Short term exposure of Lemna minor and Lemna gibba to mercury, cadmium and chromium

The effects of mercury (Hg), cadmium (Cd) and chromium (Cr) in concentrations ranging from 0.02 to 20 mg L^?1 applied for 24 h were assessed in Lemna minor and Lemna gibba by measuring changes in protein concentration, ascorbic acid, phenolics, malondialdehyde (MDA), hydrogen peroxide (H₂O₂), the activity of guaiacol peroxidase (G-POX) and catalase (CAT). Ascorbic acid, phenolics, catalase and guaiacol peroxidase played a key role in the antioxidative response of L. gibba. Inadequate activity of antioxidant enzymes in the L. minor resulted in MDA and H₂O₂ accumulation. In both used species, Hg treatment decreased protein content and increased CAT and G-POX activity, but decreased MDA and H₂O₂ levels. Cadmium and chromium had opposite impacts on two used Lemna species on almost all observed parameters. Enhanced antioxidative responses of L. gibba to lower concentrations of Hg, Cd and Cr indicated greater abiotic stress tolerance than L. minor.     

Cadmium Removal by Lemna minor and Spirodela polyrhiza

The present study investigates the ability of two genus of duckweed (Lemna minor and Spirodela polyrhiza) to phytoremediate cadmium from aqueous solution. Duckweed was exposed to six different cadmium concentrations, such as, 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 mg/L and the experiment was continued for 22 days. Water samples were collected periodically for estimation of residual cadmium content in aqueous solution. At the end of treatment period plant samples were collected and accumulated cadmium content was measured. Cadmium toxicity was observed through relative growth factor and changes in chlorophyll content. Experimental results showed that Lemna minor and Spirodela polyrhiza were capable of removing 42–78% and 52–75% cadmium from media depending upon initial cadmium concentrations. Cadmium was removed following pseudo second order kinetic model. Maximum cadmium accumulation in Lemna minor was 4734.56 mg/kg at 2 mg/L initial cadmium concentration and 7711.00 mg/kg in Spirodela polyrhiza at 3 mg/L initial cadmium concentration at the end of treatment period. Conversely in both cases maximum bio-concentration factor obtained at lowest initial cadmium concentrations, i.e., 0.5 mg/L, were 3295.61 and 4752.00 for Lemna minor and Spirodela polyrhiza respectively. The present study revealed that both Lemna minor and Spirodela polyrhiza was potential cadmium accumulator.     

Induction of metal binding compounds and antioxidative defence in callus cultures of two black poplar (<Emphasis Type="Italic">P. nigra</Emphasis>) clones with different tolerance to cadmium

Callus cultures of two parental clones of Populus nigra L., Poli and 58-861, originating from contrasting environments, were exposed to different cadmium concentrations (0, 150 and 250 μM CdSO₄). Clones showed different growth responses to cadmium, evaluated by the tolerance index (Ti), with Poli being more tolerant to the metal at both concentrations. The cadmium concentration at the end of the treatment was very similar between clones at 150 μM CdSO₄, while a higher value in 58-861 compared to Poli was detected at 250 μM CdSO₄. The bioconcentration factor evidenced the lowest value in Poli at 250 μM CdSO₄. Unlike 58-861, cadmium provoked a strong induction of thiols and phytochelatins in clone Poli. In both clones, organic acid concentration differed notably in untreated calli and cadmium treatment induced a general lowering of these compounds. A notably higher antioxidant enzyme activity (ascorbate peroxidase, APX; catalase, CAT; guaiacol peroxidase, GPX) was measured in control calli of clone Poli compared to 58-861. Cadmium induced a remarkable enhancement of APX and CAT, but not GPX, activity at 150 μM CdSO₄ in Poli. Conversely, in 58-861 at 150 μM CdSO₄, and in both clones at 250 μM CdSO₄, a decrease in the antioxidant activity occurred. This investigation provided evidence that these two contrasting genotypes of P. nigra are characterised by a different response to cadmium in callus cultures. In particular, in Poli, the higher tolerance to cadmium is associated with a higher activity of antioxidative enzymes and the ability to strongly increase thiol and PC concentration in response to metal exposure.     

Effect of cadmium on growth and the activity of H2O2 scavenging enzymes in Colocassia esculentum

Cadmium is a non-essential heavy metal that can be harmful even at low concentrations to plants. Colocassia esculentum (Araceae) plant was studied to know its tolerance capability to cadmium. Colocassia esculentum plants grown in pots containing different concentrations of cadmium (Cd) were analyzed for dry matter, fresh weight and total metal content. Cadmium depressed dry matter production of the plant up to 33%. Plant accumulated larger portion of the heavy metal in the roots followed by stem and leaf. Chlorophyll content of the plant declined on treatment with heavy metal while the activity of antioxidative enzymes catalase and peroxidase increased. Colocassia esculentum also showed an increase in total protein along with greater A₂₅₀/A₂₈₀ value suggesting an increase in metal protein complexes. Cadmium elicited anatomical changes in the root of C. esculentum. These changes under heavy metal stress indicate the adaptive properties of this plant species.     

Seedlings growth and antioxidative enzymes activities in leaves under heavy metal stress differ between two desert plants: a perennial (<Emphasis Type="Italic">Peganum harmala</Emphasis>) and an annual (<Emphasis Type="Italic">Halogeton glomeratus</Emphasis>) grass

The present study showed the toxicity caused by heavy metal and its detoxification responses in two desert plants: perennial Peganum harmala and annual Halogeton glomeratus. In pot experiments, 1-month-old seedlings were grown under control and three levels of combined heavy metal stress. Seedling growth as well as heavy metal accumulation, antioxidative enzymes [superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX)] activities and the contents of malondialdehyde (MDA), and hydrogen peroxide (H₂O₂) in leaves was examined after 2 months of heavy metal exposure. Compared with H. glomeratus, growth of P. harmala was more severely inhibited. In leaves, the heavy metal accumulation pattern in both the plants was dose-dependent, being more in H. glomeratus. H. glomeratus exhibited a typical antioxidative defense mechanism, as evidenced by the elevated activities of all the three enzymes tested. P. harmala exhibited a different enzyme response pattern, with a significant reduction in CAT activity, and elevated SOD and APX activities, but significantly elevated APX activity was only at the lowest heavy metal concentration. MDA and H₂O₂ contents were significantly enhanced in leaves of heavy metal-treated P. harmala, but in H. glomeratus were elevated only at the highest heavy metal treatment. These results indicated that H. glomeratus had a greater capacity than P. harmala to adapt to oxidative stress caused by heavy metal stress, and antioxidative defense in H. glomeratus might play an important role in heavy metal tolerance.     

Antioxidant defense system in leaves of Indian mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>) and rape (<Emphasis Type="Italic">Brassica napus</Emphasis>) under cadmium stress

Plant species capable of hyper-accumulating heavy metals are of considerable interest for phytoremediation, and differ in their ability to accumulate metals from environment. Using two brassica species (Brassica juncea and Brassica napus), nutrient solution experiments were conducted to study variation in tolerance to cadmium (Cd) toxicity based on (1) lipid peroxidation and (2) changes in antioxidative defense system in leaves of both plants (i.e., superoxide dismutase (SOD EC 1.15.1.1), catalase (CAT EC 1.11.1.6), ascorbate peroxidase (APX EC 1.11.1.11), guaiacol peroxidase (GPX EC 1.11.1.7), glutathione reductase (GR EC 1.6.4.2), levels of phytochelatins (PCs), non-protein thiols (NP-SH), and glutathione. Plants were grown in nutrient solution under controlled environmental conditions, and subjected to increasing concentrations of Cd (0, 10, 25 and 50 μM) for 15 days. Results showed marked differences between both species. Brassica napus under Cd stress exhibited increased level of lipid peroxidation, as was evidenced by the increased malondialdehyde (MDA) content in leaves. However, in Brassica juncea treated plants, MDA content remained unchanged. In Brassica napus, with the exception of GPX, activity levels of some antioxidant enzymes involved in detoxification of reactive oxygen species (ROS), including SOD, CAT, GR, and APX, decreased drastically at high Cd concentrations. By contrast, in leaves of Brassica juncea treated plants, there was either only slight or no change in the activities of the antioxidative enzymes. Analysis of the profile of anionic isoenzymes of GPX revealed qualitative changes occurring during Cd exposure for both species. Moreover, levels of NP-SH and PCs, monitored as metal detoxifying responses, were much increased in leaves of Brassica juncea by increasing Cd supply, but did not change in Brassica napus. These results indicate that Brassica juncea plants possess the greater potential for Cd accumulation and tolerance than Brassica napus.     

Metalliferous and non-metalliferous populations of Viola tricolor represent similar mode of antioxidative response

Heavy metal-contaminated sites are excellent areas to examine the antioxidative machinery responsible for physiological adaptations of many plant species.Superoxide dismutase (SOD), guaiacol peroxide (GPX), ascorbate peroxide (APX), catalase (CAT) activity and hydrogen peroxide (H₂O₂) content were analyzed in leaves and roots of Viola tricolor (Viola) from contaminated soils (‘Bukowno’, ‘Saturn’, ‘Warpie’ heaps), and non-contaminated soil (‘Zakopane meadow’) to examine the level of oxidative stress and antioxidative response.In leaves, six isoforms of SOD were recognized. Roots possessed two additional bands, named manganese superoxide dismutase (MnSOD)-like form (MnSODI) and Cu/ZnSOD-like form (Cu/ZnSODIV). The H₂O₂ content in leaves ranged from 554 to 5 098 μmol H₂O₂/g f.w. and was negatively correlated with CAT activity. The non-contaminated population was characterized by the lowest CAT activity combined with the highest H₂O₂ concentration. Two isoforms of CAT, CAT-1 and CAT-2, were recognized in leaves of plants from non-contaminated and contaminated sites, respectively. In roots of individuals from two heaps (‘Warpie’ and ‘Saturn’), two distinct bands for each CAT isoform were observed. A slower migrating band may be an aggregate, exhibiting CAT and MnSODs activities. Both peroxidases (APX and GPX) presented the same pattern of activity, depending on the organ, indicating that in leaves and roots APX and GPX were regulated in parallel.Differences in enzyme activities and H₂O₂ content between plants from different contaminated sites were statistically significant, but were tightly maintained at a very similar level. Prolonged and permanent heavy metal stress evoked a very similar mode of antioxidative response in specimens of analyzed metalliferous populations not causing measurable oxidative stress. Thus, our results clearly indicate that V. tricolor is a taxon well adapted to heavy metal-contaminated soils, and that differences in enzyme activities and H₂O₂ content result from adjustment of plants to a variety of conditions.     

Effects of Cadmium,Chromium and Lead on Growth,Metal Uptake and Antioxidative Capacity in <Emphasis Type="Italic">Typha angustifolia</Emphasis>

This study investigates the modulation of antioxidant defence system of Typha angustifolia after 30 days exposure of 1 mM chromium (Cr), cadmium (Cd), or lead (Pb). T. angustifolia showed high tolerance to heavy metal toxicity with no visual toxic symptom when exposed to metal stress, and Cd/Pb addition also increased plant height and biomass especially in Pb treatment. Along with increased Cr, Cd, and Pb uptake in metal treatments, there was enhanced uptake of plant nutrients including Ca and Fe, and Zn in Pb treatment. A significant increase in malondialdehyde (MDA) content and superoxide dismutase (SOD) and peroxidase (POD) activities were recorded in plants subjected to Cr, Cd, or Pb stress. Furthermore, Pb stress also improved catalase (CAT), ascorbate peroxidase (APX), and glutathione peroxidase (GPX) activities; whereas Cr stress depressed APX and GPX. The results indicate that enzymatic antioxidants and Ca/Fe uptake were important for heavy metal detoxification in T. angustifolia, stimulated antioxidative enzymes, and Ca, Fe, and Zn uptake could partially explain its hyper-Pb tolerance.     

Cd~(2+)胁迫对小桐子幼苗叶片抗氧化系统的影响

以小桐子幼苗为材料,设置不同浓度CdCl_2处理,测定Cd~(2+)胁迫对小桐子幼苗叶片中可溶性蛋白、丙二醛(MDA)含量,以及5种抗氧化酶活性和2种抗氧化剂含量的变化,探讨镉胁迫对小桐子幼苗抗氧化系统的影响。结果表明:(1)Cd~(2+)胁迫导致小桐子幼苗叶片中可溶性蛋白含量降低、MDA含量增加;(2)随着镉胁迫时间的延长,幼苗叶片中愈创木酚过氧化物酶(POD)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、抗坏血酸专一性过氧化酶(APX)、谷胱甘肽还原酶(GR)等抗氧化酶活性表现出先升高然后降低的变化趋势;(3)幼苗叶片中还原型抗坏血酸(ASA)和还原型谷胱甘肽(GSH)含量随着胁迫时间延长而降低,但其中氧化型抗坏血酸(DHA)和氧化型谷胱甘肽(GSSG)含量则升高。研究表明,镉胁迫初期能诱导小桐子幼苗抗氧化系统活性显著增强,提高其抗氧化能力,但随着胁迫时间的延长,致使其抗氧化酶的活性和抗氧物质含量下降,植株遭受明显氧化胁迫,幼苗生长受到镉的严重毒害。     

Investigations on the antioxidative defence responses to NaCl stress in a mangrove, Bruguiera parviflora: Differential regulations of isoforms of some antioxidative enzymes

Two-month-old healthy seedlings of a true mangrove, Bruguiera parviflora, raised from propagules in normal nursery conditions were subjected to varying concentrations of NaCl for 45 d under hydroponic culture conditions to investigate the defence potentials of antioxidative enzymes against NaCl stress imposed oxidative stress. Changes in the activities of the antioxidative enzymes catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POX), glutathione reductase (GR) and superoxide dismutase (SOD) were assayed in leaves to monitor the temporal regulation. Among the oxidative stress triggered chemicals, the level of H₂O₂ was significantly increased while total ascorbate and total glutathione content decreased. The ratio of reduced to oxidized glutathiones, however, increased due to decreased levels of oxidized glutathione in the leaf tissue. Among the five antioxidative enzymes monitored, the APX, POX, GR and SOD specific activities were significantly enhanced at high concentration (400 mM NaCl), while the catalase activities declined, suggesting both up and downregulations of antioxidative enzymes occurred due to NaCl imposed osmotic and ionic stress. Analysis of the stress induced alterations in the isoforms of CAT, APX, POX, GR and SOD revealed differential regulations of the isoforms of these enzymes. In B. parviflora one isoform of each of Mn-SOD and Cu/Zn-SOD while three isoforms of Fe-SOD were observed by activity staining gel. Of these, only Mn-SOD and Fe-SOD2 content was preferentially elevated by NaCl treatment, whereas isoforms of Cu/Zn-SOD, Fe-SOD1 and Fe-SOD3 remained unchanged. Similarly, out of the six isoforms of POX, the POX-1,-2,-3 and -6 were enhanced due to salt stress but the levels of POX-4 and -5 remained same as in control plants suggesting preferential upregulation of selective POX isoforms. Activity staining gel revealed only one prominent band of APX and this band increased with increased salt concentration. Similarly, two isoforms of GR (GR1 and GR2) were visualized on activity staining gel and both these isoforms increased upon salt stress. In this mangrove four CAT-isoforms were identified, among which the prominent CAT-2 isoform level was maximally reduced again suggesting differential downregulation of CAT isoforms by NaCl stress. The results presented in this communication are the first report on the resolutions of isoforms APX, POX and GR out of five antioxidative enzymes studied in the leaf tissue of a true mangrove. The differential changes in the levels of the isoforms due to NaCl stress may be useful as markers for recognizing salt tolerance in mangroves. Further, detailed analysis of the isoforms of these antioxidative enzymes is required for using the various isoforms as salt stress markers. Our results indicate that the overproduction of H₂O₂ by NaCl treatment functions as a signal of salt stress and causes upregulation of APX, POX, GR and deactivations of CAT in B. parviflora. The concentrations of malondialdehyde, a product of lipid peroxidation and lipoxygenase activity remained unchanged in leaves treated with different concentrations of NaCl, which again suggests that the elevated levels of the antioxidant enzymes protect the plants against the activated oxygen species thus avoiding lipid peroxidation during salt stress.     

Responses of <Emphasis Type="Italic">Rhodotorula</Emphasis> sp. Y11 to cadmium

Some aspects of the cellular responses to cadmium were extensively investigated in the yeast Rhodotorula sp. Y11. Scanning electron microscopy indicated that accumulation of cadmium in the Y11 did not cause any visible effects on cell morphology. More than 20% yeast cells still showed viability after 15 h of cadmium accumulation under 100 mg l⁻¹ cadmium concentration, and transmission electron microscopy analysis showed that plasmolysis and thickened cell wall were not observed in all of the cells. In the presence of cadmium, the activities of superoxide dismutase (SOD) and catalase (CAT) were all greater than the control, but the increase was in a dose-independent manner. Changes in SOD and CAT activities were also dependent on the time of exposure. Therefore, it suggests that antioxidative defenses play an important role in cadmium tolerance in Rhodotorula sp. Y11. Nondenaturing polyacrylamide gels revealed only one SOD isoforms in Y11 even under exposure to cadmium.     

Arsenic Uptake by Lemna minor in Hydroponic System

Arsenic is hazardous and causes several ill effects on human beings. Phytoremediation is the use of aquatic plants for the removal of toxic pollutants from external media. In the present research work, the removal efficiency as well as the arsenic uptake capacity of duckweed Lemna minor has been studied. Arsenic concentration in water samples and plant biomass were determined by AAS. The relative growth factor of Lemna minor was determined. The duckweed had potential to remove as well as uptake arsenic from the aqueous medium. Maximum removal of more than 70% arsenic was achieved at initial concentration of 0.5 mg/l arsenic on 15th day of experimental period of 22 days. Removal percentage was found to decrease with the increase in initial concentration. From BCF value, Lemna minor was found to be a hyperaccumulator of arsenic at initial concentration of 0.5 mg/L, such that accumulation decreased with increase in initial arsenic concentration.     

Evaluation of the effects of cadmium on rat liver

Abstact Cadmium is one of the most toxic pollutants in environment. Cadmium accumulation in blood affects the renal cortex and causes renal failure. In this study, we aimed to evaluate the effects of cadmium on rat liver tissue. Eighteen male albino rats aged ten weeks old were used in the study. 15 ppm of cadmium was administered to rats via consumption water daily. At the end of the 30th study day, the animals were killed under ether anesthesia. After the liver tissue samples were taken, histopathological and biochemical examinations were performed. Histopathologic changes have included vacuolar and granular degenerations in hepatocytes, heterochromatic nucleuses and sinusoidal and portal widenings. Central vein diameters were normal in cadmium exposed group. Whereas, there was statistically significant difference between two groups by means of sinusoidal (p< 0.001) and portal triad diameters (p< 0.01). Malondialdehyde (MDA) is an indicator of lipid peroxidation. In this study, MDA was used as a marker of oxidative stress-induced liver impairment in cadmium exposed rats. Superoxide dismutase (SOD) and catalase (CAT) activities were also measured to evaluate the changes in antioxidative system in liver tissues. Current findings showed that MDA levels were increased and SOD and CAT activities were decreased in cadmium exposed group compared to control group. The difference between two groups was statistically significant (pvalues: MDA,p< 0.01; CAT,p< 0.01 and SOD,p< 0.05). In conclusion, these findings suggest the role of oxidative mechanisms in cadmium-induced liver tissue damage     

Evaluation of heat shock protein 70 as a biomarker of environmental stress in Fucus serratus and Lemna minor

Heat shock proteins (Hsps) are known to be induced in response to short-term stress. The present study aimed to evaluate the potential of Hsp70 as a biomarker of stress produced by increased temperature, osmotic pressure, and exposure to cadmium and sodium chloride in marine macroalgae and fresh water plant species. An indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was developed with a working range of 0.025–10 μg?ml^?1 using a monoclonal antibody raised against purified Hsp70 of Phaseolus aureus (mung bean). Fucus serratus (toothed wrack), Chondrus crispus (Stackhouse or Carrageen moss), Ulva lactuca (sea lettuce) and Lemna minor (common duckweed) sample extracts were stressed for up to 24 h and then tested in the IC-ELISA. The presence of Hsp70 and cross-reactivity of the monoclonal antibody was confirmed by Western blot. The heat shock response was confirmed in each species using a 2-h 42°C treatment. Following heat shock, Hsp70 concentrations increased to a peak at 2 h (F. serratus) or 4 h (L. minor), after which concentrations decreased. Osmotic and cadmium stresses also resulted in elevated Hsp70 concentrations in samples of F. serratus and L. minor when compared with unstressed controls. In both, osmotic and metal stress, the production of Hsp70 increased to a maximum and subsequently decreased as the stressor levels increased. Results suggest that Hsp70 IC-ELISA could potentially be applied to the detection of stress in these aquatic species, although it would probably be most effective when used in conjunction with other measurements to provide a stressor-specific biomarker profile or fingerprint.     

Changes in gas exchange, proline accumulation and antioxidative enzyme activities in three olive cultivars under contrasting water availability regimes

Changes in photosynthetic performance, osmolyte accumulation and the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and polyphenol oxidase (PPO) were investigated in one-year-old olive cultivars (Chemlali, Meski and Picholine) subjected to contrasting water availability regimes under arid climatic conditions in Tunisia. Shoot elongation rates (SER) and photosynthetic performance were markedly reduced by the water deficit regime (WD) in all cultivars except for Chemlali, which proved to be superior to the other two cultivars with respect to drought tolerance. Higher photosynthetic performance (net photosynthesis (P_n), stomatal conductance (g_s) and transpiration rates (E)) in the Chemlali and Meski cvs. compared to Picholine olive allowed them to maintain better plant water status and shoot elongation rates. Under WD conditions, Chemlali showed a greater capability for proline accumulation. Leaves grown under WD conditions showed signs of oxidative stress such as reduced chlorophyll and carotenoid concentrations. Nevertheless, different cultivars developed certain antioxidative defense mechanisms, including elevated SOD, APX and CAT activities. In contrast, PPO activity decreased under WD circumstances. Comparatively, Chemlali olive displayed better antioxidative enzyme activity, and thus better protection against oxidative stress. These results show that the ability of olive trees to up-regulate the enzymatic antioxidative system might be an important attribute linked to drought tolerance. These findings demonstrate that the association of higher P_n, proline accumulation and antioxidative defenses could be effective in a water-limited environment and may be useful selection criteria in breeding programs with the objective of improving drought tolerance and growth of olive trees, at least under the described environmental conditions.     

Heat shock protein 70 regulates the abscisic acid-induced antioxidant response of maize to combined drought and heat stress

We investigated the interaction between heat shock protein 70 (HSP70) and abscisic acid (ABA)-induced antioxidant response of maize to the combination of drought and heat stress. First, the increased activities of enzymes, including superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR) and catalase (CAT), induced by drought were less than those by heat or combined drought and heat stress, except some individual cases (e.g. CAT in leaves, GR in roots). Second, both HSP70 synthesis and H₂O₂ production increased prominently under drought, heat or their combination stress; the increase in leaves induced by drought and heat combination was the highest, followed by heat and by drought, while the increase in roots had not visible difference. Third, either in leaves or roots, pretreatment with ABA inhibitor, HSP70 inhibitor and H₂O₂ scavenger, significantly arrested the stress-induced increase of antioxidant enzyme activities, and ABA inhibitor and H₂O₂ scavenger obviously suppressed HSP70 synthesis, while HSP70 inhibitor slightly heightened H₂O₂ accumulation. Finally, 100 μM ABA significantly enhanced the activities of antioxidant enzymes, HSP70 expression and H₂O₂ production under stresses in comparison with ABA-deficient mutant vp5 maize plants without pretreatment. Thus, ABA-induced H₂O₂ production enhances the HSP70 synthesis and up-regulates the activities of antioxidant enzymes, resulting in the suppression of cellular reactive oxygen species (ROS) levels. Our results suggest that HSP70 may play a crucial role in ABA-induced antioxidant defense of maize to drought and heat combination.     

Exogenous H<Subscript>2</Subscript>O<Subscript>2</Subscript> improves the chilling tolerance of manilagrass and mascarenegrass by activating the antioxidative system

The effect of foliar pretreatment by hydrogen peroxide (H₂O₂) at low concentrations of 0, 5, 10, and 15 mM on the chilling tolerance of two Zoysia cultivars, manilagrass (Zoysia matrella) and mascarenegrass (Zoysia tenuifolia), was studied. The optimal concentration for H₂O₂ pretreatment was 10 mM, as demonstrated by the lowest malondialdehyde (MDA) content and electrolyte leakage (EL) levels and higher protein content under chilling stress (7°C/2°C, day/night). Prior to initiation of chilling, exogenous 10 mM H₂O₂ significantly increased catalase (CAT), ascorbate peroxidase (APX), glutathione-dependent peroxidases (GPX), and glutathione-S-transferase (GST) activities in manilagrass, and guaiacol peroxidase (POD), APX, and glutathione reductase (GR) activities in mascarenegrass, suggesting that H₂O₂ may act as a signaling molecule, inducing protective metabolic responses against further oxidative damage due to chilling. Under further stress, optimal pretreatments alleviated the increase of H₂O₂ level and the decrease of turfgrass quality, and improved CAT, POD, APX, GR, and GPX activities, with especially significant enhancement of APX and GPX activities from the initiation to end of chilling. These antioxidative enzymes were likely the important factors for acquisition of tolerance to chilling stress in the two Zoysia cultivars. Our results showed that pretreatment with H₂O₂ at appropriate concentration may improve the tolerance of warm-season Zoysia grasses to chilling stress, and that manilagrass had better tolerance to chilling, as evaluated by lower MDA and EL, and better turfgrass quality, regardless of the pretreatment applied.     

Understanding the cellular mechanism of recovery from freeze–thaw injury in spinach: possible role of aquaporins,heat shock proteins,dehydrin and antioxidant system

Recovery from reversible freeze–thaw injury in plants is a critical component of ultimate frost survival. However, little is known about this aspect at the cellular level. To explore possible cellular mechanism(s) for post‐thaw recovery (REC), we used Spinacia oleracea L. cv. Bloomsdale leaves to first determine the reversible freeze–thaw injury point. Freeze (–4.5°C)–thaw‐injured tissues (32% injury vs <3% in unfrozen control) fully recovered during post‐thaw, as assessed by an ion leakage‐based method. Our data indicate that photosystem II efficiency (Fv/Fm) was compromised in injured tissues but recovered during post‐thaw. Similarly, the reactive oxygen species (O₂^?? and H₂O₂) accumulated in injured tissues but dissipated during recovery, paralleled by the repression and restoration, respectively, of activities of antioxidant enzymes, superoxide dismutase (SOD) (EC. 1.14.1.1), and catalase (CAT) (EC.1.11.1.6) and ascorbate peroxidase (APX) (EC.1.11.1.11). Restoration of CAT and APX activities during recovery was slower than SOD, concomitant with a slower depletion of H₂O₂ compared to O₂^??. A hypothesis was also tested that the REC is accompanied by changes in the expression of water channels [aquaporines (AQPs)] likely needed for re‐absorption of thawed extracellular water. Indeed, the expression of two spinach AQPs, SoPIP2;1 and SoδTIP, was downregulated in injured tissues and restored during recovery. Additionally, a notion that molecular chaperones [heat shock protein of 70 kDa (HSP70s)] and putative membrane stabilizers [dehydrins (DHNs)] are recruited during recovery to restore cellular homeostasis was also tested. We noted that, after an initial repression in injured tissues, the expression of three HSP70s (cytosolic, endoplasmic reticulum and mitochondrial) and a spinach DHN (CAP85) was significantly restored during the REC.     

Plant shading increases lipid peroxidation and intensifies senescence-induced changes in photosynthesis and activities of ascorbate peroxidase and glutathione reductase in wheat

Plants of spring wheat (Triticum aestivum L. cv. Saxana) were grown during the autumn. Over the growth phase of three leaves (37 d after sowing), some of the plants were shaded and the plants were grown at 100 (control without shading), 70, and 40 % photosynthetically active radiation. Over 12 d, chlorophyll (Chl) and total protein (TP) contents, rate of CO₂ assimilation (P _N), maximal efficiency of photosystem 2 photochemistry (F_V/F_P), level of lipid peroxidation, and activities of antioxidative enzymes ascorbate peroxidase (APX) and glutathione reductase (GR) were followed in the 1_st, 2_nd, and 3_rd leaves (counted according to their emergence). In un-shaded plants, the Chl and TP contents, P _N, and F_V/F_P decreased during plant ageing. Further, lipid peroxidation increased, while the APX and GR activities related to the fresh mass (FM) decreased. The APX activity related to the TP content increased in the 3_rd leaves. The plant shading accelerated senescence including the increase in lipid peroxidation especially in the 1^st leaves and intensified the changes in APX and GR activities. We suggest that in the 2_nd and 3_rd leaves a degradation of APX was slowed down, which could reflect a tendency to maintain the antioxidant protection in chloroplasts of these leaves.     

Differential biochemical responses of wheat shoots and roots to nickel stress: antioxidative reactions and proline accumulation

Wheat (Triticum aestivum L. cv. ‘Zyta’) seedlings were treated with 10, 100 and 200 μM Ni. Tissue Ni accumulation, length, relative water content (RWC), proline and H₂O₂ concentrations as well as the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POD) and glutathione S-transferase (GST) were studied in the shoots and roots after 6 days of Ni exposure. Treatment with Ni, except for its lowest concentration, resulted in a significant reduction in wheat growth. In comparison to the shoots, the roots showed greater inhibition of elongation, which corresponded with higher accumulation of Ni in these organs. Both shoots and roots responded to Ni application with a decrease in RWC and enhancement in proline concentration. Greater dehydration of the shoot tissue was accompanied by more intense accumulation of proline. Treatment of the wheat seedlings with the highest concentration of Ni led to about 60% increase in H₂O₂ concentration in both studied organs. Apart from CAT, constitutive activities of antioxidative enzymes were much higher in the roots than in the shoots. Exposure of the seedlings to Ni resulted in SOD activity decline, which was more marked in the roots. While the shoots showed a substantial decrease (up to 30%) in CAT activity, in the roots the activity of this enzyme remained unchanged. After Ni application APX, POD and GST activities increased several-fold in the shoots, whereas in the roots they were not significantly altered. The results suggest that differential antioxidative responses of the shoots and roots of wheat seedlings to Ni stress might be related to diverse constitutive levels of antioxidant enzyme activities in both organs.