Isolation of Plasmodesmata from Arabidopsis Suspension Culture Cells

Abstract

Due to their position fi rmly anchored within the plant cell wall, plasmodesmata (PD) are notoriously diffi cult to isolate from plant tissue. Yet, getting access to isolated PD represents the most straightforward strategy for the identifi cation of their molecular components. Proteomic and lipidomic analyses of such PD fractions have provided and will continue to provide critical information on the functional and structural elements that defi ne these membranous nano-pores. Here, we describe a two-step simple purifi cation procedure that allows isolation of pure PD-derived membranes from Arabidopsis suspension cells. The fi rst step of this procedure consists in isolating cell wall fragments containing intact PD while free of contamination from other cellular compartments. The second step relies on an enzymatic degradation of the wall matrix and the subsequent release of “free” PD. Isolated PD membranes provide a suitable starting material for the analysis of PD-associated proteins and lipids. Key words Plasmodesmata, Membrane, Cell wall, Cellulase, Suspension cell, Proteomic, Lipidomic, Arabidopsis thaliana.