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Plant and Cell Physiology Advance Access first published online on June 3, 2009
This version published online on June 5, 2009
Plant and Cell Physiology, doi:10.1093/pcp/pcp079
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved.
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Omics-based approaches to methionine side-chain elongation in Arabidopsis: characterization of the genes encoding methylthioalkylmalate isomerase and methylthioalkylmalate dehydrogenase

Yuji Sawada1,2, Ayuko Kuwahara1,2, Mutsumi Nagano1,2, Tomoko Narisawa1, Akane Sakata1, Kazuki Saito1,3 and Masami Yokota Hirai1,2

1RIKEN Plant Science Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.
2JST, CREST, 4-1-8 Hon-chou, Kawaguchi, Saitama 332-0012, Japan.
3Graduate School of Pharmaceutical Sciences, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba, Chiba 263-8522, Japan.

Corresponding author: Dr. Masami Yokota Hirai, RIKEN Plant Science Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan. Tel +81-45-503-9491, Fax +81-45-503-9489, e-mail myhirai{at}psc.riken.jp


  Abstract

Glucosinolates (GSLs) are secondary metabolites in Brassicaceae plants synthesized from amino acids. Met-derived GSLs (Met-GSLs) with diverse side chains of various lengths are the major GSLs in Arabidopsis. Met chain elongation enzymes are responsible for variations in chain length in Met-GSL biosynthesis. The genes encoding Met chain elongation enzymes are considered to have been recruited from the Leu biosynthetic pathway in the course of evolution. Among them, the genes encoding methylthioalkylmalate synthases and aminotransferases have been identified; however, the remaining genes that encode methylthioalkylmalate isomerase (MAM-I) and methylthioalkylmalate dehydrogenase (MAM-D) remain to be identified. In our previous study based on transcriptome coexpression analysis, we identified candidate genes for a large subunit of MAM-I and MAM-D. In this study, we confirmed their predicted functions by targeted GSL analysis of the knockout mutants, and named the respective genes MAM-IL1/AtleuC1 and MAM-D1/AtIMD1. Metabolic profiling of the knockout mutants of Met chain elongation enzymes, conducted by means of widely targeted metabolomics, implied the roles of these enzymes in controlling metabolism from Met to primary and Met-related secondary metabolites. As shown here, an omics-based approach is an efficient strategy for the functional elucidation of genes involved in metabolism.

Keywords: chain elongation - gene function - glucosinolate - high-throughput - methionine - widely targeted metabolomics

(Received April 13, 2009; Accepted June 1, 2009)
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