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Reactive oxygen species produced by NADPH oxidase regulate plant cell growth

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Abstract

Cell expansion is a central process in plant morphogenesis, and the elongation of roots and root hairs is essential for uptake of minerals and water from the soil. Ca2+ influx from the extracellular store is required for (and sets the rates of) cell elongation in roots1. Arabidopsis thaliana rhd2 mutants are defective in Ca2+ uptake and consequently cell expansion is compromised—rhd2 mutants have short root hairs2,3 and stunted roots. To determine the regulation of Ca2+ acquisition in growing root cells we show here that RHD2 is an NADPH oxidase, a protein that transfers electrons from NADPH to an electron acceptor leading to the formation of reactive oxygen species (ROS). We show that ROS accumulate in growing wild-type (WT) root hairs but their levels are markedly decreased in rhd2 mutants. Blocking the activity of the NADPH oxidase with diphenylene iodonium (DPI) inhibits ROS formation and phenocopies Rhd2-. Treatment of rhd2 roots with ROS partly suppresses the mutant phenotype and stimulates the activity of plasma membrane hyperpolarization-activated Ca2+ channels, the predominant root Ca2+ acquisition system. This indicates that NADPH oxidases control development by making ROS that regulate plant cell expansion through the activation of Ca2+ channels.

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Figure 1: Characterization of RHD2/AtrbohC gene.
Figure 2: ROS accumulation (CM-H2DCF imaging) during root-hair elongation: transmission (top) and pseudocolour fluorescent images (bottom) are displayed for WT (a), rhd2 (b) and WT (c) after treatment with DPI.
Figure 3: ROS elevates [Ca2+]c in an rhd2 root-hair bulge.
Figure 4: Activation of hyperpolarization-activated Ca2+ channels by ROS.

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Acknowledgements

We thank D. Graham and A. Dark for help with screening and cultivation, respectively; E. Ryan, P. Shaw and K. Roberts for comments on the manuscript; and P. Doerner for support. This work was funded by the BBSRC, the Gatsby Foundation, the Leverhulme Trust and the European Union.

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Author notes

  1. Miguel Angel Torres

    Present address: Department of Biology and Curriculum in Genetics and Molecular Biology, CB3280, University of North Carolina, 108 Coker Hall, Chapel Hill, NC 27599-3280, USA

  2. Julia Foreman, Vadim Demidchik, John H. F. Bothwell, Miguel Angel Torres and Jonathan D. G. Jones: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Cell and Developmental Biology, John Innes Centre, Norwich, NR4 7UH, UK

    Julia Foreman, Panagiota Mylona, Paul Linstead, Silvia Costa & Liam Dolan

  2. Department of Plant Sciences, University of Cambridge, Cambridge, CB2 3EA, UK

    Vadim Demidchik, Henk Miedema & Julia M. Davies

  3. Marine Biological Association, Citadel Hill, PL1 2PB, Plymouth, UK

    John H. F. Bothwell & Colin Brownlee

  4. Sainsbury Laboratory, John Innes Centre, Norwich, NR4 7UH, UK

    Miguel Angel Torres & Jonathan D. G. Jones

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Correspondence to Liam Dolan.

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Foreman, J., Demidchik, V., Bothwell, J. et al. Reactive oxygen species produced by NADPH oxidase regulate plant cell growth. Nature 422, 442–446 (2003). https://doi.org/10.1038/nature01485

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