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PMID: 19369275 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Highly sensitive and high-throughput analysis of plant hormones using MS-probe modification and liquid chromatography-tandem mass spectrometry: an application for hormone profiling in Oryza sativa.

Plant & cell physiology ·Vol. 50 ·No. 7 ·2009-07-00 ·Pages 1201-14

Kojima M, Kamada-Nobusada T, Komatsu H, Takei K, Kuroha T, Mizutani M, Ashikari M, Ueguchi-Tanaka M, Matsuoka M, Suzuki K, Sakakibara H

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.

MeSH Terms
Abscisic Acid/analysis Chromatography, High Pressure Liquid Cytokinins/analysis Gene Expression Profiling Gibberellins/analysis Indoleacetic Acids/analysis Oryza/chemistry Plant Growth Regulators/analysis Spectrometry, Mass, Electrospray Ionization Tandem Mass Spectrometry/methods
Chemicals
Cytokinins Gibberellins Indoleacetic Acids Plant Growth Regulators Abscisic Acid
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Kojima Mikiko
RIKEN Plant Science Center, Tsurumi, Yokohama, Japan.
Kamada-Nobusada Tomoe
Komatsu Hirokazu
Takei Kentaro
Kuroha Takeshi
Mizutani Masaharu
Ashikari Motoyuki
Ueguchi-Tanaka Miyako
Matsuoka Makoto
Suzuki Koji
Sakakibara Hitoshi
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Article Info
Journal
Plant & cell physiology
Abbr.
Plant Cell Physiol
ISSN
1471-9053
DOI
10.1093/pcp/pcp057
Published
2009-07-00
Epub
2009-00-15
Pages
1201-14
Language
English
Region
Japan
NLM ID
9430925
PMCID
PMC2709547
Subset
IM
External Links
PubMed source DOI: 10.1093/pcp/pcp057 PMC Full Text View journal details
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