Project description:To identify the genes regulated by Arabidopsis ERF19. 35S::ERF19 transgenic Arabidopsis was created, them we employ whole genome microarray expression profiling as a discovery platform to identify genes with the potential to regulae by ERF19. The RNA was isopated from the whole flower of control and 35S::ERF19 transgenic Arabidopsis.
Project description:Microarray analyses were performed on Arabidopsis thaliana plants over-expressing either AtERF5 or AtERF6, which are both transcription factors of the ethylene response factor (ERF) family. In this way it was hoped that any changes in the Arabidopsis transcriptome caused by their over-expression would provide clues as to the role(s) these genes play in vivo.<br>Arabidopsis thaliana plants were transformed with either 35S::AtERF5, 35S::AtERF6 or the empty vector pK2GW7 (used as a control). Total RNA was extracted from untreated 10-day old seedlings. Three separate arrays were performed for each of the two over-expressors, (the RNA for each slide having been extracted from an independent biological replicate).
Project description:Despite their importance, plant MAP kinase targets are still poorly elucidated. Here, the specific in vivo interaction of an ethylene response factor (ERF104) with the Arabidopsis MAP kinase, MPK6, is shown by fluorescence resonance energy transfer. The interaction, which is lost within minutes after treatment with the flagellin-derived flg22 peptide, is dependent on both MPK6 kinase activity and rapid ethylene signaling initiated downstream of MPK6 activation. ERF104 is an MPK6 substrate and phosphorylation site mutations affected its stability. ERF104 activates promoters with GCC elements. This was evident from microarray data of overexpressing transgenic plants, where promoters of up regulated genes contain GCC motifs and chromatin immunoprecipitation showing ERF104 association with PDF1.2 promoter. The ERF104 overexpressor did not affect biotrophic bacteria proliferation but was more susceptible to necrotrophic Botrytis cinerea. Microarray performed with erf104 or mpk6 revealed only a limited number of flg22-induced genes that require these elements - possibly as a result of functional redundancies. Thus, ERF104 phosphorylation by MPK6, in concert with ethylene signaling induced by pathogen-derived molecules, modulates defense in Arabidopsis.
Project description:Despite their importance, plant MAP kinase targets are still poorly elucidated. Here, the specific in vivo interaction of an ethylene response factor (ERF104) with the Arabidopsis MAP kinase, MPK6, is shown by fluorescence resonance energy transfer. The interaction, which is lost within minutes after treatment with the flagellin-derived flg22 peptide, is dependent on both MPK6 kinase activity and rapid ethylene signaling initiated downstream of MPK6 activation. ERF104 is an MPK6 substrate and phosphorylation site mutations affected its stability. ERF104 activates promoters with GCC elements. This was evident from microarray data of overexpressing transgenic plants, where promoters of up regulated genes contain GCC motifs and chromatin immunoprecipitation showing ERF104 association with PDF1.2 promoter. The ERF104 overexpressor did not affect biotrophic bacteria proliferation but was more susceptible to necrotrophic Botrytis cinerea. Microarray performed with erf104 or mpk6 revealed only a limited number of flg22-induced genes that require these elements - possibly as a; result of functional redundancies. Thus, ERF104 phosphorylation by MPK6, in concert with ethylene signaling induced by pathogen-derived molecules, modulates defense in Arabidopsis. Experiment Overall Design: Leaves of six week old Col-0, erf104, mpk6 and 35S::ERF104 plants were infiltrated with 1µM Flg22 or water and harvested four hours later. Total RNA was isolated and processed according to the Affymetrix protocol for biotin-labelled cRNA and hybridized to the Affymetrix ATH1 chip. The data Experiment Overall Design: was analyzed with Genespring GX 7.3.1 software (Agilent) with the following parameters: Filter on Flags for present or marginal in 50% of all considered experiments, Filter for reliable differentially expressed genes based on volcano plot (one way ANOVA p-value <0.05 and >3-fold change in expression). For the flg22 experiments, analysis for each genotype was separately performed and a composite list of flg22-regulated genes compiled â with the aim of including genes that may be differentially regulated in the genotype. Global expression profile was visualized by k-means clustering and condition tree (Genespring).
Project description:Microarray analyses were performed on Arabidopsis thaliana plants overexpressing ERF5. In this way it was hoped that any changes in the Arabidopsis transcriptome caused by overexpression would provide clues as to the role this gene plays in vivo.<br>Arabidopsis thaliana plants were transformed with 35S::ERF5 or the empty vector pK2GW7 (used as a control). Total RNA was extracted from untreated 10-day old seedlings. Three separate arrays were performed (the RNA for each slide having been extracted from an independent biological replicate).
Project description:Microarray analyses were performed on Arabidopsis thaliana plants overexpressing ERF6. In this way it was hoped that any changes in the Arabidopsis transcriptome caused by overexpression would provide clues as to the role this gene plays in vivo.<br>Arabidopsis thaliana plants were transformed with 35S::ERF6 or the empty vector pK2GW7 (used as a control). Total RNA was extracted from untreated 10-day old seedlings. Three separate arrays were performed (the RNA for each slide having been extracted from an independent biological replicate).
Project description:Ethylene is a gaseous plant hormone that regulates plant growth and development. Broad reprogramming of gene expression is required for ethylene responses. The primary ethylene transcription factor (TF) ETHYLENE INSENSITIVE3 (EIN3) drives expression of secondary TFs including the ETHYLENE RESPONSE DNA-BINDING FACTORS (EDFs), but the role of the EDFs within the ethylene genome regulatory network is not understood. Here, we describe an investigation into the function of the EDFs in ethylene signalling and hormonal cross-regulation. We determined the target genes and binding dynamics of EDFs 1, 2, 3 during an ethylene response and the effects of edf1234 quadruple mutation on gene expression. The EDFs and EIN3 shared a large proportion of their target genes but had different functions. The EDFs were associated with repression of target genes, but this was superseded by activation when EIN3 bound the same genes. Genes important in other hormone signalling pathways, in particular abscisic acid (ABA), were targets of the EDFs. This demonstrates how ethylene engages hormonal cross-regulation to repress genes in competing signalling pathways and prioritize itself.
Project description:Purpose: Next-generation sequencing (NGS) has been used to study the gene expression in Arabidopsis seedlings under air and ethylene treatment. The goal of this study is to uncover how TREE1 and its homologous gene DAZ3 are involved into gene expression regulation in response to ethylene.