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Experiment: Effects of the cucumber mosaic virus 2b silencing suppressor protein on inducible plant defence pathways
Experiment Description
NASCArrays Experiment Reference Number: NASCARRAYS-415
In this study, we have examined the effects of the cucumber mosaic virus 2b protein on the salicylic acid (SA) and methyl jasmonate (meJa) induced defence pathways in A. thaliana. We have also investigated the effects of the 2b protein on antimycin A-induced defence, which represents a virus-specific branch of SA-induced defence. Transgenic A. thaliana (ecotype Col 0) expressing the 2b gene from CMV (strain Fny) were treated independently with SA, meJa and AA, as were wild-type controls. After 25 hours, RNA was extracted from aerial tissue for comparison of gene expression profiles in the presence and absence of the 2b protein. The 2b protein is known to suppress SA-induced resistance to plant viruses as well as the siRNA and miRNA-mediated gene silencing pathways. It is able to suppress SA-mediated induction of alternative oxidase, a gene that functions in the AA-induced virus-specific branch of SA-induced defence (Ji and Ding, 2001, Mol. Plant Microbe Interactions 14, 715-724). Furthermore, the 2b protein is the major determinant of viral symptoms in CMV and causes symptom-like alterations in the phenotype of Arabidopsis thaliana plants expressing 2b transgenes (Lewsey et al., 2007, Plant J., in press). However, its effects on the meJa-induced defence pathway, which has crosstalk with the SA-induced defence pathway, are not known.
About the ExperimenterName: | Dr Mathew Lewsey |
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Head of Lab Name: | Dr John P Carr |
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Lab:
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Institute:
| Department of Plant Sciences |
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Address: | Molecular Virology Department of Plant Sciences Downing Street Cambridge
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Postcode:
| CB2 3EA |
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Country:
| UK |
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| Telephone Number:
| 01223330215 |
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All of the data available in this website/database is free, and you
are free to do whatever you please with it. If you intend to publish
work based on any of this data, please acknowledge us, contact the
experimenter above, and either acknowledge them or use them as
co-authors in the work.
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| About this ExperimentExperiment Type:
| compound_based_treatment_design;genetic_modification_design |
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Number of Slides: | 24 |
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| Experimental Parameters:
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parameter | compound_based_treatment |
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parameter | individual_genetic_characterisics |
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Quality Control Measures Taken:
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References:
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| Other Information:
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Slides in this Experiment
Hybridisation Set: Lewsey: Effects of the cucumber mosaic virus 2b silencing suppressor protein on inducible plant defence pathways_genome
Slide: Lewsey_1-12_Fny-CMV2b-salicylic-acid_Rep2_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
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Treatment protocol | Approximately ten plants were sprayed with water containing 1mM salicylic acid and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Salicylic acid sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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Protocols for BioSource 1 |
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Slide: Lewsey_1-11_Wild-type-salicylic-acid_Rep2_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
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Treatment protocol | Approximately ten plants were sprayed with water containing 1mM salicylic acid and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction |
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Additional Organism Information:
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Sample description | Salicylic acid sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-10_Fny-CMV2b-H2O_Rep2_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
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Treatment protocol | Approximately ten plants were sprayed with water containing 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Water sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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Protocols for BioSource 1 |
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Slide: Lewsey_1-9_Wild-type-H2O_Rep2_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
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Treatment protocol | Approximately ten plants were sprayed with water containing 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Water sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-8_Fny-CMV2b-methyl-jasmonate_Rep1_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
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Treatment protocol | Approximately ten plants were sprayed with water containing 250 micromolar methyl jasmonate and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Methyl jasmonate sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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Protocols for BioSource 1 |
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Slide: Lewsey_1-7_Wild-type-methyl-jasmonate_Rep1_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 250 micromolar methyl jasmonate and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Methyl jasmonate sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-6_Fny-CMV2b-antimycin-A_Rep1_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
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Treatment protocol | Approximately ten plants were sprayed with water containing 50 micromolar antimycin A and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Antimycin A sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants.. |
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Other Information:
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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Protocols for BioSource 1 |
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Slide: Lewsey_1-5_Wild-type-antimycinA_Rep1_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 50 micromolar antimycin A and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Antimycin A sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants.. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-4_Fny-CMV2b-salicylic-acid_Rep1_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 1mM salicylic acid and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Salicylic acid sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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Protocols for BioSource 1 |
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Slide: Lewsey_1-3_Wild-type-salicylic-acid_Rep1_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 1mM salicylic acid and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction |
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Additional Organism Information:
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Sample description | Salicylic acid sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-2_Fny-CMV2b-H2O_Rep1_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Water sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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Protocols for BioSource 1 |
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Slide: Lewsey_1-1_Wild-type-H2O_Rep1_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Water sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-13_Wild-type-antimycinA_Rep2_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 50 micromolar antimycin A and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Antimycin A sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants.. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-14_Fny-CMV2b-antimycin-A_Rep2_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 50 micromolar antimycin A and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
| |
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Sample description | Antimycin A sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants.. |
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Other Information:
| |
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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Protocols for BioSource 1 |
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Slide: Lewsey_1-15_Wild-type-methyl-jasmonate_Rep2_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 250 micromolar methyl jasmonate and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
| |
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Sample description | Methyl jasmonate sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-16_Fny-CMV2b-methyl-jasmonate_Rep2_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 250 micromolar methyl jasmonate and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
| |
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Sample description | Methyl jasmonate sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
| |
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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Protocols for BioSource 1 |
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Slide: Lewsey_1-17_Wild-type-H2O_Rep3_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
| |
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Sample description | Water sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-18_Fny-CMV2b-H2O_Rep3_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
| |
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Sample description | Water sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
| |
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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Protocols for BioSource 1 |
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Slide: Lewsey_1-19_Wild-type-salicylic-acid_Rep3_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 1mM salicylic acid and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction |
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Additional Organism Information:
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Sample description | Salicylic acid sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-20_Fny-CMV2b-salicylic-acid_Rep3_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 1mM salicylic acid and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
| |
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Sample description | Salicylic acid sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
| |
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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Protocols for BioSource 1 |
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Slide: Lewsey_1-21_Wild-type-antimycinA_Rep3_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 50 micromolar antimycin A and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
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Sample description | Antimycin A sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants.. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-22_Fny-CMV2b-antimycin-A_Rep3_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 50 micromolar antimycin A and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
| |
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Sample description | Antimycin A sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants.. |
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Other Information:
| |
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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|
Protocols for BioSource 1 |
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Slide: Lewsey_1-23_Wild-type-methyl-jasmonate_Rep3_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 250 micromolar methyl jasmonate and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
| |
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Sample description | Methyl jasmonate sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Lewsey_1-24_Fny-CMV2b-methyl-jasmonate_Rep3_ATH1 | | |
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Tissue:
| all aerial tissues |
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in vivo Treatment:
| |
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Treatment protocol | Approximately ten plants were sprayed with water containing 250 micromolar methyl jasmonate and 0.1% (v/v) ethanol, 5.5 hrs into the light cycle, when plants were 6 weeks old (growth stage 3.50 to 3.70). Plant tissues were harvested 25 hours after treatment and pooled to minimise variation. The pooled tissue was then used for RNA extraction. |
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Additional Organism Information:
| |
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Sample description | Methyl jasmonate sprayed A.thaliana plants. Aerial tissue pooled from approximately ten plants. |
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Other Information:
| |
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Allele | 35S expression of Cucumber mosaic virus 2b gene |
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|
Protocols for BioSource 1 |
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Problems? Comments? Suggestions? Contact the Affymetrix Team