Project description:We used Arabidopsis full-genome microarrays to characterize plant transcript accumulations at different stages of infection with the biotrophic oomycete downy mildew pathogen, Hyaloperonospora arabidopsidis : initiation (< 1 dpi) and maintenance of infection (> 4 dpi). In two independent experiments, cotyledons from the ecotype Wassilewskija (WS) were inoculated with water, or with Hyaloperonospora arabidopsidis to establish a compatible interaction. Affymetrix ATH1 microarrays were used to profile Arabidopsis transcript accumulations at the initiation (mixed samples at 8 and 24 hours post inoculation, hpi; early stage) and maintenance (mixed samples at 4 and 6 days post inoculation; late stage) of the compatible interaction.
Project description:UV-B (280-320 nm) exposure causes serious damage in plants, limiting their growth and survival, effects that are partly counteracted by repair mechanisms active in plants receiving accompanying visible radiation. Though no particular UV-B receptor has been identified to date, there is strong evidence to indicate that certain aspects of UV-B perception are receptor-mediated. Investigations of down-stream signalling events have thus far indicated broad similarities to pathogen-induced defence responses in plants. In order to identify genes in Arabidopsis that may be up- or down- regulated specifically in response to UV-B exposure and compare them to genes whose expression is altered in plants challenged by an avirulent isolate of Peronospora parasitica (downy mildew), we propose to analyse the transcriptional profiles for the following treatments:; 1. UV-B Responses; "A-1" Columbia (Col-0) exposed to supplementary UV-B/UV-A* with a background of low photosynthetically active radiation (PAR of 20 micromol m-2 s-1) for 1.5 photoperiods (photoperiod = 12h). [UV-B treatment]; "A-2" Col-0 exposed to supplementary UV-A and low PAR for 1.5 photoperiods [control for UV-B treatment]; "A-3" Col-0 exposed to visible light only (low PAR) (no UV) for 1.5 photoperiods [control for UV effects in general].* There are no pure sources of UV-B light available. 2. Pathogen Responses; "A-4" Col-0 spray-inoculated with P. parasitica isolate HIKS-1 (recognised by the R-gene RPP7). After spraying, plants were kept covered in plant propagators and transferred to an 18 degreeC growth chamber. !Samples for RNA extraction were taken 72h after inoculation. "A-5" The viability of spores was also checked by parallel spraying of the susceptible mutant, Col-rpp7. [pathogen treatment]; "A-6" Col-0 mock treated with water, covered and transferred to an 18 degree C growth chamber, 72h prior to sampling. [control for pathogen treatment]; In all experiments, we are using RNA from leaves taken at the same time of day (6 h into the 12 h photoperiod) from 4.5-week old plants grown under 12h photoperiod. All treatments were normalised against PR-1 expression levels to ensure comparability between UV-B and pathogen treatments. Due to the difficulty in distinguishing between local and systemic induced responses in UV-B treated plants, we are using RNA from whole rosettes for both the UV-B and pathogen treatment for better comparability among treatments. The degree of similarity between these two sets of transcriptional changes will complement and help interpret our experimental data on changes in resistance to pathogens in plants pre-treated with UV-B. Moreover, the data set obtained would allow for identification of UV-B specific changes in gene expression including cis-acting UV-B-responsive promoter elements. Experimenter name = Julia Brueggemann; Experimenter phone = 01789 470 382; Experimenter fax = 01789 470 552; Experimenter address = Horticulture Research International; Experimenter address = Wellesbourne; Experimenter address = Warwickshire; Experimenter zip/postal_code = CV35 9EF; Experimenter country = UK Experiment Overall Design: 6 samples were used in this experiment
Project description:We used Arabidopsis full-genome microarrays to characterize plant transcript accumulations in wild-type plants and pskr1-5 mutants, 3 days after water treatment and inoculation with the biotrophic oomycete downy mildew pathogen, Hyaloperonospora arabidopsidis. In two independent experiments, cotyledons from the wild-type Wassilewskija (WS) ecotype and from the pskr1-5 mutant were treated with water, or inoculated with the H. arabidopsidis isolate Emwa1 to establish a compatible interaction. Affymetrix ATH1 microarrays were used to profile Arabidopsis transcript accumulations at 3 days after onset of treatment.
Project description:Gibberellin (GA) promotes plant growth by destabilizing DELLA proteins. DELLA proteins integrate multiple hormonal and environmental stress responses. We investigated the role of GA and DELLA proteins in plant defence. We used microarrays to detail the global programme of gene expression controlled by DELLA proteins and identified distinct classes of differentially regulated genes in response to pathogens, hormones or pathogen elicitors. Experiment Overall Design: Five weeks old short day grown Arabidopsis leaf discs were used to treat with flg22 and samples were collected after 1 hour and 2 hour time points. For Alternaria brassicicola, five weeks old plants were drop inoculated with 4x 5µl droplets of Alternaria brassicicola spores (5x105 spores/ml) and samples were collected 3 days post inoculation. Five weeks old plants were infiltrated with Pst DC3000 (2x105cfu/ml) bacteria and samples were collected after 12 hours post infiltration. For methyl jasmonate treatments, five weeks old plants were sprayed with 10µM Methyl Jasmonate solution, covered with plastic bags and samples were collected after one hour.
Project description:We used Arabidopsis full-genome microarrays to characterize plant transcript accumulations in map65-3 and ugt76b1 mutants, 3 days after water treatment and inoculation with the biotrophic oomycete downy mildew pathogen, Hyaloperonospora arabidopsidis (Hpa) In two independent experiments, cotyledons from the wild-type Wassilewskija (WS) ecotype, and from the map65-3 and ugt76b1 mutants were treated with water, or inoculated with the Hpa isolate Emwa1 to establish a compatible interaction. Affymetrix ATH1 microarrays were used to profile Arabidopsis transcript accumulations at 3 days after onset of treatment. Data from the water-treated and Hpa-infected wild-type were previously deposited as GSM914964, GSM914965, GSM914966, and GSM914967. The wild-type data, and the data from the map65-3 and ugt76b1 mutant presented here were established in the same set of experiments and analyses, which also involved the previously deposited pskr1-5 mutant (GSM914968, GSM914969, GSM914970, and GSM914971).
Project description:GmMYB176 is an R1 MYB transcription factor that regulates multiple genes in the isoflavonoid biosynthetic pathway thereby affecting isoflavonoid level in soybean roots. While GmMYB176 is important for isoflavonoid synthesis, it is not sufficient for the function and requires additional cofactor (s). The aim of this study was to identify GmMYB176 interactome for the regulation of isoflavonoid biosynthesis in soybean. GmMYB176 interactome was identified by Co-Immunoprecipitation using two YFP-tagged proteins baits, N’-GmMYB176-YFP-C’ and N’-YFP-GmMYB176-C’. GmMYB176 interacting proteins were identified by LC-MS/MS.
Project description:Legumes interact with nodulating bacteria that convert atmospheric nitrogen into ammonia for plant use. This nitrogen fixation takes place within root nodules that form after infection of root hairs by compatible rhizobia. Using cDNA microarrays, we monitored gene expression in soybean (Glycine max) inoculated with the nodulating bacterium Bradyrhizobium japonicum 4, 8, and 16 days after inoculation (dai), time points that coincided with nodule development and the onset of nitrogen fixation. This experiment identified several thousand genes that were differentially expressed in response to B. japonicum inoculation. Expression of 27 genes was analyzed by qRT-PCR and their expression patterns mimicked the microarray results confirming integrity of analyses. The microarray results suggest that B. japonicum reduces plant defense responses during nodule development. In addition, the data revealed a high level of regulatory complexity (transcriptional, post-transcriptional, translational, post-translational) that is likely essential for development of the symbiosis and adjustment to an altered nutritional status. Keywords = symbiosis Keywords = nodulation Keywords = rhizobium Keywords = defense Keywords = ANOVA Keywords = plant loop design, 7 samples, 7 comparison, 2 technical repeats including dye swaps, 4 biological repeats
Project description:We report the construction of an oligo-based microarray using a total of 17,510 unigenes derived from 30,675 high-quality melon ESTs. This chip is particularly enriched with genes that are expressed in fruit and during interaction with pathogens. Hybridizations for three independent experiments allowed the characterization of global gene expression profiles during fruit ripening, as well as in response to viral and fungal infections in plant cotyledons and roots, respectively. Microarray construction, statistical analyses and validation together with functional-enrichment analysis are presented in this study.
Project description:Melon (Cucumis melo L.) is the model species of the Cucurbitaceae family and an important crop, with an estimated production at 28 million tonnes in 2020. However, its yield is primarily affected by viruses. Cucumber mosaic virus (CMV) is particularly significant due to its broad host range, capable of infecting over 100 plant families. Resistance to CMV in the melon accession Songwhan Charmi (SC) is controlled by the recessive gene cmv1, which encodes the Vacuolar Protein Sorting 41, involved in vesicle transport to the vacuole. cmv1 restricts the virus to the bundle sheath cells and impedes viral access to the phloem, and this phenotype depends on the viral movement protein (MP). However, little is known about the broader cellular changes that CMV triggers in melon or the specific biological responses that facilitate or restrict the virus' entry into the phloem in susceptible and resistant varieties. We profiled the proteomes of CMV-resistant or susceptible melon genotypes inoculated with LS or FNY strains. Analysis of co-abundance networks revealed pathways involved in CMV resistance and susceptibility at different infection stages. Upon inoculation, resistant varieties have stronger changes in proteome composition compared to susceptible varieties, including an increase in lipid signalling and upregulation of phloem defence proteins. In contrast, susceptible melon plants displayed fewer proteins related to translation, photosynthesis and intracellular transport. During the systemic infection of susceptible melon plants, proteins associated with stress responses increase, such as those involved in the ER-associated degradation (ERAD) and phenylpropanoid pathways. Collectively, our study shed new light on the interactions between CMV and melon plants.