High-Resolution Cell-Type Specific Analysis of Cytokinins in Sorted Root Cell Populations of Arabidopsis thaliana

Methods Mol Biol. 2017:1497:231-248. doi: 10.1007/978-1-4939-6469-7_19.

Abstract

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.

Keywords: Arabidopsis; Cytokinins; Fluorescence-activated cell sorting (FACS); Liquid chromatography–mass spectrometry (LC-MS); Protoplasts; Roots; Solid-phase extraction (SPE).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / metabolism*
  • Chromatography, High Pressure Liquid / methods
  • Cytokinins / metabolism*
  • Flow Cytometry / methods
  • Gene Regulatory Networks / physiology
  • Plant Growth Regulators / metabolism*
  • Plant Roots / metabolism*
  • Tandem Mass Spectrometry / methods

Substances

  • Arabidopsis Proteins
  • Cytokinins
  • Plant Growth Regulators