The secreted redox sensor roGFP2-Orp1 reveals oxidative dynamics in the plant apoplast

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The secreted redox sensor roGFP2-Orp1 reveals oxidative dynamics in the plant apoplast

Authors

Ingelfinger, J.; Zander, L.; Seitz, P. L.; Trentmann, O.; Tiedemann, S.; Sprunck, S.; Dresselhaus, T.; Meyer, A. J.; Müller-Schüssele, S. J.

Abstract

Specific generation of reactive oxygen species (ROS) is important for signalling and defence in many organisms. In plants, different types of ROS perform useful biological functions in the extracellular space (apoplast) supporting processes such as polymerisation and cell wall remodelling. In addition, increased formation of extracellular superoxide and hydrogen peroxide occurs during immune responses. Our current knowledge of apoplastic ROS dynamics is based on luminescence assays, ROS staining methods and indirect evidence via changes to intracellular redox balance. However, dynamic monitoring of extracellular redox processes in vivo remains difficult. Using two evolutionary distant land plant model species, the moss Physcomitrium patens and the flowering plant Arabidopsis thaliana, we test whether the genetically encoded redox biosensor roGFP-Orp1 can be used to assess extracellular redox dynamics. We found that secreted roGFP-Orp1 can inform about local diffusion barriers, as well as protein cysteinyl oxidation rate in the apoplast after pre-reduction. Observed re-oxidation rates were surprisingly slow within the range of hours. Comparing A. thaliana to P. patens, we found faster sensor re-oxidation that increased after triggering an immune response. Our data indicate differences in extracellular oxidative processes between species and within a species, depending on immune signalling.

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