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Fireman vs wt erwinia infection-Impact of the fireman mutation in the Erwinia amylovora response of Arabidopsis

ABSTRACT: adt09-02_fireman - fireman vs wt erwinia infection - A meta-analysis ot the CATMA transcriptomic data has shown that an orphan gene of Arabidopsis: AT5E39460, recently remaned AT5G 44585, is highly responsive to biotic stresses. An homozygous mutant of this gene, named fireman has been produced and displays less cell death symptoms in comparison to the wild type in response to Erwinia amylovora infection. The purpose of this transcriptomic analysis is to understand which pathways are altered in this mutant versus the wild-type in response to this phytopathogenic bacteria. - 3 inoculated leaves of 5 weeks old plants, per plant, have been harvested 24h after inoculation with H20 or E. amylovora, strain 1430. Keywords: normal vs disease comparison, normal vs transgenic comparaison

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE22683 | GEO | 2011/07/15

SECONDARY ACCESSION(S): PRJNA128337

REPOSITORIES: GEO

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fireman vs wt erwinia infection-Impact of the fireman mutation in the Erwinia amylovora response of Arabidopsis

Project description:adt09-02_fireman - fireman vs wt erwinia infection - A meta-analysis ot the CATMA transcriptomic data has shown that an orphan gene of Arabidopsis: AT5E39460, recently remaned AT5G 44585, is highly responsive to biotic stresses. An homozygous mutant of this gene, named fireman has been produced and displays less cell death symptoms in comparison to the wild type in response to Erwinia amylovora infection. The purpose of this transcriptomic analysis is to understand which pathways are altered in this mutant versus the wild-type in response to this phytopathogenic bacteria. - 3 inoculated leaves of 5 weeks old plants, per plant, have been harvested 24h after inoculation with H20 or E. amylovora, strain 1430. Keywords: normal vs disease comparison, normal vs transgenic comparaison 8 dye-swap - CATMA arrays

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Project description:Microarray analysis was used to identify genes that were controlled by AmyR in minimal and on immature pear fruits. Consistent with amylovoran production, an inverse correlation was observed between amyR expression and the expression level of amylovoran biosynthetic genes in liquid media. Interestingly, over-expression of AmyR suppressed the expression of type III secretion system genes including hrpA, hrpN and dspEF after pear fruit infection. Consistent with levan production and swarming motility, levasucrase and flagellar genes were both down-regulated both in the amyR mutant and over-expression strains in liquid media. Together, our results suggest that AmyR plays an important role in regulating bacterial exopolysaccharide production and virulence in E. amylovora. A total of 14 samples were analyzed in two conditions: For the in vitro condition (MBMA medium + 1% sorbitol), Erwinia amylovora wild type strain (2 replicates); E. amylovora amyR mutant strain (3 replicates); E. amylovora amyR over-expression strain (3 replicates); For the in vivo condition (on wounded immature pear fruits), Erwinia amylovora wild type strain (2 replicates); E. amylovora amyR mutant strain (2 replicates); E. amylovora amyR over-expression strain (2 replicates).

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Project description:The bacterial pathogen Erwinia amylovora is the causal agent of fire blight, an economically significant disease of apple and pear. Disease initiation by E. amylovora requires the translocation of effector proteins into host cells by the hypersensitive response and pathogenicity (hrp) type III secretion system (T3SS). The alternate sigma factor HrpL positively regulates the transcription of structural and translocated components of the T3SS via hrp promoter elements. To characterize genome-wide hrpL-dependent gene expression in E. amylovora Ea1189, wild-type and Ea1189hrpL strains were cultured in hrp-inducing minimal medium, and total RNA was compared using a custom microarray designed to represent the annotated genes of E. amylovora ATCC 49946. Results revealed 24 genes differentially-regulated in Ea1189hrpL compared to Ea1189 with fold-change expression ratios greater than 1.5; of these, the expression of 19 genes was up-regulated while five genes exhibited negative regulation. To expand our understanding of the HrpL regulon and to elucidate direct versus indirect hrpL-mediated effects on gene expression, the genome of E. amylovora ATCC 49946 was examined in silico using a hidden Markov model assembled from known Erwinia spp. hrp promoters. This technique identified 21 putative type III novel hrp promoters; of these, 8+ were validated with quantitative polymerase chain reaction based on expression analyses. In total, hrpL-regulated genes encode all known components of the hrp T3SS and 5 putative type III effectors. Eight genes displayed apparent indirect hrpL regulation suggesting that the hrpL regulon is connected to downstream signaling networks. Construction of deletion mutants of three novel hrpL-regulated genes has resulted in the identification of additional virulence factors in E. amylovora as well as mutants displaying abnormal motility and biofilm phenotypes.

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