Project description:Transcriptional time course analysis of Wheat leaves in response to exposure to the fungal toxin ToxA from Pyrenophora tritici-repentis.
Project description:Tan Spot (TS), causal agent Pyrenophora tritici-repentis (Ptr), is a major threat to wheat production due to the lack of resistant cultivars. In our previous work, we identified MAGIC population parental lines exhibiting TS resistance and susceptibility, namely Robigus and Hereward, respectively. To understand the mechanisms underlying these phenotypes, we performed RNA-seq analysis of leaves before and during Ptr interaction. When comparing mock- and Ptr-inoculated samples, differentially expressed genes (DEGs) were identified with DESeq2, leading to the targeting of 15193 DEGs. Functional annotation showed the pathways enzyme classification, solute transport, RNA biosynthesis, protein modification and homeostasis represented 49.5% of DEGs in Robigus. Cellular metabolism pathways were induced, as well as vesicle trafficking, actin polymerization and cellulose. The upregulation of these cell wall related genes along with microscopic data suggested that barrier defence is a major feature of TS resistance in Robigus. Conversely, photosynthesis was the top fifth pathway in Hereward, totalling 389 repressed genes (12.63%). Photosynthesis collapse was linked to the activation of oligosaccharide metabolism and suppression of glycolysis, TCA cycle and amino acids degradation. This may reflect mobilization of host nutrients to Ptr. Our observations could inform wheat-breeding programmes targeting TS resistance.
Project description:Pyrenophora tritici-repentis isolate PtrDW5 extracellular
Proteome mapping of fungus Pyrenophora tritici-repentis (Ptr) extracellular proteins
Sample analysis (in brief):
- extracellular proteins have been separated into 24 fractions by OFFGEL electrophoresis (IEF)
- proteins of each fraction have then been alkylated (iodoacetamide) and trypsin-digested
- peptides of each fraction have then been separated on a C18 column in an LC run and analysed by high-resolution mass spectrometry (LTQ Orbitrap Velos or XL)
