Live Monitoring of ROS-Induced Cytosolic Redox Changes with roGFP2-Based Sensors in Plants

Ugalde, José Manuel; Fecker, Lara; Schwarzländer, Markus; Müller-Schüssele, Stefanie J; Meyer, Andreas

doi:10.1007/978-1-0716-2469-2_5

Cited by 11 publications

(3 citation statements)

References 34 publications

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“…2013) while in vivo samples often displayed decreased dynamic ranges of c. 5 in confocal microscopy or c. 3 in plate reader-based sensor read-out (Ugalde et al . 2021, 2022). This difference in dynamic range between in vitro and in vivo sensor read-out can be explained by interference of plant compounds that overlay the changes in sensor absorption in the UV range (below the isosbestic point at c. 425 nm).…”

Section: Discussionmentioning

confidence: 99%

High robustness of cytosolic glutathione redox potential under combined salt and osmotic stress in barley as revealed by the biosensor Grx1-roGFP2

Bohle

Klaus

Tegethof

et al. 2022

Preprint

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Barley is a staple crop of major global importance and relatively resilient to a wide range of stress factors in the field. Transgenic reporter lines to investigate physiological parameters during stress treatments remain scarce. We generated and characterized stable homozygous barley lines (cv. Golden Promise Fast) expressing the genetically-encoded biosensor Grx1-roGFP2, which indicates the redox potential of the major antioxidant glutathione in the cytosol. Our results demonstrate functionality of the sensor in living barley plants. We determined the glutathione redox potential (EGSH) of the cytosol to be in the range of 308 to -320 mV. EGSH was robust against a combined NaCl (150 mM) and water deficit treatment (-0.8 MPa) that caused growth retardation and showed only a minor oxidation after 96 h of treatment. We conclude that the generated reporter lines are a novel resource to study stress resilience in barley.

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Section: Discussionmentioning

confidence: 99%

High robustness of cytosolic glutathione redox potential under combined salt and osmotic stress in barley as revealed by the biosensor Grx1-roGFP2

Bohle

Klaus

Tegethof

et al. 2022

Preprint

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“…For sequential perfusion of different treatments, 4- to 5-day-old seedlings were mounted in a RC-22 perfusion chamber mounted on a P1 platform using a steel anchor harp with a 1.5 mm grid mesh (Warner Instruments, Hamden, CT, USA). Imaging buffer, and the indicated solutions of DTT and H 2 O 2 , were loaded into 50 mL syringes connected to a VC-8M valve controller (Warner Instruments) with 1.5 mm polyethylene tubes ( Ugalde et al, 2022 ).…”

Section: Methodsmentioning

confidence: 99%

Endoplasmic reticulum oxidoreductin provides resilience against reductive stress and hypoxic conditions by mediating luminal redox dynamics

et al. 2022

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Oxidative protein folding in the endoplasmic reticulum (ER) depends on the coordinated action of protein disulfide isomerases and ER oxidoreductins (EROs). Strict dependence of ERO activity on molecular oxygen as the final electron acceptor implies that oxidative protein folding and other ER processes are severely compromised under hypoxia. Here, we isolated viable Arabidopsis thaliana ero1 ero2 double mutants that are highly sensitive to reductive stress and hypoxia. To elucidate the specific redox dynamics in the ER in vivo, we expressed the glutathione redox potential (EGSH) sensor Grx1-roGFP2iL-HDEL with a midpoint potential of -240 mV in the ER of Arabidopsis plants. We found EGSH values of -241 mV in wild-type plants, which is less oxidizing than previously estimated. In the ero1 ero2 mutants, luminal EGSH was reduced further to -253 mV. Recovery to reductive ER stress induced by dithiothreitol, was delayed in ero1 ero2. The characteristic signature of EGSH dynamics in the ER lumen triggered by hypoxia was affected in ero1 ero2 reflecting a disrupted balance of reductive and oxidizing inputs, including nascent polypeptides and glutathione entry. The ER redox dynamics can now be dissected in vivo, revealing a central role of EROs as major redox integrators to promote luminal redox homeostasis.

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“…To compare the antioxidant role of H2S-mediated persulfidation with that of others PTMs, such as GSH-mediated S-glutathionlyation, several features need to be taken into account, including the cellular concentration of H2S in the microenvironment. The concentrations of H2O2 and GSH in plant cells have been well documented (Cheeseman, 2006;Hasanuzzaman et al, 2017;Smirnoff and Arnaud, 2019) and measurements at a spatiotemporal resolution of H2O2 and GSH have been improved by means of genetically encoded sensors (Niemeyer et al, 2021;Nietzel et al, 2019;Ugalde et al, 2021aUgalde et al, , 2021bUgalde et al, , 2022. Unfortunately, the information on the cellular H2S content is scarce.…”

Section: Summary and Perspectivesmentioning

confidence: 99%

Hydrogen Sulfide Signaling in Plants

Huang

Xie

2023

Antioxidants & Redox Signaling

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Significance: Hydrogen sulfide (H2S) is a multitasking potent regulator that facilitates plant growth, development, and responses to environmental stimuli. Recent Advances:The important beneficial effects of H2S in various aspects of plant physiology aroused the interest of this chemical for agriculture. Protein cysteine persulfidation has been recognized as the main redox regulatory mechanism of H2S signaling. An increasing number of studies, including large-scale proteomic analyses and function characterizations, have revealed that H2S-mediated persulfidations directly regulate protein functions, altering downstream signaling in plants. To date, the importance of H2S-mediated persufidation in several abscisic acid signaling-controlling key proteins has been assessed as well as their role in stomatal movements, largely contributing to the understanding of the plant H2S-regulatory mechanism. Critical Issues:The molecular mechanisms of the H2S sensing and transduction in plants remain elusive. The correlation between H2S-mediated persulfidation with other oxidative posttranslational modifications of cysteines are still to be explored.Future Directions: Implementation of advanced detection approaches for the spatiotemporal monitoring of H2S levels in cells and the current proteomic profiling strategies for the identification and quantification of the cysteine site-specific persulfidation will provide insight into the H2S signaling in plants.

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Live Monitoring of ROS-Induced Cytosolic Redox Changes with roGFP2-Based Sensors in Plants

Cited by 11 publications

References 34 publications

High robustness of cytosolic glutathione redox potential under combined salt and osmotic stress in barley as revealed by the biosensor Grx1-roGFP2

High robustness of cytosolic glutathione redox potential under combined salt and osmotic stress in barley as revealed by the biosensor Grx1-roGFP2

Endoplasmic reticulum oxidoreductin provides resilience against reductive stress and hypoxic conditions by mediating luminal redox dynamics

Hydrogen Sulfide Signaling in Plants

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