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Successful expression of a novel bacterial gene for pinoresinol reductase and its effect on lignan biosynthesis in transgenic Arabidopsis thaliana

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Abstract

Pinoresinol reductase and pinoresinol/lariciresinol reductase play important roles in an early step of lignan biosynthesis in plants. The activities of both enzymes have also been detected in bacteria. In this study, pinZ, which was first isolated as a gene for bacterial pinoresinol reductase, was constitutively expressed in Arabidopsis thaliana under the control of the cauliflower mosaic virus 35S promoter. Higher reductive activity toward pinoresinol was detected in the resultant transgenic plants but not in wild-type plant. Principal component analysis of data from untargeted metabolome analyses of stem, root, and leaf extracts of the wild-type and two independent transgenic lines indicate that pinZ expression caused dynamic metabolic changes in stems, but not in roots and leaves. The metabolome data also suggest that expression of pinZ influenced the metabolisms of lignan and glucosinolates but not so much of neolignans such as guaiacylglycerol-8-O-4′-feruloyl ethers. In-depth quantitative analysis by liquid chromatography–tandem mass spectrometry (LC-MS/MS) indicated that amounts of pinoresinol and its glucoside form were markedly reduced in the transgenic plant, whereas the amounts of glucoside form of secoisolariciresinol in transgenic roots, leaves, and stems increased. The detected levels of lariciresinol in the transgenic plant following β-glucosidase treatment also tended to be higher than those in the wild-type plant. Our findings indicate that overexpression of pinZ induces change in lignan compositions and has a major effect not only on lignan biosynthesis but also on biosynthesis of other primary and secondary metabolites.

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Acknowledgments

The authors thank Mr. Yasuyuki Ishikawa for the preparation of a binary vector for plant transformation and the subsequent screening of transgenic lines in an earlier stage of this study. The authors also acknowledge Prof. Yoshikazu Kitano for his help in the chiral HPLC analysis and Prof. Toshiaki Umezawa for providing pinoresinol. This work was supported in part by the New Energy and Industrial Technology Development Organization (Development of Preparatory Basic Bioenergy Technology), by the Japan Science and Technology Agency (Advanced Low Carbon Technology Research and Development Program), and by the Japan Society for the Promotion of Science (Japan Advanced Plant Science Network Program).

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  1. Atsushi Okazawa

    Present address: Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Nakaku, Sakai, Osaka, 599-8531, Japan

Authors and Affiliations

  1. Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo, 184-8588, Japan

    Masayuki Tamura, Yukiko Tsuji, Kanna Sato-Izawa & Shinya Kajita

  2. Graduate School of Engineering, Osaka University, 1-1 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Tatsuya Kusunose, Atsushi Okazawa & Toshiya Muranaka

  3. Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata, 940-2188, Japan

    Naofumi Kamimura, Yuki Fukuhara, Masao Fukuda & Eiji Masai

  4. Metabolomics Research Group, RIKEN Center for Sustainable Resource Science, RIKEN, 1-7-22, Tsurumi, Kanagawa, 230-0045, Japan

    Tetsuya Mori, Ryo Nakabayashi & Kazuki Saito

  5. Forestry and Forest Products Research Institute, 1 Matsunosato, Tsukuba, Ibaraki, 305-8687, Japan

    Shojiro Hishiyama

  6. Department of Environmental Engineering and Green Technology, Malaysia-Japan International Institute of Technology, Universiti Teknologi Malaysia, Kuala Lumpur, 54100, Malaysia

    Hirofumi Hara

  7. Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8675, Japan

    Kazuki Saito

  8. College of Bioresource Sciences, Nihon University, 1866 Mameino, Fujisawa, Kanagawa, 252-0880, Japan

    Yoshihiro Katayama

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  1. Masayuki Tamura
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Correspondence to Shinya Kajita.

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Tamura, M., Tsuji, Y., Kusunose, T. et al. Successful expression of a novel bacterial gene for pinoresinol reductase and its effect on lignan biosynthesis in transgenic Arabidopsis thaliana . Appl Microbiol Biotechnol 98, 8165–8177 (2014). https://doi.org/10.1007/s00253-014-5934-x

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  • DOI: https://doi.org/10.1007/s00253-014-5934-x

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