Comparative analyses of glutathione system of vacuoles and leucoplasts isolated from the storage parenchyma cells of dormant red beetroots (Beta vulgaris L.)

Organelle-specific localization of glutathione in plants grown under different light intensities and spectra

“…Based on results reported by Zechmann ( 2014 ), Bratt et al ( 2016 ), Hajdinák et al ( 2019 ) and Pradedova et al ( 2019 )…”

“… Based on results reported by Zechmann ( 2014 ), Bratt et al ( 2016 ), Hajdinák et al ( 2019 ) and Pradedova et al ( 2019 ) apo Apoplast, cyt cytosol, cw cell wall, ER endoplasmic reticulum, GFP green fluorescent protein, GSH reduced glutathione, GSSG oxidized glutathione (glutathione disulfide), HPLC high-performance liquid chromatography, leu leucoplast, mit mitochondrium, nuc nucleus, per peroxisome, total glutathione sum of GSH and GSSG, vac vacuole …”

“…Each of the approaches evaluated here has its own specific advantages and limitations, which are summarized in Table 1 . Additional details on targets for redox-sensitive green fluorescence protein (roGFP) are included from Bratt et al ( 2016 ), as are additional details on glutathione quantification from isolated organelles from Hajdinák et al ( 2019 ) and Pradedova et al ( 2019 ). Briefly, subcellular glutathione can be bound in situ or in vivo to glutathione-specific antibodies (total glutathione = sum of GSH and GSSG), redox-sensitive dyes (monobromobimane [mbrB] or monochlorobimane [mchlB]; GSH) or roGFP probes (GSH, GSSG), visualized by transmission electron microscopy (TEM), bright-field microscopy, confocal-laser scanning microscopy (CLSM) or fluorescence microscopy, respectively, and quantified by software-assisted image analysis.…”

“…HPLC with UV detection facilitates the quantification of GSH and GSSG without the need of derivatization. Such an approach has recently been applied to leucoplasts and vacuoles isolated from storage parenchyma cells of red beet ( Beta vulgaris L.) (Pradedova et al 2019 ). Generally, great care has to be taken during sample preparation to avoid auto-oxidation of GSH, washing out of and/or redistributing of glutathione between cell compartments.…”

“…Exceptions are the ER lumen, the vacuole and the apoplast, where GR, to regenerate GSH from GSSG, is absent (Noctor et al 2012 ; Müller-Schüssele et al 2021 ). A more positive reduction potential (E h ) and evidence for the presence of GT activity, which may use GSSG in the vacuole to reduce organic hydroperoxides (Öztetik 2008 ), was recently detected in vacuoles of dormant red beet taproot cells (Pradedova et al 2019 ). The distinct glutathione (and Asc) level and its redox state in individual cell compartments allow control of the local subcellular redox environment and gradient, which affects many redox-dependent metabolic processes and is therefore considered a valuable marker for the plant stress response (Foyer and Noctor 2009 ; Noctor et al 2012 , 2013 ; Müller et al 2014 ; Zechmann 2020 ; Dorion et al 2021 ).…”