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AFB1 controls rapid auxin signalling through membrane depolarization in Arabidopsis thaliana root

Nature Plants volume 7, pages 1229–1238 (2021)Cite this article

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An Author Correction to this article was published on 07 November 2022

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Abstract

The membrane potential reflects the difference between cytoplasmic and apoplastic electrical potentials and is essential for cellular operation. The application of the phytohormone auxin (3-indoleacetic acid (IAA)) causes instantaneous membrane depolarization in various cell types1,2,3,4,5,6, making depolarization a hallmark of IAA-induced rapid responses. In root hairs, depolarization requires functional IAA transport and TIR1–AFB signalling5, but its physiological importance is not understood. Specifically in roots, auxin triggers rapid growth inhibition7,8,9 (RGI), a process required for gravitropic bending. RGI is initiated by the TIR1–AFB co-receptors, with the AFB1 paralogue playing a crucial role10,11. The nature of the underlying rapid signalling is unknown, as well as the molecular machinery executing it. Even though the growth and depolarization responses to auxin show remarkable similarities, the importance of membrane depolarization for root growth inhibition and gravitropism is unclear. Here, by combining the DISBAC2(3) voltage sensor with microfluidics and vertical-stage microscopy, we show that rapid auxin-induced membrane depolarization tightly correlates with RGI. Rapid depolarization and RGI require the AFB1 auxin co-receptor. Finally, AFB1 is essential for the rapid formation of the membrane depolarization gradient across the gravistimulated root. These results clarify the role of AFB1 as the central receptor for rapid auxin responses.

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Fig. 1: DISBAC2(3) reports membrane potential in A. thaliana roots.
Fig. 2: Auxin induces depolarization and root growth inhibition in A. thaliana root tip.
Fig. 3: Auxin transport through PIN2 and AUX1 is not essential for rapid membrane depolarization.
Fig. 4: AFB1 triggers rapid membrane depolarization in response to IAA.
Fig. 5: AFB1-induced growth inhibition drives the early stages of root gravitropic response.

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Data availability

All raw images used in this study are available on Zenodo (https://doi.org/10.5281/zenodo.4922659). All the measured datasets are available as an Excel file in the Supplementary Information. Source data are provided with this paper.

Code availability

All the script used in this study are available at https://sourceforge.net/projects/lbopsis/ and https://sourceforge.net/projects/gravifast/.

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Acknowledgements

This work was supported by the European Research Council (grant no. 803048), Charles University Primus (grant no. PRIMUS/19/SCI/09), and in the initial stages by the Czech Science Foundation (GAČR grant no. 18-10116Y). S.S. acknowledges support from the Australian Research Council and China National Distinguished Expert Project (WQ20174400441). The authors thank E. Medvecká for technical support, M. Estelle, M. Prigge and W. Gray for Arabidopsis seeds, J. Friml for the initial aux1 pin2 cross, and J. Merrin for guidance in microfluidics.

Author information

Authors and Affiliations

  1. Department of Experimental Plant Biology, Charles University, Prague, Czech Republic

    Nelson B. C. Serre, Dominik Kralík & Matyáš Fendrych

  2. Department of Chemical Engineering, University of Chemistry and Technology, Prague, Czech Republic

    Dominik Kralík & Zdeněk Slouka

  3. Tasmanian Institute of Agriculture, College of Science and Engineering, University of Tasmania, Hobart, Tasmania, Australia

    Ping Yun & Sergey Shabala

  4. New Technologies—Research Centre, University of West Bohemia, Plzeň, Czech Republic

    Zdeněk Slouka

  5. International Research Centre for Environmental Membrane Biology, Foshan University, Foshan, China

    Sergey Shabala

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Contributions

N.B.C.S. and M.F. conceived the project, performed the experimental work and analysed and interpreted the data. P.Y. and S.S. conceived the impaling electrode membrane potential experiment and P.Y. measured and analysed the data. D.K., Z.S. and M.F. conceived the microfluidic chip design and D.K. fabricated and optimized it. N.B.C.S., M.F. and S.S. wrote the manuscript.

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Correspondence to Matyáš Fendrych.

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The authors declare no competing interests.

Additional information

Peer review information Nature Plants thanks Julian Dindas and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–5 and Supplementary Methods.

Reporting Summary

Supplementary Table 1

Reported membrane potential values for various plant species and tissues under an array of physiologically relevant conditions published in the literature.

Supplementary Video 1

Membrane potential of a Col0 root growing in microfluidic channels and treated with 100 nM IAA. Scale bar, 50 μm. Treatment starts at time 0. Time is min:s.

Supplementary Video 2

Col0 root tip angle measurements over 30 min after 90° gravistimulation. Angle value between (in degrees) the orange and blue line is indicated in the top left corner. Time in minutes is indicated in the bottom left corner.

Supplementary Video 3

afb1-3 root tip angle measurements over 30 min after 90° gravistimulation. Angle value (in degrees) between the orange and blue line is indicated in the top left corner. Time in minutes is indicated in the bottom left corner.

Supplementary Data 1

Statistical tests and results.

Supplementary Data 2

Measured data.

Supplementary Data 3

Measured data.

Supplementary Data 4

Measured data.

Supplementary Data 5

Measured data.

Source data

Source Data Fig. 1

Measured data.

Source Data Fig. 2

Measured data.

Source Data Fig. 3

Measured data.

Source Data Fig. 4

Measured data.

Source Data Fig. 5

Measured data.

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Serre, N.B.C., Kralík, D., Yun, P. et al. AFB1 controls rapid auxin signalling through membrane depolarization in Arabidopsis thaliana root. Nat. Plants 7, 1229–1238 (2021). https://doi.org/10.1038/s41477-021-00969-z

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