Determination of the auxin transport parameters on the cellular level

Petrášek J., Laňková M., Zažímalová E.
Methods in Molecular Biology 1056 “Chemical Biology in Plants 1056: 241-253, 2014

Abstract: The accumulation of radioactively labelled compounds in cells is frequently used for the determination of activities of various transport systems located at the plasma membrane, including the system for carrier-mediated transport of plant hormone auxin. The measurements of auxin transport could be performed on the tissue level as well, but for more precise quantitative analysis of activity of individual auxin carriers the model of plant cell cultures represents an invaluable tool. Here we describe the method for the determination of the activities of auxin influx and efflux carriers in plant cells grown in a suspension using radiolabelled synthetic auxins 2,4-dichlorophenoxyacetic acid (2,4-D) and naphthalene-1-acetic acid (NAA). By making use of specific inhibitors of active auxin influx and efflux, as well as cell lines overexpressing or silencing particular auxin carriers, this method allows to determine kinetic parameters of auxin flow across plasma membrane and the activity of those carriers.
DOI: IEB authors: Martina Laňková, Jan Petrášek, Eva Zažímalová