Project description:Illumina HiSeq technology was used to generate mRNA profiles from Cenococcum geophilum ectomycorrhizal Pine roots, sclerotia and extramatrical mycelium compared to free-living mycelium . Ectomycorrhizal pine roots, sclerotia, extramatrical mycelium and control mycelium were harvested after 90 days and used for RNA extraction. Reads of 150bp were generated and aligned to the C. geophilum reference genome (https://genome.jgi.doe.gov/Cenge3/Cenge3.home.html).
Project description:Collectively classified as white-rot fungi, certain basidiomycetes efficiently degrade the major structural polymers of wood cell walls. A small subset of these Agaricomycetes, exemplified by Phlebiopsis gigantea, is capable of colonizing freshly exposed conifer sapwood despite its high pitch content, which retards the establishment of other fungal species. The mechanism(s) by which P. gigantea tolerates and metabolizes resinous compounds have not been explored. Here, we report the annotated P. gigantea genome and compare profiles of its transcriptome and secretome when cultured on fresh-cut versus solvent-extracted loblolly pine. The P. gigantea genome contains a conventional repertoire of hydrolase genes involved in cellulose/hemicellulose degradation, whose patterns of expression were relatively unperturbed by the absence of extractives. The expression of genes typically ascribed to lignin degradation was also largely unaffected. In contrast, genes likely involved in the transformation and detoxification of pitch were highly induced in its presence. Their products included an ABC transporter, lipases, cytochrome P450s, glutathione S-transferase and aldehyde dehydrogenase. Other regulated genes of unknown function and several constitutively expressed genes are also likely involved in P. gigantea’s pitch metabolism. These results contribute to our fundamental understanding of conifer colonization and carbon cycling processes. Phlebiopsis gigantea was cultivated in media containing one of three carbon sources: freshly harvested loblolly pine (3 replicates), acetone extracted lobollly pine (3 replicates), or glucose (2 replicates). RNA was extracted and processed for Illumina sequencing as described below.
Project description:Collectively classified as white-rot fungi, certain basidiomycetes efficiently degrade the major structural polymers of wood cell walls. A small subset of these Agaricomycetes, exemplified by Phlebiopsis gigantea, is capable of colonizing freshly exposed conifer sapwood despite its high pitch content, which retards the establishment of other fungal species. The mechanism(s) by which P. gigantea tolerates and metabolizes resinous compounds have not been explored. Here, we report the annotated P. gigantea genome and compare profiles of its transcriptome and secretome when cultured on fresh-cut versus solvent-extracted loblolly pine. The P. gigantea genome contains a conventional repertoire of hydrolase genes involved in cellulose/hemicellulose degradation, whose patterns of expression were relatively unperturbed by the absence of extractives. The expression of genes typically ascribed to lignin degradation was also largely unaffected. In contrast, genes likely involved in the transformation and detoxification of pitch were highly induced in its presence. Their products included an ABC transporter, lipases, cytochrome P450s, glutathione S-transferase and aldehyde dehydrogenase. Other regulated genes of unknown function and several constitutively expressed genes are also likely involved in P. gigantea’s pitch metabolism. These results contribute to our fundamental understanding of conifer colonization and carbon cycling processes.
2014-10-01 | GSE53112 | GEO
Project description:Ectomycorrhizal fungal communities on pine seedlings
Project description:The response mechanisms, recognition and specificity of conifer trees during interaction with pathogenic, saprotrophic or symbiotic ectomycorrhizal fungus were investigated. The roots of Pinus sylvestris were challenged for fifteen days with either Heterobasidion annosum (a pathogenic root rot fungus which attacks Norway spruce, Scots pine and broad leaf trees); Laccaria bicolor (an obligate ectomycorrhizal symbiont); or Trichoderma aureoviride (an obligate saprotroph). The gene expression data from cDNA micro-arrays consisting of 2176 Pinus taeda genes were analysed using 2-interconnected mixed linear model statistical approach. The result of the pairwise comparisons of the different treatments against un-inoculated control led to identification of genes specifically differentially expressed in the pathogenic, saprotrophic and symbiotic interactions. The results were compared with similar data obtained for two other interaction stages: 1 and 5 days post inoculation. The result of this comprehensive expression profiling will hopefully shed more light on the mechanistic basis for recognition and response of conifer trees to pathogenic and non-pathogenic fungi. Keywords: stress response
Project description:The response mechanisms, recognition and specificity of conifer trees during interaction with pathogenic, saprotrophic or symbiotic ectomycorrhizal fungus were investigated. The roots of Pinus sylvestris were challenged for five days with either Heterobasidion annosum (a pathogenic root rot fungus which attacks Norway spruce, Scots pine and broad leaf trees); Laccaria bicolor (an obligate ectomycorrhizal symbiont); or Trichoderma aureoviride (an obligate saprotroph). The gene expression data from cDNA micro-arrays consisting of 2176 Pinus taeda genes were analysed using 2-interconnected mixed linear model statistical approach. The result of the pairwise comparisons of the different treatments against un-inoculated control led to identification of genes specifically differentially expressed in the pathogenic, saprotrophic and symbiotic interactions. The results were compared with similar data obtained for two other interaction stages: 1 and 15 days post inoculation. The result of this comprehensive expression profiling will hopefully shed more light on the mechanistic basis for recognition and response of conifer trees to pathogenic and non-pathogenic fungi. Keywords: stress response
Project description:Comparative transcriptome analysis of early interaction events in Scots pine root tissues following challenge with a pathogenic, saprophytic or symbiotic fungus. Seedlings of P. sylvestris (19 days post germination) were transferred to wet, sterile filter paper on Petri-plates. Thereafter, the roots of the seedlings were inoculated with the mycelial homogenate of either Heterobasidion annosum (FP5, P-type) a pathogenic root rot fungus which attacks Norway spruce, Scots pine and broad leaf trees or Laccaria bicolor, an obligate ectomycorrhizal symbiont or Trichoderma aureoviride- an obligate saprotroph. Thereafter, incubated for 30 minutes, during which time some hyphae adhered to the roots. The inoculated seedlings (ten) were then transferred to another wet sterile filter paper placed on 1% water agar in Petri dishes. A second set of moist sterile filter paper was laid over the roots. The region of the Petri-dish containing the roots was covered with aluminium foil and the edges of the plate sealed with parafilm. The seedlings were then incubated for 24 hr under a photoperiod of 16h light at 20 ºC. Control seedlings were âinoculatedâ with sterile distilled water.