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PtdIns(3,5)P2 mediates root hair shank hardening in Arabidopsis

Nature Plantsvolume 4pages888897 (2018) | Download Citation


Root hairs elongate by tip growth and simultaneously harden the shank by constructing the inner secondary cell wall layer. While much is known about the process of tip growth1, almost nothing is known about the mechanism by which root hairs harden the shank. Here we show that phosphatidylinositol-3,5-bisphosphate (PtdIns(3,5)P2), the enzymatic product of FORMATION OF APLOID AND BINUCLEATE CELLS 1 (FAB1), is involved in the hardening of the shank in root hairs in Arabidopsis. FAB1 and PtdIns(3,5)P2 localize to the plasma membrane along the shank of growing root hairs. By contrast, phosphatidylinositol 4-phosphate 5-kinase 3 (PIP5K3) and PtdIns(4,5)P2 localize to the apex of the root hair where they are required for tip growth. Reduction of FAB1 function results in the formation of wavy root hairs while those of the wild type are straight. The localization of FAB1 in the plasma membrane of the root hair shank requires the activity of Rho-related GTPases from plants 10 (ROP10) and localization of ROP10 requires FAB1 activity. Computational modelling of root hair morphogenesis successfully reproduces the wavy root hair phenotype. Taken together, these data demonstrate that root hair shank hardening requires PtdIns(3,5)P2/ROP10 signalling.

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We thank T. Miura (Kyushu University, Japan) for his helpful comments on the computational model, T. Nakagawa (Shimane University, Japan) for providing pGWB vectors, C. Ambrose (University of Saskatchewan, Canada) for providing the GFP–MBD-expressing line, T. Hashimoto (NAIST, Japan) for providing the EB1b–GFP-expression line, Y. Jailais (Université de Lyon, France) for providing the CITRINE–2:PHPLC-expressing line and fruitful discussion, and Y. Oda (National Institute for Genetics, Japan) and S. Sakamoto and N. Mitsuda (National Institute of Advanced Industrial Science and Technology, Japan) for fruitful discussion about cell wall components. We thank T. Ando and N. Kodera (Kanazawa University) for providing us with experimental instruments, and T. Nakayama-Watanabe for critical suggestions for data analysis. We also thank K. Tamura and I. Hara-Nishimura (Kyoto University) for fruitful discussion. This work was supported by JSPS KAKENHIJP16H05068 to M.H.S., JP17K08200 to T. Hirano, JP18K06260 to H.T.-I, JP16K06260 to T.A., 16KT0170 to T.A., 17K15238 to M.K., JP16H06280, 17K19380, 18H05492, a Grant for Basic Science Research Projects from The Sumitomo Foundation (160146), and a Grant from The Canon Foundation to T. Higaki, Marie Curie Actions; Incoming Interaction Fellowship (ID: 022275) to S.T.

Author information


  1. Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Kyoto, Japan

    • Tomoko Hirano
    • , Seiji Takeda
    •  & Masa H. Sato
  2. Nano Life Science Institute, Kanazawa University, Kanazawa, Japan

    • Hiroki Konno
  3. Biotechnology Research Department, Kyoto Prefectural Agriculture Forestry and Fisheries Technology Center, Kyoto, Japan

    • Seiji Takeda
  4. Department of Plant Sciences, University of Oxford, Oxford, UK

    • Liam Dolan
  5. Institute for Chemical Research, Kyoto University, Kyoto, Japan

    • Mariko Kato
    •  & Takashi Aoyama
  6. International Research Organization for Advanced Science and Technology, Kumamoto University Kurokami, Kumamoto, Japan

    • Takumi Higaki
  7. Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    • Hisako Takigawa-Imamura


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T. Hirano, M.K., T.A. and M.H.S. conceived and designed the study. T. Hirano and S.T. performed the experiments. H.K. performed AFM. H.T.-I. and T. Higaki performed the mathematical modelling. T. Hirano, S.T., L.D., M.K., T.A., T. Higaki, H.T.-I. and M.H.S. analysed the data. T. Hirano, H.T-I., L.D. and M.H.S. wrote the manuscript. M.H.S. supervised the project.

Competing interests

The authors declare no competing interests.

Corresponding author

Correspondence to Masa H. Sato.

Supplementary information

  1. Supplementary Information

    Supplementary Figures 1–19 and Supplementary Video legends.

  2. Reporting Summary

  3. Supplementary Table 1

    List of materials used for this study.

  4. Supplementary Video 1

    Time lapse images PI(3,5)P2 and PI(4,5)P2 fluorescence of initiation step of root hair elongation.

  5. Supplementary Video 2

    Time lapse images PtdIns(3,5)P2 and PtdIns(4,5)P2 fluorescence of elongating step of root hair.

  6. Supplementary Video 3

    Time lapse images PtdIns(3,5)P2 and PtdIns(4,5)P2 fluorescence of termination step of root hair elongation.

  7. Supplementary Video 4

    Computer simulation of the growing process of the wild type root hair in the air.

  8. Supplementary Video 5

    Computer simulation of the growing process of the wild type root hair in the gel.

  9. Supplementary Video 6

    Computer simulation of the wavy root hair in the air.

  10. Supplementary Video 7

    Computer simulation of the wavy root hair in the gel.

  11. Supplementary Video 8

    Computer simulation of the swollen root hair in the air.

  12. Supplementary Video 9

    Computer simulation of the swollen root hair in the gel.

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