Unknown,Transcriptomics,Genomics,Proteomics

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Study of expression changes during RPS4-mediated resistance in Arabidopsis using a temperature-inducible system

ABSTRACT: Innate immune responses of plant cells confer the first line of defence against pathogens. Signals generated by activated receptors are integrated inside the cell and converge on transcriptional programmes in the nucleus. The Arabidopsis Toll-related intracellular receptor RPS4 operates inside nuclei to trigger resistance to Pseudomonas bacteria expressing AvrRps4 and defence gene reprogramming through the stress response regulator, EDS1. In this immune response, RPS4 cooperates genetically with RRS1 encoding a nuclear TIR-NB-LRR receptor with an additional C-terminal ‘WRKY’ DNA-binding domain. Using transgenic Arabidopsis plants constitutively expressing RPS4 (35S:RPS4), an EDS1-dependent immune response can be turned on rapidly and synchronously in leaf cells after a switch from high (28°C) to moderate (19°C) temperature. In order to determine the relative contributions of RRS1 and EDS1 to temperature-conditioned 35S:RPS4-HS transcriptional reprogramming, we performed gene expression microarray analysis of 35S:RPS4-HS, 35S:RPS4-HS rrs1-11 and 35S:RPS4-HS eds1-2 leaf mRNAs before and after temperature shift. We used transgenic Arabidopsis plants over-expressing RPS4 in EDS1 WT, eds1-2 or rrs1-11 mutant backgrounds. 35S:RPS4, 35S:RPS4 eds1-2 and 35S:RPS4 rrs1-11 plants were grown at 28°C for 3.5 weeks, and subsequently shifted to 19°C. Samples were collected before shift (0h) and 2, 8 and 24h after shift, in triplicates.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Kenichi Tsuda

PROVIDER: E-GEOD-50019 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Arabidopsis TNL-WRKY domain receptor RRS1 contributes to temperature-conditioned RPS4 auto-immunity.

Heidrich Katharina K Tsuda Kenichi K Blanvillain-Baufumé Servane S Wirthmueller Lennart L Bautor Jaqueline J Parker Jane E JE

Frontiers in plant science 20131017

In plant effector-triggered immunity (ETI), intracellular nucleotide binding-leucine rich repeat (NLR) receptors are activated by specific pathogen effectors. The Arabidopsis TIR (Toll-Interleukin-1 receptor domain)-NLR (denoted TNL) gene pair, RPS4 and RRS1, confers resistance to Pseudomonas syringae pv tomato (Pst) strain DC3000 expressing the Type III-secreted effector, AvrRps4. Nuclear accumulation of AvrRps4, RPS4, and the TNL resistance regulator EDS1 is necessary for ETI. RRS1 possesses a ...[more]

PMID: 24146667

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Project description:Arabidopsis thaliana cell suspension cultures (ACSC) were subjected to 30-min, mild chemical treatments with three different singlet oxygen elicitors at low-medium light conditions (150 µE m–2 s–1) with the aim of getting a better understanding of singlet oxygen-mediated defence responses in plants. The three elicitors Indigo Carmine (IC), Methylene Violet (MV) and Rose Bengal (RB) at a concentration of 0.5 µM were chosen because they exhibited different abilities to permeate the plasma membrane and to accumulate in the cell soma or organelles such as chloroplasts. In addition, ACSC were treated with 500 µM H2O2 for comparison. Confocal image analysis of Arabidopsis cells revealed that IC was not retained in cells, whereas MV and RB permeated the plasma membrane and accumulated in the chloroplast envelope and inside chloroplasts, respectively. As a consequence of their different cellular location, the physiological, transcriptional and photosynthetic responses of Arabidopsis cells to singlet oxygen production varied from each other and the activation of programmed cell death (PCD) was observed in ACSC treated with 0.5 µM RB, but not with the other elicitor nor with 500 µM H2O2. The role of chloroplasts in the activation of PCD was further investigated when this physiological response was analyzed in dark-grown cell cultures containing undifferentiated plastids. Interestingly, PCD was only activated in light-grown, but not in dark-grown, Arabidopsis cell cultures, suggesting that singlet oxygen-mediated defence responses were initiated inside chloroplasts. Genome-wide transcriptional profile analyses were performed as well and the results proved that there were only statistically significant changes in the transcript expression of light-grown ACSC treated with 0.5 µM RB and 500 µM H2O2, but not with IC nor with MV. Functional enrichment analyses revealed that GO/Biological process terms associated with defence responses were common in the treatments with 0.5 µM RB and 500 µM H2O2; however, resistance response to pathogen and PCD terms were only significantly over-represented in the RB treatment. Moreover, the analysis of the up-regulated transcripts in ACSC treated with 0.5 µM RB brought out that both specific markers for singlet oxygen from the conditional fluorescence (flu) mutant of Arabidopsis and transcripts with a key role in hormone-activated PCD (i.e. ethylene and jasmonic acid) were present, although there was no evidence for the up-regulation of EDS1 encoding the ENHANCED DISEASE SUSCEPTIBILITY PROTEIN 1. Finally, a co-regulation analysis proved that ACSC treated with 0.5 µM RB exhibited higher correlation with the flu family mutants than with other singlet oxygen producer mutants of Arabidopsis or wild-type plants of Arabidopsis subjected to high light treatments, where singlet oxygen was produced in photosystem II and an acclimatory response was activated instead of PCD. 18 Samples total. Six Samples are controls, 3 Samples each treated with Rose Bengal, Hydrogen Peroxide, Methyl Violet, and Indigo Carmine.

Dataset Information

Study of expression changes during RPS4-mediated resistance in Arabidopsis using a temperature-inducible system

Publications

Arabidopsis TNL-WRKY domain receptor RRS1 contributes to temperature-conditioned RPS4 auto-immunity.

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