Project description:Transcriptomic analysis of fungus Penicillium decumbens and brlA deletion strains in liquid medium and solid medium respectivelly Examination of differential gene expressions by Penicillium decumbens strains 114-2 and brlA deletion stains in liquid medium and solid medium
Project description:Digital gene expression profiling (DGE) was used to compare the responses of Penicillium decumbens strains to different carbon sources including glucose, cellulose and cellulose-wheat bran. In both wild-type strain 114-2 and cellulase hyperproducing mutant JU-A10-T, transcription of lignocellulolytic enzymes were significantly up-regulated in the presense of cellulose. Relative to 114-2, coordinated up-regulation of lignocellulolytic enzymes and down-regulation of amylases and proteases were observed in JU-A10-T, especially in the cellulose-wheat bran medium. The expression of the principal β-glucosidase BGLI gene was not elevated in JU-A10-T, like the cellulases and hemicellulases, suggesting a different regulatory mechanism for this enzyme. Functional analysis of genes up-regulated in JU-A10-T relative to 114-2 also showed enrichment of proteins involved in amino acid synthesis, protein synthesis, and post-translational modification, compatible with the higher level of production of secreted proteins in JU-A10-T.
Project description:The filamentous fungus Penicillium oxalicum can secret various enzymes for efficient saccharification of plant biomass materials. Expression of the constitutively active forms of transcriptional activators ClrB, XlnR and AraR could trigger the production of different sets of lignocellulolytic enzymes. Here, the transcriptomes of the three engineered strains were compared with that of wild type in the medium without carbon source.
Project description:The goals of this study are to compare differential gene expressions for Penicillium oxalicum wild type strain (WT), and laeA knockout strain (ΔlaeA) in different development phase. The deletion of laeA downregulated genes involved in oxidation- reduction process, alkaloid metabolic process, and transmembrane transport. We find the expression levels of seven secondary metabolism gene clusters (totally 28 clusters) were silenced inΔlaeA. This study provides the information that laeA function are required in conidiation and hydrolase activity of P. oxalicum. Examination of differential gene expressions by digital gene expression tag profiling in Penicillium oxalicum wild type strain and laeA knockout mutant strains in 24h and 60h in modified Czapek culture medium with 2% glucose as carbon resource. qRT–PCR validation was performed using SYBR Green assays.
Project description:The recent discovery of a velvet complex containing several regulators of secondary metabolism in the model fungus Aspergillus nidulans raises the question whether similar type complexes direct fungal development in genera other than Aspergillus. Penicillium chrysogenum is the industrial producer of the antibiotic penicillin, whose biosynthetic regulation is barely understood. Here we provide a functional analysis of two major homologues of the velvet complex in P. chrysogenum, that we have named PcvelA and PclaeA. Data from array analysis using a ?PcvelA deletion strain indicate a significant role of PcvelA on the expression of biosynthesis and developmental genes, including PclaeA. Northern hybridization and HPLC quantifications of penicillin titres clearly show that both PcvelA and PclaeA play a major role in penicillin biosynthesis. Both regulators are further involved in different and distinct developmental processes. While PcvelA deletion leads to light independent conidial formation, dichotomous branching of hyphae and pellet formation in shaking cultures, a ?PclaeA strain shows a severe impairment in conidiophore formation in both the light and dark. Bimolecular fluorescence complementation assays finally provide evidence for a velvet-like complex in Penicillium chrysogenum, with structurally conserved components that have distinct developmental roles, illustrating the functional plasticity of these regulators within filamentous ascomycetes. Transcriptomes of PcvelA- and PclaeA- deletion mutants were compared with expression data from recipient strain deltaPcku70 and reference strain P2niaD18 as a control
