High-Resolution Cell-Type Specific Analysis of Cytokinins in Sorted Root Cell Populations of Arabidopsis thaliana
We describe a method combining fluorescence-activated cell sorting (FACS) with one-step miniaturized isolation and accurate quantification of cytokinins (CKs) using ultra-high performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) to measure these phytohormones in specific cell types of Arabidopsis thaliana roots. The methodology provides information of unprecedented resolution about spatial distributions of CKs, and thus should facilitate attempts to elucidate regulatory networks involved in root developmental processes.
Key wordsCytokinins Fluorescence-activated cell sorting (FACS) Solid-phase extraction (SPE) Liquid chromatography–mass spectrometry (LC-MS) Roots Arabidopsis Protoplasts
This work was supported by the Ministry of Education, Youth and Sports of the Czech Republic (the National Program for Sustainability I No. LO1204 and the “Návrat” program LK21306) and by the Czech Science Foundation (No. GA14-34792S), the European Molecular Biology Organization (EMBO ASTF 297-2013), the Erasmus program (European Commission: Lifelong Learning Programme), Development-The Company of Biologists (DEVTF2012), the Plant Fellows programme (http://www.plantfellows.ch/), the Swedish Governmental Agency for Innovation Systems (VINNOVA), and the Swedish Research Council (VR).