HiSeq technology was used to generate mRNA profiles from Paxillus involutus ectomycorrhizal roots compared to mycelium patches . Mycorrhizal roots were harvested after 4 weeks, pooled and used for RNA extraction. Reads of 2X100bp were generated and aligned to Paxillus involutus (http://genome.jgi-psf.org/Paxin1/Paxin1.home.html) using CLC Genomics Workbench 6.
Project description:Illumina HiSeq technology was used to generate mRNA profiles from Paxillus involutus ectomycorrhizal roots compared to mycelium patches . Mycorrhizal roots were harvested after 4 weeks, pooled and used for RNA extraction. Reads of 2X100bp were generated and aligned to Paxillus involutus (http://genome.jgi-psf.org/Paxin1/Paxin1.home.html) using CLC Genomics Workbench 6. mRNA profiles from Paxillus involutus ectomycorrhizal roots and mycelium patches were generated by paired-end (2x100bp) Illumina HiSeq2000 sequencing. Two biological replicates were sequenced for mycorrhizal and mycelium samples.
Project description:Variations in gene content and sequence that could be associated with symbiotic adaptations of the ectomycorrhizal fungus Paxillus involutus were investigated by analyses of strains showing various abilities to form mycorrhiza. Five strains of Paxillus involutus (ATCC 200175, Maj; Nau, Pi01SE, and Pi08BE) and one strain of Paxillus filamentosus (Pf01De) were analyzed by comparative genomic hybridizations using cDNA microarrays. Two batches of arrays were used containing 1,076 unique fungal reporters. DNA was prepared from each strain, and after fragmentation and labelling used for dual-label microarray hybridizations. The experimental design includes 16 arrays (CGH_01 -- CGH_16), of which 12 arrays represent dye-swapped and direct contrasts between the sample strains and the reference strain ATCC 200175. Two arrays represent dye-swapped self-self hybridizations of the reference strain ATCC 200175 (CGH_01 and CGH_02). The remaining two arrays represent dye-swapped and direct contrasts between the sample strains Maj and Nau (CGH_06 and CGH_07).
Project description:Transcriptional analyses of the ectomycorrhizal (ECM) fungus Paxillus involutus during growth on various lignocellulose soil substrates
Project description:Purpose: Fungi actively weather silicate minerals to extract nutrients, using different mechanisms. However, it is not clear whether the same mechanism(s) is used for all minerals. The question addressed here is whether distinct minerals can stimulate different weathering mechanisms and lead to a shift in fungal genes expression. Methods: Microcosm experiments were set up with one fungus and three minerals containing the same targeted metal nutrient (K), but with a variable degree of resistance to dissolution. The experiment used transcriptomics to compare the active gene expression of the ectomycorrhizal basidiomycetes Paxillus involutus fungus in response to developing on each of the three minerals as sole K sources. The selected minerals, with a decreasing resistance to K extraction, were: muscovite, phlogopite, and K-exchanged vermiculite. Results: Gene expression analysis after the experiments indicated that the level of K-deprivation stress was muscovite > phlogopite >> K-vermiculite. Genes and functions overexpressed in the experiments indicated meaningful metabolic activities including K extraction and transport. Conclusions: The fungus Paxillus involutus activated different metabolic pathways and, accordingly, switched on or off different genes depending on the minerals from which it was forced to obtain potassium, a fundamental nutrient for its growth. The differential expression of the fungal genes generated alternative chemical attacks on the minerals, resulting in a tailored kind of dissolution and selective uptake of chemical elements. When exposed to K-vermiculite, the fungus overexpressed gene sets related to the organisation of the cytoskeleton and also to vesicle-mediated transport. If phlogopite was supplied, the fungus overexpressed gene sets involved with organic acid and oxoacid metabolic processes. When the only source of potassium was muscovite, the fungus overexpressed monooxygenase activity and oxidoreductase activity. These changing strategies, which resulted in a different impact on the fungal uptake of the elements, always granted some potassium to the fungus.