Plant extracellular ATP signalling by plasma membrane NADPH oxidase and Ca2+ channels

Authors

  • Vadim Demidchik,

    1. Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK
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    • Present addresses: VD; Department of Biology, University of Essex, Colchester, CO43SQ, United Kingdom. ZS; College of Life Science, Hebei Normal University, Yuhua East Road, Shijiazhang 050016, Hebei, China. RS; Riken Plant Science Center, Yokohama City, Kanagawa 230-0045, Japan. LR; Departamento de Biología Vegetal, Facultad de Ciencias, Universidad de Málaga, Campus de Teatinos, 29071 Málaga, Spain.

    • These authors contributed equally to this work.

  • Zhonglin Shang,

    1. Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK
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    • Present addresses: VD; Department of Biology, University of Essex, Colchester, CO43SQ, United Kingdom. ZS; College of Life Science, Hebei Normal University, Yuhua East Road, Shijiazhang 050016, Hebei, China. RS; Riken Plant Science Center, Yokohama City, Kanagawa 230-0045, Japan. LR; Departamento de Biología Vegetal, Facultad de Ciencias, Universidad de Málaga, Campus de Teatinos, 29071 Málaga, Spain.

    • These authors contributed equally to this work.

  • Ryoung Shin,

    1. Donald Danforth Plant Science Center, St Louis, MO 63132, USA
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    • Present addresses: VD; Department of Biology, University of Essex, Colchester, CO43SQ, United Kingdom. ZS; College of Life Science, Hebei Normal University, Yuhua East Road, Shijiazhang 050016, Hebei, China. RS; Riken Plant Science Center, Yokohama City, Kanagawa 230-0045, Japan. LR; Departamento de Biología Vegetal, Facultad de Ciencias, Universidad de Málaga, Campus de Teatinos, 29071 Málaga, Spain.

    • These authors contributed equally to this work.

  • Elinor Thompson,

    1. Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK
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  • Lourdes Rubio,

    1. Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK
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    • Present addresses: VD; Department of Biology, University of Essex, Colchester, CO43SQ, United Kingdom. ZS; College of Life Science, Hebei Normal University, Yuhua East Road, Shijiazhang 050016, Hebei, China. RS; Riken Plant Science Center, Yokohama City, Kanagawa 230-0045, Japan. LR; Departamento de Biología Vegetal, Facultad de Ciencias, Universidad de Málaga, Campus de Teatinos, 29071 Málaga, Spain.

  • Anuphon Laohavisit,

    1. Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK
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  • Jennifer C. Mortimer,

    1. Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK
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  • Stephen Chivasa,

    1. School of Biological and Biomedical Sciences, University of Durham, Durham DH1 3LE, UK
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  • Antoni R. Slabas,

    1. School of Biological and Biomedical Sciences, University of Durham, Durham DH1 3LE, UK
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  • Beverley J. Glover,

    1. Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK
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  • Daniel P. Schachtman,

    1. Donald Danforth Plant Science Center, St Louis, MO 63132, USA
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  • Sergey N. Shabala,

    1. School of Agricultural Sciences, University of Tasmania, Hobart, Tas. 7001, Australia
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  • Julia M. Davies

    Corresponding author
    1. Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK
      *(fax +44 (0) 1223 333 953; e-mail jmd32@cam.ac.uk).
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*(fax +44 (0) 1223 333 953; e-mail jmd32@cam.ac.uk).

Summary

Extracellular ATP regulates higher plant growth and adaptation. The signalling events may be unique to higher plants, as they lack animal purinoceptor homologues. Although it is known that plant cytosolic free Ca2+ can be elevated by extracellular ATP, the mechanism is unknown. Here, we have studied roots of Arabidopsis thaliana to determine the events that lead to the transcriptional stress response evoked by extracellular ATP. Root cell protoplasts were used to demonstrate that signalling to elevate cytosolic free Ca2+ is determined by ATP perception at the plasma membrane, and not at the cell wall. Imaging revealed that extracellular ATP causes the production of reactive oxygen species in intact roots, with the plasma membrane NADPH oxidase AtRBOHC being the major contributor. This resulted in the stimulation of plasma membrane Ca2+-permeable channels (determined using patch-clamp electrophysiology), which contribute to the elevation of cytosolic free Ca2+. Disruption of this pathway in the AtrbohC mutant impaired the extracellular ATP-induced increase in reactive oxygen species (ROS), the activation of Ca2+ channels, and the transcription of the MAP kinase3 gene that is known to be involved in stress responses. This study shows that higher plants, although bereft of purinoceptor homologues, could have evolved a distinct mechanism to transduce the ATP signal at the plasma membrane.

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