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


Author(s): Simon, Sibu S; Skůpa, Petr; Dobrev, Petre I; Petrášek, Jan; Zažímalová, Eva; Friml, Jiří
Article/Chapter Title: Analyzing the in vivo status of exogenously applied auxins: A HPLC-based method to characterize the intracellularly localized auxin transporters
Title Series: Methods in Molecular Biology
Affiliation IST Austria
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 classes of compounds which are often applied exogenously. Nevertheless, plants possess a 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 them with some of the regulatory events in plant developmental processes. The in vivo status of applied compounds can be measured by incubating plants with radiolabeled compounds, followed by extraction, purification, and HPLC metabolic profiling of plant extracts. Recently we have used this method to characterize the intracellularly localized PIN protein, PIN5. Here we explain the method in detail, with a focus on general application.
Keywords: Auxin; PIN proteins; Auxin metabolism; BY-2 tobacco cells; HPLC; Plant cell cultures
Book Title: Plant Chemical Genomics
Volume: 1056
ISBN: 978-1-62703-591-0
Publisher: Springer  
Date Published: 2014-01-01
Start Page: 255
End Page: 264
DOI: 10.1007/978-1-62703-592-7_23
Open access: no