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Research article2015Peer reviewedOpen access

Cell-type specific metabolic profiling of Arabidopsis thaliana protoplasts as a tool for plant systems biology

Petersson, Sara; Lindén, Pernilla; Moritz, Thomas; Ljung, Karin

Abstract

Flow cytometry combined with cell sorting of protoplasts has previously been used successfully for transcript profiling of the Arabidopsis thaliana root. We have developed the technique further, and in this paper we present a robust and reliable method for metabolite profiling in specific cell types isolated from Arabidopsis roots. The method uses a combination of fluorescence-activated cell sorting and gas chromatography-time of flight-mass spectrometry analysis. Cortical and endodermal cells from the green fluorescent protein (GFP)-expressing enhancer trap line J0571 were analysed and compared with non-GFP-expressing cells and intact root tissue. Of the metabolites identified, several showed significant differences in concentration between cell types. Multivariate statistical analysis was used to compare metabolite patterns between cell and tissue types, showing that the patterns differed substantially. Isolation of specific cell populations combined with highly sensitive MS-analysis will be a powerful tool for future studies of plant metabolism, and can also be combined with transcript and protein profiling for in-depth analyses of cellular processes.

Keywords

Metabolite profiling; Untargeted metabolomics; Gas chromatography-mass spectrometry; Arabidopsis thaliana; Flow cytometry; Multivariate statistical analysis

Published in

Metabolomics
2015, Volume: 11, number: 6, pages: 1679-1689
Publisher: SPRINGER