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Experiment: Transcriptome changes of Arabidopsis during pathogen and insect attack
Experiment Description
NASCArrays Experiment Reference Number: NASCARRAYS-330
Plant defenses against pathogens and insects are regulated differentially by cross-communicating signaling pathways in which salicylic acid (SA), jasmonic acid (JA) and ethylene (ET) play key roles. To understand how plants integrate pathogen- and insect-induced signals into specific defense responses, we monitored the dynamics of SA, JA, and ET signaling in Arabidopsis after attack by a set of microbial pathogens and herbivorous insects with different modes of attack. Arabidopsis plants were exposed to a pathogenic leaf bacterium (Pseudomonas syringae pv. tomato), a pathogenic leaf fungus (Alternaria brassicicola), tissue-chewing caterpillars (Pieris rapae), cell-content-feeding thrips (Frankliniella occidentalis), or phloem-feeding aphids (Myzus persicae). Monitoring the signal signature in each plant-attacker combination showed that the kinetics of SA, JA, and ET production varies greatly in both quantity and timing. Analysis of global gene expression profiles demonstrated that the signal signature characteristic of each Arabidopsis-attacker combination is orchestrated into a surprisingly complex set of transcriptional alterations in which, in all cases, stress-related genes are overrepresented. Comparison of the transcript profiles revealed that consistent changes induced by pathogens and insects with very different modes of attack can show considerable overlap. Of all consistent changes induced by A. brassicicola, P. rapae, and F. occidentalis, more than 50% were also induced consistently by P. syringae. Notably, although these four attackers all stimulated JA biosynthesis, the majority of the changes in JA-responsive gene expression were attacker-specific. All together our study shows that SA, JA, and ET play a primary role in the orchestration of the plant's defense response, but other regulatory mechanisms, such as pathway cross-talk or additional attacker-induced signals, eventually shape the highly complex attacker-specific defense response.
In the experimental set up we intended to select for genes that showed a more than 2-fold change in the same direction(up or down)at two time points after pathogen or insect attack.
The results are described in the following paper:
De Vos, M., Van Oosten, V.R., Van Poecke, R.M.P., Van Pelt, J.A., Pozo, M.J., Mueller, M.J., Buchala, A.J., Metraux, J.-P., Van Loon, L.C., Dicke, M. and Pieterse, C.M.J. (2005). Signal signature and transcriptome changes in Arabidopsis upon pathogen and insect attack. Molecular Plant-Microbe Interactions, in press.
About the ExperimenterName: | Prof. Corne Pieterse |
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Head of Lab Name: | Prof Corne Pieterse |
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Lab:
| Section Phytopathology |
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Institute:
| Utrecht University |
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Address: | Department of Biology Utrecht University Sorbonnelaan 16 Utrecht
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Postcode:
| 3584CA |
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Country:
| The Netherlands |
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| Telephone Number:
| +31 30 253 3013 |
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Fax Number:
| +31 30 251 8366 |
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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: | 14 |
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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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References:
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reference | De Vos, M., Van Oosten, V.R., Van Poecke, R.M.P., Van Pelt, J.A., Pozo, M.J., Mueller, M.J., Buchala, A.J., Métraux, J.-P., Van Loon, L.C., Dicke, M. and Pieterse, C.M.J. (2005). Signal signature and transcriptome changes in Arabidopsis upon pathogen and insect attack. Molecular Plant-Microbe Interactions, in press. |
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| Other Information:
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GEO accession | GSE5525 |
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Slides in this Experiment
Hybridisation Set: Pieterse_set
Slide: Pieterse_1-1_Control-12h_Rep1_ATH1 | | |
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Diseased:
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Separation Technique:
| Rosette was cut with a razor blade from the root system. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-2_Control-24h_Rep1_ATH1 | | |
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Diseased:
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-3_Control-48h_Rep1_ATH1 | | |
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Diseased:
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-4_Control-72h_Rep1_ATH1 | | |
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Diseased:
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-5_avrPstDC3000-12h_Rep1_ATH1 | | |
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Diseased:
| Normal |
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in vivo Treatment:
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Treatment | P. syringae pv. tomato DC3000 with the plasmid pV288 carrying avirulence gene avrRpt2 (Kunkel et al., 1993) was cultured overnight at 28°C in liquid King's medium B, supplemented with 25 mg.L-1 kanamycin to select for the plasmid. Subsequently, bacterial cells were collected by centrifugation and resuspended in 10 mM MgSO4 to a final density of 107 cfu.mL-1. Wild-type Col-0 plants were inoculated by pressure infiltrating a suspension of P. syringae pv. tomato DC3000(avrRpt2) at 107 cfu.mL-1 into all fully expanded leaves of 5-week-old plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-6_avrPstDC3000-24h_Rep1_ATH1 | | |
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Diseased:
| Normal |
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in vivo Treatment:
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Treatment | P. syringae pv. tomato DC3000 with the plasmid pV288 carrying avirulence gene avrRpt2 (Kunkel et al., 1993) was cultured overnight at 28°C in liquid King's medium B, supplemented with 25 mg.L-1 kanamycin to select for the plasmid. Subsequently, bacterial cells were collected by centrifugation and resuspended in 10 mM MgSO4 to a final density of 107 cfu.mL-1. Wild-type Col-0 plants were inoculated by pressure infiltrating a suspension of P. syringae pv. tomato DC3000(avrRpt2) at 107 cfu.mL-1 into all fully expanded leaves of 5-week-old plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-7_Abrassicicola-24h_Rep1_ATH1 | | |
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Diseased:
| Normal |
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in vivo Treatment:
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Treatment | Bioassays with the fungal leaf pathogen Alternaria brassicicola strain MUCL 20297 were carried out as described by Ton et al. (MPMI, 2002). Briefly, A. brassicicola was grown on potato dextrose agar plates for 2 weeks at 22°C. Subsequently, conidia were collected as described by Broekaert et al. (1990). Five-week-old susceptible pad3-1 plants were challenge inoculated by applying 3-µL drops of 10 mM MgSO4 containing 106 spores per mL onto all fully expanded leaves of 5-week-old plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-8_Abrassicicola-48h_Rep1_ATH1 | | |
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Diseased:
| Normal |
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in vivo Treatment:
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Treatment | Bioassays with the fungal leaf pathogen Alternaria brassicicola strain MUCL 20297 were carried out as described by Ton et al. (MPMI, 2002). Briefly, A. brassicicola was grown on potato dextrose agar plates for 2 weeks at 22°C. Subsequently, conidia were collected as described by Broekaert et al. (1990). Five-week-old susceptible pad3-1 plants were challenge inoculated by applying 3-µL drops of 10 mM MgSO4 containing 106 spores per mL onto all fully expanded leaves of 5-week-old plants. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-9_Prapae-12h_Rep1_ATH1 | | |
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Diseased:
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in vivo Treatment:
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Treatment | Tissue-chewing larvae of the small cabbage white butterfly Pieris rapae were reared on Brussels sprout plants (Brassica oleracea gemmifera cv. Cyrus) in a growth chamber with a 16-h day and 8-h night cycle (21°C; 50-70% relative humidity), as described previously (Van Poecke et al., 2001). Infestation of Arabidopsis Col-0 plants was carried out by transferring five first-instar larvae of P. rapae to each plant using a fine paintbrush. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-10_Prapae-24h_Rep1_ATH1 | | |
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Diseased:
| Normal |
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in vivo Treatment:
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Treatment | Tissue-chewing larvae of the small cabbage white butterfly Pieris rapae were reared on Brussels sprout plants (Brassica oleracea gemmifera cv. Cyrus) in a growth chamber with a 16-h day and 8-h night cycle (21°C; 50-70% relative humidity), as described previously (Van Poecke et al., 2001). Infestation of Arabidopsis Col-0 plants was carried out by transferring five first-instar larvae of P. rapae to each plant using a fine paintbrush. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-11_Foccidentalis-24h_Rep1_ATH1 | | |
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Diseased:
| Normal |
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in vivo Treatment:
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Treatment | The population of the Western flower thrips Frankliniella occidentalis originated from a greenhouse infestation on chrysanthemum. This virus-free population was reared on Phaseolus vulgaris cv. Prelude pods, supplied with Pinus pollen, in glass jars that were placed at 25°C in a growth chamber with a 16-h day and 8-h night cycle as described (Kindt et al., 2003). Thrips infestations were performed by transferring 20 larvae of F. occidentalis to each Arabidopsis Col-0 plant. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-12_Foccidentalis-48h_Rep1_ATH1 | | |
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Diseased:
| Normal |
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in vivo Treatment:
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Treatment | The population of the Western flower thrips Frankliniella occidentalis originated from a greenhouse infestation on chrysanthemum. This virus-free population was reared on Phaseolus vulgaris cv. Prelude pods, supplied with Pinus pollen, in glass jars that were placed at 25°C in a growth chamber with a 16-h day and 8-h night cycle as described (Kindt et al., 2003). Thrips infestations were performed by transferring 20 larvae of F. occidentalis to each Arabidopsis Col-0 plant. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-13_Mpersicae-48h_Rep1_ATH1 | | |
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Diseased:
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in vivo Treatment:
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Treatment | Phloem-feeding green peach aphids (Myzus persicae) were maintained on Brassica chinensis L. cv. Granaat under greenhouse conditions (25°C; 50-70% relative humidity). The 16-h light period prevented sexual reproduction, keeping the population clonal. Arabidopsis Col-0 plants were infested with M. persicae by transferring 40 nymphs and apterous adults to each plant (Van Poecke et al., 2003). |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pieterse_1-14_Mpersicae-72h_Rep1_ATH1 | | |
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Diseased:
| Normal |
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in vivo Treatment:
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Treatment | Phloem-feeding green peach aphids (Myzus persicae) were maintained on Brassica chinensis L. cv. Granaat under greenhouse conditions (25°C; 50-70% relative humidity). The 16-h light period prevented sexual reproduction, keeping the population clonal. Arabidopsis Col-0 plants were infested with M. persicae by transferring 40 nymphs and apterous adults to each plant (Van Poecke et al., 2003). |
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Other Information:
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Protocols for BioSource 1 |
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