Plant Chemical Genomics

0429994 - ÚEB 2015 RIV US eng M - Monography Chapter
Petrášek, Jan - Laňková, Martina - Zažímalová, Eva
Determination of Auxin Transport Parameters on the Cellular Level.
Plant Chemical Genomics. Totowa: Humana Press, 2014 - (Hicks, G.; Robert, S.), s. 241-253. Methods in Molecular Biology. ISBN 978-1-62703-591-0
Institutional support: RVO:61389030
Keywords : Auxin transport assay * Plant cell cultures * Auxin influx
Subject RIV: ED - Physiology

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 the determination of kinetic parameters of auxin flow across the plasma membrane and the activity of those carriers.
Permanent Link: http://hdl.handle.net/11104/0234937