Project description:When studying filamentous fungi in terms of adaptation to environmental conditions, a reference organism is generally missing. This is mainly obstructing in the research of pathogens (well defined model organisms are non-pathogenic). Here, we investigated the mechanism underlying the ability of N. crassa to respond to various environmental stimuli, including the presence of azole and echinocandin antifungals. Our molecular analysis revealed that the Δccg8 deletion impacts the fungus on a broad physiological level, including dysregulation of ribosome biogenesis and fatty acid β-oxidation (as demonstrated by proteomic profiling), and supporting changes in glycosylated proteins and fatty acid and ergosterol composition. To bridge the gap between in vitro and in vivo conditions, we successfully applied Galleria mellonella larval model. Despite being non-pathogenic, when injected with conidia of tested filamentous fungi, the immune system cleared conidia of Δccg8 mutant strain more effectively than wild-type strain, mirroring in vitro observations and demonstrating a reproducible immune reaction. This work not only provides molecular insights into the fungal stress adaptation but also establishes N. crassa as a viable non-pathogenic organism for in vivo analysis, substantially broadening its application, reaching into dominion of research of pathogenic filamentous fungi.
Project description:Transcriptional profiling with next-generation sequencing methods demonstrated that a Neurospora crassa mutant with the three most highly expressed beta-glucosidase genes deleted had a transcriptional response to cellobiose similair to that of wild type N. crassa exposed to cellulose. N. crassa was pregrown in Sucrose and transferred to Avicel (cellulose), Cellobiose, Sucrose or media with no carbon added. Biological triplicates used to identify differentially expressed genes in WT on Avicel. Single libraries for mutant strains identify which genes show similair expression on cellobiose as in the WT on cellulose.