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The Bowman–Birk Trypsin Inhibitor IBP1 Interacts with and Prevents Degradation of IDEF1 in Rice

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Abstract

To acquire slightly soluble iron from the rhizosphere, plants transcriptionally induce genes involved in iron acquisition in response to low iron availability. The transcription factor IDEF1 plays an important role in the regulation of this response in graminaceous plants. By yeast two-hybrid screening, we identified IDEF1-binding proteins, designated IBP1.1 and IBP1.2, as two homologous Bowman–Birk trypsin inhibitors. Interaction between IDEF1 and IBP1.1 was also confirmed by pull-down assay. IBP1.1 and IBP1.2 expression showed induction in response to iron deficiency and IDEF1 dependence. IBP1.1 localized to the nucleus and the cytoplasm when transiently expressed in onion epidermal cells. Transgenic rice plants overexpressing IBP1.1 showed enhanced expression of the Fe(II)-nicotianamine transporter gene OsYSL2. IDEF1 protein is degraded in a 26S proteasome-dependent manner and this degradation was prevented by IBP1.1. These results suggest that induced expression of IBP1.1 under Fe-deficient conditions contributes to the IDEF1-mediated iron deficiency response by preventing the degradation of IDEF1.

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Acknowledgments

We thank Prof. Hiroshi Takatsuji, Dr. Haruhiko Inoue and Dr. Chang-Jie Jiang of the National Institute of Agrobiological Sciences for providing the facilities for the pull-down and protoplast transfection experiments, as well as for valuable discussion and help. We thank Dr. Hiroaki Ichikawa of the National Institute of Agrobiological Sciences for providing the seeds of FOX lines (K33309), Prof. Tsuyoshi Nakagawa of Shimane University for providing the gateway vectors pGWB541 and pGWB542, and Dr. Haruhiko Inoue for providing the gateway vectors, modified pSAT4-Pubi-ADH-HA-NRluc and pSAT4-Pubi-ADH-Myc-CRluc. We also thank Dr. Tomoko Nozoye and Dr. Takeshi Senoura for valuable discussion.

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Authors and Affiliations

  1. Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan

    Lixia Zhang, Reiko Nakanishi Itai, Takashi Yamakawa & Hiromi Nakanishi

  2. Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, 1-308 Suematsu, Nonoichi, Ishikawa, 921-8836, Japan

    Naoko K. Nishizawa & Takanori Kobayashi

  3. PRESTO, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama, 332-0012, Japan

    Takanori Kobayashi

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  1. Lixia Zhang
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  2. Reiko Nakanishi Itai
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  3. Takashi Yamakawa
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  4. Hiromi Nakanishi
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  5. Naoko K. Nishizawa
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Correspondence to Takanori Kobayashi.

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Zhang, L., Nakanishi Itai, R., Yamakawa, T. et al. The Bowman–Birk Trypsin Inhibitor IBP1 Interacts with and Prevents Degradation of IDEF1 in Rice. Plant Mol Biol Rep 32, 841–851 (2014). https://doi.org/10.1007/s11105-013-0695-8

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