Nanoscale regulation of ROS signaling at the plasma membrane tunes the plant response to osmotic stress

A Arthur Poitout (Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro) J José M. Ugalde (Institut für Nutzpflanzenwissenschaften und Ressourcenschutz-Chemical Signalling, University of Bonn) L Lucille Gorgues (Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro) A Armelle Dongois (Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro) P Patricia Scholz P Philippe Nacry (Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro) C Carine Alcon (Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro) J Jean-Bernard Fiche (Centre de Biochimie Structurale, CNRS Unité Mixte de Recherche 5048, Institut National de la Santé et de la Recherche Médicale U1054, Université de Montpellier) X Xavier Dumont (Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro) M Marcelo Nollman (Centre de Biochimie Structurale, CNRS Unité Mixte de Recherche 5048, Institut National de la Santé et de la Recherche Médicale U1054, Université de Montpellier) K Komal Jhala (Department of Environmental Sciences, Institute of Biochemical Plant Pathology, Helmholtz Zentrum München) A Anton R. Schäffner (Department of Environmental Sciences, Institute of Biochemical Plant Pathology, Helmholtz Zentrum München) Y Yvon Jaillais L Lionel Verdoucq (Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro) A Andreas J. Meyer (Institut für Nutzpflanzenwissenschaften und Ressourcenschutz-Chemical Signalling, University of Bonn) A Alexandre Martinière (Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro)

Abstract

The spatiotemporal organization of proteins and lipids within membranes is crucial for ensuring proper cellular signaling. While the segregation of proteins and lipids into membrane nanodomains is well established, it remains unclear whether nanodomains can generate gradients of small diffusible molecules. In plants, reactive oxygen species (ROS), especially hydrogen peroxide (H 2 O 2 ), act as key signaling molecules in response to environmental stimuli such as osmotic stress. However, how extracellular H 2 O 2 affects intracellular signaling has remained unknown. Here, we show that osmotic stimulation induces the formation of localized, H 2 O 2 -rich nanoenvironments at the cytoplasmic face of the plasma membrane (PM) in Arabidopsis root cells. Using a PM-tethered H 2 O 2 biosensor, we found that these oxidized nanodomains arise from the clustering of RESPIRATORY BURST OXIDASE HOMOLOGs (RBOHs) and RHO OF PLANTS 6 (ROP6), in coordination with aquaporin-mediated H 2 O 2 transport via the PLASMA MEMBRANE INTRINSIC PROTEIN2;7 (PIP2;7). These local redox hotspots at the PM create a feedforward loop in which H 2 O 2 enhances ROP6 nanoclustering, thereby amplifying ROS signaling. Disruption of H 2 O 2 production or transport dampens both ROP6 clustering and anisotropic cell expansion, indicating a crucial role for spatially confined redox signaling in regulating plant growth under osmotic stress. Our findings propose a model in which ROP6/RBOHD-F/PIP2;7 nanodomains function as discrete redox signaling units, redefining ROS signaling at the PM as a structured, signal-specific, and compartmentalized process.

Article Details

Volume / Issue Vol. 123, Issue 26
Published June 30, 2026
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (16)

A

Arthur Poitout

Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro

J

José M. Ugalde

Institut für Nutzpflanzenwissenschaften und Ressourcenschutz-Chemical Signalling, University of Bonn

L

Lucille Gorgues

Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro

A

Armelle Dongois

Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro

P

Patricia Scholz

P

Philippe Nacry

Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro

C

Carine Alcon

Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro

J

Jean-Bernard Fiche

Centre de Biochimie Structurale, CNRS Unité Mixte de Recherche 5048, Institut National de la Santé et de la Recherche Médicale U1054, Université de Montpellier

X

Xavier Dumont

Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro

M

Marcelo Nollman

Centre de Biochimie Structurale, CNRS Unité Mixte de Recherche 5048, Institut National de la Santé et de la Recherche Médicale U1054, Université de Montpellier

K

Komal Jhala

Department of Environmental Sciences, Institute of Biochemical Plant Pathology, Helmholtz Zentrum München

A

Anton R. Schäffner

Department of Environmental Sciences, Institute of Biochemical Plant Pathology, Helmholtz Zentrum München

Y

Yvon Jaillais

L

Lionel Verdoucq

Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro

A

Andreas J. Meyer

Institut für Nutzpflanzenwissenschaften und Ressourcenschutz-Chemical Signalling, University of Bonn

A

Alexandre Martinière

Institute for Plant Sciences of Montpellier, Univ Montpellier, CNRS, Institut national de recherche pour l‘agriculture, l‘alimentation et l‘environnement, Institut Agro