Project description:We isolated a bacterial identified as the genus of Mycetocola from the infected Flammulina velutipes samples collected from mushroom house in Hebei, North China. The stipe of F. velutipes appeared water-soaked, soften and browning after infected, finally, the whole plant rotten. The incidence of the disease can reach more than 80% in the serious areas, resulting in an obvious decline in the yield and quality of the F. velutipes. Furthermore, the pathogenic bacteria may occurrence in large areas by accumulated in the annual cultivation process. Therefore, the most effective measures to control this bacterial disease is screening resistant strains of F. velutipes with high yield and good agronomic traits. We obtained a white F. velutipes strain JK502 which has strong resistance to this bacterium by using the double-layer plate method combined with the sporozoite hybridization. JK501, which can early fruited, was screened using the same parent SH9 by multi spore hybridization. In order to investigate the differences of gene expression between the parent strain and the selected strains, we completed the transcriptome sequencing by Illumina HiSeq X system.
Project description:Twelve Flammulina velutipes strains originating from Poland were identified using internal transcribed spacer (ITS) region sequencing. Based on the sequences obtained, the genomic relationship of the analyzed strains was determined. All F. velutipes strains were also characterized using Biolog FF MicroPlates to obtain data on C-substrate utilization and mitochondrial activity. The ability to decompose various substrates differed among the F. velutipes strains up to five times. The highest catabolic activities were characteristic for only two strains with capabilities to decompose up to 22 carbon sources. The correlation between carbon repression and laccase production by F. velutipes was analyzed based on glucose assimilation by these strains. Moreover, the influence of metal ions (Cu(2+), Cd(2+)), veratric and ferulic acids, and temperature on laccase activities in the analyzed strains was determined. The results obtained proved that all the inducers influenced laccase expression in almost all the analyzed strains. However, the degree of induction depended not only on the strain used but also on the day of the induction.
Project description:Arabinogalactan proteins are proteoglycans found on the cell surface and in the cell walls of higher plants. The carbohydrate moieties of most arabinogalactan proteins are composed of β-1,3-galactan main chains and β-1,6-galactan side chains, to which other auxiliary sugars are attached. For the present study, an endo-β-1,3-galactanase, designated FvEn3GAL, was first purified and cloned from winter mushroom Flammulina velutipes. The enzyme specifically hydrolyzed β-1,3-galactan, but did not act on β-1,3-glucan, β-1,3:1,4-glucan, xyloglucan, and agarose. It released various β-1,3-galactooligosaccharides together with Gal from β-1,3-galactohexaose in the early phase of the reaction, demonstrating that it acts on β-1,3-galactan in an endo-fashion. Phylogenetic analysis revealed that FvEn3GAL is member of a novel subgroup distinct from known glycoside hydrolases such as endo-β-1,3-glucanase and endo-β-1,3:1,4-glucanase in glycoside hydrolase family 16. Point mutations replacing the putative catalytic Glu residues conserved for enzymes in this family with Asp abolished activity. These results indicate that FvEn3GAL is a highly specific glycoside hydrolase 16 endo-β-1,3-galactanase.
Project description:White-rot basidiomycete fungi are potent degraders of plant biomass with the ability to mineralize all lignocellulose components. Recent comparative genomics studies showed that these fungi use a wide diversity of enzymes for wood degradation. In order to improve our understanding on the enzymatic mechanisms leading to lignocellulose breakdown, we analysed the early response of the white-rot fungus Pycnoporus coccineus CIRM-BRFM310 to various lignocellulosic substrates at two time points; Day 3 and Day 7.
Project description:Methionine oxydation level was monitored by tandem mass spectrometry for secreted proteins and intracellular proteins from the white-rot fungus Pycnoporus cinnabarinus grown on aspen wood.