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Laser microdissection of Arabidopsis cells at the powdery mildew infection site
Mary C Wildermuth, University of California Berkeley, Plant and Microbial Biology Department (mwildermuth@berkeley.edu)
Experiment design (16 hybridizations)
genotype
•Col-0 •eds16
tissue type name
•laser microdisected •whole leaf •whole leaf amplified •leaf scrape
pathogen inoculation
•G-orontii •uninoculated

This experiment has been imported by PLEXdb from NCBI GEO (GSE15680) Series_summary: To elucidate host processes and components required for t...[complete overview]

Experiment     Expression     Hybridizations & Samples     Quality Control     Compare Treatments     Downloads    

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Experiment Name: Laser microdissection of Arabidopsis cells at the powdery mildew infection site
Accession No: AT80
Microarray: ATH1-121501
Visibility: public
Experiment Type:
Experiment Factor(s):
genotype
•Col-0   •eds16
tissue type name
•laser microdisected   •whole leaf   •whole leaf amplified   •leaf scrape
pathogen inoculation
•G-orontii   •uninoculated
Quality Control: biological replicates
Treatment summary:
 genotype  tissue type name  pathogen inoculation  # replicates
 Col-0  laser microdisected  G-orontii  2
 Col-0  laser microdisected  uninoculated  2
 Col-0  whole leaf  G-orontii  2
 Col-0  whole leaf  uninoculated  2
 Col-0  whole leaf amplified  G-orontii  2
 Col-0  whole leaf amplified  uninoculated  2
 Col-0  leaf scrape  G-orontii  2
 Col-0  leaf scrape  uninoculated  0
 eds16  laser microdisected  G-orontii  2
 eds16  laser microdisected  uninoculated  0
 eds16  whole leaf  G-orontii  0
 eds16  whole leaf  uninoculated  0
 eds16  whole leaf amplified  G-orontii  0
 eds16  whole leaf amplified  uninoculated  0
 eds16  leaf scrape  G-orontii  0
 eds16  leaf scrape  uninoculated  0
Total hybridizations: 16
Description: This experiment has been imported by PLEXdb from NCBI GEO (GSE15680)

Series_summary:
To elucidate host processes and components required for the sustained growth and reproduction of the obligate biotrophic fungus Golovinomyces orontii on Arabidopsis thaliana, laser microdissection was used to isolate cells at the site of infection at 5 days postinfection for downstream global Arabidopsis expression profiling. Site-specific profiling increased sensitivity dramatically, allowing us to identify specific host processes, process components, and their putative regulators hidden in previous whole-leaf global expression analyses. For example, 67 transcription factors exhibited altered expression at the powdery mildew (PM) infection site, with subsets of these playing known or inferred roles in photosynthesis, cold/dehydration responses, defense, auxin signaling, and the cell cycle. Using integrated informatics analyses, we constructed putative regulatory networks for a subset of these processes and provided strong support for host cell cycle modulation at the PM infection site. Further experimentation revealed induced host endoreduplication occurred exclusively at the infection site and led us to identify MYB3R4 as a transcriptional regulator of this process. Induced endoreduplication was abrogated in myb3r4 mutants, and G. orontii growth and reproduction were reduced. This suggests that, by increasing gene copy number, localized endoreduplication serves as a mechanism to meet the enhanced metabolic demands imposed by the fungus, which acquires all its nutrients from the plant host.
Keywords: endoreduplication, cell cycle, obligate biotroph

Series_overall_design:
Arabidopsis whole leaves from wild type Columbia-0 and enhanced disease susceptibility mutant eds16-1, a null isochorismate synthase 1 (At1g74710) mutant were harvested at 5 days after Golovinomyces orontii infection, microwave-fixed, paraffin-embedded and sectioned. Groups of epidermal and mesophyll cells (~20 cells/group) surrounding the G. orontii infected epidermal cell were cut using a Leica AS laser microdissection (LMD) system. In parallel, groups of epidermal and mesophyll cells were collected from uninfected leaves at 5 days from wild type Arabidopsis. LMD-isolated cells, whole leaf, whole leaf amplified and tissue scrape samples were used for RNA extraction and hybridization to Affymetrix Arabidopsis ATH1 microarrays. Gene expression profiles were obtained for wild type from all samples and for ics1 mutant from LMD infected samples. The experiment includes 2 biological replicates.

PLEXdb Curator's Note:

We want the users to be aware of the fact that some of the replications within a treatment have low correlation after MAS5.0 normalization, e.g., H13-H14 and H15-H16. One can see these plots by using the Treatmentrs Tab and then Show 'MAS Scatterplot'.

Publication: 'Laser microdissection of Arabidopsis cells at the powdery mildew infection site reveals site-specific processes and regulators', Chandran D, Inada N, Hather G, Kleindt CK et al
Proc Natl Acad Sci U S A 2010 Jan 5;107(1):460-5
pubmed: 20018666
Created: 2010-08-24 12:15:23
Last Update: 2010-08-24 13:16:22
Released: 2010-09-07
GEO Accession GSE15680
Submitter: PLEXdb Curator
Name: Mary C Wildermuth
Institution: University of California Berkeley, Plant and Microbial Biology Department
Head of Laboratory: --
email(s):
Homepage:

 
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