Highly sensitive and high-throughput analysis of plant hormones using ms-probe modification and liquid chromatographytandem mass spectrometry: An application for hormone profiling in oryza sativa

Mikiko Kojima, Tomoe Kamada-Nobusada, Hirokazu Komatsu, Kentaro Takei, Takeshi Kuroha, Masaharu Mizutani, Motoyuki Ashikari, Miyako Ueguchi-Tanaka, Makoto Matsuoka, Koji Suzuki, Hitoshi Sakakibara

Research output: Contribution to journalArticlepeer-review

287 Citations (Scopus)

Abstract

We have developed a highly sensitive and high-throughput method for the simultaneous analysis of 43 molecular species of cytokinins, auxins, ABA and gibberellins. This method consists of an automatic liquid handling system for solid phase extraction and ultra-performance liquid chromatography (UPLC) coupled with a tandem quadrupole mass spectrometer (qMS/MS) equipped with an electrospray interface (ESI; UPLC-ESI-qMS/MS). In order to improve the detection limit of negatively charged compounds, such as gibberellins, we chemically derivatized fractions containing auxin, ABA and gibberellins with bromocholine that has a quaternary ammonium functional group. This modification, that we call 'MS-probe', makes these hormone derivatives have a positive ion charge and permits all compounds to be measured in the positive ion mode with UPLC-ESI-qMS/MS in a single run. Consequently, quantification limits of gibberellins increased up to 50-fold. Our current method needs <100 mg (FW) of plant tissues to determine phytohormone profiles and enables us to analyze >180 plant samples simultaneously. Application of this method to plant hormone profiling enabled us to draw organ distribution maps of hormone species in rice and also to identify interactions among the four major hormones in the rice gibberellin signaling mutants, gid1-3, gid2-1 and slr1. Combining the results of hormone profiling data with transcriptome data in the gibberellin signaling mutants allows us to analyze relationships between changes in gene expression and hormone metabolism.

Original languageEnglish
Pages (from-to)1201-1214
Number of pages14
JournalPlant and Cell Physiology
Volume50
Issue number7
DOIs
Publication statusPublished - 2009 Jul

Keywords

  • ABA
  • Auxins
  • Cytokinins
  • Gibberellins
  • Mass spectrometry
  • Oryza sativa

ASJC Scopus subject areas

  • Physiology
  • Plant Science
  • Cell Biology

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