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Expression and functional analyses of a putative phenylcoumaran benzylic ether reductase in Arabidopsis thaliana

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A candidate gene for phenylcoumaran benzylic ether reductase in Arabidopsis thaliana encodes a peptide with predicted functional activity and plays a crucial role in secondary metabolism.

Abstract

Phenylcoumaran benzylic ether reductase (PCBER) is thought to be an enzyme crucial in the biosynthesis of 8–5′-linked neolignans. Genes of the enzyme have been isolated and characterized in several plant species. In this study, we cloned cDNA and the 5′-untranslated region of one PCBER candidate gene (At4g39230, designated AtPCBER1) from Arabidopsis thaliana. At the amino acid level, AtPCBER1 shows high sequence identity (64–71 %) with PCBERs identified from other plant species. Expression analyses of AtPCBER1 by reverse transcriptase-polymerase chain reaction and histochemical analysis of transgenic plants harboring the 5′-untranslated region of AtPCBER1 linked with gus coding sequence indicate that expression is induced by wounding and is expressed in most tissues, including flower, stem, leaf, and root. Catalytic analysis of recombinant AtPCBER1 with neolignan and lignans in the presence of NADPH suggests that the protein can reduce not only the 8–5′-linked neolignan, dehydrodiconiferyl alcohol, but also 8–8′ linked lignans, pinoresinol, and lariciresinol, with lower activities. To investigate further, we performed metabolomic analyses of transgenic plants in which the target gene was up- or down-regulated. Our results indicate no significant effects of AtPCBER1 gene regulation on plant growth and development; however, levels of some secondary metabolites, including lignans, flavonoids, and glucosinolates, differ between wild-type and transgenic plants. Taken together, our findings indicate that AtPCBER1 encodes a polypeptide with PCBER activity and has a critical role in the biosynthesis of secondary metabolites in A. thaliana.

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Acknowledgments

This work was supported in part by the Japan Science and Technology Agency (Advanced Low Carbon Technology Research and Development Program) and by Japan Society for the Promotion of Science (Japan Advanced Plant Science Network Program).

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  1. Yukiko Tsuji

    Present address: Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, 53726-4084, USA

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

    Nuoendagula, Yukiko Tsuji & Shinya Kajita

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

    Naofumi Kamimura & Eiji Masai

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

    Tetsuya Mori, Ryo Nakabayashi & Kazuki Saito

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

    Shojiro Hishiyama

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  1. Nuoendagula
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  2. Naofumi Kamimura
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Correspondence to Shinya Kajita.

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Communicated by H. Ebinuma.

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Nuoendagula, Kamimura, N., Mori, T. et al. Expression and functional analyses of a putative phenylcoumaran benzylic ether reductase in Arabidopsis thaliana . Plant Cell Rep 35, 513–526 (2016). https://doi.org/10.1007/s00299-015-1899-1

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