Systematic monitoring of 2-Cys peroxiredoxin-derived redox signals unveiled its role in attenuating carbon assimilation rate

Abstract

Transmission of reductive and oxidative cues from the photosynthetic electron transport chain to redox regulatory protein networks plays a crucial role in coordinating chloroplast metabolism and photosynthetic activities. The tight balance between these two signals dictates the cellular response to changing light conditions. The highly abundant stromal peroxidase, 2-Cys peroxiredoxin (2-Cys Prx), derives light-dependent oxidative signals by transferring oxidizing equivalents of photosynthetically produced H2O2 to redox-regulated proteins. However, a comprehensive physiological understanding of the role of oxidative signals under changing light intensities is still lacking, mainly due to the difficulties in accurately and nondestructively monitoring them. Here, we introduced chl-roGFP2-PrxΔCR, a 2-Cys Prx-based biosensor, into Arabidopsis thaliana chloroplasts to monitor the dynamic changes in photosynthetically-derived oxidative signaling. We showed that chl-roGFP2-PrxΔCR oxidation states reflected the similar oxidation patterns of endogenous 2-Cys Prx under varying light conditions, as determined by both redox western blots and fluorescence measurements. Furthermore, systematic monitoring of probe oxidation throughout the day led to the identification of the dynamic light intensity range of 2-Cys Prx-mediate redox signaling and demonstrated the induction of quantifiable redox signals under low-light conditions. The presented data highlights the role of oxidative signals under nonstressed conditions and suggests 2-Cys Prx-based biosensor as a powerful tool to explore their dynamic in higher plants.

Early detection of late blight in potato by whole‐plant redox imaging

“…High spatiotemporal‐resolution‐monitoring of glutathione redox potential ( E GSH ) can be achieved using redox‐sensitive green fluorescent proteins (roGFPs; Meyer et al., 2007; Meyer & Dick, 2010), which carry two engineered cysteine residues that form an intramolecular disulfide bridge that impacts its fluorescence characteristics (Dooley et al., 2004; Hanson et al., 2004). roGFP‐based biosensors are effective tools for exploring redox dynamics in subcellular compartments at high spatiotemporal resolution in plants (Bratt et al., 2016; Gutscher et al., 2008; Jiang et al., 2006; Lampl et al., 2022; Meyer, 2008; Meyer & Dick, 2010; Nietzel et al., 2019; Schwarzländer et al., 2016; Ugalde et al., 2021; Ugalde et al., 2022). Genetically encoded biosensors have also been introduced into crop plants, including potato (Chamusco et al., 2022; Hipsch et al., 2021; Huang et al., 2014; Rattanawong et al., 2021), and spatially resolved mapping of stress‐induced long‐term redox perturbations has recently been demonstrated by whole‐plant imaging of potato plants expressing roGFP2 in their chloroplasts (Hipsch et al., 2021).…”

Early detection of late blight in potato by whole-plant redox imaging

“…High spatiotemporal-resolution-monitoring of glutathione redox potential ( E GSH ) can be achieved using redox-sensitive green fluorescent proteins (roGFPs) (Meyer et al, 2007; Meyer & Dick, 2010), which carry two engineered cysteine residues that form an intramolecular disulfide bridge which impacts its fluorescence characteristics (Dooley et al, 2004; Hanson et al, 2004). roGFP-based biosensors are effective tools for exploring redox dynamics in subcellular compartments at high spatiotemporal resolution in plants (Bratt et al, 2016; Gutscher et al, 2008; Jiang et al, 2006; Lampl et al, 2022; Meyer, 2008; Meyer & Dick, 2010; Nietzel et al, 2019; Schwarzländer et al, 2016; Ugalde et al, 2020, 2022). Spatially resolved mapping of stress-induced long-term redox perturbations has recently been demonstrated by whole-plant imaging of potato plants expressing roGFP2 in their chloroplasts (Hipsch et al, 2021b).…”