Ultra-Rapid Auxin Metabolite Profiling for High-Throughput Arabidopsis Mutant Screening
J Exp Bot. 2018; 69 (10):2569-2579

Pencík A, Casanova-Sáez R, Pilarová V, Žukauskaite A, Pinto R, Luis Micol J, Ljung K, Novák O

Abstract
Auxin (indole-3-acetic acid, IAA) plays fundamental roles as a signalling molecule during numerous plant growth and development processes. The formation of local auxin gradients and auxin maxima/minima, which is very important for these processes, is regulated by auxin metabolism (biosynthesis, degradation and conjugation) as well as transport. When studying auxin metabolism pathways it is crucial to combine data obtained from genetic investigations with the identification and quantification of individual metabolites. Thus, to facilitate efforts to elucidate auxin metabolism and its roles in plants, we have developed a high-throughput method for simultaneously quantifying IAA and its key metabolites in minute samples (<10 mg fresh weight) of Arabidopsis thaliana tissuesby intip micro solid-phase extraction and fast liquid chromatography-tandem mass spectrometry. As a proof of concept, we applied the method to a collection of Arabidopsis mutant lines and identified lines with altered IAA metabolite profiles using multivariate data analysis. Finally, we explored correlation of IAA metabolite profiles with IAA-related phenotypes. The developed rapid analysis of large numbers of samples (more than 100 samples per day) is a valuable tool to screen for novel regulators of auxin metabolism and homeostasis among large collections of genotypes.

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