Project description:This SuperSeries is composed of the following subset Series: GSE37942: Comparison of gene expression during meiosis between wild-type and rad50-deficient strains of Coprinopsis cinerea GSE37943: Comparison of gene expression during meiosis between wild-type and msh5-deficient strains of Coprinopsis cinerea Refer to individual Series
Project description:We have recently shown that the coprophilous model mushroom Coprinopsis cinerea transcribes a broad array of genes encoding defense proteins in the vegetative mycelium and fruiting bodies that target bacterial competitors and animal predators challenging the respective tissues of this fungus. In addition, we have demonstrated in previous work that two nematotoxic defense proteins from Coprinopsis, CGL1 and CGL2, were induced in vegetative mycelium challenged with the predatory nematode Aphelenchus avenae; however, the specificity and broadness of this response remained unclear. In order to resolve these issues, we sequenced the poly(A)-positive transcriptome of vegetative mycelium of C. cinerea confronted with nematode predation, hyphal mechanical damage or bacterial co-culture.
Project description:Coprinopsis cinerea vegetative dikaryotic mycelia compared to vegetative monokaryotic mycelia Vegetative dikaryon cDNA was comparitively hybridized with monokaryotic vegetative cDNA. Labelled cDNA was hybridized to arrays in a 2 channel reaction.
Project description:Sexual reproduction is an ancient trait that evolved shortly after the appearance of the first eukaryotic cell. In order to study the transcriptional circuitries driving sexual reproduction in basidiomycota, we sequenced the poly(A)-positive transcriptome of stage 1 primordia and vegetative mycelia from the self-compatible dikaryotic basidiomycete Coprinopsis cinerea A43mutB43mut. Please note Okayama7 samples not included in this submission.
Project description:The fungal toxin-encoding genes are highly upregulated in the vegetative mycelium upon challenge with the predator. Our recent studies in microfluidics have shown that latter induction is spatially restricted to parts of the vegetative mycelium that is in direct contact with the predator. In order to dissect the defensome of a multicellular fungus against a predator, here, we performed RNA - sequencing of mushroom Coprinopsis cinerea upon challenged with fungivorous nematode Aphelenchus avenae in Microfluidics device at three different time points. We analyzed hyphae that were collected from a microfluidics device where they have been in direct contact with or cultivated without A. avenae.
Project description:Bioactivities of fungal peptides are of interest for basic research and therapeutic drug development. Some of these peptides are derived from “KEX2-processed repeat proteins” (KEPs), a recently defined class of precursor proteins that contain multiple peptide cores flanked by KEX2 protease cleavage sites. Genome mining has revealed that KEPs are widespread in the fungal kingdom. Their functions are largely unknown. Here, we present the first in-depth structural and functional analysis of KEPs in a basidiomycete. We bioinformatically identified KEP-encoding genes in the genome of the model agaricomycete Coprinopsis cinerea and established a detection protocol for the derived peptides by overexpressing the C. cinerea KEPs in the yeast Pichia pastoris. Using this protocol, which includes peptide extraction and mass spectrometry with data analysis using the search engine Mascot, we confirmed the presence of several KEP-derived peptides in C. cinerea as well as in the edible mushrooms Lentinula edodes, Pleurotus ostreatus and Pleurotus eryngii. Processing by the dipeptidyl aminopeptidase STE13 is likely involved in the biosynthesis of some of these peptides. While CRISPR-mediated knockouts of the C. cinerea kep genes did not result in any detectable phenotype, knockout of kex genes caused defects in mycelial growth and fruiting body formation. These results suggest that KEP-derived peptides may play a role in the interaction of C. cinerea with the biotic environment and that the KEP-processing KEX proteases target a variety of substrates in agaricomycetes, including some important for mycelial growth and differentiation. In the past, fungal RiPPs and KEP-derived peptides were mostly studied using forward genetics, where the peptides were first isolated and characterized and only then the corresponding precursor genes were identified (e.g., ustiloxins, phomopsins, candidalysin, Rep1, omphalotin). In this paper, we established a protocol for the identification of KEP-derived peptides in fungal samples by a combination of reverse genetics and peptidomics. The protocol was developed using the model agaricomycete Coprinopsis cinerea, but is also applicable to other fungi. In a first step, we bioinformatically screened the predicted proteome of C. cinerea for KEP-encoding genes. Second, we expressed six different KEP genes from C. cinerea in the yeast Pichia pastoris to (1) establish a protocol for extraction and detection of KEP-derived peptides by mass spectrometry in P. pastoris culture supernatants known for their low complexity in terms of proteins and peptides (Higgins 1995); (2) obtain an indication that the detected KEPs were indeed processed to peptides, and (3) use the structures of the detected peptides as a guide for which peptides to expect in C. cinerea in terms of peptide length and peptide modifications. In a third step, we applied the established peptide extraction and detection method to detect KEP-derived peptides in culture supernatants and tissue samples of the agaricomycetes C. cinerea, Lentinula edodes, Pleurotus ostreatus and Pleurotus eryngii.
2022-10-25 | PXD036934 | Pride
Project description:Small RNA sequencing of Coprinopsis cinerea