Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

Pavement cells distinguish touch from letting go


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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

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.

Similar content being viewed by others

Data availability

Additional videos of calcium responses can be found at 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


  1. Coutand, C. Mechanosensing and thigmomorphogenesis, a physiological and biomechanical point of view. Plant Sci. 179, 168–182 (2010).

    Article  CAS  Google Scholar 

  2. Chehab, E. W., Eich, E. & Braam, J. Thigmomorphogenesis: a complex plant response to mechano-stimulation. J. Exp. Bot. 60, 43–56 (2009).

    Article  CAS  PubMed  Google Scholar 

  3. Braam, J. & Davis, R. W. Rain-, wind-, and touch-induced expression of calmodulin and calmodulin-related genes in Arabidopsis. Cell 60, 357–364 (1990).

    Article  CAS  PubMed  Google Scholar 

  4. Hamant, O. & Haswell, E. S. Life behind the wall: sensing mechanical cues in plants. BMC Biol. 15, 59 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  5. Landrein, B. & Ingram, G. Connected through the force: mechanical signals in plant development. J. Exp. Bot. 70, 3507–3519 (2019).

    Article  CAS  PubMed  Google Scholar 

  6. Kurusu, T. et al. Plasma membrane protein OsMCA1 is involved in regulation of hypo-osmotic shock-induced Ca2+ influx and modulates generation of reactive oxygen species in cultured rice cells. BMC Plant Biol. 12, 11 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Yuan, F. et al. OSCA1 mediates osmotic-stress-evoked Ca2+ increases vital for osmosensing in Arabidopsis. Nature 514, 367–371 (2014).

    Article  CAS  PubMed  Google Scholar 

  8. Basu, D. & Haswell, E. S. The mechanosensitive ion channel MSL10 potentiates responses to cell swelling in Arabidopsis seedlings. Curr. Biol. 30, 2716–2728 (2020).

    Article  CAS  PubMed  Google Scholar 

  9. Finkler, A., Ashery-Padan, R. & Fromm, H. CAMTAs: calmodulin-binding transcription activators from plants to human. FEBS Lett. 581, 3893–3898 (2007).

    Article  CAS  PubMed  Google Scholar 

  10. Whalley, H. J. et al. Transcriptomic analysis reveals calcium regulation of specific promoter motifs in Arabidopsis. Plant Cell 23, 4079–4095 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Xu, Y. et al. Mitochondrial function modulates touch signalling in Arabidopsis thaliana. Plant J. 97, 623–645 (2019).

    Article  CAS  PubMed  Google Scholar 

  12. Darwish, C. et al. Touch signaling and thigmomorphogenesis are regulated by complementary CAMTA3- and JA-dependent pathways. Sci. Adv. 8, eabm2091 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Matsumura, M. et al. Mechanosensory trichome cells evoke a mechanical stimuli-induced immune response in Arabidopsis thaliana. Nat. Commun. 13, 1216 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Liu, H. et al. Gradient mechanical properties facilitate Arabidopsis trichome as mechanosensor. ACS Appl. Mater. Interf. 8, 9755–9761 (2016).

    Article  CAS  Google Scholar 

  15. Zhou, L. H. et al. The Arabidopsis trichome is an active mechanosensory switch. Plant Cell Environ. 40, 611–621 (2017).

    Article  CAS  PubMed  Google Scholar 

  16. Symonds, V. V. et al. Mapping quantitative trait loci in multiple populations of Arabidopsis thaliana identified natural allelic variation for trichome density. Genetics 169, 1649–1658 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Sato, Y., Shimizu-Inatsugi, R., Yamazaki, M., Shimizu, K. K. & Nagano, A. J. Plant trichomes and a single gene GLABRA1 contribute to insect community composition on field-grown Arabidopsis thaliana. BMC Plant Biol. 19, 163 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  18. Zhao, Y. et al. An expanded palette of genetically encoded Ca2+ indicators. Science 333, 1888–1891 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Wendler, S. & Zimmermann, U. Compartment analysis of plant cells by means of turgor pressure relaxation. II. Experimental results on Chara corallina. J. Membr. Biol. 85, 133–142 (1985).

    Article  Google Scholar 

  20. Steudle, E. Water flow in plants and its coupling to other processes: an overview. Methods Enzymol. 174, 183–225 (1989).

    Article  CAS  Google Scholar 

  21. Fricke, W. Water movement between epidermal cells of barley leaves—a symplastic connection? Plant Cell Environ. 23, 991–997 (2000).

    Article  Google Scholar 

  22. Hamilton, E. S., Schlegel, A. M. & Haswell, E. S. United in diversity: mechanosensitive ion channels in plants. Annu. Rev. Plant Biol. 66, 113–137 (2015).

    Article  CAS  PubMed  Google Scholar 

  23. Tatsumi, H. et al. Mechanosensitive channels are activated by stress in the actin stress fibres, and could be involved in gravity sensing in plants. Plant Biol. 16, 18–22 (2014).

    Article  PubMed  Google Scholar 

  24. Bellandi, A. et al. Diffusion and bulk flow of amino acids mediate calcium waves in plants. Sci. Adv. 8, eabo6693 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Johns, S., Hagihara, T., Toyota, M. & Gilroy, S. The fast and the furious: rapid long-range signaling in plants. Plant Physiol. 185, 694–706 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Waadt, R., Krebs, M., Kudla, J. & Schumacher, K. Multiparameter imaging of calcium and abscisic acid and high‐resolution quantitative calcium measurements using R‐GECO1‐mTurquoise in Arabidopsis. N. Phytol. 216, 303–320 (2017).

    Article  CAS  Google Scholar 

  27. Toyota, M. et al. Glutamate triggers long-distance, calcium-based plant defense signaling. Science 361, 1112–1115 (2018).

    Article  CAS  PubMed  Google Scholar 

  28. An, G. High efficiency transformation of cultured tobacco cells. Plant Physiol. 79, 568–570 (1985).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Pratt, A. I., Knoblauch, J. & Kunz, H. H. An updated pGREEN-based vector suite for cost-effective cloning in plant molecular biology. microPubl. Biol. 2020, 000317 (2020).

    Google Scholar 

  30. Feynman, R. P., Leighton, R. B. & Sands, M. The Feynman Lectures on Physics: The New Millennium Edition—Mainly Electromagnetism and Matter Vol. 2 (Basic Books, 2011).

  31. Tomos, A. D. & Leigh, R. A. The pressure probe: a versatile tool in plant cell physiology. Annu. Rev. Plant Biol. 50, 447–472 (1999).

    Article  CAS  Google Scholar 

  32. Motulsky, H. & Christopoulos, A. Fitting Models to Biological Data Using Linear and Nonlinear Regression (Oxford Univ. Press, 2004).

Download references


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.

Author information

Authors and Affiliations



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.

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

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

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Howell, A.H., Völkner, C., McGreevy, P. et al. Pavement cells distinguish touch from letting go. Nat. Plants 9, 877–882 (2023).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing