Analyzing the in vivo status of exogenously applied auxins: a HPLC-based method to characterize the intracellularly localized auxin transporters.

Simon S., Skůpa P., Dobrev P.I., Petrášek J., Zažímalová E., Friml J.
Methods in Molecular Biology 1056 “Chemical Biology in Plants“ Springer-Verlag 1056: 255-264, 2014

Abstract: Exogenous application of biologically important molecules for plant growth promotion and/or regulation is very common both in plant research and horticulture. Plant hormones, such as auxins and cytokinins are class of compounds, which are often applied exogenously. Nevertheless, plants possess well established machinery to regulate the active pool of exogenously applied compounds by converting them to metabolites and conjugates. Consequently, it is often very useful to know the in vivo status of applied compounds to connect it with some of the regulatory events in plant developmental processes. The in vivo status of the applied compound can be measured by incubating plants with radiolabeled compounds, followed by extraction, purification and HPLC metabolic profiling of plant extracts. Recently we used this method to characterize the intracellularly localized PIN protein, PIN5 and here we explain the method in detail.
DOI: IEB authors: Petre I. Dobrev, Jan Petrášek, Petr Skůpa, Eva Zažímalová