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Transcription profiling of poplar WRKY23 reveals a regulatory role in defense: WRKY23-RNAi

ABSTRACT: To investigate the function of poplar WRKY23, we generated PtWRKY23-overexpressing and -underexpressing (RNAi) plants. Transgenic plants were inoculated with Melampsora rust or mock-inoculated for assessment of rust-resistance and for gene expression profiling using the poplar Affymetrix GeneChipÂ® to study the consequences of PtWRKY23 overexpression and underexpression. Transcriptome analysis of PtWRKY23 overexpressors revealed a significant overlap with the Melampsora-infection response. Transcriptome analysis also indicated that PtWRKY23 affects redox homeostasis and cell wall-related metabolism. Experiment Overall Design: Leaf discs from wildtype, WRKY23-overexpressor and WRKY23-RNAi plants were inoculated with Melampsora medusae f. sp. tremuloidae (Mmt) or were mock-inoculated (control). At five days post-inoculation, tissues were harvested for RNA extraction and hybridization on Affymetrix GeneChipsÂ®. Three biological replications were performed for each line and each inoculation or mock-inoculation. A total of 15 Affymetrix GeneChipsÂ® were used in this study. Data for the Mmt-inoculated WRKY23-RNAi plants were processed separately from the data for the WRKY23-overexpressor plants. The raw data for the Mmt-inoculated wildtype plants were the same in both cases. Normalized data for the Mmt-inoculated wildtype plants that were processed with the Mmt-inoculated WRKY23-RNAi plants are denoted with an "a" suffix. Experiment Overall Design: This part of the study includes 6 RNA samples consisting of three biological replicates for the Mmt-inoculated WT and WRKY23-RNAi (underexpressing) plants.

ORGANISM(S): Populus tremula x Populus alba

SUBMITTER: Armand SÃ©guin

PROVIDER: E-GEOD-16419 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Transcription profiling of poplar WRKY23 reveals a regulatory role in defense: WRKY23-overexpressor

Project description:To investigate the function of poplar WRKY23, we generated PtWRKY23-overexpressing and -underexpressing (RNAi) plants. Transgenic plants were inoculated with Melampsora rust or mock-inoculated for assessment of rust-resistance and for gene expression profiling using the poplar Affymetrix GeneChipÂ® to study the consequences of PtWRKY23 overexpression and underexpression. Transcriptome analysis of PtWRKY23 overexpressors revealed a significant overlap with the Melampsora-infection response. Transcriptome analysis also indicated that PtWRKY23 affects redox homeostasis and cell wall-related metabolism. Experiment Overall Design: Leaf discs from wildtype, WRKY23-overexpressor and WRKY23-RNAi plants were inoculated with Melampsora medusae f. sp. tremuloidae (Mmt) or were mock-inoculated (control). At five days post-inoculation, tissues were harvested for RNA extraction and hybridization to Affymetrix GeneChipsÂ®. Three biological replications were performed for each line and each inoculation or mock-inoculation. A total of 15 Affymetrix GeneChipsÂ® were used in this study. Data for the WRKY23-overexpressor plants were processed separately from the data for the Mmt-inoculated WRKY23-RNAi plants. The raw data for the Mmt-inoculated wildtype plants were the same in both cases. Experiment Overall Design: This part of the study includes 12 RNA samples consisting of three biological replicates for each of the mock-inoculated or Mmt-inoculated WT and WRKY23-overexpressing plants.

2009-08-09 | E-GEOD-16417 | biostudies-arrayexpress

Expression profiling and functional analysis of poplar WRKY23 reveals a regulatory role in defense: WRKY23-RNAi

2009-08-01 | GSE16419 | GEO

Expression profiling and functional analysis of poplar WRKY23 reveals a regulatory role in defense: WRKY23-overexpressor

2009-08-01 | GSE16417 | GEO

Transcription analysis by RNAseq of adult wheat after inoculation with Puccinia graminis f. sp. tritici

Project description:The RNA sequencing analysis was undertaken to investigate the transcriptomic changes in adult wheat inoculated with Puccinia graminis f. sp. tritici the causal agent of stem rust disease. The project firstly aims to compare gene expression in one susceptible wheat line with two wheat lines exhibiting adult plant resistance to the stem rust. Secondly, the project aims to examine the temporal changes in gene expression in wheat after inoculation. Wheat plants was grown until maturity under greenhouse conditions. Plants were inoculated with Puccinia graminis f. sp. tritici and the flag leaf sheath sampled for RNA sequencing. The project aims to give essential insight into the adult plant resistance response in wheat to Puccinia graminis f. sp. tritici inoculation.

2022-10-30 | E-MTAB-9681 | biostudies-arrayexpress

Identification of rust-induced genes in poplar using Populus 2.0 NimbleGen oligoarray

Project description:Oligoarray expression profiling was carried out in poplar leaves upon infection with rust in order to identify genes expressed during tree defense response. For this purpose, we inoculated detached leaves of the interamerican hybrid poplar Populus trichocarpa x Populus deltoides 'Beaupré' grown in greenhouse either with spores of avirulent strain 93ID6 (incompatible interaction I48) or spores of virulent strain 98AG31 (compatible interaction C48) of the pathogenic rust fungus Melampsora larici-populina. Besides, we mock-inoculated 'Beaupré' leaves with water (control condition, T48). Detached leaves were maintained in vitro in controled conditions to allow fungal infection and colonization of plant tissue. Leaves were sampled 48 hours post-inoculation after that the fungus attempt to penetrate plant cells in mesophyll. Keywords: Plant tissue infection, Plant defense response, Oligonucleotide array

2007-03-15 | GSE7098 | GEO

Target genes associated to siRNA mediated repression of Pea3 and Erm transcription factors in mammary cancerous cells

Project description:Pea3 and Erm are transcription factors belonging to the ETS family which are involved in tumorigenesis, particularly mammary oncogenesis. To elucidate the mechanism of Pea3/Erm induced tumorigenesis in mouse mammary cancerous MMT cells (ATCC CCL-51), we have down-regulated Pea3 or Erm expression in this cell line by using small interfering RNA and performed a microarray analysis using Applied Biosystems AB1700 technology to define target associated genes. Keywords: Comparative transcriptome hybridization, small interfering RNA, mouse mammary cancerous cells Mouse mammary cancerous MMT cells (ATCC CCL-51) have been transfected with a siRNA control, an erm-directed siRNA or an pea3-directed siRNA. MMT cells were also used to determine the modifications due to the transfection protocol. These conditions correspond to four sample. Two biological replicate were used for the MMT cells (control sample), three biological replicates were used for the siRNA control (control sample) and erm-directed siRNA (reference sample) MMT cells and four biological replicates were used for the pea3-directed siRNA (reference sample) MMT cells. Transcriptome profiles (28.218 validated mouse genes) were acquired using Applied Biosystems AB1700 technology, which make use of chemiluminescence-based detection chemistry.

2008-08-01 | E-GEOD-6588 | biostudies-arrayexpress

Identification of rust-induced genes in poplar using Populus PICME 28K cDNA microarrays

Project description:Microarray expression profiling was carried out in poplar leaves upon infection with rust in order to identify genes expressed during tree defense response. For this purpose, we inoculated detached leaves of the interamerican hybrid poplar Populus trichocarpa x Populus deltoides 'Beaupré' grown in greenhouse with spores of avirulent strain 93ID6 of the pathogenic rust fungus Melampsora larici-populina (incompatible interaction, I48). Besides, we mock-inoculated 'Beaupré' leaves with water (control condition, T48). Detached leaves were maintained in vitro in controled conditions to allow fungal infection and colonization of plant tissue. Leaves were sampled 48 hours post-inoculation after that the fungus attempt to penetrate plant cells in mesophyll. Competitive hybridization between transcripts of incompatible interaction (I48) and control condition (T48) was done on Populus PICME 28K cDNA microarray. Keywords: Time-course infection of plant tissue, defense response, cDNA microarray

2007-03-15 | GSE7049 | GEO

Dynamic Reconfiguration of Switchgrass Proteomes in Response to Rust (Puccinia novopanici) Infection

Project description:Switchgrass (Panicum virgatum) can be infected by the rust pathogen (Puccinia novopanici) resulting in lowering biomass yields and quality. Here, two cultivars with divergent rust resistance were used to evaluate the dynamic changes in proteomes following rust inoculation. Label-free quantitative proteomics was conducted on leaf extracts harvested from a susceptible cultivar Summer at 7, 11, and 18 DAI, and leaves collected at 18 DAI from controls and infected plants of the more resistant cultivar Kanlow.

2023-10-24 | PXD036978 | Pride

Identification of rust-induced genes in poplar using Populus 4.6K cDNA macroarray

Project description:cDNA macroarray expression profiling was carried out in poplar leaves upon infection with rust in order to identify genes expressed during tree defense response. For this purpose, we inoculated detached leaves of the interamerican hybrid poplar Populus trichocarpa x Populus deltoides 'Beaupré' grown in greenhouse either with spores of avirulent strain 93ID6 (incompatible interaction, I) or spores of virulent strain 98AG31 (compatible interaction, C) of the pathogenic rust fungus Melampsora larici-populina. Besides, we mock-inoculated 'Beaupré' leaves with water (control condition, T). Detached leaves were maintained in vitro in controled conditions to allow fungal infection and colonization of plant tissue. Leaves were sampled at 12, 24 and 48 hours post-inoculation (hpi) in a time-course experiment before (12 hpi) and after (24 and 48 hpi) that the fungus attempt to penetrate plant cells in mesophyll. Keywords: Plant tissue infection, Plant defense response, cDNA macroarray

2007-03-15 | GSE7121 | GEO

Network analysis combining proteomics and metabolomics reveals new insights into early responses of Eucalyptus grandis during rust infection.

Project description:Eucalyptus rust is caused by the biotrophic fungus, Austropuccinia psidii, which affects commercial plantations of Eucalyptus, a major raw material for the pulp and paper industry in Brazil. Aiming to uncover the molecular mechanisms involved in rust resistance and susceptibility in Eucalyptus grandis, we used epifluorescence microscopy to follow the fungus development inside the leaves of two contrasting half-sibling genotypes (rust-resistance and rust-susceptible), to determine the time-course for comparative metabolomic and proteomic analyses in plantlets artificially inoculated with rust. Within 24 hours of complete fungal invasion, a total of 709 plant metabolites showed that the rust-resistant genotype suppressed many metabolites 6 hours after inoculation (hai), with responses being progressively induced after 12 hai. In contrast, the rust-susceptible genotype displayed an alternated metabolite response to infection, which culminated in a strong suppression at 24 hai. Multivariate analyses of genotypes and time points were used to select 16 differential metabolites chemically classified as flavonoids, benzenoids and other compounds. Applying the Weighted Gene Co-Expression Network Analysis (WGCNA), rust-resistant and rust-susceptible genotypes had, respectively, 871 and 852 proteins grouped into 14 and 13 modules, of which 10 and 7 protein modules were significantly correlated to the selected metabolites. Functional analyses revealed roles for oxidative-dependent responses leading to temporal activity of metabolites and proteins after 12 hai in rust-resistance, while the initial over-accumulation of metabolites and correlated proteins caused a lack of progressive response after 12 hai in rust-susceptible genotype. This study provides a brief understand on the temporal divergences of resistant and susceptible molecular responses of E. grandis plants to rust.

2021-09-09 | PXD021280 | Pride

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