Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of Arabidopsis thaliana Col‑0 wild-type, Col-0 nrpe1-11, and Col-0 ros1-4 at 48 and 72 hours after inoculation with Hyaloperonospora arabidopsidis strain WACO9.

ABSTRACT: The experiment was designed to gain more insight into the role of DNA (de)methylation in the regulation of Arabidopsis thaliana gene transcription during a compatible interaction with the downy mildew pathogen Hyaloperonospora arabidopsidis strain WACO9 (Hpa). For this study, comparisons were made between Col-0 (wild-type), nrpe1-11 (which is affected in DNA methylation and globally hypo-methylated) and ros1-4 (impaired in DNA de-methylation and globally hyper-methylated). The latter two mutants have opposite phenotypes with respect to basal resistance and salicylic acid-dependent defence against Hpa. Samples were taken at 48 and 72 hours after spray-inoculation with either 10^5 spores ml^-1 Hyaloperonospora arabidopsidis strain WACO9 in water or water only (mock).

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Joost Stassen

PROVIDER: E-MTAB-3963 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Plants have evolved strong innate immunity mechanisms, but successful pathogens evade or suppress plant immunity via effectors delivered into the plant cell. Hyaloperonospora arabidopsidis (Hpa) causes downy mildew on Arabidopsis thaliana, and a genome sequence is available for isolate Emoy2. Here, we exploit the availability of genome sequences for Hpa and Arabidopsis to measure gene-expression changes in both Hpa and Arabidopsis simultaneously during infection. Using a high-throughput cDNA tag sequencing method, we reveal expression patterns of Hpa predicted effectors and Arabidopsis genes in compatible and incompatible interactions, and promoter elements associated with Hpa genes expressed during infection. By resequencing Hpa isolate Waco9, we found it evades Arabidopsis resistance gene RPP1 through deletion of the cognate recognized effector ATR1. Arabidopsis salicylic acid (SA)-responsive genes including PR1 were activated not only at early time points in the incompatible interaction but also at late time points in the compatible interaction. By histochemical analysis, we found that Hpa suppresses SA-inducible PR1 expression, specifically in the haustoriated cells into which host-translocated effectors are delivered, but not in non-haustoriated adjacent cells. Finally, we found a highly-expressed Hpa effector candidate that suppresses responsiveness to SA. As this approach can be easily applied to host-pathogen interactions for which both host and pathogen genome sequences are available, this work opens the door towards transcriptome studies in infection biology that should help unravel pathogen infection strategies and the mechanisms by which host defense responses are overcome. Three weeks old Arabidopsis (Col-0) plants were inoculated with either the avirulent isolate Emoy2 (incompatible interaction) or the virulent isolate Waco9 (compatible interaction) of Hpa, and infected plants were harvested at 1, 3 and 5 days post-inoculation (dpi) for total RNA extraction. mRNA profiles were generated by deep sequencing on Illumina GAIIx using EXPRSS tag-seq protocol. Each biological replicate set of samples were sequenced as one batch (biorep1, biorep2 and biorep3 in filenames) and each batch was sequenced in 4 individual Illumina flowcell lanes (lane1 to lane4 in filenames). Four sequecing lanes of each three biological repliates resulted in 12 sequencing libraries. Emoy2 1, 3 and 5 dpi samples and Waco9 1 dpi samples were made two different codes (lib1 and lib2 in filenames) to generate higher depth of sequencing data. Each biological replicate set of samples were sequenced as one batch and each batch was sequenced in 4 individual Illumina flowcell lanes. Four sequecing lanes of each three biological repliates resulted in 12 sequencing libraries. 'The unassigned read_*' samples are for 'nobarcode_biorep[x]_lanne[x].fq' file containing sequences unassigned to any barcode from respective bioreplicate sequencing lanes.

Dataset Information

Transcription profiling of Arabidopsis thaliana Col‑0 wild-type, Col-0 nrpe1-11, and Col-0 ros1-4 at 48 and 72 hours after inoculation with Hyaloperonospora arabidopsidis strain WACO9.

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