Determination of Auxin Transport Parameters on the Cellular Level
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.
Key wordsAuxin transport assay Plant cell cultures Auxin influx Auxin efflux Radiolabelled auxin
This work was supported by the Czech Science Foundation, projects GAP305/11/2476 (JP) and GAP305/11/0797 (EZ).
- 4.Goldsmith MHM (1977) Polar transport of auxin. Annu Rev Plant Physiol Plant Mol Biol 28:439–478Google Scholar
- 8.Nagata T, Nemoto Y, Hasezawa S (1992) Tobacco BY-2 cell-line as the Hela-cell in the cell biology of higher-plants. Int Rev Cytol 132:1–30Google Scholar
- 12.Petrášek J, Zažímalová E (2006) The BY-2 cell line as a tool to study auxin transport. In: Biotechnology in Agriculture and Forestry. Tobacco BY-2 Cells: From Cellular Dynamics to Omics, Nagata, T., Matsuoka, K., Inzé, D. (eds.), Springer-Verlag, Berlin Heidelberg, 58:107–115Google Scholar