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:Mushroom-forming fungi (Agaricomycetes) are emerging as pivotal players in several fields, as drivers of nutrient cycling, sources of novel applications, or as the group that includes the most morphologically complex fungi. Genomic data for Agaricomycetes are accumulating at a steady pace, however, this is not paralleled by improvements in the quality of genome sequence and associated functional gene annotations, which leaves gene function notoriously poorly understood in comparison with other fungi and model eukaryotes. We set out to improve our functional understanding of the model mushroom Coprinopsis cinerea by integrating a new, chromosome-level assembly with high-quality gene predictions and functional information derived from gene-expression profiling data across 67 developmental, stress, and light conditions. The new annotation includes 5′- and 3′-untranslated regions (UTRs), polyadenylation sites (PAS), upstream ORFs (uORFs), splicing isoforms, conserved sequence motifs (e.g., TATA and Kozak boxes) and microexons. We found that alternative polyadenylation is widespread in C. cinerea, but that it is not specifically regulated across the various conditions used here. Transcriptome profiling allowed us to delineate core gene sets corresponding to carbon starvation, light-response, and hyphal differentiation, and uncover new aspects of the light-regulated phases of life cycle. As a result, the genome of C. cinerea has now become the most comprehensively annotated genome among mushroom-forming fungi, which will contribute to multiple rapidly expanding fields, including research on their life history, light and stress responses, as well as multicellular development.
Project description:Coprinopsis cinerea exhibits synchronised meiosis in the gill tissue of the fungus, which is 75% meiotic. The mushroom develops from a dikaryon, which contains two separate nuclei. These nucleifuse in the basidia (karyogamy). After karyogamy, the nuclei enter an extended meiotic prophase, in which pahcytene occurs at 6 hours post karyogamy (K+6). The tetrads produced by the second meiotic division are present 12 hours after karyogamy (K+12). To examine a comprehensive timecourse of meiosis in this organism, we took samples over a 15-hour period, 3 hours apart: K-3, K, K+3, K+6, K+9, K+12. Keywords: time course
Project description:Coprinopsis cinerea exhibits synchronised meiosis in the gill tissue of the fungus, which is 75% meiotic. The mushroom develops from a dikaryon, which contains two separate nuclei. These nucleifuse in the basidia (karyogamy). After karyogamy, the nuclei enter an extended meiotic prophase, in which pahcytene occurs at 6 hours post karyogamy (K+6). The tetrads produced by the second meiotic division are present 12 hours after karyogamy (K+12). To examine a comprehensive timecourse of meiosis in this organism, we took samples over a 15-hour period, 3 hours apart: K-3, K, K+3, K+6, K+9, K+12. Keywords: time course Four biological replicate samples of each timepoint were taken. Labelled cDNA was hybridized to arrays in a 2 channel reaction. The reference sample was a mixture of timepoint samples.
Project description:Meiosis was compared in msh5-22 and wild type strains of C. cinerea at 6 time points spanning the meiotic timecourse. Abstract: The basidiomycete Coprinopsis cinerea is well-suited to studies of meiosis because meiosis progresses synchronously in ten million cells within each mushroom cap. Approximately 20% of C. cinerea genes exhibit changing expression during meiosis, but meiosis and mushroom development happen concurrently so differentially expressed genes might not be directly involved in meiotic processes. Using microarrays, we examined global gene expression across a meiotic time course in two mutants in which meiosis arrests but mushrooms develop normally. Genes differentially expressed in the mutants compared to wild type are likely to be involved in meiosis and sporulation as opposed to mushroom development. In rad50-1, which arrests in late prophase, RNA abundance for a group of early meiotic genes remains high, while the expression of a group of late meiotic genes is never induced. In contrast, in msh5-22 (which fails to undergo pre-meiotic DNA replication), both early and late meiotic genes are underexpressed relative to wild type at late meiotic time points as the cells die. Genes that are differentially expressed in both mutants are particularly strong candidates for playing roles in meiosis and sporulation.
Project description:Meiosis was compared in rad50-1 and wild type strains of C. cinerea at 6 time points spanning the meiotic timecourse. Abstract: The basidiomycete Coprinopsis cinerea is well-suited to studies of meiosis because meiosis progresses synchronously in ten million cells within each mushroom cap. Approximately 20% of C. cinerea genes exhibit changing expression during meiosis, but meiosis and mushroom development happen concurrently so differentially expressed genes might not be directly involved in meiotic processes. Using microarrays, we examined global gene expression across a meiotic time course in two mutants in which meiosis arrests but mushrooms develop normally. Genes differentially expressed in the mutants compared to wild type are likely to be involved in meiosis and sporulation as opposed to mushroom development. In rad50-1, which arrests in late prophase, RNA abundance for a group of early meiotic genes remains high, while the expression of a group of late meiotic genes is never induced. In contrast, in msh5-22 (which fails to undergo pre-meiotic DNA replication), both early and late meiotic genes are underexpressed relative to wild type at late meiotic time points as the cells die. Genes that are differentially expressed in both mutants are particularly strong candidates for playing roles in meiosis and sporulation.
Project description:Meiosis was compared in rad50-1 and wild type strains of C. cinerea at 6 time points spanning the meiotic timecourse. Abstract: The basidiomycete Coprinopsis cinerea is well-suited to studies of meiosis because meiosis progresses synchronously in ten million cells within each mushroom cap. Approximately 20% of C. cinerea genes exhibit changing expression during meiosis, but meiosis and mushroom development happen concurrently so differentially expressed genes might not be directly involved in meiotic processes. Using microarrays, we examined global gene expression across a meiotic time course in two mutants in which meiosis arrests but mushrooms develop normally. Genes differentially expressed in the mutants compared to wild type are likely to be involved in meiosis and sporulation as opposed to mushroom development. In rad50-1, which arrests in late prophase, RNA abundance for a group of early meiotic genes remains high, while the expression of a group of late meiotic genes is never induced. In contrast, in msh5-22 (which fails to undergo pre-meiotic DNA replication), both early and late meiotic genes are underexpressed relative to wild type at late meiotic time points as the cells die. Genes that are differentially expressed in both mutants are particularly strong candidates for playing roles in meiosis and sporulation. Five time points were analyzed, with four biological replicate rad50-1 samples used for each timepoint. Reference wild-type samples consisted of pooled RNA from ten samples at the appropriate timepoint.
Project description:Meiosis was compared in msh5-22 and wild type strains of C. cinerea at 6 time points spanning the meiotic timecourse. Abstract: The basidiomycete Coprinopsis cinerea is well-suited to studies of meiosis because meiosis progresses synchronously in ten million cells within each mushroom cap. Approximately 20% of C. cinerea genes exhibit changing expression during meiosis, but meiosis and mushroom development happen concurrently so differentially expressed genes might not be directly involved in meiotic processes. Using microarrays, we examined global gene expression across a meiotic time course in two mutants in which meiosis arrests but mushrooms develop normally. Genes differentially expressed in the mutants compared to wild type are likely to be involved in meiosis and sporulation as opposed to mushroom development. In rad50-1, which arrests in late prophase, RNA abundance for a group of early meiotic genes remains high, while the expression of a group of late meiotic genes is never induced. In contrast, in msh5-22 (which fails to undergo pre-meiotic DNA replication), both early and late meiotic genes are underexpressed relative to wild type at late meiotic time points as the cells die. Genes that are differentially expressed in both mutants are particularly strong candidates for playing roles in meiosis and sporulation. Six time points were analyzed, with four biological replicate msh5-22 samples used for each timepoint. Reference wild-type samples consisted of pooled RNA from ten samples at the appropriate timepoint.
Project description:To identify genes express in earliest stage of fruiting body initiation in Coprinopsis cinerea, strain #326 (AmutBmut haploid fruiting strain), genes expressed at 0, 0.5, 1, 3h after light exposure were compared by Super-SAGE Approximately, 1 million tags were sequenced from each mycelial sample to obtain 65,535 unique tags. First, we made matches between the tags and predicted genes of the WT strain, and 24.4% tags matched to predicted genes. Tags of 11.4% genes that did not match with predicted genes matched with the genome sequence of strain #326, and 64.1% of unique tags (2.5~4.5% of total tag counts) were not identified in the #326 genome sequence. In tags that matched to #326 strain genome, we obtained reads 1,000 bp upstream of the genome sequence from the tags, and found that 81% of 1000 bp upstream genome sequences contain predicted genes. The tag counts that matched the same predicted genes were incremented, and fisher’s exact test was carried out using IDEG6