Determination of Auxin Transport Parameters on the Cellular Level

  • Jan Petrášek
  • Martina Laňková
  • Eva Zažímalová
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1056)

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 the determination of kinetic parameters of auxin flow across the plasma membrane and the activity of those carriers.

Key words

Auxin transport assay Plant cell cultures Auxin influx Auxin efflux Radiolabelled auxin 

Notes

Acknowledgement

This work was supported by the Czech Science Foundation, projects GAP305/11/2476 (JP) and GAP305/11/0797 (EZ).

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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Jan Petrášek
    • 1
  • Martina Laňková
    • 1
  • Eva Zažímalová
    • 1
  1. 1.Institute of Experimental Botany, ASCPragueCzech Republic

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