Project description:The Arabidopsis thaliana mutant wrky33 is highly susceptible to the necrotrophic fungus Botrytis cinerea. We identified by ChIP-seq >1680 high-confidence WRKY33 binding sites associated with 1576 genes within the Arabidopsis genome, with all of them being dependent on rapid activation of WRKY33 expression by Botrytis cinerea strain 2100. Genome-wide transcriptional analysis defined 318 genes as direct functional targets at 14 h post inoculation. Comparison between resistant wild-type Columbia-0 and susceptible wrky33 mutant plants revealed that expression of 75% of all WRKY33 regulated targets were down-regulated upon infection, indicating that WRKY33 predominately acts as a repressor. However, WRKY33 appears to possess dual functionality acting either as a repressor or as an activator in a promoter-context dependent manner. Our genome-wide analysis confirmed known WRKY33 targets involved in ethylene and jasmonic acid hormone signaling and phytoalexin biosynthesis, but also uncovered a previously unknown role of abscisic acid (ABA) biosynthesis in the complex regulatory circuitry affecting resistance towards Botrytis. Analysis of transgenic plants expressing WRKY33-HA under its native promoter post inoculation with spores of Botrytis cinerea 2100
Project description:The Arabidopsis thaliana mutant wrky33 is highly susceptible to the necrotrophic fungus Botrytis cinerea. Comparing the expression profiles of B. cinerea-infected wrky33 and WT plants we identified 2765 differentially expressed genes dependent on WRKY33, of which 1675 were up-regulated in the mutant (termed WRKY33-repressed genes) and 1090 were down-regulated in the mutant. Combined with ChIP-seq data 318 genes were identified as direct functional targets of WRKY33 at 14 h post inoculation with spores of Botrytis cinerea 2100. Comparison of altered gene expression in Arabidopsis WT and wrky33 mutant plants 14 hours post inoculation with Botrytis cinerea 2100.
Project description:The Arabidopsis thaliana mutant wrky33 is highly susceptible to the necrotrophic fungus Botrytis cinerea. We identified by ChIP-seq >1680 high-confidence WRKY33 binding sites associated with 1576 genes within the Arabidopsis genome, with all of them being dependent on rapid activation of WRKY33 expression by Botrytis cinerea strain 2100. Genome-wide transcriptional analysis defined 318 genes as direct functional targets at 14 h post inoculation. Comparison between resistant wild-type Columbia-0 and susceptible wrky33 mutant plants revealed that expression of 75% of all WRKY33 regulated targets were down-regulated upon infection, indicating that WRKY33 predominately acts as a repressor. However, WRKY33 appears to possess dual functionality acting either as a repressor or as an activator in a promoter-context dependent manner. Our genome-wide analysis confirmed known WRKY33 targets involved in ethylene and jasmonic acid hormone signaling and phytoalexin biosynthesis, but also uncovered a previously unknown role of abscisic acid (ABA) biosynthesis in the complex regulatory circuitry affecting resistance towards Botrytis.
Project description:The Arabidopsis thaliana mutant wrky33 is highly susceptible to the necrotrophic fungus Botrytis cinerea. Comparing the expression profiles of B. cinerea-infected wrky33 and WT plants we identified 2765 differentially expressed genes dependent on WRKY33, of which 1675 were up-regulated in the mutant (termed WRKY33-repressed genes) and 1090 were down-regulated in the mutant. Combined with ChIP-seq data 318 genes were identified as direct functional targets of WRKY33 at 14 h post inoculation with spores of Botrytis cinerea 2100.
Project description:Next generation sequencing was performed to identify genes changed in Arabidopsis thaliana upon Botrytis cinerea infection. The goal of the work is to find interesting genes involved in plant defense. The object is to reveal the molecular mechanism of plant defense.
Project description:Arabidopsis thaliana exhibits differential susceptibility to the fungal pathogen Botrytis cinerea depending on the time of day that infection occurs. We hypothesised that this is driven by teh circadian clock and that differences in the amplitude or speed of the plant defence response will underlie the difference in susceptiblity. A major component of the defence response is transcriptional reprogramming, hence we investigated whether the transcriptional response to B. cinerea infection differs following inoculation at subjective dawn or night (the points of greatest difference in susceptiblity) under constant light conditions.
Project description:The goal of the microarray experiment was to identify defense genes that were differentially expressed in the Arabidopsis mutant elp2 and wild type in response to infection of the necrotrophic fungal pathogen Botrytis cinerea. Results indicated that, compared with the wild type, the WRKY33/OCTADECANOID-RESPONSIVE ARABIDOPSIS AP2/ERF59 (ORA59)/ETHYLENE RESPONSE FACTOR1 transcriptional cascades are down-regulated, whereas the MYC2 transcriptional cascade is up-regulated in the elp2 mutant. Three biological replicates with leaves from 8-12 plants per sample were collected at 0, 3, 6, 12, 24, and 48 hours after inoculation with the necrotrophic fungal pathogen Botrytis cinerea. After extraction, RNA concentration was determined on a NanoDrop Spectrophotometer (Thermofisher Scientific, Waltham, MA) and sample quality was assessed using the 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). Equal amount of RNA from the 3 biological replicates were used for microarray analysis. The channels of the dual-channel arrays were analyzed independently.