Project description:The grey mould fungus Botrytis cinerea is a typical necrotrophic fungus that can infect hundreds of host plants, including high-value crops such as grapevine, strawberry and tomato. In order to find new important genes involved in the fungal virulence, a mutant library was generated by random insertional mutagenesis, using Agrobacterium tumefaciens mediated transformation. Ten mutants exhibiting total loss of virulence towards different host plants were considered. Their molecular characterization revealed a single T-DNA insertion in unique and different loci. Using a proteomics approach, the secretome of four of these strains was compared to that of the parental strain and a common profile of reduced lytic enzymes was recorded. Significant variations in this profile, notably proteases and hemicellulases deficiencies, were observed and validated by biochemical tests. They were also a hallmark of the remaining six non pathogenic strains, suggesting the importance of these secreted proteins in the infectious process.
Project description:The selective advantage of bioluminescence in bacterial cells that do not form symbiotic relationships with aquatic animals is still not known. Some evidence suggests that bioluminescence plays a role in DNA repair by a photoreactivation process (Czyz 2000) and that non-bioluminescent strains are less virulent than their bioluminescent isogenic counterparts (Ruwandeepika 2010). All hypotheses to date suggest bioluminescence associated or mediated changes in gene expression, yet the evidence for this does not exist. In this study, we generated an in-frame luxAB deletion mutant (the two contiguous genes that encode for bacterial luciferase) and compared its mid-log phase gene expression profile with that of the wild type spontaneous streptomycin resistant (STR) V. campbellii BAA-1116 parental strain from which it was derived. Both mid-log phase transcriptomes were elucidated using custom designed whole genome microarrays (520694F, Affymetrix) to determine the effect luciferase has on V. campbellii gene expression. The virulence phenotypes of both strains were also subsequently tested in Artemia franciscana challenge experiments.
Project description:The eight-subunit COP9 signalosome (CSN complex), controls the exchange of E3 ubiquitin cullin RING ligase receptors through its deneddylase activity, providing specificity to eukaryotic protein degradation. The conserved eight-subunit CSN complex is required for multicellular development of the filamentous fungus Aspergillus nidulans. The human, as well as the fungal CSN complex assembles through a heptameric subcomplex (pre-CSN complex) that is activated by the integration of the eighth subunit, the catalytic CsnE/5 deneddylase. The focus of this work was to unveil the assembly of the native fungal pre-CSN complex via combined genetic and biochemical approaches. Interactomes of various functional GFP-Csn subunit fusions, produced in pre-CSN deficient fungal double csn subunit gene deletion strains, were compared by affinity purifications coupled to mass spectrometry analyses. With this approach, two distinct heterotrimeric CSN subcomplexes were identified as pre-CSN assembly intermediates, namely CsnA/1-C/3-H/8 and CsnD/4-F/6-G/7 that form independently of CsnB/2 that connects the heterotrimers to a heptameric subcomplex and enables subsequent integration of CsnE/5, yielding in the enzymatically active CSN holocomplex. Surveillance mechanisms control accurate Csn subunit abundances and correct cellular localization for sequential assembly, since deprivation of Csn subunits change the abundance and location of the remaining Csn subunits.
2023-08-10 | PXD043205 | Pride
Project description:Genomic comparison of Mucor strains including four new genomic sequences
Project description:The aim of this experiment was to determine if the development of resistance to antibiotics can be driven by the concentration and speciation of Cu. Experimental setup was designed to investigate two hypotheses for which two strains of Gram- bacteria have been selected: - Do TE enhance AR in resistant bacteria? Resistant strain: Bioluminescent Pseudomonas aeruginosa PAO1 (Xen41, Tetracycline resistant) - Do TE induce AR in sensitive bacteria? Sensitive strain: Pseudomonas aeruginosa PAO1 (Wild Type)
Project description:The aim of this experiment was to determine if the development of resistance to antibiotics can be driven by the concentration and speciation of Cu. Experimental setup was designed to investigate two hypotheses for which two strains of Gram- bacteria have been selected: - Do TE enhance AR in resistant bacteria? Resistant strain: Bioluminescent Pseudomonas aeruginosa PAO1 (Xen41, Tetracycline resistant) - Do TE induce AR in sensitive bacteria? Sensitive strain: Pseudomonas aeruginosa PAO1 (Wild Type)
Project description:Wine biological aging is a wine making process used to produce specific beverages in several countries in Europe, including Spain, Italy, France, and Hungary. This process involves the formation of a velum at the surface of the wine. Here, we present the first large scale comparison of all European flor strains involved in this process. We inferred the population structure of these European flor strains from their microsatellite genotype diversity and analyzed their ploidy. We show that almost all of these flor strains belong to the same cluster and are diploid, except for a few Spanish strains. Comparison of the array hybridization profile of six flor strains originating from these four countries, with that of three wine strains did not reveal any large segmental amplification. Nonetheless, some genes, including YKL221W/MCH2 and YKL222C, were amplified in the genome of four out of six flor strains. Finally, we correlated ICR1 ncRNA and FLO11 polymorphisms with flor yeast population structure, and associate the presence of wild type ICR1 and a long Flo11p with thin velum formation in a cluster of Jura strains. These results provide new insight into the diversity of flor yeast and show that combinations of different adaptive changes can lead to an increase of hydrophobicity and affect velum formation.
Project description:Invasion of host tissue by the human fungal pathogen, Candida albicans is an important step during many forms of candidosis. However, not all C. albicans strains possess the same invasive and virulence properties. It is known for example that the two clinical isolates SC5314 and ATCC10231 differ in their ability to invade into host tissue and to cause infections. Strain SC5314 is invasive whereas strain ATCC10231 is non-invasive and strongly attenuated in virulence as compared to SC5314. In this study we compare the in vitro transcriptional profiles and the genotypic profiles of these two widely used laboratory strains in order to determine the principal biological and genetic properties which may govern the different potential for invasiveness and virulence. Keywords: transcriptional profiling, comparative genomic hybridisation, invasive vs. non-invasive C. albicans strain Genomic DNA from C. albicans strains SC5314 and ATCC10231 hybridisations were done in duplicate including one dye swap. Total RNA from C. albicans strains SC5314 and ATCC10231 strains were compared in triplicate including one biological replicate and one dye swap.
Project description:We report the transcriptomic comparisions between ku70 control and ste12 mutant strains in nematode-trapping fungus Arthrobotrys oligospora when induced with nematodes. Fungal Ste12 transcription factor and the upstream MAPK cascade are highly conserved and plays a role in host sensing and pathogenesis in various fungal pathogens. Identification of Ste12-dependent in A. oligospora may provide further insights into the molecular mechanisms of nematode-sensing and trap morphogenesis. The reference assemly used for remapping is A. oligospora TWF154 (GenBank assembly accession: GCA_004768765.1)