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Plant and Cell Physiology Advance Access originally published online on February 5, 2007
Plant and Cell Physiology 2007 48(3):550-554; doi:10.1093/pcp/pcm018
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Short Communication

Molecular Cloning of N-methylputrescine Oxidase from Tobacco

Akira Katoh, Tsubasa Shoji and Takashi Hashimoto*

Nara Institute of Science and Technology, Graduate School of Biological Sciences, 8916-5 Takayama, Ikoma, Nara, 630-0192 Japan

*Corresponding author: E-mail, hasimoto{at}bs.naist.jp; Fax, +81-743-72-5529.


  Abstract

Nicotine biosynthesis in Nicotiana species requires an oxidative deamination of N-methylputrescine, catalyzed by N-methylputrescine oxidase (MPO). In a screen for tobacco genes that were down-regulated in a tobacco mutant with altered regulation of nicotine biosynthesis, we identified two homologous MPO cDNAs which encode diamine oxidases of a particular subclass. Tobacco MPO genes were expressed specifically in the root, and up-regulated by jasmonate treatment. Recombinant MPO protein expressed in Escherichia coli formed a homodimer and deaminated N-methylputrescine more efficiently than symmetrical diamines. These results indicate that MPO evolved from general diamine oxidases to function effectively in nicotine biosynthesis.

Keywords: Diamine oxidase - Nicotiana tabacum (tobacco) - Nicotine - N-methylputrescine

Abbreviations: GC-MS, gas chromatography–mass spectrometry; GST, glutathione S-transferase; MPO, N-methylputrescine oxidase; PMT, putrescine N-methyltransferase

(Received January 9, 2007; Accepted January 31, 2007)
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