Skip Navigation


Plant and Cell Physiology Advance Access originally published online on April 15, 2009
Plant and Cell Physiology 2009 50(7):1201-1214; doi:10.1093/pcp/pcp057
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow Supplementary Data
Right arrowOA All Versions of this Article:
50/7/1201    most recent
pcp057v3
pcp057v2
pcp057v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Google Scholar
Right arrow Articles by Kojima, M.
Right arrow Articles by Sakakibara, H.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kojima, M.
Right arrow Articles by Sakakibara, H.
Agricola
Right arrow Articles by Kojima, M.
Right arrow Articles by Sakakibara, H.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved.
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and the Japanese Society of Plant Physiologists are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

This article appears in the following Plant and Cell Physiology issue: Special Issue Articles: Omics and Bioinformatics [View the issue table of contents]

Special Issue - Regular Paper

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

Mikiko Kojima1, Tomoe Kamada-Nobusada1, Hirokazu Komatsu2,5, Kentaro Takei1, Takeshi Kuroha1,6, Masaharu Mizutani3, Motoyuki Ashikari4, Miyako Ueguchi-Tanaka4, Makoto Matsuoka4, Koji Suzuki2 and Hitoshi Sakakibara1,*

1RIKEN Plant Science Center, 1-7-22, Suehiro, Tsurumi, Yokohama, 230-0045 Japan
2Faculty of Science and Technology, Keio University, Kohoku, Yokohama, 223-8522 Japan
3Graduate School of Agricultural Science, Kobe University, Rokkodai, Nada, Kobe, 657-8501 Japan
4Bioscience and Biotechnology Center, Nagoya University, Nagoya, 464-8601 Japan

*Corresponding author: E-mail, sakaki{at}riken.jp; Fax, +81-45-503-9609.


  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.

Keywords: ABA - Auxins - Cytokinins - Gibberellins - Mass spectrometry - Oryza sativa

Abbreviations: AAO, aldehyde oxidase; ASA, anthranilate synthase {alpha} subunit; ASB, anthranilate synthase β subunit; CKX, cytokinin oxidase/dehydrogenase; cZ, cis-zeatin; DZ, dihydrozeatin; DZR, DZ riboside; ESI, electrospray interface; EUIL, EUI-like; GA2ox, gibberellin 2-oxidase; GA20ox, gibberellin 20-oxidase; IA-Ala, indole-3-acetyl-L-alanine; IA-Asp, indole-3-acetyl-L-aspartic acid; IA-Ile, indole-3-acetyl-L-isoleucine; IA-Leu, indole-3-acetyl-L-leucine; IA-Phe, indole-3-acetyl-L-phenylalanine; iP, N6-({Delta}2-isopentenyl)adenine; iP7G, iP-7-N-glucoside; iP9G, iP-9-N-glucoside; iPR, iP riboside; iPRMP, iPR 5'-monophosphate; iPRDP, iPR 5'-diphosphate; iPRTP, iPR 5'-triphosphate; iPRPs, iPR 5'-phosphates; KAO, ent-kaurenoic acid oxidase; KO, ent-kaurene oxidase; LOG, lonely guy; NCED, 9-cis-epoxycarotenoid dioxygenase; NIT, nitrilase; qMS/MS, tandem quadrupole mass spectrometer; TAA, tryptophan aminotransferase; tZ, trans-zeatin; tZ7G, tZ-7-N-glucoside; tZ9G, tZ-9-N-glucoside; tZOG, tZ-O-glucoside; tZR, tZ riboside; tZROG, tZR-O-glucoside; tZRMP, tZR 5'-monophosphate; tZRPs, tZR 5'-phosphates; UPLC, ultra-performance liquid chromatography; ZEP, zeaxanthin epoxidase.

5Present address: Kyoto City Collaboration of Regional Entities for the Advancement of Technological Excellence, Kyoto, 615-8510 Japan.

6Present address: Department of Biology, University of Washington, Seattle, WA 98195, USA.

(Received March 8, 2009; Accepted April 12, 2009)
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Plant Cell PhysiolHome page
J. Preston, K. Tatematsu, Y. Kanno, T. Hobo, M. Kimura, Y. Jikumaru, R. Yano, Y. Kamiya, and E. Nambara
Temporal Expression Patterns of Hormone Metabolism Genes during Imbibition of Arabidopsis thaliana Seeds: A Comparative Study on Dormant and Non-Dormant Accessions
Plant Cell Physiol., October 1, 2009; 50(10): 1786 - 1800.
[Abstract] [Full Text] [PDF]

Home page
 Plant Cell PhysiolHome page
K. Shinozaki and H. Sakakibara
Omics and Bioinformatics: An Essential Toolbox for Systems Analyses of Plant Functions Beyond 2010
Plant Cell Physiol., July 1, 2009; 50(7): 1177 - 1180.
[Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.