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Pavement cells distinguish touch from letting go

Nature Plants volume 9, pages 877–882 (2023)Cite this article

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Abstract

A micro-cantilever technique applied to individual leaf epidermis cells of intact Arabidopsis thaliana and Nicotiana tabacum synthesizing genetically encoded calcium indicators (R-GECO1 and GCaMP3) revealed that compressive forces induced local calcium peaks that preceded delayed, slowly moving calcium waves. Releasing the force evoked significantly faster calcium waves. Slow waves were also triggered by increased turgor and fast waves by turgor drops in pressure probe tests. The distinct characteristics of the wave types suggest different underlying mechanisms and an ability of plants to distinguish touch from letting go.

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Fig. 1: Non-invasively evoked responses of cytosolic calcium to mechanical stimulation in pavement cells of the Arabidopsis abaxial leaf epidermis.
Fig. 2: Invasively evoked cytosolic calcium responses to mechanostimulation in pavement cells of the abaxial Arabidopsis leaf epidermis, and comparison of propagation rates.

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Data availability

Additional videos of calcium responses can be found at https://doi.org/10.5281/zenodo.6909354. Source data are provided with this paper.

Code availability

The algorithm created for determining average fluorescence intensities at defined distances from the point of impact is available at https://github.com/patrickmcgreevy/geco_data_processing.git.

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Acknowledgements

We thank V. V. Vasina for assistance with generating stable Nicotiana tabacum pUBQ10::R-GECO1 transformants, M. Toyoda and K. Tanaka for sharing 35S::GCaMP3 Arabidopsis seeds, S. Mühlbauer (LMU Munich) for help with figure design, and C. Cody and A. Linskey for plant care. Special thanks to D. L. Mullendore for troubleshooting and support. A.H.H. and C.V. acknowledge support from the Washington State University Elling and Higinbotham scholarship programme and a Washington State University Franceschi training grant. M.K. was funded by National Science Foundation (NSF) grant no. IOS-1656769. H.-H.K. was funded by an NSF Career Award (no. IOS-1553506), and S.G. was supported by NSF grant no. MCB2016177. We acknowledge technical support from the Franceschi Microscopy and Imaging Center at Washington State University, Pullman.

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Authors and Affiliations

  1. School of Biological Sciences, Washington State University, Pullman, WA, USA

    Alexander H. Howell, Carsten Völkner, Hans-Henning Kunz, Winfried S. Peters & Michael Knoblauch

  2. Department of Plant Biochemistry, Ludwig Maximilian Universität München, Planegg-Martinsried, Germany

    Carsten Völkner & Hans-Henning Kunz

  3. School of Electrical Engineering and Computer Science, Washington State University, Pullman, WA, USA

    Patrick McGreevy

  4. Department of Physics, Technical University of Denmark, Lyngby, Denmark

    Kaare H. Jensen

  5. Institute of Plant Biology and Biotechnology, Westfälische Wilhelms-Universität Münster, Münster, Germany

    Rainer Waadt

  6. Department of Botany, University of Wisconsin–Madison, Madison, WI, USA

    Simon Gilroy

  7. Department of Biology, Purdue University Fort Wayne, Fort Wayne, IN, USA

    Winfried S. Peters

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Contributions

A.H.H., K.H.J., M.K., H.-H.K., W.S.P. and C.V. designed the experiments, and A.H.H. conducted the experiments. A.H.H. and C.V. generated the stable Nicotiana tabacum transformants. R.W. generated the stable Arabidopsis thaliana pUBQ10::R-GECO1 plants. A.H.H., P.M., W.S.P. and C.V. analysed the primary data, and all authors discussed and interpreted the results. W.S.P. wrote the draft manuscript with input from A.H.H., H.-H.K. and C.V.; all authors discussed, augmented and improved the draft. M.K. coordinated the project.

Corresponding author

Correspondence to Michael Knoblauch.

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Nature Plants thanks the anonymous reviewers for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–6.

Reporting Summary

Supplementary Video 1

A cantilever placed on an Arabidopsis pUBQ10::R-GECO1 pavement cell at 01:33 and removed at 06:53 induces a slow wave after placement and a fast wave upon removal.

Supplementary Video 2

Cantilever placement on an Arabidopsis p35S::GCAMP3 pavement cell at 01:33 and its removal at 06:45 induce a slow and a fast wave, respectively.

Supplementary Video 3

A cantilever placed on a Nicotiana pUBQ10::R-GECO1 leaf epidermis at 02:12 and removed at 07:16 triggers a slow wave after placement and a fast wave after removal.

Supplementary Video 4

Pressure probe impalement of an Arabidopsis pUBQ10::R-GECO1 pavement cell at 00:49 and subsequent pressure increase (indicated in atmospheres). Pressure was reduced to ambient at 07:37.

Supplementary Video 5

Glass needles with closed tips but otherwise resembling pressure probes were used to test the response to impalement as such without turgor increase. Impalement of an Arabidopsis pUBQ10::R-GECO1 pavement cell at 00:54 and removal of the glass needle at 05:28 both triggered fast waves.

Supplementary Video 6

Original micrographs on which the analysis in Supplementary Fig. 4 (slow wave) is based. The large analysed cell is marked.

Supplementary Video 7

Original micrographs on which the analysis in Supplementary Fig. 4 (fast wave) is based. The large analysed cell is marked.

Supplementary Data 1

Numerical source data for the supplementary figures.

Source data

Source Data Figs. 1 and 2

Numerical source data for Figs. 1 and 2.

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Howell, A.H., Völkner, C., McGreevy, P. et al. Pavement cells distinguish touch from letting go. Nat. Plants 9, 877–882 (2023). https://doi.org/10.1038/s41477-023-01418-9

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