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J Exp Bot. 2010 Mar;61(5):1321-35. doi: 10.1093/jxb/erq002. Epub 2010 Feb 11.

Metabolic profiling of Arabidopsis thaliana epidermal cells.

Author information

1
Max-Planck-Institute of Molecular Plant Physiology, Campus Golm, Am Mühlenberg 1, D-14476 Potsdam OT Golm, Germany.

Abstract

Metabolic phenotyping at cellular resolution may be considered one of the challenges in current plant physiology. A method is described which enables the cell type-specific metabolic analysis of epidermal cell types in Arabidopsis thaliana pavement, basal, and trichome cells. To achieve the required high spatial resolution, single cell sampling using microcapillaries was combined with routine gas chromatography-time of flight-mass spectrometry (GC-TOF-MS) based metabolite profiling. The identification and relative quantification of 117 mostly primary metabolites has been demonstrated. The majority, namely 90 compounds, were accessible without analytical background correction. Analyses were performed using cell type-specific pools of 200 microsampled individual cells. Moreover, among these identified metabolites, 38 exhibited differential pool sizes in trichomes, basal or pavement cells. The application of an independent component analysis confirmed the cell type-specific metabolic phenotypes. Significant pool size changes between individual cells were detectable within several classes of metabolites, namely amino acids, fatty acids and alcohols, alkanes, lipids, N-compounds, organic acids and polyhydroxy acids, polyols, sugars, sugar conjugates and phenylpropanoids. It is demonstrated here that the combination of microsampling and GC-MS based metabolite profiling provides a method to investigate the cellular metabolism of fully differentiated plant cell types in vivo.

PMID:
20150518
PMCID:
PMC2837255
DOI:
10.1093/jxb/erq002
[Indexed for MEDLINE]
Free PMC Article

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