Project description:This SuperSeries is composed of the following subset Series: GSE27941: Lignocellulose-induced regulation of Phanaerochaete chrysosporium genes GSE29656: Postia placenta MAD-698 gene expression in ball-milled aspen or ball-milled pine medium Refer to individual Series
Project description:Transcript profiles of Postia placenta grown on media containing ball-milled aspen or ball-milled pine as the sole carbon source were analyzed. Array design was based on the DoE's Joint Genome Institute's gene models for P. placenta version 1. The research goal is to identtify genes essential for cellulose depolymerization.
Project description:Transcript profiles of Phanerochaete chrysosporium grown on ball-milled aspen or ball-milled pine were analyzed. Array design based on the DoE's Joint Genome Institute's v2.1 annotation. Goal is to define genes involved in lignocellulose degradation.
Project description:Transcript profiles of Postia placenta grown on media containing ball-milled aspen or ball-milled pine as the sole carbon source were analyzed. Array design was based on the DoE's Joint Genome Institute's gene models for P. placenta version 1. The research goal is to identtify genes essential for cellulose depolymerization. From a data set of 12,438 unique alleles, each NimbleGen (Madison, WI) array featured 10 unique 60mers per gene, all in triplicate. The dataset was manually annotated to include only the ‘best allelic model’ among CAZY-encoding genes. Total RNA was purified from medium containing ball-milled aspen or ball-milled pine as the sole carbon source. Three biological replicates per medium were used (6 separate arrays). RNA was converted to double-strand cDNA and labeled with the Cy3 fluorophore sample for hybridization to the Postia placenta MAD-698 whole genome expression array by Roche NimbleGen (Iceland). In brief, 10ug of total RNA was incubated with 1X first strand buffer, 10 mM DTT, 0.5mM dNTPs, 100 pM oligo T7 d(T)24 primer, and 200 units of SuperScript II (Invitrogen) for 60 min at 42°C. Second strand cDNA was synthesized by incubation with 1X second strand buffer, 0.2mM dNTPs, 0.07 units per ul DNA ligase (Invitrogen), 0.27 units per ul DNA polymerase I (Invitrogen), 0.013 units per ul RNase H (Invitrogen), at 16°C for 2 hours. Immediately following, 10 units T4 DNA polymerase (Invitrogen) was added for additional 5 minute incubation at 16°C. Double-stranded cDNA was treated with 27ng/ul of RNase A (EpiCenter Technologies) for 10 minutes at 37°C. Treated cDNA was purified using an equal volume of phenol:chloroform:isoamyl alcohol (Ambion), ethanol precipitated, washed with 80% ethanol, and resuspended in 20ul water. One ug of each cDNA sample was amplified and labeled with 1 unit per ul of Klenow Fragment (New England BioLabs) and 1 O.D unit of Cy3 fluorophore (TriLink Biotechnologies, Inc.) for 2 hours at 37°C. Array hybridization was carried out with 6ug of labeled cDNA suspended in NimbleGen hybridization solution for 17 hours at 42°C.
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:This SuperSeries is composed of the following subset Series: GSE14734: Cellulose-induced regulation of Phanaerochaete chrysosporium genes GSE14735: Nutrient limitation-induced regulation of Phanaerochaete chrysosporium genes Refer to individual Series
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.