Project description:Using 21K spruce microarray (that contains 21.8 thousand unique transcripts) we performed analysis of the transcriptome response of lodgepole pine (Pinus contorta) inoculated with the mountain pine beetle (Dendroctonus ponderosae) vectored fungal pathogen Grosmannia clavigera or treated with wounding. This microarray analysis revealed large transcriptome reorganization with close to 2000 transcripts (10% of the studied transcriptome) differentially expressed within two weeks of treatment, with the wounding response affecting close to 2% of the lodgepole pine transcriptome. RNA was isolated from the bark of lodgepole pine inoculated with Grosmannia clavigera, treated with wounding, or untreated control for three time points (6h, 2days and 2 weeks). Three independent biological replicates were included for each treatment and each time point. Three hybridizations were performed for each comparison of different treatments (fungal, wounding, control) within each time point (6 hours, 2 days, 2 weeks) and one hybridization was performed for the comparison of the same treatments between time points (total 36 hybridizations/slides).
Project description:Gc, a Mountain pine beetle associated pathogen, can survive from highly abundant pine chemicals ( e.g terpenes) and use some unflavored compound’s as carbon source. using RNA_Seq, we analyzed the transcriptome of Gc when it grew on limonene, mannose oliver-oil, oleic acid as carbon source, as well as when it survived from high concentration of limonene or heptane. We profiled the expression of some interesting genes ( ABC transporters, P450s) potentially involved in the tree-pathogen interaction. An ABC-G group transporter gene (GcABC-G1) was one of the most highly induced genes and characterized as a mono-terpene specific efflux transporter with genetic and molecular tools. RNA- seq also indicated Gc utilize limonene and oleic acid through the same beta-oxidation pathway. However the degradation of limonene is more complex and multiple pathways contributed to the survival/utilization.
Project description:Using 21K spruce microarray (that contains 21.8 thousand unique transcripts) we performed analysis of the transcriptome response of lodgepole pine (Pinus contorta) inoculated with the mountain pine beetle (Dendroctonus ponderosae) vectored fungal pathogen Grosmannia clavigera or treated with wounding. This microarray analysis revealed large transcriptome reorganization with close to 2000 transcripts (10% of the studied transcriptome) differentially expressed within two weeks of treatment, with the wounding response affecting close to 2% of the lodgepole pine transcriptome.
Project description:Gc, a Mountain pine beetle associated pathogen, can survive from highly abundant pine chemicals ( e.g terpenes) and use some unflavored compound’s as carbon source. using RNA_Seq, we analyzed the transcriptome of Gc when it grew on limonene, mannose oliver-oil, oleic acid as carbon source, as well as when it survived from high concentration of limonene or heptane. We profiled the expression of some interesting genes ( ABC transporters, P450s) potentially involved in the tree-pathogen interaction. An ABC-G group transporter gene (GcABC-G1) was one of the most highly induced genes and characterized as a mono-terpene specific efflux transporter with genetic and molecular tools. RNA- seq also indicated Gc utilize limonene and oleic acid through the same beta-oxidation pathway. However the degradation of limonene is more complex and multiple pathways contributed to the survival/utilization. mRNA was extracted from Gc mycelium under various conditions and cDNA libraries were generated for pair-end sequencing. The 70-100bp illumina sequence read was mapping to reference genome and RNA-Seq was carried out in CLC genomic work bench.