Exogenous GSH Research Articles

Evaluation of Antibacterial Activity of Exogenous Glutathione and its Synergism with Meropenem against Clinical Isolates of Acinetobacter baumannii

Introduction: Acinetobacter baumannii (A. baumannii) is the major nosocomial pathogen especially associated with ventilator associated pneumonia. It often poses significant challenge in treatment as it is resistant to almost all classes of antibiotics. Glutathione (GSH), a tripeptide antioxidant has been demonstrated with antibacterial activity in various bacteria. Aim: To detect the antibacterial activity of exogenous GSH against clinical isolates of A. baumannii and its synergism with meropenem. Materials and Methods: This prospective cross- sectional study was conducted at Department of Microbiology, Pondicherry Institute of Medical sciences, Puducherry, India. Sixty clinical isolates of A. baumannii from various clinical specimens between August to September 2018 were included in the study. The antibacterial activity of GSH at concentrations 10-20 mmol/L and meropenem were determined by broth dilution method. To determine the synergism, onto Muller Hinton Agar (MHA) with GSH (at Minimum Inhibitory Concentration (MIC)), Epsilometer (E) strips containing meropenem were placed over the lawn cultures. Fractional Inhibitory Concentration (FIC) of the isolates was also determined to demonstrate synergism of GSH with meropenem. The results were analysed by percentages and proportions. Results: Levels of MIC ranged from 16-18 mmol/L of exogenous GSH and >32 μg/mL for meropenem. The MIC of meropenem in the presence of GSH for all the isolates were <0.25 μg and the FIC <0.5, suggesting GSH synergistically interacts with meropenem. Conclusion: The GSH showed antibacterial activity against Acinetobacter baumannii and also acted synergistically with meropenem to reduce its MIC. Hence this study demonstrates the potential therapeutic utility of exogenous GSH against A. baumannii.

Mutation of SlSBPASE Aggravates Chilling-Induced Oxidative Stress by Impairing Glutathione Biosynthesis and Suppressing Ascorbate-Glutathione Recycling in Tomato Plants.

Sedoheptulose-1,7-bisphosphatase (SBPase) is a crucial enzyme for photosynthetic carbon assimilation in the Calvin-Benson cycle. Previous studies have shown that overexpression of SBPase is advantageous to chilling tolerance in plants; however, the mechanisms of SBPase acting in the improvement of chilling tolerance remain largely unknown. In the present study, we aimed to uncover the essential role of SBPase in the response of tomato plants to oxidative stress induced by low temperature. To fulfill that, we performed an array of comparative studies between slsbpase mutant plants that we previously generated using CRISPR/Cas9 genome editing system and their wild-type counterparts under chilling stress. It was observed that following a 24 h chilling treatment, slsbpase mutant plants accumulated higher levels of reactive oxygen species (ROS) than wild-type plants and consequently, more severe lipid peroxidation occurred in slsbpase plants. Activity assay of antioxidant enzymes showed that mutation in SlSBPASE significantly decreased activities of peroxidase (POD) and ascorbate peroxidase (APX), but surprisingly did not significantly alter activities of superoxide dismutase (SOD) and catalase (CAT) under the chilling condition. Notably, mutation in SlSBPASE reduced the contents of total ascorbate (AsA) and total glutathione (GSH) and suppressed the recycling of AsA and GSH in chilling-stressed tomato plants. In addition, activities of two GSH biosynthetic enzymes (gamma-glutamylcysteine synthetase and glutathione synthetase) and transcript abundance of their coding genes (GSH1 and GSH2) were markedly reduced in slsbpase mutant plants in comparison with those in wild-type plants under chilling stress. Furthermore, exogenous GSH remarkably mitigated chilling damage in slsbpase plants. Collectively, these results support that mutation in SlSBPASE aggravates chilling-induced oxidative stress by suppressing GSH biosynthesis and AsA-GSH recycling and suggest that SBPase is required for optimal response to chilling stress in tomato plants. The findings also shed light on the idea to mitigate chilling-induced damages by genetically manipulating a photosynthetic enzyme in plants.

Enhanced vacuole compartmentalization of cadmium in root cells contributes to glutathione-induced reduction of cadmium translocation from roots to shoots in pakchoi (Brassica chinensis L.)

Our previous studies showed that exogenous glutathione (GSH) decreased cadmium (Cd) concentration in shoots and alleviated the growth inhibition in pakchoi (Brassica chinensis L.) under Cd stress. Nevertheless, it is largely unknown how GSH decreases Cd accumulation in edible parts of pakchoi. This experiment mainly explored the mechanisms of GSH-induced reduction of Cd accumulation in shoot of pakchoi. The results showed that compared with sole Cd treatment, Cd + GSH treatment remarkably increased the expression of BcIRT1 and BcIRT2, and further enhanced the concentrations of Cd and Fe in root. By contrast, GSH application declined the concentration of Cd in the xylem sap. However, these results were not caused by xylem loading process because the expression of BcHMA2 and BcHMA4 had not significant difference between sole Cd treatment and Cd + GSH treatment. In addition, exogenous GSH significantly enhanced the expression of BcPCS1 and promoted the synthesis of PC2, PC3 and PC4 under Cd stress. At the same time, exogenous GSH also significantly improved the expression of BcABCC1 and BcABCC2 in the roots of seedling under Cd stress, suggesting that more PCs-Cd complexes may be sequestrated into vacuoles by ABCC1 and ABCC2 transporters. The results showed that exogenous GSH could up-regulate the expression of BcIRT1/2 to increase the Cd accumulation in root, and the improvement of PCs contents and the expression of BcABCC1/2 enhanced the compartmentalization of Cd in root vacuole of pakchoi under Cd stress. To sum up, exogenous GSH reduce the concentration of free Cd2+ in the cytoplast of root cells and then dropped the loading of Cd into the xylem, which eventually given rise to the reduction of Cd accumulation in edible portion of pakchoi.

Enhanced glutathione content improves lateral root development and grain yield in rice plants.

Enhanced glutathione content improves lateral root development by positively regulating the transcripts of root development genes responsive to glutathione treatment, thereby increasing the overall productivity of rice plants. Glutathione is primarily known as a cellular antioxidant molecule, but its role in lateral root development in rice plants has not been elucidated. Here, we have investigated its role in lateral root development of rice Oryza sativa L. Exogenous glutathione (GSH) promoted both the number and length of lateral roots in rice, and the GSH biosynthesis inhibitor buthionine sulfoximine (BSO) significantly reduced these parameters, compared to untreated plants. The inhibition by BSO was reversed with exogenous GSH. Transcript profiling by RNA-seq revealed that expression of the transcription factor genes DREB and ERF and the hormone-related genes AOS, LOX, JAZ, and SAUR were significantly downregulated in the BSO-treated plants and, in contrast, upregulated in plants treated with GSH and with GSH and BSO together. We generated OsGS-overexpressing transgenic plants in which the transgene is controlled by the abiotic-stress-inducible OsRab21 promoter to study the effect of endogenously increased GSH levels. In cold stress, transgenic rice plants enhanced stress tolerance and lateral root development by maintaining redox homeostasis and improving upregulating the expression of transcription factors and hormone-related genes involved in lateral root development. We observed improved root growth of OsGS-overexpressing plants in paddy fields compared to the wild-type controls. These traits may have alleviated transplanting stress during early growth in the field and accounted for the increased productivity. These results provide information and perspectives on the role of GSH in gene expression, lateral root development, and grain yield in rice.

Glutathione improves rice tolerance to submergence: insights into its physiological and biochemical mechanisms

Complete submergence (Sub) imposes detrimental effects on growth and survival of crop plants, including rice. Here, we investigated the beneficial effects of reduced glutathione (GSH) in mitigating Sub-induced adverse effects in two high-yielding rice cultivars BRRI dhan29 and dhan52. Both cultivars experienced growth defects, severe yellowing, necrosis and chlorosis, when they were completely immersed in water for 14 days. The poor growth performance of these cultivars was linked to biomass reduction, decreased levels of photosynthetic pigments and proline, increased levels of H2O2 and malondialdehyde, and declined activities of enzymatic antioxidants like superoxide dismutase, ascorbate peroxidase, peroxidase, catalase, glutathione peroxidase and glutathione S-transferase. Pretreatment with exogenous GSH led to significant growth restoration in both cultivars exposed to Sub. The elevated Sub-tolerance promoted by GSH could partly be attributed to increased levels of chlorophylls, carotenoids, soluble proteins and proline. Exogenous GSH also mitigated Sub-induced oxidative damage, as evidenced from reduced levels of H2O2 and malondialdehyde in accordance with the increased activities of antioxidant enzymes. Results revealed that dhan52 was more tolerant to Sub-stress than dhan29, and GSH successfully rescued both cultivars from the damage of Sub-stress. Collectively, our findings provided an insight into the GSH-mediated active recovery of rice from Sub-stress, thereby suggesting that external supply of GSH may be an effective strategy to mitigate the adverse effects of Sub in rice.

Exogenous glutathione revealed protection to bacterial spot disease: Modulation of photosystem II and H2O2 scavenging antioxidant enzyme system in Capsicum annum L

In a greenhouse, parallel two experiments were carried out for the estimation of the impact of exogenous GSH (reduced glutathione) on photosynthetic properties, the efficiency of the photosystem II, and the H2O2-scavenging mechanism in chilli (Capsicum annum L.) seedlings. On the bases of pathogenicity, resistant and susceptible verities were selected. The application of GSH increased the chlorophyll fluorescence parameters such as photochemical quantity yield (YII). It declined the excitation pressure (1-qP) of photosystem and production of non-regulated energy dissipation [Y (NO)] in Xanthomonas campestris treated chilli leaves. In the addition of GSH, accumulation of reactive oxygen species was down and maintained the activity of ROS-scavenging enzymes (enzymatic antioxidants) in resistant and susceptible plants. Consequently, GSH supplementation relieves inhibition of production of bacterial toxin and photosynthesis primarily by overwhelming stomatal limitations, enhancing the performance of photosystem, and modulating the antioxidant protection mechanism to defend chloroplasts from oxidation. The Present study confirmed that seed priming with GSH in chilli plants to relieve biotic stress.

Exogenous Glutathione Alleviation of Cd Toxicity in Italian Ryegrass (Lolium multiflorum) by Modulation of the Cd Absorption, Subcellular Distribution, and Chemical Form.

Subcellular fractions and the chemical forms of cadmium (Cd) reflect its level of toxicity to plants; however, these effects of exogenous glutathione (GSH) are poorly understood. We exposed two Italian ryegrass (Lolium multiflorum) cultivars (IdyII and Harukaze) to 50 µM Cd or 200 µM GSH to investigate the effect of GSH on the Cd uptake, subcellular compartments, and chemical forms. Cd significantly inhibited the plant growth, while GSH supplementation decreased this inhibition. The application of GSH significantly improved the Cd concentration in the roots but reduced that in the shoots and decreased the Cd translocation from root to shoot. The Cd concentration of the root in the cell wall was increased while the concentration in the soluble fraction was decreased when supplied with GSH. The inorganic form (80% ethanol for Cd extraction) in the roots was significantly reduced when treated with GSH. The Cd form extracted by 2% acetic acid (HAC) with low toxicity and immobility were greatly increased. In leaves, the application GSH decreased in any form of Cd form extracted. In conclusion, exogenous GSH decreased the translocation of Cd and alleviated Italian ryegrass Cd toxicity by accumulating more Cd in the root cell wall and immobilizing more Cd in lower toxicity fractions.

The critical role of the shoot base in inhibiting cadmium transport from root to shoot in a cadmium-safe rice line (Oryza sativa L.)

Cadmium (Cd) is harmful to rice and human, thus screening and understanding the mechanism of Cd-safe rice lines, which accumulate little Cd in brown rice, is necessary. D62B was screened as a Cd-safe rice line with low Cd translocation from roots to shoots, and there must be a switch restricting Cd transport from roots to shoots. Here we found that shoot base played the role as switch. Cd concentration in the shoot base of D62B was 1.57 times higher compared with a high Cd-accumulating rice line (Wujin4B) and lower Cd translocation under Cd stress. Glutathione (GSH) and phytochelatins (PCs) were important in this process. GSH and PCs concentrations in the shoot bases of D62B were 1.01– 1.83 times higher than Wujin4B as well as the glutathione S-transferase (GST) and phytochelatin synthase (PCS) concentrations, keeping in consistent with up-regulation of the genes OsGST and OsPCS1. PCs synthesis was further promoted by exogenous GSH. Our results prove the role of shoot bases as switch for restricting Cd transport in D62B due to its great potential for GSH and PCs biosynthesis, and thereby Cd chelation. This could be considered a key mechanism for low Cd accumulation in brown rice of the Cd-safe rice line.

A simple highly selective probe for discriminative visualization of endogenous cysteine, homocysteine and glutathione in living cells via three separated fluorescence channels

Biothiols, as basic biological reactive sulfur species, perform vital actions in many critical physiological processes. Simultaneous detection and direct visualization of intracellular biothiols has great significance to figure out their metabolic mechanisms and cellular functions in living organism. However, it is an enormous challenge to selectively sense biothiols, troubled by their similar chemical structures and properties. In this work, we reported a novel chlorinated coumarin based multi-signal fluorescent probe (CC) for discriminative detection of cysteine (Cys), homocysteine (Hcy) and glutathione (GSH) through three different emission channels. CC exhibited favorable sensing properties towards biothiols such as large fluorescence enhancement, low limits of detection (6, 3 and 200 nM for Cys, Hcy and GSH), fast response speed (15 min) and pretty good water solubility. Moreover, it was also utilized for discriminative visualization of endogenous and exogenous Cys, Hcy and GSH in A375 cells and Tcam-2 cells successfully.

Tn-Seq reveals hidden complexity in the utilization of host-derived glutathionein Francisellatularensis.

Host-derived glutathione (GSH) is an essential source of cysteine for the intracellular pathogen Francisella tularensis. In a comprehensive transposon insertion sequencing screen, we identified several F. tularensis genes that play central and previously unappreciated roles in the utilization of GSH during the growth of the bacterium in macrophages. We show that one of these, a gene we named dptA, encodes a proton-dependent oligopeptide transporter that enables growth of the organism on the dipeptide Cys-Gly, a key breakdown product of GSH generated by the enzyme γ-glutamyltranspeptidase (GGT). Although GGT was thought to be the principal enzyme involved in GSH breakdown in F. tularensis, our screen identified a second enzyme, referred to as ChaC, that is also involved in the utilization of exogenous GSH. However, unlike GGT and DptA, we show that the importance of ChaC in supporting intramacrophage growth extends beyond cysteine acquisition. Taken together, our findings provide a compendium of F. tularensis genes required for intracellular growth and identify new players in the metabolism of GSH that could be attractive targets for therapeutic intervention.

Astrocyte glutathione maintains endothelial barrier stability

Blood-brain barrier (BBB) impairment clearly accelerates brain disease progression. As ways to prevent injury-induced barrier dysfunction remain elusive, better understanding of how BBB cells interact and modulate barrier integrity is needed. Our metabolomic profiling study showed that cell-specific adaptation to injury correlates well with metabolic reprogramming at the BBB. In particular we noted that primary astrocytes (AC) contain comparatively high levels of glutathione (GSH)-related metabolites compared to primary endothelial cells (EC). Injury significantly disturbed redox balance in EC but not AC motivating us to assess 1) whether an AC-EC GSH shuttle supports barrier stability and 2) the impact of GSH on EC function. Using an isotopic labeling/tracking approach combined with Time-of-Flight Mass Spectrometry (TOF-MS) we prove that AC constantly shuttle GSH to EC even under resting conditions - a flux accelerated by injury conditions in vitro. In correlation, co-culture studies revealed that blocking AC GSH generation and secretion via siRNA-mediated γ-glutamyl cysteine ligase (GCL) knockdown significantly compromises EC barrier integrity. Using different GSH donors, we further show that exogenous GSH supplementation improves barrier function by maintaining organization of tight junction proteins and preventing injury-induced tight junction phosphorylation. Thus the AC GSH shuttle is key for maintaining EC redox homeostasis and BBB stability suggesting GSH supplementation could improve recovery after brain injury.

Characteristics of oxidative stress and antioxidant defenses by a mixed culture of acidophilic bacteria in response to Co2+ exposure.

During bioleaching of Cobalt from waste lithium-ion batteries, the biooxidation activity of acidophilic bacteria is inhibited by a high concentration of Co ion in the liquid phase. However, the mechanism for Co2+ toxicity to acidophilic bacteria has not been fully elucidated. In this study, the effects of Co2+ concentration on the biooxidation activity for Fe2+, intracellular reactive oxygen species (ROS) level and antioxidant defense systems in a mixed-culture of acidophilic bacteria (MCAB) were investigated. The results showed that the biooxidation activity of the MCAB was inhibited by Co2+. Furthermore, it was indicated that the intracellular ROS contents of the MCAB under conditions of 0.4M and 0.6M Co2+ were 2.60 and 3.34 times higher than that under the condition of 0M Co2+. The increase in intracellular malondialdehyde content indicated that the oxidative damage was induced by Co2+. Moreover, the antioxidant systems in MCAB were affected by Co2+. It was observed that the Co2+ exposure increased the catalase and glutathione peroxidase activities while reducing the superoxide dismutase activity and the intracellular glutathione (GSH) content. It was found that an exogenous GSH supplementation eliminated excess intracellular ROS and improved the biooxidation activity of the MCAB.

The ROS-associated programmed cell death causes the decline of pollen viability recovered from cryopreservation in Paeonia lactiflora.

After cryopreservation, the occurrence of apoptosis-like programmed cell death events induced by the accumulation of ROS reduces pollen viability. Cryopreservation, as a biotechnological means for long-term preservation of pollen, has been applied to many species. However, after cryopreservation, the viability of pollen significantly decreases via a mechanism that is not completely clear. In this study, the pollen of Paeonia lactiflora 'Zi Feng Chao Yang', which exhibits significantly reduced viability after liquid nitrogen (LN2) storage, was used to study the relationship among pollen viability, programmed cell death (PCD) and reactive oxygen species (ROS). The apoptosis rate was increased significantly in pollen with decreased viability after cryopreservation, and the changes in ROS generation and hydrogen peroxide (H2O2) were consistent with the apoptosis rate. Correlation analysis results showed that the apoptosis rate is positively correlated with ROS generation and H2O2 content. In addition, ascorbic acid (AsA), glutathione (GSH) and ascorbic acid reductase (APX) levels were significantly correlated with ROS and H2O2. After LN2 preservation for 8months, the exogenous antioxidants AsA and GSH at appropriate concentrations significantly decreased H2O2 content, inhibited PCD indicator levels, and increased cryopreserved pollen viability. These observations suggest that PCD occurred in pollen during LN2 preservation for 1-8months and was induced by the accumulation of ROS in pollen after cryopreservation, thus explaining the main reasons for the reduction in pollen viability after cryopreservation in LN2.

A novel fluorescent probe for detection of Glutathione dynamics during ROS-induced redox imbalance

Monitoring cellular GSH dynamics is of great value to understand its complex biological functions and related diseases. It still remains challenging to understand the mechanism of GSH dynamics in complex intracellular environment. Herein, a novel fluorescent probe featured with chlorinated coumarin-TCF was exploited for sensing of GSH with high selectivity and high sensitivity. Large fluorescence enhancement at 471 nm was observed upon addition of GSH with large emission distance (Δλem = 219 nm). This novel probe was successfully applied to monitor endogenous and exogenous GSH dynamics without interference of other thiols via fluorescence imaging. More importantly, using this probe, low dose reactive oxygen species induced GSH increasement through nuclear factor (erythroid-derived 2)-like 2 (Nrf2) signal pathway in BEL-7402 cells was observed. This novel fluorescent probe has the potential for quantitative monitoring of cellular GSH, which will promote further understanding of its physiological and pathological roles in biological systems.

Effects of exogenous glutathione on antioxidant system of cucumber seedlings under cadmium stress

With the development of social economy, environmental protection has become a hot topic. Excessive emissions of agricultural chemicals increase the accumulation of the heavy metal cadmium (Cd) in soil. When plants are stressed by high concentrations of heavy metals, their antioxidant capacity and detoxification capacity will decrease, which will lead to damage the cell mechanism of plants. Glutathione (GSH), as an important water-soluble antioxidant in the organism, can enhance plant resistance in stress. This study used cucumber seedlings as test material, using hydroponics, to research the effects of exogenous GSH on antioxidant system under Cd stress. The study showed that exogenous GSH could enhance the antioxidant capacity of cucumber leaves under Cd stress and increase the activity of SOD and POD, and reduce the content of MDA. However, the contents of reduced GSH and GSSH were reduced when exogenous glutathione was applied under Cd stress.

Insights into the Authentic Active Ingredients and Action Sites of Oral Exogenous Glutathione in the Treatment of Ischemic Brain Injury Based on Pharmacokinetic-Pharmacodynamic Studies.

Glutathione (GSH) has been reported to be closely related to various diseases of the central nervous system, yet its authentic active ingredients and action sites remain unclear. In the present study, oral exogenous GSH significantly alleviated ischemic brain injury, but this result was inconsistent with its low bioavailability and blood-brain barrier (BBB) permeability. To ascertain the exposure of GSH-derived ingredients, including GSH, cysteine (CYS), glutamate (Glu), glycine (GLY), CYS-GLY, and γ-glutamylcysteine (γ-GC) were systematically studied both in vitro and in vivo. The outcomes demonstrated that oral GSH not only increases the GSH and CYS levels in rat striatum and cortex, but it also can decrease the rise of intracerebral Glu concentration caused by ischemia/reperfusion surgery. Then the influence of GSH on the BBB was investigated via measuring IgG leakage, intracerebral endotoxin, and tight-junction proteins. All indicators showed that GSH dosing can repair the destroyed BBB. Oral GSH greatly enhances the exposure of GSH, CYS, CYS-GLY, and γ-GC in rat duodenum, jejunum, ileum, and colon. Accumulating evidence reveals a close link between brain injury and gastrointestinal dysfunction. Our findings further suggest that oral GSH significantly improves intestinal inflammatory damage and barrier disruptions. In conclusion, oral GSH can have a direct therapeutic role in brain injury by stabilizing intracerebral levels of GSH, CYS, and Glu. It can also play an indirect therapeutic role by enhancing the intestinal exposure of GSH, CYS, CYS-GLY, and γ-GC and improving intestinal barrier disruptions. SIGNIFICANCE STATEMENT: The authentic active ingredients and action sites of exogenous glutathione (GSH) in the treatment of ischemic brain injury are unclear. We have shown that oral exogenous GSH not only stabilizes intracerebral levels of GSH, cysteine (CYS), and glutamate (Glu) to act directly on brain injury, but it can also exert an indirect therapeutic role by improving intestinal barrier disruptions. These findings have great significance for revealing the therapeutic effect of GSH on ischemic brain injury and for promoting its further development and clinical application.

The Arabidopsis APR2 positively regulates cadmium tolerance through glutathione-dependent pathway

Cadmium (Cd) is a dangerous environmental pollutant with high toxicity to plants. The adenosine 5′-phosphosulfate reductase 2 (APR2) is the dominant APRs in Arabidopsis and plays an important role in reductive sulfate assimilation pathway. However, whether the involvement of plant APRs in Cd stress response is largely unclear. Herein, we report that APR2 functions in Cd accumulation and tolerance in Arabidopsis. The transcript levels of APR2 were markedly induced by Cd exposure. Transgenic plants overexpressing APR2 improved Cd tolerance, whereas knockout of APR2 reduced Cd tolerance. APR2-overexpressing plants with increased Cd accumulation and tolerance showed higher glutathione (GSH) and phytochelatin (PC) levels than the wild type and apr2 mutant plants, but lower H2O2 and TBARS contents upon Cd exposure. Moreover, exogenous GSH application effectively rescued Cd hypersensitivity in APR2-knockout plants. Further analysis showed that buthionine sulfoximine (BSO, an inhibitor of GSH synthesis) treatment completely eliminated the enhanced Cd tolerance phenotypes of APR2-overexpressing plants, implying that APR2-mediated enhanced Cd tolerance is GSH dependent. In addition, over-expression of the APR2 led to elevated expressions of the GSH/PC synthesis-related genes under Cd stress. Taken together, our results indicated that APR2 regulated Cd accumulation and tolerance possibly through modulating GSH-dependent antioxidant capability and Cd-chelation machinery in Arabidopsis. APR2 could be exploited for engineering heavy metal-tolerant plants in phytoremediation.

Effects of enhancing endogenous and exogenous glutathione in roots on cadmium movement in Arabidopsis thaliana

Glutathione (GSH) is a thiol-containing compound involved in many aspects of plant metabolism. In the present study, we investigated how enhancing endogenous and exogenous GSH affects cadmium (Cd) movement and distribution in Arabidopsis plants cultured hydroponically. Transgenic Arabidopsis plants with a strong ability to synthesize GSH in roots were generated by transforming the gene encoding the bifunctional γ-glutamylcysteine synthetase-glutathione synthetase enzyme from Streptococcus thermophiles (StGCS-GS). Enhancing endogenous and exogenous GSH decreased the Cd translocation ratio in different ways. Only exogenous GSH significantly inhibited Cd translocation from roots to shoots in wild-type and transgenic Arabidopsis plants. Our study demonstrated that GSH mainly functions outside root cells to inhibit Cd translocation from roots to shoots.

A simple ratiometric fluorescent sensor selectively compatible of different combinations of characteristic groups for identification of glutathione, cysteine and homocysteine

Simultaneous detection of biothiols would be helpful for understanding their biological functions and pathological study of related diseases. The structural similarities of biothiols present a great challenge to achieve their selective detection. Herein, we report a simple acrylate-substituted fluorescent sensor BTHA with one recognition site and an assisted neighboring hydroxy group for this purpose. BTHA exhibited an emission band at 507 nm with a 147 nm Stokes shift. Upon addition of glutathione (GSH) or cysteine (Cys), deacrylate product was obtained and new fluorescent band at 459 nm occurred. The addition reaction product with decreased fluorescence was achieved as BTHA was treated with homocysteine (Hcy). BTHA acts with the mercapto group of Cys, the mercapto and amino group of Hcy, and the mercapto and carboxyl group of GSH, respectively. Correspondingly, different ratiometric fluorescence changes (I459/I507) with these biothiols was produced. Using the two-dimensional map with two channel signals, BTHA can simultaneous distinguish GSH, Cys and Hcy. The sensor BTHA can also be used for exogenous and endogenous GSH imaging with ratiometric fluorescence response in live cells.

Exogenous Glutathione Increases Arsenic Translocation Into Shoots and Alleviates Arsenic-Induced Oxidative Stress by Sustaining Ascorbate-Glutathione Homeostasis in Rice Seedlings.

Glutathione (GSH) plays diverse roles in the physiological processes, stress defense, growth, and development of plants. This study investigated the effects of exogenous GSH on the biochemical responses of reactive oxygen species and antioxidant levels in rice (Oryza sativa L. cv. Dasan) seedlings under arsenic (As) stress. As treatment inhibited growth; increased the level of superoxide, hydrogen peroxide, and malondialdehyde; and enhanced the uptake of As by the roots and shoots in hydroponically grown 14-day-old seedlings. Furthermore, it reduced GSH content and GSH redox ratios, which have been correlated with the decrease in ascorbate (AsA) redox state. Whereas the exogenous application of GSH in As-treated seedlings reduced As-induced oxidative stress, improved antioxidant defense systems by maintaining antioxidant and/or redox enzyme homeostasis, and increased the AsA and GSH contents, the GSH application also increased the As translocation from the roots to the shoots. These results indicated that the increase in GSH redox state can be linked to an increase in the AsA redox ratio via the induction of the AsA–GSH cycle. Therefore, the results suggest that exogenous GSH application should be a promising approach to enhance As stress resistance in rice plants.