In this protocol, we present a noninvasive in planta bioimaging technique for the analysis of hydrogen peroxide (H2O2) and glutathione redox potential in adult Arabidopsis thaliana plants. The technique is based on the use of stereo fluorescence microscopy to image A. thaliana plants expressing the two genetically encoded fluorescent sensors roGFP2-Orp1 and Grx1-roGFP2. We provide a detailed step-by-step protocol for performing low magnification imaging with mature plants grown in soil or hydroponic systems. This protocol aims to serve the scientific community by providing an accessible approach to noninvasive in planta bioimaging and data analysis.

Noninvasive In Planta Live Measurements of H2O2 and Glutathione Redox Potential with Fluorescent roGFPs-Based Sensors / S. Buratti, M. Grenzi, G. Tortora, S.P. Nastasi, E. Dell'Aglio, A. Bassi, A. Costa (METHODS IN MOLECULAR BIOLOGY). - In: ROS Signaling in Plants : Methods and Protocols / [a cura di] Corpas, F.J., Palma, J.M.. - Cham : Springer, 2024. - ISBN 9781071638255. - pp. 45-64 [10.1007/978-1-0716-3826-2_4]

Noninvasive In Planta Live Measurements of H2O2 and Glutathione Redox Potential with Fluorescent roGFPs-Based Sensors

S. Buratti
Primo
;
M. Grenzi
Secondo
;
S.P. Nastasi;E. Dell'Aglio;A. Costa
Ultimo
2024

Abstract

In this protocol, we present a noninvasive in planta bioimaging technique for the analysis of hydrogen peroxide (H2O2) and glutathione redox potential in adult Arabidopsis thaliana plants. The technique is based on the use of stereo fluorescence microscopy to image A. thaliana plants expressing the two genetically encoded fluorescent sensors roGFP2-Orp1 and Grx1-roGFP2. We provide a detailed step-by-step protocol for performing low magnification imaging with mature plants grown in soil or hydroponic systems. This protocol aims to serve the scientific community by providing an accessible approach to noninvasive in planta bioimaging and data analysis.
Data analysis pipeline; Fluorescent biosensors; Glutathione redox status; Hydrogen peroxide; Image analysis; Noninvasive bioimaging; Salt stress
Settore BIO/04 - Fisiologia Vegetale
   DEcoding Metabolic Effectors in plant sTress Response and Acclimation" (DEMEtRA)
   DEMEtRA
   EUROPEAN COMMISSION
   101059695
2024
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/1044715
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