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Experiment: Systemic response to avirulent bacterial infection
Experiment Description
NASCArrays Experiment Reference Number: NASCARRAYS-403
In the absence of adaptive immunity displayed by animals, plants respond locally to biotic challenge via inducible basal defense networks activated through recognition and response toconserved pathogen associated molecular patterns (PAMPs). In addition, immunity can be induced in tissues remote from infection sites via systemic acquired resistance (SAR), initiated following gene-for-gene recognition between plant resistance proteins and microbial effectors.The nature of the mobile signal and remotely activated networks responsible for establishing SAR remain unclear.
Here we show that despite the absence of PAMP contact, systemically responding leaves rapidly activate a SAR transcriptional signature with strong similarity to local basal defense. Jasmonates have previously been implicated in systemic signalling in response to wounding and plant herbivory but not SAR. We present several lines of evidence that suggest jasmonates may also be central to SAR. Jasmonic acid (JA) rapidly accumulates in phloem exudates of leaves challenged with an avirulent strain of Pseudomonas syringae. In systemically responding leaves transcripts associated with jasmonate biosynthesis are upregulated and JA increases transiently. SAR can be mimicked by foliar JA application and is abrogated in mutants impaired in jasmonate synthesis or response.
We conclude that, jasmonate signalling appears to mediate long-distance information transmission. Moreover, the systemic transcriptional response shares extraordinary overlap with local herbivory and wounding responses, indicating that jasmonates may be central to an evolutionarily conserved signalling network, which decodes multiple abiotic and biotic stress signals.
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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Institute:
| University of Exeter |
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Address: | School of Biosciences Geoffrey Pope Building Stocker Road Exeter Devon
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Postcode:
| EX4 4QD |
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Country:
| UK |
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| Telephone Number:
| +44 (0)1392 263789 |
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Fax Number:
| +44 (0)1392 263434 |
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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:
| pathogenicity_design |
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Number of Slides: | 9 |
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| Experimental Parameters:
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parameter | infect |
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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: Truman: Systemic response to avirulent bacterial infection_genome
Slide: Truman_1-1_Pst-DC3000-4hpi_Rep1_ATH1 | | |
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Tissue:
| whole leaves from the inner rosette (systemic/ uninoculated leaves) |
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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 4hpi |
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Separation Technique:
| The systemic leaves were removed with dissection scissors |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_1-2_Pst-DC3000(hrpA)-4hpi_Rep1_ATH1 | | |
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Tissue:
| whole leaves from the inner rosette (systemic/ uninoculated leaves) |
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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 4hpi |
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Separation Technique:
| The systemic leaves were removed with dissection scissors |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Truman_1-3_Pst-DC3000(avrRpm1)-4hpi_Rep1_ATH1 | | |
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Tissue:
| whole leaves from the inner rosette (systemic/ uninoculated leaves) |
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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 4hpi |
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Separation Technique:
| The systemic leaves were removed with dissection scissors |
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Other Information:
| |
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Protocols for BioSource 1 |
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Slide: Truman_1-4_Pst-DC3000-4hpi_Rep2_ATH1 | | |
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Tissue:
| whole leaves from the inner rosette (systemic/ uninoculated leaves) |
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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 4hpi |
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Separation Technique:
| The systemic leaves were removed with dissection scissors |
---|
Other Information:
| |
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|
Protocols for BioSource 1 |
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Slide: Truman_1-5_Pst-DC3000(hrpA)-4hpi_Rep2_ATH1 | | |
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Tissue:
| whole leaves from the inner rosette (systemic/ uninoculated leaves) |
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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 4hpi |
---|
Separation Technique:
| The systemic leaves were removed with dissection scissors |
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Other Information:
| |
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|
Protocols for BioSource 1 |
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Slide: Truman_1-6_Pst-DC3000(avrRpm1)-4hpi_Rep2_ATH1 | | |
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Tissue:
| whole leaves from the inner rosette (systemic/ uninoculated leaves) |
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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 4hpi |
---|
Separation Technique:
| The systemic leaves were removed with dissection scissors |
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Other Information:
| |
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|
Protocols for BioSource 1 |
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Slide: Truman_1-7_Pst-DC3000-4hpi_Rep3_ATH1 | | |
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Tissue:
| whole leaves from the inner rosette (systemic/ uninoculated leaves) |
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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. |
---|
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 4hpi |
---|
Separation Technique:
| The systemic leaves were removed with dissection scissors |
---|
Other Information:
| |
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|
Protocols for BioSource 1 |
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Slide: Truman_1-8_Pst-DC3000(hrpA)-4hpi_Rep3_ATH1 | | |
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Tissue:
| whole leaves from the inner rosette (systemic/ uninoculated leaves) |
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in vivo Treatment:
| |
---|
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. |
---|
Additional Organism Information:
| |
---|
sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested 4hpi |
---|
Separation Technique:
| The systemic leaves were removed with dissection scissors |
---|
Other Information:
| |
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|
Protocols for BioSource 1 |
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Slide: Truman_1-9_Pst-DC3000(avrRpm1)-4hpi_Rep3_ATH1 | | |
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Tissue:
| whole leaves from the inner rosette (systemic/ uninoculated leaves) |
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in vivo Treatment:
| |
---|
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. |
---|
Additional Organism Information:
| |
---|
sample description | Systemic, uninfected leaves from the inner rosette of developing, not yet fully expanded, leaves were harvested 4hpi |
---|
Separation Technique:
| The systemic leaves were removed with dissection scissors |
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Other Information:
| |
---|
|
Protocols for BioSource 1 |
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Problems? Comments? Suggestions? Contact the Affymetrix Team