High-Throughput Confocal Imaging in Intact Live Tissue Enables Quantification of Membrane Trafficking in Arabidopsis

Publication Type:Journal Article
Year of Publication:2010
Authors:Salomon, S, Grunewald, D, Stüber, K, Schaaf, S, MacLean, D, Schulze-Lefert, P, Robatzek, S
Journal:Plant Physiology
Volume:154
Issue:3
Date Published:2010
ISBN Number:00320889
Keywords:cotyledon
Abstract:

Membrane compartmentalization and trafficking within and between cells is considered an essential cellular property of higher eukaryotes. We established a high-throughput imaging method suitable for the quantitative detection of membrane compartments at subcellular resolution in intact epidermal tissue. Whole Arabidopsis (Arabidopsis thaliana) cotyledon leaves were subjected to quantitative confocal laser microscopy using automated image acquisition, computational pattern recognition, and quantification of membrane compartments. This revealed that our method is sensitive and reliable to detect distinct endomembrane compartments. We applied quantitative confocal laser microscopy to a transgenic line expressiong GFP-2xFYVE as a marker for endosomal compartments during biotic or abiotic stresses, and detected markedly quantitative adaptations in response to changing environments. Using a transgenic line expressing the plasma membrane-resident syntaxin GFP-PEN1, we quantified the pathogen-inducible extracellular accumulation of this fusion protein at fungal entry sites. Our protocol provides a platform to study the quantitative and dynamic changes of endomembrane trafficking, and potential adaptations of this machinery to physiological stress.

URL:http://www.jstor.org/stable/25758660
Short Title:Plant Physiology
Taxonomic name: 
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