Activities Of Ascorbate-glutathione Cycle Enzymes Research Articles

Physiological responses to water deficiency in bread wheat (Triticum aestivum L.) lines with genetically different leaf pubescence.

Studying the relationship between leaf pubescence and drought resistance is important for assessing Triticum aestivum L. genetic resources. The aim of the work was to assess resistance of common wheat genotypes with different composition and allelic state of genes that determine the leaf pubescence phenotype. We compared the drought resistance wheat variety Saratovskaya 29 (S29) with densely pubescent leaves, carrying the dominant alleles of the Hl1 and Hl3 genes, and two near isogenic lines, i: S29 hl1, hl3 and i: S29 Hl2aesp, with the introgression of the additional pubescence gene from diploid species Aegilops speltoides. Under controlled conditions of the climatic chamber, the photosynthetic pigments content, the activity of ascorbate-glutathione cycle enzymes and also the parameters of chlorophyll fluorescence used to assess the physiological state of the plants photosynthetic apparatus were studied in the leaves of S29 and the lines. Tolerance was evaluated using the comprehensive index D, calculated on the basis of the studied physiological characteristics. The recessive state of pubescence genes, as well as the introduction of the additional Hl2aesp gene, led to a 6-fold decrease in D. Under the water deficit influence, the fluorescence parameters profile changed in the lines, and the viability index decreased compared with S29. Under drought, the activity of ascorbate peroxidase, glutathione reductase and dehydroascorbate reductase in the line i: S29 hl1, hl3 decreased 1.9, 3.3 and 2.3 times, in the line i: S29 Hl2aesp it decreased 1.8, 3.6 and 1.8 times respectively, compared with S29. In a hydroponic greenhouse, line productivity was studied. Compared with S29, the thousand grains mass in the line i: S29 hl1, hl3 under water deficit was reduced. The productivity of the line i: S29 Hl2aesp was significantly reduced regardless of water supply conditions in comparison with S29. Presumably, the revealed effects are associated with violations of cross-regulatory interactions between the proteins of the trichome formation network and transcription factors that regulate plant growth and stress response.

Influence of exogenous jasmonic acid on the functioning of a defense system of potato seedlings (<i>Solanum tuberosum</i> L.) grown on artificial ion exchange substrate and infected with potato virus X

The content of reactive oxygen species, the activities of ascorbate-glutathione cycle enzymes, the content of low molecular weight antioxidants: ascorbate and glutathione, the activity of phenolic peroxidase, the content of water-soluble phenols and the expression level of the hypersensitive response marker gene (HSR) and PR protein genes (Chit, Glu and TLP) in potato seedlings grown on an ion-exchange substrate in the presence of jasmonic acid at a concentration of 10 -8 and 10 -6 M and infected with potato virus X were studied. Accumulation of reactive oxygen species, an increase in the content of reduced ascorbate and an intensive consumption of reduced glutathione upon action of jasmonic acid, as well as a sharp increase in the expression of HSR, Chit, Glu, and TLP genes in the leaves of potato seedlings grown on a substrate with the addition of 10 -6 M jasmonic acid before infection were shown. In control plants, the expression of the HSR and TLP genes increased only after infection with potato virus X.

Role of Ascorbate and Ascorbate–Glutathione Cycle for Photosynthetic Protection in Selected Indigenous Rice Landraces Under Drought Stress

From precise drought tolerance screening of local rice landraces of Koraput, six rice landraces were identified as superior drought tolerant capacity in our earlier study. However, little information is available on their physiological response of redox regulation and photochemical activity under drought stress. The present work evaluates their redox regulations driven by ascorbate–glutathione cycle and photochemical activity of photosystem II under simulated drought stress by polyethylene glycol 6000. Leaf chlorophyll content, electron transport rate and effective quantum yield of PSII were significantly higher in studied indigenous rice genotypes in comparison with susceptible check variety under drought condition. Further, greater quantities of ascorbate and elevated levels of ascorbate–glutathione cycle enzyme activities were observed under drought treatment in studied indigenous rice genotypes in compared with susceptible IR64. The findings suggested that maintenance of better photochemical activity in studied rice genotypes under water-deficit condition may be due to improved redox regulation of ascorbate and efficient ascorbate regeneration pathways. This study thus shows a new light into drought stress response of indigenous rice of Koraput with global implications.

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.

Effects of 1-methylcyclopropene on ascorbate-glutathione cycle enzyme activities of postharvest Dendrobium ‘Khao Sanan’

Effects of 1-methylcyclopropene on ascorbate-glutathione cycle enzyme activities of postharvest <i>Dendrobium</i> ‘Khao Sanan’

Exogenous silicon alters ascorbate-glutathione cycle in two salt-stressed indica rice cultivars (MTU 1010 and Nonabokra).

Silicon is widely available in soil and is known to mitigate both biotic and abiotic stress in plants. Very low doses of silicon are becoming increasingly essential in rice for biofortification and preventing water loss. Soil salinity is a matter of grave concern in various parts of the world, and silicon is a suitable candidate to mitigate salinity-induced stress of important plants in affected areas. The present study investigates the protective capability of exogenously applied silicon in ameliorating NaCl-induced toxicity in two rice (Oryza sativa L.) cultivars, the salt-sensitive MTU 1010, and salt-tolerant Nonabokra. Rice seedlings were treated with three doses of NaCl (25, 50, and 100mM), initially alone and subsequently in combination with 2mM sodium silicate (Na2SiO3, 9H2O). After 21days, these plants were examined to determine levels of reduced glutathione, ascorbic acid, cysteine, and activities of different enzymes involved in the ascorbate-glutathione cycle, viz., glutathione reductase (GR), ascorbate peroxidase (APX), glutathione peroxidase (GPx), and glutathione S-transferase (GST). Though ROS levels increased in both the cultivars with increasing NaCl concentrations, cv. MTU 1010 accumulated comparatively higher amounts. A differential response of NaCl-induced toxicity on the two cultivars was observed with respect to the various enzymatic and non-enzymatic antioxidants. APX and GST activities, as well as, cysteine contents, increased concomitantly with salt concentrations, whereas GR activity declined at increasing salt concentrations, in both cultivars. Activity of GPx increased in cv. Nonabokra but declined in cv. MTU 1010, under similar NaCl concentrations. Reduced glutathione (GSH) contents decreased in both cultivars, whereas ascorbate contents declined in only the sensitive cultivar. Application of silicon, along with NaCl, in the test seedlings of both the cultivars, reduced ROS accumulation and boosted antioxidant defense mechanism, through enhancing ascorbate and GSH levels, and activities of ascorbate-glutathione cycle enzymes as well. However, amelioration of salt-induced damages in the sensitive cv. MTU 1010 was more pronounced upon silicon administration, than the tolerant cv. Nonabokra. Thus, cv. MTU 1010 was found to be more responsive to applied silicon. Hence, this study was instrumental in realizing a successful strategy in silicon-mediated amelioration of salinity stress in plants.

Interactive effects of Salicylic acid and nitric oxide in alleviating zinc toxicity of Safflower (Carthamus tinctorius L.)

The purpose of this study was to assess the possible protective role of exogenous salicylic acid (SA), sodium nitroprusside (SNP), a donor of nitric oxide, and their combination on 21-day-old safflower (Carthamus tinctorius L.) seedlings grown under zinc (Zn) stress. The results revealed that exposure to 500 µM ZnSO4.7H2O for 10 days markedly reduced the root and shoot dry weights in Zn-treated plants, while the application of SA, SNP and specially SA + SNP significantly increased the root and shoot dry weights in seedlings subjected to Zn stress. Addition of SA, SNP and SA + SNP interestingly reduced root-to-shoot translocation of zinc and increased significantly the level of glutathione (GSH) and ascorbate (ASC) in leaves of Zn-stressed plants. The Zn-treated plants supplemented with SA and SNP revealed an improved activity of ascorbate-glutathione cycle enzymes and those enzymes which are involved in glyoxalase system as compared to the plants treated with Zn only. However, no significant relationship was found between SA or SNP supplementation and glutathione S-transferase activity in Zn-stressed plants. These findings demonstrate that exogenous application of SA or SNP could ameliorate the negative effects of Zn on safflower plants probably by stimulation of antioxidant defense and glyoxalase systems.

Sex‐related adaptive responses to interaction of drought and salinity in Populus yunnanensis

We used Populus yunnanensis Dode., a native dioecious species in southwestern China, as a model species to study morphological, physiological, biochemical and ultrastructural responses to drought, salinity and their combination. Females exhibited more growth inhibition, gas exchange rate depression and reactive oxygen species (ROS) accumulation; higher lipid peroxide levels, lower osmotic adjustment capacity and ascorbate-glutathione cycle enzyme activities; and more damage to cell organelles than did males under drought, salinity and especially under their combination. In addition, we found sex-specific responses in total chlorophyll content (TC), carotenoid concentration and carbon isotope composition under different osmotic stresses. Our results indicated that: (1) females are more sensitive and suffer from greater negative effects than do males under drought, salinity and especially under their combination; (2) sexual differences in adaptive responses to drought, salinity and their combination are context dependent; and (3) sex-specific reactions under a combination of stresses are distinct from single-stress responses. Thus, these results provide evidence for adaptive differentiation between sexes in responses to osmotic stresses and in the sensitivity to environmental change.

Proline and Glycinebetaine Confer Cadmium Tolerance on Tobacco Bright Yellow-2 Cells by Increasing Ascorbate-Glutathione Cycle Enzyme Activities

Cadmium (Cd) stress significantly decreased membrane integrity and impaired the ascorbate (ASC)-glutathione (GSH) cycle in tobacco Bright Yellow-2 cells. Exogenous application of proline and glycinebetaine (betaine) significantly restored the membrane integrity and increased the activities of ASC-GSH cycle enzymes under Cd stress without maintenance of the rich ASC or GSH pools. Moreover, proline offered more efficient protection against Cd stress than betaine.

The effect of acute high light and low temperature stresses on the ascorbate–glutathione cycle and superoxide dismutase activity in two Dunaliella salina strains

Dunaliella species accumulate carotenoids and their role in protection against photooxidative stress has been investigated extensively. By contrast, the role of other antioxidants in this alga, has received less attention. Therefore, the components of the ascorbate-glutathione cycle, along with superoxide dismutase (E.C. 1.15.1.1) and peroxidase (E.C. 1.11.1.11) activity were compared in two strains of Dunaliella salina. Strain IR-1 had two-fold higher chlorophyll and beta-carotene concentration than Gh-U. IR-1 had around four-fold higher superoxide dismutase, ascorbate peroxidase and pyrogallol peroxidase activities than Gh-U on a protein basis. Ascorbate and glutathione concentrations and redox state did not differ between strains and there was little difference in the activity of ascorbate-glutathione cycle enzymes (monodehydroascorbate reductase [E.C. 1.6.5.4], dehydroascorbate reductase [E.C. 1.8.5.1] and glutathione reductase [E.C. 1.8.1.7]). The response of these antioxidants to high light and low temperature was assessed by transferring cells from normal growth conditions (28 degrees C, photon flux density of 100 micromol m(-2) s(-1))to 28 degrees C/1200 micromol m(-2) s(-1); 13 degrees C/100 micromol m(-2) s(-1); 13 degrees C/1200 micromol m(-2) s(-1) and 28 degrees C/100 micromol m(-2) s(-1) for 24 h. Low temperature and combined high light-low temperature decreased chlorophyll and beta-carotene in both strains indicating that these treatments cause photooxidative stress. High light, low temperature and combined high light-low temperature treatments increased the total ascorbate pool by 10-50% and the total glutathione pool by 20-100% with no consistent effect on their redox state. Activities of ascorbate-glutathione cycle enzymes were not greatly affected but all the treatments increased superoxide dismutase activity. It is concluded that D. salina can partially adjust to photooxidative conditions by increasing superoxide dismutase activity, ascorbate and glutathione.

Metabolism of Reactive Oxygen Species Is Attenuated in Leghemoglobin-Deficient Nodules of Lotus japonicus

Leghemoglobins together with high rates of respiration are believed to be major sources of reactive oxygen species (ROS) in root nodules of leguminous plants. High capacities of antioxidative systems apparently protect this organ from oxidative damage. Using leghemoglobin-RNA interference (LbRNAi) lines of Lotus japonicus, we found that loss of leghemoglobin results in significantly lower H(2)O(2) levels in nodules. Transcript levels and catalytic activities of ascorbate-glutathione cycle enzymes involved in H(2)O(2) detoxification as well as concentrations of reduced ascorbate were also altered in LbRNAi nodules. Thus, symbiotic leghemoglobins contribute significantly to ROS generation in functional nodules.

Photosynthesis and photoprotection in Nicotiana tabacumL. in vitro-grown plantlets

Nicotiana tabacum L. plantlets were cultured in vitro photoautotrophically (0% sucrose) and photomixotrophically (3% or 5% sucrose) at two irradiances (80 or 380 mumol m-2 s-1) with the aim of investigating the effect of these culture conditions on photosynthetic parameters and on protective systems against excess excitation energy. In plantlets grown photoautotrophically under higher irradiance photoinhibition was demonstrated. These plantlets had a decreased chlorophyll (Chl) a + b content and Chl a/b ratio, an increased content of xanthrophyll cycle pigments and a higher deepoxidation state, a decreased maximum photochemical efficiency of photosystem II (PS II) and actual photochemical efficiency of PS II, and an increased non-photochemical quenching. In the photoautotrophically grown plantlets and those photomixotrophically grown with 3% sucrose, the increase of growth irradiance from 80 to 380 mumol m-2 s-1 stimulated the activities of ascorbate-glutathione cycle enzymes with the exception of ascorbate peroxidase. Ascorbate peroxidase activity was not affected by the increase in growth irradiance but a significant decrease with increasing sucrose concentration was evident. The higher concentration of sucrose in the medium (5%) in combination with the higher irradiance inhibited photosynthesis (decrease in Chl a + b content and net photosynthetic rate) but no significant changes in activities of ascorbate-glutathione cycle enzymes were found. These results suggest that exogenous sucrose added to the medium improved high irradiance and oxidative stress resistance of the plantlets but the effect of sucrose is concentration dependent.

The Effect of Habitat Conditions on the Activity of Enzymes and Content of Metabolites of the Ascorbate-Glutathione Cycle in Plantago media Leaves

The ascorbate-glutathione cycle (AGC) is a metabolic pathway that detoxifies H2O2, which is a reactive oxygen species produced as a waste product in metabolism. The cycle involves the antioxidant metabolites – ascorbate, glutathione, and NADPH, as well as enzymes linking them. We studied the effect of habitat conditions on the activity of enzymes and contents of metabolites of the ascorbate-glutathione cycle in Plantago media leaves. The experimental plants grew on the floodplain meadow: on the sparsely vegetated coastal edge (Site 1) and in the grass stand in the central part of the meadow (Site 2). The hoary plantain plants growing in Site 1 received twice more light than the plants in Site 2. The ascorbate and glutathione concentrations in leaves of the well-lit plants were 2–3 times higher than in shaded plants. The maximal levels of these metabolites were observed at midday, when light intensity and air temperature were increased, and relative humidity was decreased. The activity of AGC enzymes was changing similarly to the metabolite contents. As a result, the leaves of hoary plantain plants from the sites with different light levels did not significantly differ in their hydrogen peroxide concentrations. Our data suggest that the environmental conditions and, above all, the light intensity fine-tune the operation of AGC in plant leaves

Antioxidant systems in the leaf apoplast compartment of Pinus pinaster Ait. and Pinus radiata D. Don. Plants exposed to SO2

Summary.The activities of guaiacol peroxidase (GuPOD), ascorbate peroxidase (ASAp), superoxide dismutase (SOD) and ascorbate/glutathione cycle (AGC) enzymes, together with ascorbate (ASC) and glutathione contents, were determined in apoplastic‐fluid and cell‐wall fractions of needles of Pinus pinaster Ait. and Pinus radiata D. Don. exposed for up to 6 months to SO2 (0.01 ppm or 0.30 ppm) in fumigation chambers. AGC enzyme activities (monodehydroascorbate reductase, dehydroascorbate reductase and glutathione reductase) were in all cases undetectable, as was glutathione content. In needles of P. pinaster plants exposed to SO2, ascorbate content and all enzyme activities considered (except AGC enzymes) increased. The increases were most marked in response to the higher SO2 concentration. In needles of P. radiata, similar but less marked responses were observed. These findings suggest a) that enzyme activities and ascorbate contents increase in order to deal with the reactive oxygen intermediates produced during long‐term contamination with SO2, and b) that P. pinaster has more effective defences against contamination of this type than P. radiata.