Ascorbate-glutathione Cycle Research Articles

Nitric Oxide and Its Interaction with Hydrogen Peroxide Enhance Plant Tolerance to Low Temperatures by Improving the Efficiency of the Calvin Cycle and the Ascorbate–Glutathione Cycle in Cucumber Seedlings

The present study was aimed to assess the effect of nitric oxide (NO) and the interaction of NO with hydrogen peroxide (H2O2) on plant tolerance to low temperatures in cucumber seedlings. Exogenous NO significantly increased the endogenous NO content, initial and total activities of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), RuBisCO carboxylation rate (Vc,max), RuBP regeneration rate (Jmax) and the transcript levels of related genes in cucumber seedlings under low temperatures (11 °C/7 °C); however, the effect of NO was blocked by PTIO (NO scavenger). In addition, the SNP treatment significantly improved the contents of glucose, fructose, sucrose, starch, and the activities of sucrose phosphate synthase (SPS), acid invertase (AI), sucrose synthase (SS), as well as the expression levels of SUCROSE PHOSPHATE SYNTHASE 1&2 (SPS1, SPS2), SUCROSE TRANSPORTER 1&2 (SUT2, SUT4), β-starch hydrolase (BAM), and invertase gene (INVERTASE) in cucumber leaves under low temperatures, and the positive effect of NO was impaired by PTIO. Furthermore, we found that the H2O2, induced by NO, participated in NO-induced elevation of ascorbic acid (AsA), glutathione (GSH), and increased activities of related enzymes in the AsA-GSH cycle at low temperatures. However, the positive effect of NO was blocked by l-NAME (NOS inhibitor), PTIO, DPI (inhibitor of NADPH oxidase), and DMTU (reactive oxygen species scavenger). Taken together, our findings indicate that NO increased the low temperature tolerance of cucumber seedlings via H2O2 by improving the efficiency of the Calvin cycle, which in turn increased the carbohydrates content and accelerated the AsA-GSH cycle to enhance ROS scavenging.

Changes in subcellular localization of enzymes involved in ascorbate-glutathione cycle during the maturation of sweet pepper (Capsicum annuum L.) fruit

Changes in subcellular localization of enzymes involved in ascorbate-glutathione cycle during the maturation of sweet pepper (<i>Capsicum annuum</i> L.) fruit

CBF-responsive pathway and phytohormones are involved in melatonin-improved photosynthesis and redox homeostasis under aerial cold stress in watermelon

Due to the insulation provided by soil, plant shoots always suffer cold stress prior to roots when ambient temperatures fall rapidly. Melatonin can enhance cold tolerance of shoots via systemic signaling from roots; however, the underlying mechanisms driving the process remain unclear. This study reports that root-pretreatment with 1.5 μM melatonin alleviated aerial cold-induced inhibition of photosystem II and oxidative stress. The fortification of the photosystem by melatonin was accompanied by increased thermal dissipation, while the melatonin-induced alleviation of oxidative stress was attributed to the improved efficiency of the ascorbate–glutathione cycle under aerial cold stress. Melatonin significantly upregulated the expressions of C-repeat binding factor 1 (CBF1), inducer of CBF expressions 1, and four cold-responsive genes after aerial cold stress. It suggests that melatonin-promoted cold tolerance might be related to the CBF-responsive pathway, which plays an important role in improving photosynthesis and redox homeostasis. Additionally, rhizospheric melatonin application increased both jasmonic acid (JA) and indoleacetic acid (IAA) levels but decreased abscisic acid (ABA) levels in leaves by regulating biosynthetic or catabolic genes under aerial cold stress. Thus, melatonin might function synergistically with JA and IAA, while antagonistically with ABA in regulating the responses of plants to cold stress. These results suggest the involvement of both CBF-responsive pathway and phytohormones in melatonin-mediated systemic tolerance against cold stress.

Nitric oxide and hydrogen sulfide modulate the NADPH-generating enzymatic system in higher plants.

Nitric oxide (NO) and hydrogen sulfide (H2S) are two key molecules in plant cells that participate, directly or indirectly, as regulators of protein functions through derived post-translational modifications, mainly tyrosine nitration, S-nitrosation, and persulfidation. These post-translational modifications allow the participation of both NO and H2S signal molecules in a wide range of cellular processes either physiological or under stressful circumstances. NADPH participates in cellular redox status and it is a key cofactor necessary for cell growth and development. It is involved in significant biochemical routes such as fatty acid, carotenoid and proline biosynthesis, and the shikimate pathway, as well as in cellular detoxification processes including the ascorbate-glutathione cycle, the NADPH-dependent thioredoxin reductase (NTR), or the superoxide-generating NADPH oxidase. Plant cells have diverse mechanisms to generate NADPH by a group of NADP-dependent oxidoreductases including ferredoxin-NADP reductase (FNR), NADP-glyceraldehyde-3-phosphate dehydrogenase (NADP-GAPDH), NADP-dependent malic enzyme (NADP-ME), NADP-dependent isocitrate dehydrogenase (NADP-ICDH), and both enzymes of the oxidative pentose phosphate pathway, designated as glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH). These enzymes consist of different isozymes located in diverse subcellular compartments (chloroplasts, cytosol, mitochondria, and peroxisomes) which contribute to the NAPDH cellular pool. We provide a comprehensive overview of how post-translational modifications promoted by NO (tyrosine nitration and S-nitrosation), H2S (persulfidation), and glutathione (glutathionylation), affect the cellular redox status through regulation of the NADP-dependent dehydrogenases.

Antioxidant Profile of Pepper (Capsicum annuum L.) Fruits Containing Diverse Levels of Capsaicinoids.

Capsicum is the genus where a number of species and varieties have pungent features due to the exclusive content of capsaicinoids such as capsaicin and dihydrocapsaicin. In this work, the main enzymatic and non-enzymatic systems in pepper fruits from four varieties with different pungent capacity have been investigated at two ripening stages. Thus, a sweet pepper variety (Melchor) from California-type fruits and three autochthonous Spanish varieties which have different pungency levels were used, including Piquillo, Padrón and Alegría riojana. The capsaicinoids contents were determined in the pericarp and placenta from fruits, showing that these phenyl-propanoids were mainly localized in placenta. The activity profiles of catalase, total and isoenzymatic superoxide dismutase (SOD), the enzymes of the ascorbate–glutathione cycle (AGC) and four NADP-dehydrogenases indicate that some interaction with capsaicinoid metabolism seems to occur. Among the results obtained on enzymatic antioxidants, the role of Fe-SOD and the glutathione reductase from the AGC is highlighted. Additionally, it was found that ascorbate and glutathione contents were higher in those pepper fruits which displayed the greater contents of capsaicinoids. Taken together, all these data indicate that antioxidants may contribute to preserve capsaicinoids metabolism to maintain their functionality in a framework where NADPH is perhaps playing an essential role.

Crosstalk of hydrogen sulfide and nitric oxide requires calcium to mitigate impaired photosynthesis under cadmium stress by activating defense mechanisms in Vigna radiata

Hydrogen sulfide (H2S) and nitric oxide (NO) have been known to affect vast number of processes in plants under abiotic stresses. Also, calcium (Ca) works as a second messenger in plants, which underpins the abiotic stress-induced damage. However, the sequence of action of these signaling molecules against cadmium (Cd)-induced cellular oxidative damage remains unidentified. Therefore, we studied the synergistic actions and/or relationship of signaling molecules and Ca-dependent activation of tolerance mechanisms in Vigna radiata seedlings under Cd stress. The present study shows that exogenous Ca supplemented to Cd-stressed V. radiata seedlings reduced Cd accumulation and improved the activity of nitrate reductase, and L/D-cysteine desulfhydrase (LCD/DCD) that resulted in improved synthesis of NO and H2S content. Application of Ca also elevated the level of cysteine (Cys) by upregulating the activity of Cys-synthesizing enzymes serine acetyltransferase and O-acetylserine(thiol)lyase in Cd-stressed seedlings. Maintenance of Cys pool under Cd stress contributed to improved H2S content which together with Ca and NO improved antioxidant enzymes and components of ascorbate-glutathione (AsA-GSH) cycle. All these collectively regulated the activity of NADPH oxidase and glycolate oxidase, resulting in the inhibition of Cd-induced generation of reactive oxygen species. The elevated level of Cys also assisted the Cd-stressed seedlings in maintaining GSH pool which retained normal functioning of AsA-GSH cycle and led to enhanced content of phytochelatins coupled with reduced Cd content. The positive effect of these events manifested in an enhanced rate of photosynthesis, carbohydrate accumulation, and growth attributes of the plants. On the contrary, addition of NO scavenger cPTIO [2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide], H2S scavenger HT (Hypotaurine) and Ca-chelator EGTA (Ethylene glycol-bis(b-aminoethylether)-N,N,Nʹ,Nʹ-tetraacetic acid) again developed a condition similar to stress and positive effect of the signaling molecules was abolished. The findings of the study postulate that Ca in association with NO and H2S mitigates Cd-induced impairment and enhances the tolerance of the V. radiata plants against Cd stress. The results of the study also substantiate that Ca acts both upstream as well as downstream of NO signals whereas, H2S acts downstream of Ca and NO during Cd-stress responses of the plants.

Local and Systemic Changes in Photosynthetic Parameters and Antioxidant Activity in Cucumber Challenged with Pseudomonas syringae pv lachrymans.

We studied changes in gas exchange, photochemical activity and the antioxidant system in cucumber leaves locally infected with Pseudomonas syringae pv lachrymans and in uninfected systemic ones. Infection-induced declined net photosynthesis rate and the related changes in transpiration rate, the intracellular CO2 concentration, and prolonged reduction in maximal PSII quantum yield (Fv/Fm), accompanied by an increase in non-photochemical quenching (NPQ), were observed only in the infected leaves, along with full disease symptom development. Infection severely affected the ROS/redox homeostasis at the cellular level and in chloroplasts. Superoxide dismutase, ascorbate, and tocopherol were preferentially induced at the early stage of pathogenesis, whereas catalase, glutathione, and the ascorbate–glutathione cycle enzymes were activated later. Systemic leaves retained their net photosynthesis rate and the changes in the antioxidant system were partly like those in the infected leaves, although they occurred later and were less intense. Re-balancing of ascorbate and glutathione in systemic leaves generated a specific redox signature in chloroplasts. We suggest that it could be a regulatory element playing a role in integrating photosynthesis and redox regulation of stress, aimed at increasing the defense capacity and maintaining the growth of the infected plant.

The Co-regulation of Ethylene Biosynthesis and Ascorbate-Glutathione Cycle by Methy Jasmonate Contributes to Aroma Formation of Tomato Fruit during Postharvest Ripening.

Currently, many fruits are always harvested at the early ripening stage to reduce postharvest losses followed by 1-methylcyclopropene (1-MCP) or ethephon treatment. However, harvesting at the early ripening stage adversely affects fruit quality, especially for the aroma. Methyl jasmonate (MeJA) treatment could induce the biosynthesis of bioactive compounds and maintain postharvest fruit quality. In the present work, the contributions of MeJA to tomato fruit quality during postharvest ripening were studied. The results showed that MeJA treatment significantly promoted the accumulation of volatile organic components (VOCs) by inducing the activities of enzymes related to lipoxygenase pathway and ethylene biosynthesis, whereas 1-MCP treatment largely inhibited the accumulation of VOCs by inhibiting activities of those enzymes. Although the application of ethephon also induced activities of the above enzymes in comparison with control, no significant differences were observed between the VOCs contents of the control and ethephon-treated fruit. Further study revealed that the ethephon treatment resulted in the enhancement of electrical conductivity and malondialdehyde content. Conversely, MeJA treatment inhibited the superoxide anion radical and hydrogen peroxide by regulating the ascorbate-glutathione cycle and further inhibited the enhancement of electrical conductivity and malondialdehyde content, which might be one of the most important reasons why the VOCs contents in fruit treated with ethephon were lower than those in MeJA-treated fruit. Thus, it is considered that MeJA treatment may be an effective and promising strategy to regulate postharvest tomato fruit quality, especially for the aroma, by regulating the ascorbate-glutathione cycle and ethylene biosynthesis.

Exogenous Melatonin Improves Drought Tolerance in Maize Seedlings by Regulating Photosynthesis and the Ascorbate–Glutathione Cycle

Melatonin is known to exert protective effects in maize against drought stress, but the knowledge regarding interaction among the melatonin, photosynthetic efficiency, ascorbate–glutathione cycle still remains elusive. Two contrasting maize (Zea mays L.) genotypes, SD609 (drought-tolerant) and SD902 (drought-sensitive), were subjected to moderate and severe drought stress along with 100 μM melatonin treatment. Exogenous melatonin increased fluorescence curve levels for both genotypes under drought stress, and higher in SD609 than SD902. In both genotypes, melatonin application also significantly increased the photochemical efficiency of photosystem II (PSII), the effective quantum yield of PSII and photosystem I (PSI), and the electron transport rate between PSII and PSI under drought stress, but it decreased the quantum yield of energy dissipation and the donor side and acceptor side impairment of PSI under drought stress, regardless of stress severity. Exogenous melatonin also contributed to the reduction of membrane damage under drought stress, as reflected by significantly decreased levels of MDA, superoxide anions and H2O2 of both genotypes exposed to drought. Furthermore, melatonin treatment increased enzyme activity in AsA–GSH cycle, and upregulated the genes expression of related enzymes in AsA–GSH cycle. The effects of melatonin were more pronounced in SD609 than SD902. Collectively, these results indicated that exogenous melatonin application increased photosynthetic electron transport rate and accelerated the AsA–GSH cycle, which are factors that play an important role in drought tolerance in maize. The beneficial effect of melatonin on the drought-tolerance SD609 was more obvious.

Silicon tackles butachlor toxicity in rice seedlings by regulating anatomical characteristics, ascorbate-glutathione cycle, proline metabolism and levels of nutrients

Reckless use of herbicides like butachlor (Buta) in the fields represents a serious threat to crop plants, and hence to their productivity. Silicon (Si) is well known for its implication in the alleviation of the effects of abiotic stresses; however, its role in mitigating Buta toxicity is not yet known. Therefore, this study was carried out to explore the role of Si (10 µM) in regulating Buta (4 µM) toxicity in rice seedlings. Buta reduced growth and photosynthesis, altered nitric oxide (NO) level and leaf and root anatomy, inhibited enzyme activities of the ascorbate-glutathione cycle (while transcripts of associated enzymes, increased except OsMDHAR), as well as its metabolites (ascorbate and glutathione) and uptake of nutrients (Mg, P, K, S, Ca, Fe, etc. except Na), while addition of Si reversed Buta-induced alterations. Buta stimulated the expression of Si channel and efflux transporter genes- Lsi1 and Lsi2 while the addition of Si further greatly induced their expression under Buta toxicity. Buta increased free proline accumulation by inducing the activity of Δ1-pyrroline-5-carboxylate synthetase (P5CS) and decreasing proline dehydrogenase (PDH) activity, while Si reversed these effects caused by Buta. Our results suggest that Si-governed mitigation of Buta toxicity is linked with favorable modifications in energy flux parameters of photosynthesis and leaf and root anatomy, up-regulation of Si channel and transporter genes, ascorbate-glutathione cycle and nutrient uptake, and lowering in oxidative stress. We additionally demonstrate that NO might have a crucial role in these responses.

Regulation of ascorbate-glutathione cycle by exogenous nitric oxide and hydrogen peroxide in soybean roots under arsenate stress

The role of nitric oxide (NO) and hydrogen peroxide (H2O2) is well known for regulating plant abiotic stress responses. However, underlying mechanisms are still poorly understood. Therefore, the present study investigated the involvement of NO and H2O2 signalling in the regulation of arsenate toxicity (AsV) in soybean roots employing a pharmacological approach. Results show that AsV toxicity declined root length and biomass due to greater As accumulation in the cell wall and cellular organelles. Arsenate induced cell death due to enhanced levels of reactive oxygen species, lipid and protein oxidation and down-regulation in ascorbate-glutathione cycle and redox states of ascorbate and glutathione. These results correlate with lower endogenous level of NO. Interestingly, addition of L-NAME increased AsV toxicity. However, addition of SNP reverses effect of L-NAME, suggesting that endogenous NO has a role in mitigating AsV toxicity. Exogenous H2O2 also demonstrated capability of alleviating AsV stress, while NAC reversed the protective effect of H2O2. Furthermore, DPI application further increased AsV toxicity, suggesting that endogenous H2O2 is also implicated in mitigating AsV stress. SNP was not able to mitigate AsV toxicity in the presence of DPI, suggesting that H2O2 might have acted downstream of NO in accomplishing amelioration of AsV toxicity.

Modulation of ascorbate-glutathione cycle by selenate and sulphate treatments in the seedlings of two rice (Oryza sativa L.) cultivars

The trace element Selenium (Se) has a dual role in the growth and metabolism of plants. Low concentration of selenium (2 ?M selenate) promotes growth and counteracts the detrimental effects of abiotic stress as opposed to higher levels (?10 ?M) where it acts as a pro-oxidant. We focused on both individual and interactive influence of selenate and sulphate on thiol metabolism in seedlings of rice cultivars, satabdi and khitish. Inhibition of ascorbate contents by about 17% on an average, in the test seedlings treated with Se correlated with increased activities of ascorbate peroxidase and ascorbate oxidase in the cultivars. The glutathione levels also increased significantly, on an average by about 102% in roots and 74% in shoots of cv. satabdi compared to a rise, by about 49% in roots and 56% in shoots of cv. khitish. The elevated level of glutathione coincided with the stimulatory influence of Se on its regulatory enzymes. Concomitantly the levels of ?-tocopherol and phytochelatins were also induced in both the test cultivars. Increase in ?-tocopherol activity reached a maximum by about 47% in roots and 80% in shoots of cv. satabdi whereas it increased by about 36% in roots and about 64% in shoots of cv. khitish. Substantive increase in the levels of PC4 followed by PC2 and PC3 was also noted. The effects were found to be less conspicuous in shoots than in roots. Rice seedlings exposed to combined Se and 10mM sulphate treatments showed improved growth and development as a result of better thiol metabolism due to amelioration of the adverse effects caused by selenium alone on all the parameters tested.

Salicylic acid-induced hydrogen sulphide improves lead stress tolerance in pepper plants by upraising the ascorbate-glutathione cycle.

The contribution of hydrogen sulphide (H2 S) to salicylic acid (SA) induced lead (Pb) stress tolerance modulated by the ascorbate-glutathione (AsA-GSH) cycle was examined in pepper (Capsicum annuum L.) plants. One week after germination, pepper seedlings were sprayed with 0.5 mM SA once a day for a week. Thereafter, seedlings were grown under control (no Pb) or Pb stress (Pb-S treatment consisting of 0.1 mM PbCl2 ) for a further 2 weeks. Lead stress reduced plant growth and leaf water status as well as the activities of dehydroascorbate reductase and monodehydroascorbate reductase. However, lead stress elevated leaf Pb, the proline contents, oxidative stress, activities of glutathione reductase and ascorbate peroxidase, as well as the endogenous H2 S content. Supplements of SA resulted in improvements in growth parameters, biomass, leaf water status and AsA-GSH cycle-related enzyme activities, as well as increasing the H2 S content. The positive effect of SA was further enhanced when sodium hydrosulphide was added. However, 0.1 mM hypotaurine (HT) treatment reversed the beneficial effect of SA by reducing the plant H2 S content. Application of NaHS in combination with SA + HT suppressed the adverse effect of HT mainly by restoring the plant H2 S content, suggesting that higher H2 S content, induced by exogenous SA supply, resulted in elevated regulation of the AsA-GSH cycle.

Reactive Oxygen Species (ROS) Metabolism and Nitric Oxide (NO) Content in Roots and Shoots of Rice (Oryza sativa L.) Plants under Arsenic-Induced Stress

Arsenic (As) is a highly toxic metalloid for all forms of life including plants. Rice is the main food source for different countries worldwide, although it can take up high amounts of As in comparison with other crops, showing toxic profiles such as decreases in plant growth and yield. The induction of oxidative stress is the main process underlying arsenic toxicity in plants, including rice, due to an alteration of the reactive oxygen species (ROS) metabolism. The aim of this work was to gain better knowledge on how the ROS metabolism and its interaction with nitric oxide (NO) operate under As stress conditions in rice plants. Thus, physiological and ROS-related biochemical parameters in roots and shoots from rice (Oryza sativa L.) were studied under 50 μM arsenate (AsV) stress, and the involvement of the main antioxidative systems and NO in the response of plants to those conditions was investigated. A decrease of 51% in root length and 27% in plant biomass was observed with 50 μM AsV treatment, as compared to control plants. The results of the activity of superoxide dismutase (SOD) isozymes, catalase, peroxidase (POD: total and isoenzymatic), and the enzymes of the ascorbate–glutathione cycle, besides the ascorbate and glutathione contents, showed that As accumulation provoked an overall significant increase of most of them, but with different profiles depending on the plant organ, either root or shoot. Among the seven identified POD isozymes, the induction of the POD-3 in shoots under As stress could help to maintain the hydrogen peroxide (H2O2) redox homeostasis and compensate the loss of the ascorbate peroxidase (APX) activity in both roots and shoots. Lipid peroxidation was slightly increased in roots and shoots from As-treated plants. The H2O2 and NO contents were enhanced in roots and shoots against arsenic stress. In spite of the increase of most antioxidative systems, a mild oxidative stress situation appears to be consolidated overall, since the growth parameters and those from the oxidative damage could not be totally counteracted. In these conditions, the higher levels of H2O2 and NO suggest that signaling events are simultaneously occurring in the whole plant.

Involvement of L-Cysteine Desulfhydrase and Hydrogen Sulfide in Glutathione-Induced Tolerance to Salinity by Accelerating Ascorbate-Glutathione Cycle and Glyoxalase System in Capsicum.

The aim of this study is to assess the role of l-cysteine desulfhydrase (l-DES) and endogenous hydrogen sulfide (H2S) in glutathione (GSH)-induced tolerance to salinity stress (SS) in sweet pepper (Capsicum annuum L.). Two weeks after germination, before initiating SS, half of the pepper seedlings were retained for 12 h in a liquid solution containing H2S scavenger, hypotaurine (HT), or the l-DES inhibitor dl-propargylglycine (PAG). The seedlings were then exposed for three weeks to control or SS (100 mmol L−1 NaCl) and supplemented with or without GSH or GSH+NaHS (sodium hydrosulfide, H2S donor). Salinity suppressed dry biomass, leaf water potential, chlorophyll contents, maximum quantum efficiency, ascorbate, and the activities of dehydroascorbate reductase, monodehydroascorbate reductase, and glyoxalase II in plants. Contrarily, it enhanced the accumulation of hydrogen peroxide, malondialdehyde, methylglyoxal, electrolyte leakage, proline, GSH, the activities of glutathione reductase, peroxidase, catalase, superoxide dismutase, ascorbate peroxidase, glyoxalase I, and l-DES, as well as endogenous H2S content. Salinity enhanced leaf Na+ but reduced K+; however, the reverse was true with GSH application. Overall, the treatments, GSH and GSH+NaHS, effectively reversed the oxidative stress and upregulated salt tolerance in pepper plants by controlling the activities of the AsA-GSH and glyoxalase-system-related enzymes as well as the levels of osmolytes.

Nitrate reductase is required for salicylic acid-induced water stress tolerance of pepper by upraising the AsA-GSH pathway and glyoxalase system.

A trial was conducted to evaluate whether nitrate reductase (NR) participates in salicylic acid (SA)-improved water stress (WS) tolerance in pepper (Capsicum annuum L.) plants. Before starting WS treatment, 0.5 mM SA was applied to half of the well-watered (WW) plants as well as to WS-plants as a foliar spray once a day for a week. The soil water holding capacity was maintained at 40 and 80% of the full water storing capacity for WS and and well-watered (WW) plants, respectively. Water stress caused substantial decreases in total plant dry weight, Fv /Fm , chlorophyll a and b, relative water content, leaf water potential (ΨI) by 53, 37, 49, 21, 36 and 33%, respectively relative to control, but significant increases in malondialdehyde (MDA), hydrogen peroxide (H2 O2 ), electrolyte leakage (EL), methylglyoxal (MG), proline, key antioxidant enzymes' activities, NO and NR activity. The SA reduced oxidative stress, but improved antioxidant defence system, ascorbate-glutathione (AsA-GSH) cycle enzymes, glyoxalase system-related enzymes, glyoxalase I (Gly I) and glyoxalase II (Gly II), plant growth, photosynthetic traits, NO, NR and proline. SA-induced WS tolerance was further improved by supplementation of sodium nitroprusside (SNP), a donor of NO. NR inhibitor, sodium tungstate (ST) was applied in conjunction with SA and SA + SNP to the WW and WS-plants to assess whether NR contributes to SA-improved WS tolerance. ST abolished the beneficial effects of SA by reducing NO and NR activity in WS-pepper, but the application of SNP along with SA + ST reversed negative effects of ST, showing that NO and NR are jointly needed for SA-induced WS tolerance of pepper plants.

Ascorbate glutathione-dependent H2O2 scavenging is an important process in axillary bud outgrowth in rosebush.

Branching is an important mechanism of plant shape establishment and the direct consequence of axillary bud outgrowth. Recently, hydrogen peroxide (H2O2) metabolism, known to be involved in plant growth and development, has been proposed to contribute to axillary bud outgrowth. However, the involvement of H2O2 in this process remains unclear. We analysed the content of H2O2 during bud outgrowth and characterized its catabolism, both at the transcriptional level and in terms of its enzymatic activities, using RT-qPCR and spectrophotometric methods, respectively. In addition, we used in vitro culture to characterize the effects of H2O2 application and the reduced glutathione (GSH) synthesis inhibitor l-buthionine sulfoximine (BSO) on bud outgrowth in relation to known molecular markers involved in this process. Quiescent buds displayed a high content of H2O2 that declined when bud outgrowth was initiated, as the consequence of an increase in the scavenging activity that is associated with glutathione pathways (ascorbate-glutathione cycle and glutathione biosynthesis); catalase did not appear to be implicated. Modification of bud redox state after the application of H2O2 or BSO prevented axillary bud outgrowth by repressing organogenesis and newly formed axis elongation. Hydrogen peroxide also repressed bud outgrowth-associated marker gene expression. These results show that high levels of H2O2 in buds that are in a quiescent state prevents bud outgrowth. Induction of ascorbate-glutathione pathway scavenging activities results in a strong decrease in H2O2 content in buds, which finally allows bud outgrowth.

Effects of salt stress on antioxidant and ascorbate glutathione cycle enzyme activities in Pokkali rice varieties – Vytilla 1-9

The enzymatic and non-enzymatic antioxidant levels in the released salt tolerant Pokkali, (vytilla, VTL 1-9) varieties were studied under different NaCl concentrations (0-150 mM NaCl). The specific activity of superoxide dismutase (SOD), catalase (CAT) and ascorbate-glutathione cycle enzymes and non-enzymatic antioxidants like superoxide (O2-), hydrogen peroxide (H2O2), malondialdehyde (MDA), glutathione (GSH) and ascorbic acid (AsA) was determined in plants exposed to salt stress. IR-28 was used as positive control and the VTL varieties were used as negative control. The H2O2 and superoxide (O2-) contents were higher in IR-28 at all the applied concentrations of NaCl. The VTL varieties without salt treatment did not evoke any response substantiating the role of salt priming in antioxidant signalling. The MDA contents were higher in the positive and negative control. MDA content was reduced in the NaCl treated VTL varieties. In the positive and negative control varieties, the quantity of ascorbate and glutathione contents were lesser and upregulated in salt treated VTL varieties. Highest H2O2 content was observed in 150 mM NaCl treatment. The H2O2 contents decreased with the increase in all concentrations of NaCl and lowest H2O2 contents was observed in VTL-1 and highest in VTL-2 and VTL-8 treated with 150 mM NaCl. Superoxide contents varied in all the nine varieties depending on the salt concentration. The SOD levels in all the varieties showed a positive correlation with the superoxide and H2O2 content. Lesser quantities of SOD, CAT and the ascorbate - glutathione cycle enzymes were expressed in the positive and negative control. The increased NaCl concentration (25-150 mM) upregulated antioxidant and ascorbate-glutathione cycle enzymes in the VTL varieties. The APX activity was lower in the control and salt treated plants. The GR activity increased linearly in all the varieties with respect to salt concentrations. The MDHAR and DHAR activities showed marginally linear increase, with all concentrations of NaCl. The APX activity was similar or lower to MDHAR activity while DHAR activity was similar to MDHAR activity. The results of the present study reveals the higher levels of enzymatic and non-enzymatic antioxidants under salt stress reflect the salt tolerance potential of pokkali varieties mediated by the up regulation of ROS scavenging enzymes.

Zinc Oxide Nanoparticles Application Alleviates Arsenic (As) Toxicity in Soybean Plants by Restricting the Uptake of as and Modulating Key Biochemical Attributes, Antioxidant Enzymes, Ascorbate-Glutathione Cycle and Glyoxalase System

Accumulation of arsenic (As) in soils is increasing consistently day-by-day, which has resulted in increased toxicity of this element in various crop plants. Arsenic interferes with several plant metabolic processes at molecular, biochemical and physiological levels, which result in reduced plant productivity. Hence, the introduction of novel ameliorating agents to combat this situation is the need of the hour. The present study was designed to examine the effect of zinc oxide nanoparticles (ZnO–NPs) in As-stressed soybean plants. Various plant growth factors and enzymes were studied at varying concentrations of As and ZnO–NPs. Our results showed that with the application of ZnO–NPs, As concentration declined in both root and shoot of soybean plants. The lengths of shoot and root, net photosynthetic rate, transpiration, stomatal conductance, photochemical yield and other factors declined with an increase in external As level. However, the application of ZnO–NPs to the As-stressed soybean plants resulted in a considerable increase in these factors. Moreover, the enzymes involved in the ascorbate–glutathione cycle including superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) showed a significant increase in their activity with the application of ZnO–NPs to the As-stressed plants. Hence, our study confirms the significance of ZnO–NPs in alleviating the toxicity of As in soybean plants.

Окислительный стресс и низкомолекулярные антиоксиданты в побегах тритикале при хлоридном засолении

We studied the alterations in oxidative stress indicators (hydrogen peroxide, superoxide radical, lipid peroxidation – LPO) and the alterations in the content of low molecular weight metabolites (ascorbic acid, glutathione, proline, chlorophyll and carotenoids) in the shoots of triticale (Triticosecale) under short-term (0-96 h) sodium chloride stress (120 mM) with statistical methods: principal component analysis (PCA) and cluster analysis. An analysis of the alterations in oxidative stress indicators allowed us to calculate the correlation coefficients for the pairs: peroxide – superoxide (0.52), peroxide – LPO (0.62), superoxide – LPO (0.23). The inclusion in the analysis of data on alterations in the content of low molecular weight antioxidants showed that the PCA method forms three main groups for all the studied characteristics: (1) LPO and hydrogen peroxide, (2) chlorophyll and carotenoids, (3) glutathione and ascorbate. The correlation coefficients were calculated for pairs: ascorbate – glutathione (0.71), ascorbate – proline (0.81), glutathione – proline (0.28). Such a value of the coefficient of the first pair suggests that ascorbic acid also performs numerous other functions, in addition to participating in the ascorbate-glutathione cycle. The high correlation between ascorbate and proline can be explained by the similar nature of alterations in their content in triticale shoots under conditions of short-term sodium chloride stress. Negative correlation coefficients in pairs of hydrogen peroxide – chlorophyll (-0.73), peroxide – carotenoids (-0.75), ascorbic acid – LPO (-0.70), LPO – proline (-0.69) give reason to talk about the possible protective role of photosynthesis pigments from accumulating hydrogen peroxide, as well as the potential role of ascorbic acid and proline in protecting membranes from lipid peroxidation. The application of the cluster analysis method showed that first and second order clusters between ascorbate, proline and glutathione reflect their known antioxidant role. The results obtained may also indicate that pigments have a much lower protective function.