Project description:LC-MS/MS data were collected from uninfected and parallel Golovinomyces orontii MGH1- infected Arabidopsis thaliana leaf tissue (leaves 7-9) at 12 days post inoculation to understand the manipulation of host lipid metabolism by the powdery mildew.
Project description:We performed RNA-sequencing of Golovinomyces orontii-infected Arabidopsis leaves of wild type, the double or triple mutants of AtMLKLs to examine the role of AtMLKLs in response to the powdery mildew fungus.
Project description:Obligate biotrophs such as the virulent powdery mildew Golovinomyces orontii alter plant host cellular architecture, metabolism, and defense in order to acquire nutrients while suppressing cell death and senescence. G. orontii exclusively infects epidermal cells of Arabidopsis with clearly defined stages of infection. Host factors mediating the powdery mildew (PM) interaction are often expressed in the mesophyll cells underlying the infected epidermal cells. Therefore, in order to identify Arabidopsis processes and regulators mediating this PM interaction, we used UV laser microdissection to isolate cells at the PM infection site for global expression profiling. As part of this process, we optimized and/or developed novel tissue preparation, RNA extraction and amplification, and quality control protocols resulting in highly correlated biological replicate data. We focused on the growth and reproduction stage of the PM infection (5 days post infection) when the number of reproductive structures, conidiophores, can be quantified. Site-specific profiling increased our sensitivity dramatically, allowing us to identify specific processes, process components, and their putative regulators hidden in previous whole leaf global expression analyses. For example, the known cell cycle regulator MYB3R4 exhibits altered expression at the site of infection, as do a subset of cell-cycle-associated genes. Furthermore, null myb3r4 mutants exhibit enhanced resistance to PM with reduced conidiophores per colony, suggesting cell cycle control plays an important role in the PM interaction. Experiment Overall Design: Arabidopsis whole leaves from wild type Columbia-0 and enhanced disease susceptibility mutant eds16-1, a null isochorismate synthase 1 (At1g74710) mutant were harvested at 5 days after Golovinomyces orontii infection, microwave-fixed, paraffin-embedded and sectioned. Groups of epidermal and mesophyll cells (~20 cells/group) surrounding the G. orontii infected epidermal cell were cut using a Leica AS laser microdissection (LMD) system. In parallel, groups of epidermal and mesophyll cells were collected from uninfected leaves at 5 days from wild type Arabidopsis. LMD-isolated cells, whole leaf, whole leaf amplified and tissue scrape samples were used for RNA extraction and hybridization to Affymetrix Arabidopsis ATH1 microarrays. Gene expression profiles were obtained for wild type from all samples and for ics1 mutant from LMD infected samples. The experiment includes 2 biological replicates.
Project description:Salicylic acid (SA) is a critical molecule mediating plant innate immunity with an important role limiting the growth and reproduction of the virulent powdery mildew (PM) Golovinomyces orontii on Arabidopsis thaliana. To investigate this later phase of the PM interaction, and the role played by SA, we performed replicated global expression profiling for wild type and SA biosynthetic mutant ics1 Arabidopsis from 0 to 7 days post infection. We found that ICS1-impacted genes comprise 3.8% of profiled genes with known molecular markers of Arabidopsis defense ranked very highly by the multivariate empirical Bayes statistic (T2 statistic ((Tai and Speed, 2006)). Functional analyses of T2-selected genes identified statistically significant PM-impacted processes including photosynthesis, cell wall modification, and alkaloid metabolism that are ICS1-independent. ICS1-impacted processes include redox, vacuolar transport/secretion, and signaling. Our data also supports a role for ICS1 (SA) in iron and calcium homeostasis and identifies components of SA crosstalk with other phytohormones. Through our analysis, 39 novel PMâimpacted transcriptional regulators were identified. Insertion mutants in one of these regulators, PUX2, results in significantly reduced reproduction of the powdery mildew in a cell death independent manner. Though little is known about PUX2, PUX1 acts as a negative regulator of Arabidopsis CDC48 (Rancour et al., 2004; Park et al., 2007), an essential AAA-ATPase chaperone that mediates diverse cellular activities including homotypic fusion of ER and Golgi membranes, ER-associated protein degradation, cell cycle progression, and apoptosis. Future work will elucidate the functional role of the novel regulator PUX2 in PM resistance. Experiment Overall Design: Arabidopsis whole leaves were harvested at 6h, 1 day, 3 days, 5 days and 7 days after Golovinomyces orontii infection for RNA extraction and hybridization to Affymetrix Arabidopsis ATH1 microarrays. Gene expression profiles were obtained for wild type Columbia-0 and enhanced disease susceptibility mutant eds16-1, a null isochorismate synthase 1 (At1g74710) mutant. In parallel, uninfected samples were collected at 0 hr and 7days from wild type and mutant plants. The experiment includes 4 biological replicates.
Project description:Comparative transcriptomic analysis of Arabidopsis thaliana yda11 plants (in Col-0 background), and wild-type plants (Col-0) non-infected or infected with the necrotrophic fungal pathogen Plectosphaerella cucumerina BMM (PcBMM)
Project description:The edr1 mutant of Arabidopsis thaliana displays enhanced resistance to the powdery mildew Golovinomyces cichoracearum, resulting in cell death and an absence of visible disease symptoms. To better characterize and understand the defense response of edr1, a time course of early signaling responses was performed after inoculation with powdery mildew and compared to the responses of wild-type Col-0. These time points represent early stages in the infection process, before any signs of susceptibility or resistance are visible.
Project description:Here, we sought to describe qualitative and quantitative changes in the global gene expression profiles of susceptible Arabidopsis plants supporting the development of G. cichoracearum haustoria. We analyzed the features of compatibility at this infection stage, and further evaluated the contribution of the SA- and JA/ET-dependent defense signaling pathways in the pathogen-induced responses by comparing responses in infected wild-type, npr1-1, and jar1-1 plants. Our findings collectively contribute to knowledge regarding early host cell alterations generated in response to attack by this virulent obligate biotrophic fungus. Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Disease State: plant infected/uninfected with Golovinomyces orontii Genotype: wt/mutant strains Keywords: Logical Set
Project description:Here, we sought to describe qualitative and quantitative changes in the global gene expression profiles of susceptible Arabidopsis plants supporting the development of G. cichoracearum haustoria. We analyzed the features of compatibility at this infection stage, and further evaluated the contribution of the SA- and JA/ET-dependent defense signaling pathways in the pathogen-induced responses by comparing responses in infected wild-type, npr1-1, and jar1-1 plants. Our findings collectively contribute to knowledge regarding early host cell alterations generated in response to attack by this virulent obligate biotrophic fungus. Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Disease State: plant infected/uninfected with Golovinomyces orontii Genotype: wt/mutant strains Keywords: Logical Set Computed
Project description:LC-MS/MS data were collected from uninfected and parallel Golovinomyces orontii MGH1- infected Arabidopsis thaliana leaf tissue (leaves 7-9) at 12 days post inoculation to understand the manipulation of host lipid metabolism by the powdery mildew.
Project description:rs12-08_cyp715a1 - col-0 vs cyp715a1 - The microarray analysis is part of a project aimed at characterizing the function of the cytochrome P450 CYP715A1 in Arabidopsis thaliana. - Flower buds of Arabidopsis Col-0 (wild-type) and cyp715A1 mutant were harvested for a comparative analysis of their transcriptomes.