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Plant and Cell Physiology Advance Access published online on October 4, 2007
Plant and Cell Physiology, doi:10.1093/pcp/pcm125
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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 e-mail: journals.permissions@oxfordjournals.org

2-Hydroxyisoflavanone dehydratase is a critical determinant of isoflavone productivity in hairy root cultures of Lotus japonicus

Masayuki Shimamura1, Tomoyoshi Akashi1, Nozomu Sakurai2, Hideyuki Suzuki2, Kazuki Saito3, Daisuke Shibata2, Shin-ichi Ayabe1 and Toshio Aoki1,*

1Department of Applied Biological Sciences, Nihon University, Fujisawa, Kanagawa, 252-8510 Japan
2Kazusa DNA Research Institute, Kisarazu, Chiba, 292-0818 Japan
3Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 263-8522, Japan

Corresponding author: Toshio Aoki, Department of Applied Biological Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-8510, Japan, phone +81-466-84-3939; facsimile +81-466-84-3353; e-mail taoki{at}brs.nihon-u.ac.jp


  Abstract

Hairy root cultures of a model legume, Lotus japonicus, were established to characterize two heterologous cDNAs encoding enzymes in the isoflavone biosynthesis, i.e., licorice 2-hydroxyisoflavanone synthase (IFS) and soybean 2-hydroxyisoflavanone dehydratase (HID) catalyzing sequential reactions to yield isoflavones. While the control and the IFS overexpressor did not accumulate detectable isoflavones, the HID overexpressors did accumulate daidzein and genistein, showing that HID is a critical determinant of isoflavone productivity. Production of coumestrol in all the genotypes and isoliquiritigenin/liquiritigenin in IFS+HID overexpressing lines was also noted. These results provide insight into the regulatory mechanism that controls the isoflavonoid biosynthesis.

Keywords: biosynthesis - hairy root - 2-hydroxyisoflavanone synthase - 2-hydroxyisoflavanone dehydratase - isoflavone - Lotus japonicus

(Received July 18, 2007; Accepted September 28, 2007)
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