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Measuring surface potential changes on leaves

Nature Protocols volume 9, pages 1997–2004 (2014)Cite this article

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Abstract

We provide here a detailed protocol for studying the changes in electrical surface potential of leaves. This method has been developed over the years by plant physiologists and is currently used in different variants in many laboratories. The protocol records surface potential changes to measure long-distance electrical signals induced by diverse stimuli such as leaf wounding or current injection. This technique can be used to determine signaling speeds, to measure the connectivity between different plant organs and—by exploiting mutant plants—to identify transporters and ion channels involved in electrical signaling. The approach can be combined with the analysis of mRNA expression and of metabolite concentrations to correlate electrical signaling to specific physiological events. We describe how to use this protocol on Arabidopsis, looking at the effects of leaf wounding; however, it is broadly applicable to other plants and can be used to study other aspects of plant physiology. After wound infliction, surface potential recording takes ∼20 min per plant.

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Figure 1: Immobilization of an Arabidopsis leaf for wounding with forceps.
Figure 2: Ground and recording electrodes.
Figure 3: Current injection into leaves.
Figure 4: Traces from a typical recording of wound-induced surface potential changes.
Figure 5: Traces illustrating technical problems.

References

  1. Hille, B. Ionic Channels of Excitable Membranes 2nd edn. (Sinauer Associates, 1992).

  2. Bean, B.P. The action potential in mammalian central neurons. Nat. Rev. Neurosci. 8, 451–465 (2007).

    Article  CAS  Google Scholar 

  3. Stankovic, B., Witters, D.L., Zawadzki, T. & Davies, E. Action potentials and variation potentials in sunflower: an analysis of their relationships and distinguishing characteristics. Physiol. Plantarum 103, 51–58 (1998).

    Article  CAS  Google Scholar 

  4. Fromm, J. & Lautner, S. Electrical signals and their physiological significance in plants. Plant Cell Environ. 30, 249–257 (2007).

    Article  CAS  Google Scholar 

  5. Brenner, E.D. et al. Plant neurobiology: an integrated view of plant signaling. Trends Plant Sci. 11, 413–419 (2006).

    Article  CAS  Google Scholar 

  6. Stahlberg, R. Historical introduction to plant electrophysiology. in Plant Electrophysiology (ed. Volkov, A.G.) 3–14 (Springer, 2006).

  7. Pickard, B.G. Action potentials in higher plants. Bot. Rev. 39, 172–201 (1973).

    Article  Google Scholar 

  8. Sibaoka, T. Physiology of rapid movements in higher plants. Annu. Rev. Plant. Physiol. 20, 165–184 (1969).

    Article  CAS  Google Scholar 

  9. Stahlberg, R. & Cosgrove, D.J. Rapid alterations in growth-rate and electrical potentials upon stem excision in pea-seedlings. Planta 187, 523–531 (1992).

    Article  CAS  Google Scholar 

  10. Malone, M. Wound-induced hydraulic signals and stimulus transmission in Mimosa pudica L. New Phytol. 128, 49–56 (1994).

    Article  Google Scholar 

  11. Davies, E. Electrical signals in plants: facts and hypotheses. in Plant Electrophysiology (ed. Volkov, A.G.) 407–422 (Springer, 2006).

  12. Stahlberg, R., Cleland, R.E. & Van Volkenburgh, E. in Slow Wave Potentials: A Propagating Electrical Signal Unique to Higher Plants. Communication in Plants (eds. Baluska, F., Mancuso, S., & Volkmann, D.) 291–308 (Springer, 2006).

  13. Zimmermann, M.R., Maischak, H., Mithofer, A., Boland, W. & Felle, H.H. System potentials, a novel electrical long-distance apoplastic signal in plants, induced by wounding. Plant Physiol. 149, 1593–1600 (2009).

    Article  CAS  Google Scholar 

  14. Mousavi, S.A., Chauvin, A., Pascaud, F., Kellenberger, S. & Farmer, E.E. GLUTAMATE RECEPTOR-LIKE genes mediate leaf-to-leaf wound signalling. Nature 500, 422–426 (2013).

    Article  CAS  Google Scholar 

  15. Felle, H.H. & Zimmermann, M.R. Systemic signalling in barley through action potentials. Planta 226, 203–214 (2007).

    Article  CAS  Google Scholar 

  16. Fensom, D.S. The bioelectric potentials of plants and their functional significance. V. Some daily and seasonal changes in the electrical potential and resistance of living trees. Can. J. Botany 41, 831–851 (1963).

    Article  CAS  Google Scholar 

  17. Fromm, J. & Spanswick, R. Characteristics of action-potentials in willow (Salix viminalis L.). J. Exp. Bot. 44, 1119–1125 (1993).

    Article  Google Scholar 

  18. Favre, P. & Degli Agosti, R. Voltage-dependent action potentials in Arabidopsis thaliana. Physiol Plant 131, 263–272 (2007).

    CAS  PubMed  Google Scholar 

  19. Carpaneto, A. et al. Cold transiently activates calcium-permeable channels in Arabidopsis mesophyll cells. Plant Physiol. 143, 487–494 (2007).

    Article  CAS  Google Scholar 

  20. Minorsky, P.V. Temperature sensing by plants: a review and hypothesis. Plant Cell Environ. 12, 119–135 (1989).

    Article  CAS  Google Scholar 

  21. Krol, E., Dziubinska, H. & Trebacz, K. Low-temperature-induced transmembrane potential changes in mesophyll cells of Arabidopsis thaliana, Helianthus annuus and Vicia faba. Physiol. Plant 120, 265–270 (2004).

    Article  CAS  Google Scholar 

  22. Newman, I.A. Ion transport in roots: measurement of fluxes using ion-selective microelectrodes to characterize transporter function. Plant Cell Environ. 24, 1–14 (2001).

    Article  CAS  Google Scholar 

  23. Maffei, M., Bossi, S., Spiteller, D., Mithofer, A. & Boland, W. Effects of feeding Spodoptera littoralis on Lima bean leaves. I. Membrane potentials, intracellular calcium variations, oral secretions, and regurgitate components. Plant Physiol. 134, 1752–1762 (2004).

    Article  CAS  Google Scholar 

  24. Schaller, A. & Oecking, C. Modulation of plasma membrane H+-ATPase activity differentially activates wound and pathogen defense responses in tomato plants. Plant Cell 11, 263–272 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Wildon, D.C. et al. Electrical signaling and systemic proteinase-inhibitor induction in the wounded plant. Nature 360, 62–65 (1992).

    Article  CAS  Google Scholar 

  26. Stahlberg, R. & Cosgrove, D.J. The propagation of slow wave potentials in pea epicotyls. Plant Physiol. 113, 209–217 (1997).

    Article  CAS  Google Scholar 

  27. Farmer, E.E., Mousavi, S.A. & Lenglet, A. Leaf numbering for experiments on long distance signalling in Arabidopsis. Protoc. Exchange doi:10.1038/protex.2013.071 (2013).

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Acknowledgements

We thank M. Blanchard and F. Pascaud for help with the electrophysiology approaches. This research was supported by a Faculty of Biology and Medicine Interdisciplinary grant (to S.K. and E.E.F.) and by Swiss National Science Foundation grant no. 31003A-138235 (to E.E.F.).

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Authors and Affiliations

  1. Department of Plant Molecular Biology, University of Lausanne, Lausanne, Switzerland

    Seyed A R Mousavi, Chi Tam Nguyen & Edward E Farmer

  2. Department of Pharmacology and Toxicology, University of Lausanne, Lausanne, Switzerland

    Stephan Kellenberger

Authors
  1. Seyed A R Mousavi
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  2. Chi Tam Nguyen
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  3. Edward E Farmer
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  4. Stephan Kellenberger
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Contributions

S.A.R.M. developed the technique together with S.K. and E.E.F. and provided some figures; C.T.N. applied the technique, drafted the manuscript and provided most figures; and S.K. and E.E.F. wrote the manuscript. All authors contributed to the final manuscript.

Corresponding author

Correspondence to Stephan Kellenberger.

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The authors declare no competing financial interests.

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Mousavi, S., Nguyen, C., Farmer, E. et al. Measuring surface potential changes on leaves. Nat Protoc 9, 1997–2004 (2014). https://doi.org/10.1038/nprot.2014.136

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