Short-term effect of elevated CO2 concentration and high irradiance on the antioxidant enzymes in bean plants

The effect of short-term exposure to elevated CO₂ concentration and high irradiance on the activity of superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidases (GPX) and catalase (CAT), and on the extent of the lipid peroxidation was studied in bean (Phaseolus vulgaris L.) plants. Plants were exposed for 4 d (8 h a day) to irradiance of 100 (LI) or 1000 (HI) μmol m⁻² s⁻¹ at ambient (CA, 350 μmol mol⁻¹) or elevated (CE, 1300 μmol mol⁻¹) CO₂ concentration. Four-day exposure to CE increased the leaf dry mass in HI plants and RuBPC activity and chlorophyll content in LI plants. Total soluble protein content, leaf dry matter and RuBPC activity were higher in HI than in LI plants, although the HI and CE increased the contents of malonyldialdehyde and H₂O₂. Under CA, exposure to HI increased the activity of APX and decreased the total SOD activity. Under CE, HI treatment also activated APX and led to reduction of both, SOD and GPX, enzymes activities. CE considerably reduced the CAT activity at both irradiances, possibly due to suppressed rate of photorespiration under CE conditions.     

Response of antioxidant systems to NaCl stress in the halophyte Cakile maritima

The effects of salt stress on antioxidative activities were investigated in a coastal halophyte, Cakile maritima . Two Tunisian accessions, Jerba and Tabarka, were compared. Plants were subjected to 100, 200, or 400 m M NaCl for 20 days. Parameters of oxidative stress [malondialdehyde (MDA), electrolyte leakage (EL), and hydrogen peroxide (H₂O₂) concentration], activities of several enzymes [superoxide dismutase (SOD), catalase (CAT), peroxydase (POD), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), and glutathione reductase (GR)], and antioxidant molecules (ascorbate, ASC, and glutathione, GSH) were determined. Growth of Jerba plants was improved at 100 m M NaCl as compared to that of control. Tabarka growth was inhibited by salt at all NaCl concentrations. The relative salt tolerance of Jerba was associated with high antioxidant enzyme activities and glutathione content, together with low MDA content, EL, and H₂O₂ concentration. Lower antioxidant activities and higher MDA content, EL, and H₂O₂ concentration were found in Tabarka. As a whole, these data suggest that the capacity to limit oxidative damage is important for salt tolerance of C. maritima .     

不同浓度铵态氮对镉胁迫轮叶黑藻生长及抗氧化酶系统的影响

用含有不同浓度NH₄⁺-N (0、0.5、2.0和4.0 mg·L^-1)和10 mg·L^-1Cd的1/10 Hoagland营养液培养沉水植物轮叶黑藻,研究了氨态氮对Cd胁迫下轮叶黑藻的生长及抗氧化酶系统的影响,探讨富营养化污染水体沉水植物退化机理.结果表明,10 mg·L^-1Cd对轮叶黑藻能产生明显的胁迫作用,叶绿素合成量明显降低;超氧化物岐化酶(SOD)与过氧化物酶(POD)活性呈先升高后下降的趋势.NH₄⁺-N能加速Cd对植物的胁迫作用,单因子Cd作用3 h时SOD明显升高,而Cd和NH₄⁺-N共同作用0.5 h SOD就明显升高.Cd与NH₄⁺-N共同作用时,相对于叶绿素和蛋白质,抗氧化酶是早期敏感指标,并且SOD比POD更敏感.本试验条件下,NH₄⁺-N与Cd共同作用2 d后,对轮叶黑藻的胁迫作用主要取决于Cd,NH₄⁺-N的作用几乎可以忽略.     

Response of growth and antioxidant enzymes in Azolla plants (Azolla pinnata and Azolla filiculoides) exposed to UV-B

Effect of ultravilolet-B (0.4 Wm(-2)) irradiation on growth, flavonoid content, lipid peroxidation, proline accumulation and activities of superoxide dismutase and peroxidase was comparatively analysed in Azolla pinnata and Azolla filiculoides. Growth measured as increment in dry weight reduced considerably due to all UV-B treatments. However, the reduction was found to be severe in A. filiculoides as compared to A. pinnata. The level of UV-absorbing compound flavonoids increased significantly in A. pinnata plants whereas only a slight increase in the flavonoid content was observed in A. filiculoides. UV-B exposure led to enhanced production of malondialdehyde (MDA) and electrolyte leakage in A. filiculoides than A. pinnata. Proline accumulation also showed a similar trend. Marked differences in the activity of antioxidant enzymes such as superoxide dismutase (SOD) and peroxidase (POD) was noticed in both the plants exposed to UV-B. Our comparative studies indicate A. pinnata to be better tolerant to UV-B as compared with A. filiculoides which appears to be sensitive.     

Response to NaCl of alfalfa plants regenerated from non-saline callus cultures

Salinity restricts crop productivity in many arid environments. Inadvertent selection for tolerance to osmotic stress may occur under cell or tissue culture conditions and could affect the performance of regenerated plants. The effect of NaCl on forage produced by alfalfa (Medicago sativa L.) plants regenerated from non-saline callus cultures was examined in this study. Plants of Regen-S, which was selected for improved callus growth and regeneration in non-saline cultures, had higher forage weight when grown on SHII medium at NaCl levels up to 100 mM compared to its parental cultivars, Saranac and DuPuits. Five additional original-regenerant plant pairs, each derived from non-saline callus cultures of different alfalfa plants, were evaluated in a solid (soil-like) substrate under saline and non-saline conditions. Weight of forage produced by rooted stem cuttings of regenerated plants was 33% higher at 50 mM NaCl compared to cuttings of explant donor plants. Self progenies from four of five regenerants had higher relative forage weight at 100 mM NaCl (percent of 0 NaCl treatment) than the original plants indicating increased NaCl tolerance.     

Response of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii to salt-dependent oxidative stress: increased activities of antioxidant enzymes in root plastids

Root plastids of the cultivated tomato Lycopersicon esculentum (Lem) exhibited salt-induced oxidative stress as indicated by the increased H 2 O 2 and lipid peroxidation levels which were accompanied with increased contents of the oxidized forms of ascorbate and glutathione. In contrast, H 2 O 2 level decreased, lipid peroxidation level slightly decreased and the levels of the reduced forms of ascorbate and glutathione increased in plastids of L. pennellii (Lpa) species in response to salinity. This better protection of Lpa root plastids from salt-induced oxidative stress was correlated with increased activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidases (POD), monodehydroascorbate reductase (MDHAR), glutathione peroxidase (GPX), glutathione- S -transferase (GST) and phospholipid hydroperoxide glutathione peroxidase (PHGPX). In the plastids of both species, activities of SOD, APX, and POD could be resolved into several isozymes. In Lem plastids two Cu/ZnSOD isozymes were found whereas in Lpa an additional FeSOD type could also be detected. In response to salinity, activities of selected SOD, APX, and POD isozymes were increased in Lpa, while in Lem plastids the activities of most of SOD and POD isozymes decreased. Taken together, it is suggested that plastids play an important role in the adaptation of Lpa roots to salinity.     

Response of antioxidant enzymes to intermittent and continuous hyperbaric oxygen

Rats and guinea pigs were exposed to O2 at 2.8 ATA (HBO) delivered either continuously or intermittently (repeated cycles of 10 min of 100% O2 followed by 2.5 min of air). The O2 time required to produce convulsions and death was increased significantly in both species by intermittency. To determine whether changes in brain and lung superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSHPx) correlated with the observed tolerance, enzyme activities were measured after short or long HBO exposures. For each exposure duration, one group received continuous and one intermittent HBO; O2 times were matched. HBO had marked effects on these enzymes: lung SOD increased (guinea pigs 47%, rats 88%) and CAT and GSHPx activities decreased (33%) in brain and lung. No differences were seen in lung GSHPx or brain CAT in rats or brain SOD in either species. In guinea pigs, but less so in rats, the observed changes in activity were usually modulated by intermittency. Increases in hematocrit, organ protein, and lung DNA, which may also reflect ongoing oxidative damage, were also slowed with intermittency in guinea pigs. Intermittency benefited both species by postponing gross symptoms of toxicity, but its modulation of changes in enzyme activities and other biochemical variables was more pronounced in guinea pigs than in rats, suggesting that there are additional mechanisms for tolerance.     

Responses of amino acid metabolizing enzymes from plants differing in salt tolerance to NaCl

Summary This paper reports the effects of NaCl on the in vivo activity of glutamate dehydrogenase (GDH) and glutamic-oxaloacetic transaminase (GOT) and on the in vitro activity of GDH, both enzymes having been isolated from plants differing in salt tolerance. The plants investigated were Vicia faba (salt-sensitive), Atriplex nitens and Atriplex calotheca (more or less salt-tolerant), and Atriplex halimus (halophyte) grown at various NaCl concentrations. GDH and GOT isolated from various salt-tolerant plants grown at low NaCl concentrations were inhibited in a similar way. At high NaCl concentrations, the enzyme activities remain at constant values only in the Atriplex species. GOT was more impaired by NaCl than GDH. In the case of GOT, the double reciprocal plot indicated the type of a noncompetitive inhibition. The in vitro effect of NaCl on the activity of GDH from the differentially salt-tolerant plants was of a different kind, i.e. GDH isolated from V. faba was clearly inhibited by NaCl, whereas NaCl stimulated the activity of GDH from all Atriplex species investigated. Kinetic analysis showed that substrate inhibition of GDH from A. nitens and A. calotheca grown at non-saline conditions could be removed by NaCl. Inhibition by high NaCl concentrations at low substrate concentrations was removable by increasing substrate concentrations. Moreover, the inhibition at low substrate concentrations was shown to be competitive. GDH lost this regulatory property when the plants were pretreated with 500 mM NaCl. GDH from A. halimus also possessed this control, but in contrast to A. nitens and A. calotheca, activity and control of GDH isolated from A. halimus were stimulated by pretreating the plants with 500 mM NaCl. The results showed that DDH isolated from the salt-tolerant Atriplex species was adapted to high NaCl concentrations of the tissue. Possible mechanisms of the interactions between GDH from salt-tolerant Atriplex species and NaCl are discussed.     

Antioxidative enzymes in cultivars of pepper plants with different sensitivity to cadmium

The effect of growing five different cultivars of pepper plants (Capsicum annuum L.) with CdCl₂ concentrations ranging from 0.125 to 0.5 mM on different physiological parameters, and antioxidative enzyme activities of leaves was studied. On the basis of growth parameters, pepper plants were relatively tolerant to cadmium, although metal concentrations higher than 0.125 mM produced a significant inhibition of growth, net photosynthesis, and water use efficiency. Different sensitivities to Cd⁺⁺ ions were observed among cultivars, Abdera being the most resistant to cadmium stress, while Mondo and Herminio were the most sensitive cultivars. Cadmium concentrations of 0.5 mM produced an increase in the activity of glutathione reductase, and guaiacol peroxidase in most cultivars, while catalase and superoxide dismutase (SOD) were slightly depressed. The analysis of the SOD activity pattern by native-PAGE showed the presence in most cultivars of four SODs which were identified as Mn–SOD, Fe–SOD, CuZn–SOD I and CuZn–SOD II. However, the two CuZn–SODs were absent in the Cd-sensitive cv. Herminio. The growth of pepper plants with 0.5 mM cadmium inhibited the activity of CuZn–SODs in all cultivars, while the activity of Mn- and Fe–SOD was enhanced. The activity of NADPH-dehydrogenases (glucose-6-P-dehydrogenase, 6-phosphogluconate dehydrogenase, NADP–isocitrate dehydrogenase and malic enzyme) showed a Cd-dependent enhancement in most cultivars, the highest increase being observed in the tolerant cv. Abdera. These results suggest that in pepper plants the tolerance to Cd toxicity is more dependent on the availability of NADPH than on its antioxidant capacity.     

Enhanced tolerance to oxidative stress in transgenic tobacco plants expressing three antioxidant enzymes in chloroplasts

The effect of simultaneous expression of genes encoding three antioxidant enzymes, copper zinc superoxide dismutase (CuZnSOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11), and dehydroascorbate (DHA) reductase (DHAR, EC 1.8.5.1), in the chloroplasts of tobacco plants was investigated under oxidative stress conditions. In previous studies, transgenic tobacco plants expressing both CuZnSOD and APX in chloroplast (CA plants), or DHAR in chloroplast showed enhanced tolerance to oxidative stresses, such as paraquat and salt. In this study, in order to develop transgenic plants that were more resistant to oxidative stress, we introduced the gene encoding DHAR into CA transgenic plants. Mature leaves of transgenic plants expressing all three antioxidant genes (CAD plants) had approximately 1.6–2.1 times higher DHAR activity, and higher ratios of reduced ascorbate (AsA) to DHA, and oxidized glutathione (GSSG) to reduced glutathione (GSH) compared to CA plants. CAD plants were more resistant to paraquat-induced stress, exhibiting only 18.1% reduction in membrane damage relative to CA plants. In addition, seedlings of CAD plants had enhanced tolerance to NaCI (100 mM) compared to CA plants. These results indicate that the simultaneous expression of multiple antioxidant enzymes, such as CuZnSOD, APX, and DHAR, in chloroplasts is more effective than single or double expression for developing transgenic plants with enhanced tolerance to multiple environmental stresses.     

Production of reactive oxygen species and antioxidant enzymes of pea and soybean plants under hypoxia and high CO<Subscript>2</Subscript> concentration in medium

Generation of reactive oxygen species (ROS) and activities of antioxidant enzymes (catalase, peroxidase, ascorbate peroxidase) in pea (Pisum sativum L.) and soybean (Glycine max L.) under hypoxia (3–24 h) and high CO₂ concentration in medium were studied. In sensitive to hypoxia pea seedlings, hypoxia enhanced markedly production of superoxide anion-radical, hydroperoxides, and especially hydrogen peroxide. In more tolerant soybean plants, these changes were less pronounced. During first hours of hypoxia, activity of lipoxygenase in plant cells increased. This allows a suggestion that this enzyme is involved in the processes of hydroperoxide accumulation in plant tissues under oxygen deficit. In pea and soybean plants, a correlation between tolerance to hypoxia, the rate of ROS generation, and antioxidant enzyme activities was established. During the first hours of hypoxia, the catalase activity in soybean plants increased stronger than in sensitive to hypoxia pea plants. At longer exposure to hypoxia (24 h), peroxidases started to play the higher role in cell defense against hypoxia, but only in soybean plants. The medium with the higher CO₂ content induced higher changes in the processes of ROS accumulation and activities of lipoxygenase and antioxidant enzymes. This permits us to refer CO₂, accumulated as a product of respiration in the cells, to low-molecular signal molecules switching on plant adaptation to hypoxic stress.     

Active-site titration of enzymes at high concentration. Application to myosin ATPase

The number of active sites of soluble and filamentous myosin and of its subfragments, heavy meromyosin and subfragment-1, has been determined. The titration involves steady-state kinetic measurements at a high enzyme concentration and varying substrate concentrations (or vice versa), in the presence of a substrate-regenerating system. Some practical and theoretical conditions for its execution are given, and, in particular, the effect of a possible heterogeneity of the active sites on the titration curves is analysed. Under the experimental conditions of the study, the number of active sites is close to that of myosin heads, and the heads seem to be functionally identical; the catalytic constants kcat and Km characterizing each active site are similar within some limits (1-2 for the ratio of kcat values; 1-5 for that of Km values).     

The activities of antioxidant enzymes in response to oxidative stresses and hormones in paraquat-tolerant Rehmannia glutinosa plants

All members of R. glutinosa show the unique characteristic of intrinsic tolerance to paraquat (PQ). Antioxidant enzymes have been proposed to be the primary mechanism of PQ resistance in several plant species. Therefore, the antioxidant enzyme systems of R. glutinosa were evaluated by comparatively analyzing cellular antioxidant enzyme levels, and their responses of oxidative stresses and hormones. The levels of ascorbate peroxidase (APX), glutathione reductase (GR), non-specific peroxidase (POX), and superoxide dismutase (SOD) were 7.3-, 4.9-, 2.7- and 1.6-fold higher in PQ-tolerant R. glutinosa than in PQ-susceptible soybeans. However, the activity of catalase (CAT) was about 12-fold higher in the soybeans. The activities of antioxidant enzymes reduced after PQ treatment in the two species, with the exception of POX and SOD in R. glutinosa, which increased by about 40 %. Interestingly, the activities of APX, SOD and POX in R. glutinosa, relative to those in soybeans, were further increased by 49, 67 and 93 % after PQ treatment. The considerably higher intrinsic levels, and increases in the relative activities of antioxidant enzymes in R. glutinosa under oxidative stress support the possible role of these enzymes in the PQ tolerance of R. glutinosa. However, the relatively lower levels of SOD versus PQ tolerance, and the mixed responses of antioxidant enzymes to stresses and hormones, suggest a possible alternative mechanism(s) for PQ tolerance in R. glutinosa.     

Response of liver antioxidant system to taurine in rats fed high fructose diet

Fructose-fed rats were more susceptible to peroxidative damage as measured by thiobarbituric acid reactive species. The concentrations of lipid peroxides, diene conjugates, lipofuscin and hydroperoxides were significantly higher. The levels of enzymic antioxidants such as vitamin C, vitamin E and glutathione and activities of antioxidant enzymes were significantly lower in fructose-fed rats. When these rats received taurine in drinking water, peroxidative damage was minimal in both plasma and liver. Taurine was effective in inducing the antioxidant potential in fructose-fed rats. Increased peroxidative damage in liver is likely to be associated with fructose dependent pathology, which could be reduced by taurine by enhancing the antioxidant potential.     

The effects of NaCl on antioxidant enzyme activities in callus tissue of salt-tolerant and salt-sensitive cotton cultivars (Gossypium hirsutum L.)

Summary To determine NaCl effects on callus growth and antioxidant activity, callus of a salt-tolerant and a salt-sensitive cultivar of cotton was grown on media amended with 0, 75, and 150 mM NaCl. Callus of the salt-tolerant cultivar, Acala 1517-8 8, grown at 150 mM NaCl, showed significant increases in superoxide dismutase, catalase, ascorbate peroxidase, peroxidase and glutathione reductase activities compared to callus tissue grown at 0 mM NaCl. In contrast, callus tissue of the salt-sensitive cultivar, Deltapine 50, grown at 0, 75, and 150 mM NaCl, showed no difference in the activities of these enzymes. At the 150 mM NaCl treatment, peroxidase was the only antioxidant enzyme from Deltapine 50 with an activity as high as that observed in Acala 1517-88. The NaCl-induced increase in the activity of these enzymes in Acala 1517-88 indicates that callus tissue from the more salt-tolerant cultivar has a higher capacity for scavenging and dismutating superoxide, an increased ability to decompose H₂O₂, and a more active ascorbate-glutathione cycle when grown on media amended with NaCl.     

抗旱性不同的小麦幼苗对水分和NaCl胁迫的反应

分别测定抗旱小麦的８１３９（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．ｃｖ．８１３９）和干旱敏感品种甘麦８号（Ｔ．ａｅｓｔｉｖｕｍ Ｌ．ｃｖ．Ｇａｎｍａｉ Ｎｏ．８）在２０％ＰＥＧ６０００和１．２％ＮａＣｌ胁迫下的生长、光合作用、蒸腾作用及抗氧化保护系统的变化。结果表明,抗旱小麦８１３９对ＰＥＧ６０００有较强的抗性,但对ＮａＣｌ胁迫的抗性较差。ＮａＣｌ胁迫下,两种小麦根的生长均受到严重抑制,而在ＰＥ