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Double-stranded RNA-binding protein DRB3 negatively regulates anthocyanin biosynthesis by modulating PAP1 expression in Arabidopsis thaliana

  • JPR Symposium
  • Expanding plant non-coding RNA world
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Abstract

The model plant Arabidopsis thaliana has five double-stranded RNA-binding proteins (DRB1-DRB5), two of which, DRB1 and DRB4, are well characterized. In contrast, the functions of DRB2, DRB3 and DRB5 have yet to be elucidated. In this study, we tried to uncover their functions using drb mutants and DRB-over-expressed lines. In over-expressed lines of all five DRB genes, the over-expression of DRB2 or DRB3 (DRB2ox or DRB3ox) conferred a downward-curled leaf phenotype, but the expression profiles of ten small RNAs were similar to that of the wild-type (WT) plant. Phenotypes were examined in response to abiotic stresses. Both DRB2ox and DRB3ox plants exhibited salt-tolerance. When these plants were exposed to cold stress, drb2 and drb3 over-accumulated anthocyanin but DRB2ox and DRB3ox did not. Therefore, the over-expression of DRB2 or DRB3 had pleiotropic effects on host plants. Microarray and deep-sequencing analyses indicated that several genes encoding key enzymes for anthocyanin biosynthesis, including chalcone synthase (CHS), dihydroflavonol reductase (DFR) and anthocyanidin synthase (ANS), were down-regulated in DRB3ox plants. When DRB3ox was crossed with the pap1-D line, which is an activation-tagged transgenic line that over-expresses the key transcription factor PAP1 (Production of anthocyanin pigmentation1) for anthocyanin biosynthesis, over-expression of DRB3 suppressed the expression of PAP1, CHS, DFR and ANS genes. DRB3 negatively regulates anthocyanin biosynthesis by modulating the level of PAP1 transcript. Since two different small RNAs regulate PAP1 gene expression, a possible function of DRB3 for small RNA biogenesis is discussed.

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Acknowledgements

We are grateful to Drs. Kazuki Saito and Keiko Yonekura-Sakakibara, at The RIKEN Center for Sustainable Resource Science, for fruitful discussion. The authors also thank Dr. Richard A. Dixon and the Arabidopsis Biological Resource Center for kindly providing Arabidopsis seeds. This work was supported by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan [a Grant-in-Aid for Scientific Research (C) (No. 24570044) and Scientific Research on Innovative Area (No. 16H06435) to T.F.].

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Authors and Affiliations

  1. Departments of Applied Biological Sciences, Tokyo University of Agriculture and Technology, 3-5-8 Saiwaicho, Fuchu, Tokyo, 183-8509, Japan

    Hikaru Sawano, Takuma Matsuzaki, Tomoyuki Usui, Midori Tabara, Akihito Fukudome, Akihiro Kanaya, Daichi Tanoue, Toshinori Kozaki, Kazuo Ishii, Hiromitsu Moriyama & Toshiyuki Fukuhara

  2. Molecular and Environmental Plant Sciences, Department of Horticultural Sciences, Vegetable and Fruit Development Center, Texas A&M University, College Station, TX, 77843, USA

    Akihito Fukudome

  3. Department of Clinical Plant Science, Hosei University, 3-7-2 Kajino-cho, Koganei, Tokyo, 184-8584, Japan

    Akihiro Hiraguri

  4. Department of Life Science and Research Center for Life Science, College of Science, Rikkyo University, Toshima, Tokyo, Japan

    Gorou Horiguchi

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

    Misato Ohtani & Taku Demura

  6. RIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa, 230-0045, Japan

    Misato Ohtani & Taku Demura

  7. Innovation Advancement Organization, Tokyo University of Agriculture and Technology, 3-5-8 Saiwaicho, Fuchu, Tokyo, 183-8509, Japan

    Toshinori Kozaki

  8. Institute of Global Innovation Research, Tokyo University of Agriculture and Technology, 3-5-8 Saiwaicho, Fuchu, Tokyo, 183-8509, Japan

    Toshiyuki Fukuhara

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  1. Hikaru Sawano
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Correspondence to Toshiyuki Fukuhara.

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Sawano, H., Matsuzaki, T., Usui, T. et al. Double-stranded RNA-binding protein DRB3 negatively regulates anthocyanin biosynthesis by modulating PAP1 expression in Arabidopsis thaliana . J Plant Res 130, 45–55 (2017). https://doi.org/10.1007/s10265-016-0886-0

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  • DOI: https://doi.org/10.1007/s10265-016-0886-0

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