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Experiment: Systemic responses to Pseudomonas syringae infection
Experiment Description
NASCArrays Experiment Reference Number: NASCARRAYS-437
We used three different strains of Pseudomonas syringae pv tomato DC3000 to investigate systemic responses to infection in Arabidopsis and the development of SAR. Wildtype DC3000, the hrpA mutant and DC3000 carrying the avirulence gene avrRpm1 were syringe infiltrated into 4 week old plants at a concentration of 10e8 cfu/ml. At least 5 leaves per plant were infiltrated and at least 10 plants were pooled for each sample. Systemic, uninfected tissue was then harvested at 8, 12 and 21h after inoculation. Three independent experiments were carried out to give three biological replicates for each timepoint.
About the ExperimenterName: | Dr William Truman |
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Head of Lab Name: | Prof Murray Grant |
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Lab:
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Address: | School of Biosciences University of Exeter Geoffrey Pope Building Stocker Road Exeter
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Postcode:
| EX44QD |
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Country:
| UK |
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| Telephone Number:
| 01392269162 |
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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:
| time_series_design; pathogenicity_design |
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Number of Slides: | 27 |
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| Experimental Parameters:
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parameter | infect |
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parameter | timepoint |
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Quality Control Measures Taken:
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no-plants-pooled | 5 plants |
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References:
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| Other Information:
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Slides in this Experiment
Hybridisation Set: Truman: Systemic responses to Pseudomonas syringae infection_genome
Slide: Truman_2-1_8h-hrpA_Rep1_ATH1 | | |
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Tissue:
| leave |
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in vivo Treatment:
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treatment | Leaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-2_8h-DC3000_Rep1_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-3_8h-avrRpm1_Rep1_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-4_12h-hrpA_Rep1_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-5_12h-DC3000_Rep1_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-6_12h-avrRpm1_Rep1_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-7_21h-hrpA_Rep1_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-8_21h-DC3000_Rep1_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-9_21h-avrRpm1_Rep1_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-10_8h-hrpA_Rep2_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-11_8h-DC3000_Rep2_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-12_8h-avrRpm1_Rep2_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-13_12h-hrpA_Rep2_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-14_12h-DC3000_Rep2_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-15_12h-avrRpm1_Rep2_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-16_21h-hrpA_Rep2_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-17_21h-DC3000_Rep2_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-18_21h-avrRpm1_Rep2_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-19_8h-hrpA_Rep3_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-20_8h-DC3000_Rep3_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-21_8h-avrRpm1_Rep3_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-22_12h-hrpA_Rep3_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-23_12h-DC3000_Rep3_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 lg ml)1) and kanamycin (25 lg ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-24_12h-avrRpm1_Rep3_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-25_21h-hrpA_Rep3_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with Pseudomonas syringae pv tomato DC3000::hrpA- mutant. DC3000::hrpA- cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000::hrpA- containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin(50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-26_21h-DC3000_Rep3_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with virulent Pseudomonas syringae pv tomato DC3000. DC3000 cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the empty vector plasmid pVSP61 (Innes et al., 1993) Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_2-27_21h-avrRpm1_Rep3_ATH1 | | |
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in vivo Treatment:
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treatment | Leaves were inoculated with avirulent Pseudomonas syringae pv tomato DC3000(avrRpm1). DC3000(avrRpm1) cultures were grown overnight in 10 ml of Kings B solution (King et al., 1954) supplemented with antibiotics. Plants were inoculated with DC3000 containing the vector plasmid pVSP61 (Innes et al., 1993) carrying the avirulence gene avrRpm1. Antibiotics rifampicin (50 g ml)1) and kanamycin (25 g ml)1) were used for selection of the bacterium and plasmid, respectively. Overnight cultures were washed once in 10 mM MgCl2 and final cell densities were adjusted to OD600 0.2. Both sides of a leaf were infiltrated on their abaxial surface with a needleless 1ml syringe. At least 5 leaves per plant were inoculated and 16 plants were pooled to make this sample. |
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Additional Organism Information:
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sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested |
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Other Information:
| |
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|
Protocols for BioSource 1 |
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