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Spatial dissection of the Arabidopsis thaliana transcriptional response to downy mildew using fluorescence activated cell sorting
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Coker, Timothy L. R., Cevik, Volkan, Beynon, Jim and Gifford, Miriam L. (2015) Spatial dissection of the Arabidopsis thaliana transcriptional response to downy mildew using fluorescence activated cell sorting. Frontiers in plant science, Volume 6 . pp. 1-13. Article number 527. doi:10.3389/fpls.2015.00527 ISSN 1664-462X.
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Official URL: http://dx.doi.org/10.3389/fpls.2015.00527
Abstract
Changes in gene expression form a crucial part of the plant response to infection. In the last decade, whole-leaf expression profiling has played a valuable role in identifying genes and processes that contribute to the interactions between the model plant Arabidopsis thaliana and a diverse range of pathogens. However, with some pathogens such as downy mildew caused by the biotrophic oomycete pathogen Hyaloperonospora arabidopsidis (Hpa), whole-leaf profiling may fail to capture the complete Arabidopsis response encompassing responses of non-infected as well as infected cells within the leaf. Highly localized expression changes that occur in infected cells may be diluted by the comparative abundance of non-infected cells. Furthermore, local and systemic Hpa responses of a differing nature may become conflated. To address this we applied the technique of Fluorescence Activated Cell Sorting (FACS), typically used for analyzing plant abiotic responses, to the study of plant-pathogen interactions. We isolated haustoriated (Hpa-proximal) and non-haustoriated (Hpa-distal) cells from infected seedling samples using FACS, and measured global gene expression. When compared with an uninfected control, 278 transcripts were identified as significantly differentially expressed, the vast majority of which were differentially expressed specifically in Hpa-proximal cells. By comparing our data to previous, whole organ studies, we discovered many highly locally regulated genes that can be implicated as novel in the Hpa response, and that were uncovered for the first time using our sensitive FACS technique.
Item Type: | Journal Article | ||||||||
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Subjects: | S Agriculture > SB Plant culture | ||||||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||||||
Library of Congress Subject Headings (LCSH): | Plant-pathogen relationships, Oomycetes, Cytofluorometry | ||||||||
Journal or Publication Title: | Frontiers in plant science | ||||||||
Publisher: | Frontiers Media S.A. | ||||||||
ISSN: | 1664-462X | ||||||||
Official Date: | 10 July 2015 | ||||||||
Dates: |
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Volume: | Volume 6 | ||||||||
Number of Pages: | 13 | ||||||||
Page Range: | pp. 1-13 | ||||||||
Article Number: | Article number 527 | ||||||||
DOI: | 10.3389/fpls.2015.00527 | ||||||||
Status: | Peer Reviewed | ||||||||
Publication Status: | Published | ||||||||
Access rights to Published version: | Open Access (Creative Commons) | ||||||||
Description: | All microarray data have been deposited in GEO (Series GSE58046), released upon publication. |
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Date of first compliant deposit: | 30 December 2015 | ||||||||
Date of first compliant Open Access: | 30 December 2015 | ||||||||
Funder: | Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), Engineering and Physical Sciences Research Council (EPSRC), University of Warwick. Systems Biology Doctoral Training Centre | ||||||||
Grant number: | BB/H109502/1 (BBSRC), BB/F001347 (BBSRC) | ||||||||
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