Project description:Five years old Ulmus minor plants from three different genotypes, two tolerant and one sensitive to Dutch Elm disease, were inoculated with an aggressive local strain of Ophiostoma novo-ulmi (Z-BU1) while the other half were inoculated with sterile and distilled water as control treatment. following the procedure described by Solla et al. (2005). A healthy 3-year-old branch located around 2 meters tall, from both inoculated and control plants, were collected at 1, 3, 7, 14 and 21 days after inoculation. The stem from the branch was individualized and RNA isolated to hybridize two colors microarrays.
Project description:6-year-old ramets of U. minor (Atinian elm clone) were inoculated with O. novo-ulmi in order to evaluate molecular responses activated during plant colonization. To elucidate the different genes involved in the U. minor immune system and the molecular changes suffered after inoculation, oligomicroarrays were constructed using the data from the transcriptome available in the Dryad database (Perdiguero et al., 2015; http://dx.doi.org/10.5061/dryad.ps837), and hybridized with cDNA obtained from the ramets over a time course following inoculation. Three biological replicates for control and inoculated plants for each sampling point (1, 3, 7, 14 and 21 days post-inoculation) were hybridised using two colors (inoculated vs control).
Project description:Quorum sensing (QS) is a complex cell-cell communication mechanism that coordinates population-level behaviors in microbes. In eukaryotes, this phenomenon has been extensively described in the dimorphic yeast Candida albicans as its main QS molecule, the sesquiterpene alcohol farnesol, is responsible for various phenotypic (i.e., inhibition of yeast-to-hyphae transition, biofilm formation and, hence, pathogenesis) and metabolic (i.e., induction of oxidative stress and apoptosis) changes. Ophiostoma piceae CECT 20416 is a dimorphic saprotrophic ascomycete with biotechnological interest that also produces farnesol as a QS molecule, but in this case, the alcohol promotes the morphological transition to the mycelial form, biofilm formation, enzyme secretion, and melanin production. Here, we characterized the physiological response of Ophiostoma piceae to farnesol, and the molecular components of the QS system of this fungus have been investigated using a ‘multiomics’ approach that involved genomic, transcriptomic, and proteomic analyses. Some genes identified in this work are proposed as key factors in farnesol transport and signaling. We have also cataloged the genes undergoing major transcriptional changes triggered by the presence of the autoinducer, such as cell-wall remodeling, ROS protection, and melanin biosynthesis, using self-organizing maps (SOMs). This analysis could be useful for applications in the forestry industry, for enzyme production, and for the valorization of residues. Furthermore, it might as well help to investigate the QS mechanisms of clinically relevant fungi phylogenetically related to Ophiostoma.