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Plant and Cell Physiology Advance Access originally published online on October 30, 2009
Plant and Cell Physiology 2009 50(11):1988-1993; doi:10.1093/pcp/pcp141
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use distribution, and reproduction in any medium, provided the original work is properly cited.

Short Communication

A Highly Sensitive, Quick and Simple Quantification Method for Nicotianamine and 2'-Deoxymugineic Acid from Minimum Samples Using LC/ESI-TOF-MS Achieves Functional Analysis of These Components in Plants

Yusuke Kakei1, Isomaro Yamaguchi2, Takanori Kobayashi1, Michiko Takahashi1,4, Hiromi Nakanishi1, Takashi Yamakawa1 and Naoko K. Nishizawa1,3,*

1Department of Global Agricultural Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657 Japan
2Maebashi Institute of Technology, Maebashi, Gunma, 371-0816 Japan
3Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, 1-308 Suematsu, Nonoichi-machi, Ishikawa, 921-8836 Japan

*Corresponding author: E-mail, annaoko{at}mail.ecc.u-tokyo.ac.jp; Fax, +81-3-5841-7514.


  Abstract

A highly sensitive quantitative method for assaying nicotianamine (NA) and 2'-deoxymugineic acid (DMA) using liquid chromatography/electrospray ionization time-of-flight mass spectrometry (LC/ESI-TOF-MS) was developed. The amino and hydroxyl groups of NA and DMA were derivatized using 9-fluorenylmethoxycarbonyl chloride. The amounts of NA and DMA in 10 µl of xylem sap from rice cultivated under iron (Fe)-sufficient and Fe-deficient conditions were quantified without concentration. In Fe-sufficient plants, the concentrations of NA and DMA were almost equal to that of Fe. In Fe-deficient plants, the concentration of NA did not change significantly, whereas that of DMA increased markedly.

Keywords: 2'-Deoxymugineic acid - Fe deficiency - Nicotianamine - Xylem transport

Abbreviations: BPDS, bathophenanthrolinedisulfonic acid; CE, capillary electrophoresis; DMA, 2'-deoxymugineic acid; FMOC-Cl, 9-fluorenyl methoxycarboxyl chloride; GC/MS, gas chromatography/mass spectrometry; ICP/MS, inductively coupled plasma/mass spectrometry; LC, liquid chromatography; LC/MS, liquid chromatography/mass spectrometry; LC/ESI-TOF-MS, liquid chromatography/electrospray ionization time-of-flight mass spectrometry; MAs, mugineic acid family phytosiderophores; MS, mass spectrometry; NA, nicotianamine; NL, N{varepsilon}-nicotyl-lysine.

4Present address: Department of Bioproductive Science, Utsunomiya University, 350 Mine, Utsunomiya, Tochigi, 321-8505 Japan.

(Received June 22, 2009; Accepted October 7, 2009)
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