Project description:Certain wood decay basidiomycetes, collectively referred to as brown-rot fungi rapidly depolymerize cellulose while leaving behind the bulk of cell wall lignin as a modified residue. The mechanism(s) employed are unclear, but considerable evidence implicates the involvement of diffusible oxidants, particularly hydroxyl radical. Toward a better understanding of this process, we have examined the transcriptome and secretome of Wolfiporia cocos when cultivated on media containing glucose, purified crystalline cellulose, aspen (Populus grandidentata) or lodgepole pine (Pinus contorta) as sole carbon source. Compared to glucose, 39, 331 and 357 genes exhibited 4-fold increases in transcript levels in cellulose, aspen and lodgepole pine, respectively. Mass spectrometry identified peptides corresponding to 64 glycoside hydrolase (GH) proteins and, of these, 17 corresponded to transcripts upregulated on one or both woody substrates. Most of these genes were broadly categorized as hemicellulases or chitinases. Consistent with an important role for ·OH in cellulose depolymerization, high transcript levels and upregulation were observed for genes involved in iron homeostasis, iron reduction and extracellular peroxide generation. These patterns of regulation differ markedly from the closely related brown rot fungus, Postia placenta, and expand the number of enzymes potentially involved in the oxidative depolymerization of cellulose. Medium containing glucose, microcrystalline cellulose, ground aspen or ground lodgepole pine was inoculated with W. cocos. RNA was purified from cultures. Single read 100 bp Illumina runs were performed.
Project description:Certain wood decay basidiomycetes, collectively referred to as brown-rot fungi rapidly depolymerize cellulose while leaving behind the bulk of cell wall lignin as a modified residue. The mechanism(s) employed are unclear, but considerable evidence implicates the involvement of diffusible oxidants, particularly hydroxyl radical. Toward a better understanding of this process, we have examined the transcriptome and secretome of Wolfiporia cocos when cultivated on media containing glucose, purified crystalline cellulose, aspen (Populus grandidentata) or lodgepole pine (Pinus contorta) as sole carbon source. Compared to glucose, 39, 331 and 357 genes exhibited 4-fold increases in transcript levels in cellulose, aspen and lodgepole pine, respectively. Mass spectrometry identified peptides corresponding to 64 glycoside hydrolase (GH) proteins and, of these, 17 corresponded to transcripts upregulated on one or both woody substrates. Most of these genes were broadly categorized as hemicellulases or chitinases. Consistent with an important role for ·OH in cellulose depolymerization, high transcript levels and upregulation were observed for genes involved in iron homeostasis, iron reduction and extracellular peroxide generation. These patterns of regulation differ markedly from the closely related brown rot fungus, Postia placenta, and expand the number of enzymes potentially involved in the oxidative depolymerization of cellulose.