Biological responses of wheat (Triticum aestivum) plants to the herbicide chlorotoluron in soils

Chlorotoluron is a phenylurea herbicide that is widely used for controlling grass weeds in the land of cereal, cotton and fruit production. However, extensive use of this herbicide may lead to its accumulation in ecosystems, thus inducing the toxicity to crops and vegetables. To assess chlorotoluron-induced toxicity in plants, we performed the experiment focusing on the metabolic adaptation of wheat plants (Triticum aestivum) to the chlorotoluron-induced oxidative stress. The wheat plants were cultured in the soils with chlorotoluron at concentrations of 0-25mg/kg. Chlorotoluron accumulation in plants was positively correlated with the external chlorotoluron concentrations, but negatively with the plant growth. Treatment with chlorotoluron induced the accumulation of O(2)(-) and H(2)O(2) in leaves and resulted in the peroxidation of plasma membrane lipids in the plant. We measured the endogenous proline level and found that it accumulated significantly in chlorotoluron-exposed roots and leaves. To understand the biochemical responses to the herbicide, activities of the antioxidant enzymes, such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were assayed. Analysis of SOD activity by non-denaturing polyacrylamide gel electrophoresis (PAGE) revealed that there were three isoforms in the roots and leaves, but the isoforms in the tissues showed different patterns. Also, using the native PAGE, 6 isoforms of root POD and 10 in leaves were detected. The total activity of POD in roots was significantly enhanced. Activities of APX in roots and leaves showed a similar pattern. The CAT activities were generally suppressed under the chlorotoluron exposure.     

Biological detection and analysis of mercury toxicity to alfalfa (Medicago sativa) plants

Mercury has become one of the major causes of toxic metal pollution in agricultural lands. Accumulation of mercury by plants may disrupt many cellular functions and block growth and development. To assess mercury toxicity, we performed an experiment focusing on the responses of alfalfa (Medicago sativa) to Hg(2+)-induced oxidative stress. Alfalfa plants were treated with 0-40microM HgCl(2) for 7d. The concentrations of Hg(2+) were positively correlated with the generation of O2- and H(2)O(2) in leaves. Treatment with Hg(2+) increased the activities of NADH oxidase and lipoxygenase (LOX) and damaged the biomembrane lipids. To understand biochemical responses under Hg stress, activities of several antioxidant enzymes, superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) were assayed. Analysis of SOD activity by non-denaturing polyacrylamide gel electrophoresis revealed five isoforms in leaves, but they showed different patterns. Also, eight isoenzymes of APX and seven of POD in leaves were detected. However, only one isoform of CAT was visualized. The total activities of APX, POD and CAT were generally enhanced. We also measured several antioxidative metabolites such as ascorbate and glutathione (GSH), and found that both differentially accumulated in leaves. These results indicate that the increased levels of O2- and H(2)O(2) under Hg stress were closely linked to the improved capacity of antioxidant enzymes. The data not only provide the important information for better understanding of the toxic and tolerance mechanisms, but as well can be used as a bio-indicator for soil contamination by Hg.     

Impact of fungicides on active oxygen species and antioxidant enzymes in spring barley (Hordeum vulgare L.) exposed to ozone.

Two modern fungicides, a strobilurin, azoxystrobin (AZO), and a triazole, epoxiconazole (EPO), applied as foliar spray on spring barley (Hordeum vulgare L. cv. Scarlett) 3 days prior to fumigation with injurious doses of ozone (150-250 ppb; 5 days; 7 h/day) induced a 50-60% protection against ozone injury on leaves. Fungicide treatments of barley plants at growth stage (GS) 32 significantly increased the total leaf soluble protein content. Additionally, activities of the antioxidative enzymes superoxide dismutase (SOD), catalase (CAT), ascorbate-peroxidase (APX) and glutathione reductase (GR) were increased by both fungicides at maximal rates of 16, 75, 51 and 144%, respectively. Guiacol-peroxidase (POX) activity was elevated by 50-110% only in AZO treated plants, while this effect was lacking after treatments with EPO. This coincided with elevated levels of hydrogen peroxide (H2O2) only in EPO and not in AZO treated plants. The enhancement of the plant antioxidative system by the two fungicides significantly and considerably reduced the level of superoxide (O2*-) in leaves. Fumigation of barley plants for 4 days with non-injurious ozone doses (120-150 ppb, 7 h/day) markedly and immediately stimulated O2*- accumulation in leaves, while H2O2 was increased only after the third day of fumigation. Therefore, O2*- itself or as precursor of even more toxic oxyradicals appears to be more indicative for ozone-induced leaf damage than H2O2. Ozone also induced significant increases in the activity of antioxidant enzymes (SOD, POX and CAT) after 2 days of fumigation in fungicide untreated plants, while after 4 days of fumigation these enzymes declined to a level lower than in unfumigated plants, due to the oxidative degradation of leaf proteins. This is the first report demonstrating the marked enhancement of plant antioxidative enzymes and the enhanced scavenging of potentially harmful O2*- by fungicides as a mechanism of protecting plants against noxious oxidative stress from the environment. The antioxidant effect of modern fungicides widely used in intense cereal production in many countries represents an important factor when evaluating potential air pollution effects in agriculture.     

Role of salicylic acid in alleviating oxidative damage in rice roots (Oryza sativa) subjected to cadmium stress

Time-dependent changes in enzymatic and non-enzymatic antioxidants, and lipid peroxidation were investigated in roots of rice (Oryza sativa) grown hydroponically with Cd, with or without pretreatment of salicylic acid (SA). Exposure to 50 microM Cd significantly decreased root growth, and activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), but increased the concentrations of H(2)O(2), malondialdehyde (MDA), ascorbic acid (AsA), glutathione (GSH) and non-protein thiols (NPT). However, pretreatment with 10 microM SA enhanced the activities of antioxidant enzymes and the concentrations of non-enzymatic antioxidants, but lowered the concentrations of H(2)O(2) and MDA in the Cd-stressed rice compared with the Cd treatment alone. Pretreatment with SA alleviated the Cd-induced inhibition of root growth. The results showed that pretreatment with SA enhanced the antioxidant defense activities in Cd-stressed rice, thus alleviating Cd-induced oxidative damage and enhancing Cd tolerance. The possible mechanism of SA-induced H(2)O(2) signaling in mediating Cd tolerance was discussed.     

Role of cation structure in the phytotoxicity of ionic liquids: growth inhibition and oxidative stress in spring barley and common radish

The present study determines the influence of three ionic liquids (ILs) containing cations with diversified structure on the growth and development of spring barley seedlings and common radish leaves. Increasing amounts of 1-butyl-1-methylpyrrolidinium hexafluorophosphate [Pyrrol][PF₆], 1-butyl-1-methylpiperidinium hexafluorophosphate [Piper][PF₆], and 1-butyl-4-methylpyridinium hexafluorophosphate [Pyrid][PF₆] were added to the soil on which both plants were cultivated. The results of this studies showed that the applied ILs were highly toxic for plants, demonstrated by the inhibition of length of plant shoots and roots, decrease of fresh mass, and increase of dry weight content. Common radish turned out to be the plant with higher resistance to the used ILs. The differences in the cation structure did not influence phytotoxity of ILs for spring barley. Furthermore, all ILs led to a decrease of photosynthetic pigments, which was directly followed by decreased primary production in plants. Oxidative stress in plants occurred due to the presence of ILs in the soil, which was demonstrated by the increase of malondialdehyde (MDA) content, changes in the H₂O₂ level, and antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD). The changes in the chlorophyll contents and the increase of POD activity turned out to be the most significant oxidative stress biomarkers in spring barley and common radish. Both spring barley and radish exposed to ILs accumulated a large amount of fluoride ion.

     

Effect of iron on lipid peroxidation, and enzymatic and non-enzymatic antioxidants and bacoside-A content in medicinal plant Bacopa monnieri L

The effect of Fe was investigated in medicinally important plant, Bacopa monnieri L. and the response on malondialdehyde (MDA) content, superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX) was found different in roots and leaves of the metal treated plants. Iron induced stress was observed as indicated by high level of lipid peroxidation, being more steep increase in leaves than roots. In roots, SOD activity was found to increase in metal treated plants except 80 and 160 microM at 72 h, whereas, it decreased in leaves except 10 and 40 microM after 48 h as compared to their respective controls. Among H2O2 eliminating enzymes, POD activity increased in roots, however, it decreased in leaves except at 10 and 40 microM Fe after 48 h as compared to control. At 24 and 48 h, APX activity and ascorbic acid content followed the similar trend and were found to increase in both parts of the metal treated plants as compared to their respective controls. The level of cysteine content in the roots increased at initial period of exposure; however, no marked change in its content was noticed in leaves. In both roots and leaves, non-protein thiol content was found to increase except at higher metal concentrations at 72 h. The data of proline content have shown significant (p<0.01) increase at 40 microM onwards in both part of the plants after 48 and 72 h. Correlation coefficient was evaluated between metal accumulations with various parameters and also between different antioxidant parameters with MDA. Since the level of bacoside-A (active constituent) content in metal treated plants increases, therefore, it is advisable to assess the biological activity of the plants before using for medicinal purposes, particularly in developing countries.     

A comparison of biochemical responses to oxidative and metal stress in seedlings of barley, Hordeum vulgare L

Biochemical responses on the bases of activities of antioxidant enzymes; peroxidase, catalase, superoxide dismutase and glutathione reductase as well as estimations of total protein, lipid peroxidation and thiols in the form of protein, non-protein, glutathione and phytochelatin measured in growing seedlings of barley, Hordeum vulgare L., from Day 2 through 8 were compared following treatment of seeds for 2 h with oxidative agents, paraquat 5 x 10(-5), 10(-4), 10(-3) M, H2O2 10(-3), 5 x 10(-3), 10(-2) M and a metal salt, CdSO4 10(-5), 10(-4), 10(-3) M. A significant induction of all antioxidant enzymes along with an increase in the levels of protein, lipid peroxidation and glutathione was noted in response to oxidative stress, CdSO4 induced significant peroxidase and catalase activities but not superoxide dismutase. In a marked contrast from oxidative stress, CdSO4 decreased glutathione reductase activity as well as glutathione levels but increased phytochelatin level. The differential biochemical responses thus underlined the crucial involvement of glutathione and phytochelatin in the oxidative and metal-induced adaptive responses, respectively.     

Distribution of peroxidase activity and polyamine pattern of Heliotropium europeum in potassium deficiency conditions

Polyamines, especially putrescine, are reported to increase in higher plants in response to various stressors, in particular mineral deficiency, low pH and different atmospheric pollutants. Peroxidases are another stress indicator in plants. The aim of this study is to establish the relationship between polyamine content and peroxidase activity in potassium deficiency conditions. In this study, potassium deficiency increased the free and bound Put and Spd contents. We estimated more stimulation in free Put synthesis (300%) in potassium deficiency. We also determined increment in POD activity in whole plant, especially root POD activity increased much more compared to stem and leaf.     

Phytoremediation of phenol using Vicia sativa L. plants and its antioxidative response

Common vetch (Vicia sativa L.) is a legume species with an extensive agricultural use. However, the phytoremediation potentiality of this species has not been sufficiently explored because little is known about its resistance to inorganic and organic pollutants. In the present work, phenol tolerance of common vetch was assayed at different stages of growth. Germination index and germination rate decreased only at high phenol concentrations (250 and 500 mg L(-1)), whereas 30-day-old plants were able to tolerate this pollutant, with high removal efficiencies. The activities of antioxidative enzymes, such as peroxidase (POD) and ascorbate peroxidase, increased significantly with the highest phenol concentration, whereas superoxide dismutase activity, malondialdehyde, and H(2)O(2) levels remained unaltered. Besides, an increase in two basic isoforms of POD was observed in plants treated with phenol. The results suggested that common vetch has an efficient protection mechanism against phenol-induced oxidative damage. Moreover, it could tolerate and remove high phenol concentrations, avoiding serious phytotoxic effects. Thus, V. sativa could be considered an interesting tool in the field of phytoremediation.     

Electrochemical oxidation of bisphenol A. Application to the removal of bisphenol A using a carbon fiber electrode

Leachate samples with a high strength of ammonium-nitrogen (NH4+-N) were collected from a local landfill site in Hong Kong. Two experiments were carried out to study (1) the inhibition of microbial activity of activated sludge by NH4+-N and (2) the chemical precipitation of NH4+-N from leachate as a preliminary treatment prior to the activated sludge process. The experimental results demonstrated that the efficiency of COD removal decreased from 97.7% to 78.1%, and the dehydrogenase activity of activated sludge decreased from 9.29 to 4.93 microg TF/mg MLSS, respectively, when the NH4+-N concentration increased from 53 to 800 mg/l. The experiment also demonstrated that the NH4+-N in the leachate can be quickly precipitated as MgNH4PO4 x 6H2O after addition of MgCl2 x 6H2O + Na2HPO4 x 12H2O. The NH4+-N concentration was reduced from 5618 to 112 mg/l within 15 min when a molar ratio of Mg2+:NH+:PO4(3-) = 1:1:1 was used. The optimum pH to reach the minimum solubility of MgNH4PO4 x 6H2O was found to be in the range of 8.5-9.0. Attention should be given to the high salinity formed in the treated leachate by using MgCl2 x 6H2O + Na2HPO4 x 12H2O, which may affect microbial activity in the following biological treatment processes. Using two other combinations of chemicals [MgO + 85%H3PO4 and Ca(H2PO4)2 x H2O + MgSO4 x 7H2O] could minimise salinity generation after precipitation, while they were less efficient for NH4+-N removal.     

Effect of heavy metal stress on antioxidative enzymes and lipid peroxidation in leaves and roots of two mangrove plant seedlings (Kandelia candel and Bruguiera gymnorrhiza)

The effects of multiple heavy metal stress on the activity of antioxidative enzymes and lipid peroxidation were studied in leaves and roots of two mangrove plants, Kandelia candel and Bruguiera gymnorrhiza, grown under control (10 per thousand NaCl nutrient solution) or five levels of multiple heavy metal stress (10 per thousand NaCl nutrient solution containing different concentration of Pb2+, Cd2+, and Hg2+). Leaves and roots of control and heavy metal-stressed plants were harvested after two months. In leaves of heavy metal-stressed plants superoxide dismutase (SOD) and peroxidase (POD) activities fluctuated in different stress levels compared to the control, while catalase (CAT) activity increased with stress levels in K. candel, but remained unchanged in leaves of B. gymnorrhiza. In comparison with the control, the dynamic tendency of SOD, CAT, and POD activities in roots of heavy metal-stressed plants all ascended, and then declined. The increase in enzyme activities demonstrated that K. candel is more tolerant to heavy metals than B. gymnorrhiza. Lipid peroxidation was enhanced only in leaves of heavy metal-stressed B. gymnorrhiza. These results indicate that in heavy-metal stress antioxidative activities may play an important role in K. candel and B. gymnorrhiza and that cell membrane in leaves and roots of K. candel have greater stability than those of B. gymnorrhiza. For pollution monitoring purposes, POD activity in roots and leaves maybe serve as a biomarker of heavy metal stress in K. candel, while lipid peroxidation maybe serve as biomarker in B. gymnorrhiza.     

Development and validation of a terrestrial biotic ligand model predicting the effect of cobalt on root growth of barley (Hordeum vulgare)

A Biotic Ligand Model was developed predicting the effect of cobalt on root growth of barley (Hordeum vulgare) in nutrient solutions. The extent to which Ca(2+), Mg(2+), Na(+), K(+) ions and pH independently affect cobalt toxicity to barley was studied. With increasing activities of Mg(2+), and to a lesser extent also K(+), the 4-d EC50(Co2+) increased linearly, while Ca(2+), Na(+) and H(+) activities did not affect Co(2+) toxicity. Stability constants for the binding of Co(2+), Mg(2+) and K(+) to the biotic ligand were obtained: logK(CoBL)=5.14, logK(MgBL)=3.86 and logK(KBL)=2.50. Limited validation of the model with one standard artificial soil and one standard field soil showed that the 4-d EC50(Co2+) could only be predicted within a factor of four from the observed values, indicating further refinement of the BLM is needed.     

Effects of air pollutants on the growth and antioxidative system of Norway spruce exposed in open-top chambers

Grafted Norway spruce trees were subjected to exposure beginning in April 1988, to one of four different air treatments in open-top chambers: Charcoal filtered air (CF), non-filtered air (NF), non-filtered air with the addition of O(3) during summer (NFO), and SO(2) plus NO(2) during winter (NFOSN). CF trees were considered as the reference group. No effects on growth parameters were observed. Samples of the two youngest needle year classes were taken late in November 1989 for enzyme determinations. The activity of ascorbic acid peroxidase (A-POD) increased the same level in all treatments, and activities of catalase and dehydroascorbic acid reductase (DHA-R) increased only in NF and NFO treatments. A higher level of activity in the NFOSN treatment was observed only for glucose-6-phosphate-dehydrogenase (Glc-6-P-DH) and non-specific peroxidase (POD). Isoelectric focusing of POD showed a changed pattern in the NFOSN treatment. Neither activity nor isoelectric focusing of superoxidase dismutase (SOD) was changed in any of the treatments.     

Effects of tetrabromobisphenol A as an emerging pollutant on wheat (Triticum aestivum) at biochemical levels

Biochemical responses of wheat (Triticum aestivum) to the stress of tetrabromobisphenol A (TBBPA) as an emerging pollutant were examined. The results indicated that reduction of the chlorophyll (CHL) content in wheat leaves could be observed. However, the changes in the CHL content with the increasing TBBPA concentration from 50 to 5000 mg kg(-1) were insignificant (p>0.05). Increased malondialdehyde levels were detected in wheat leaves after both 7-d and 12-d exposures. The changes in the activity of superoxide dismutases (SOD), peroxidases (POD) and catalases (CAT) in wheat leaves irregularly fluctuated with time as the TBBPA concentration increased. However, significant (p<0.05) decrease in the activity of POD and CAT treated with 500 and 5000 mg kg(-1) TBBPA could be observed. Our data also showed that the plant has the capacity to tolerate the oxidative stress, but the capacity would be lost with prolonged exposure and increasing TBBPA concentration. There were no dose-response effects in the changes between the activity of antioxidant enzymes (SOD, POD and CAT) and the concentration of TBBPA. The decrease in the activity of POD and CAT could be considered as good biomarkers of serious stress by TBBPA in soil environment.     

The effect of excess Zn on mineral nutrition and antioxidative response in rapeseed seedlings

Zinc (Zn) is a necessary element for plants, but excess Zn can be detrimental. To investigate Zn toxicity, rapeseed (Brassica napus) seedlings were treated with 0.07–1.12 mM Zn for 7 d. Inhibition of plant growth along with root damage, chlorosis and decreased chlorophyll (a and b) content in newly expanded leaves (the second and third leaves formed following cotyledons) were found under Zn stress. The Zn content increased in plants under external Zn stress, while concentrations of phosphorus, copper, iron, manganese and magnesium reduced significantly, especially in roots. Meanwhile, increased lipid peroxidation was detected biochemically and histochemically. Compared with controls, NADH oxidase and peroxidase (POD) activity increased in leaves and roots of plants under high Zn, but superoxide dismutase (SOD), catalase and ascorbate peroxidase activities decreased. The changes in glutathione S-transferase activity and in ascorbic acid, dehydroascorbate, non-protein thiols and glutathione contents were also measured under Zn stress. Isoforms of SOD and POD were separated using non-denaturing polyacrylamide gel electrophoresis and their activities were analyzed. Our results suggested that excess Zn exerts its toxicity partially through disturbing nutrient balance and inducing oxidative stress in plants. These data will be helpful for better understanding of toxicity of Zn and the adaptive mechanism in Zn non-hyperaccumulator plants.     

Coupled oxidation of aromatic hydrocarbons by horseradish peroxidase and hydrogen peroxide

The oxidation capability of horseradish peroxidase (HRP) and hydrogen peroxide (H2O2) coupled oxidation of aromatic hydrocarbons (o-xylene-d10 and naphthalene-d8) was investigated. Batch experiments were conducted using horseradish peroxidase prepared in potassium phosphate buffer in the presence of H2O2. The oxidation of aromatic hydrocarbon was tested as a function of HRP at a fixed concentration of H2O2, and as a function of the concentration of H2O2 at a constant HRP activity (4000 units/ml). The mass removal of o-xylene-d10 and naphthalene-ds increased with increasing HRP enzymatic activity, and up to 54% and 51% of mass removal were observed for o-xylene-d10 and naphthalene-d8, respectively. Increasing the concentration of H2O2 resulted in increased mass removal of aromatic hydrocarbons.     

Phytotoxicity of nickel in a range of European soils: influence of soil properties, Ni solubility and speciation

We investigated the influence of soil properties on Ni toxicity to barley root and tomato shoot growth, using 16 European soils. The effective concentration of added Ni causing 50% inhibition (EC(50)) ranged from 52 to 1929mgkg(-1) and from 17 to 920mgkg(-1) for the barley and tomato test, respectively, representing 37- and 54-fold variation among soils. Soil cation exchange capacity was the best single predictor for the EC(50). The EC(50) based on either the Ni concentration or free Ni(2+) activity in soil solution varied less among soils (7-14 fold) than that based on the total added Ni, suggesting that solubility of Ni is a key factor influencing its toxicity to plants. The EC(50) for free Ni(2+) activity from the barley test decreased with increasing pH, indicating a protective effect of protons. The results can be used in the risk assessment of Ni in the terrestrial environment.     

Mitigation of peroxidative stress for barley exposed to cadmium in the presence of water-extractable organic matter from compost-like materials

The effects of water-extractable organic matter (WEOM) from compost-like materials on peroxidative stress were investigated for hydroponic culture of barley exposed to Cd. In the presence of WEOM, lipoxygenase activity and malondialdehyde, indices of peroxidative stress in barley, were significantly reduced, compared to those with Cd alone (5 μM) for a 30-d culture (p < 0.05). In addition, Cd uptake in the presence of WEOM samples was significantly lower than that in their absence (p < 0.05). These results indicate that the addition of WEOM can be effective in mitigating the peroxidative stress in barley exposed to Cd. Of the total Cd in the solution, 7–8% was complexed with WEOM, indicating that the complexation of Cd with WEOM is a minor factor in reducing Cd-induced stress in barley. The WEOM sample was purified by cation-exchange column and ultrafiltration to remove the nutrient minerals, such as Ca, Mg and Fe. When the purified WEOM was employed for hydroponic culture in the presence of Cd, significant decreases in peroxidative stress and Cd uptake were observed (p < 0.05). These results show that the organic components in WEOM contribute to the mitigation of peroxidative stress in barley exposed to Cd.     

Photo-oxidation of cork manufacturing wastewater

Several photo-activated processes have been investigated for oxidation of a cork manufacturing wastewater. A comparative activity study is made between different homogeneous (H2O2/UV-Vis and H2O2/Fe2+/UV-Vis) and heterogeneous (TiO2/UV-Vis and TiO2/H2O2/UV-Vis) systems, with degradation performances being evaluated in terms of total organic carbon (TOC) removal. Results obtained in a batch photo-reactor show that photo-catalysis with TiO2 is not suitable for this kind of wastewater while the H2O2/UV-Vis oxidation process, for which the effect of some operating conditions was investigated, allows to remove 39% of TOC after 4 h of operation (for C(H2O2)=0.59 M, pH=10 and T=35 degrees C). The combined photo-activated process, i.e., using both TiO2 and H2O2, yields an overall TOC decrease of 46% (for C(TiO2)=1.0 gl(-1)). The photo-Fenton process proved to be the most efficient, proceeds at a much higher oxidation rate and allows to achieve 66% mineralization in just 10 min of reaction time (for C(H2O2)=0.31 M, T=30 degrees C, Fe2+:H2O2=0.12 (mol) and pH=3.2).     

Lead induced changes in antioxidant metabolism of horsegram (Macrotyloma uniflorum (Lam.) Verdc.) and bengalgram (Cicer arietinum L.)

One-month old horsegram (Macrotyloma uniflorum (Lam.) Verdc. cv VZM1) and bengalgram (Cicer arietinum L. cv Annogiri) were exposed to different regimes of lead stress as Pb(NO3)2 at 0, 200, 500 and 800 ppm concentrations. The extent of oxidative damage as the rate of lipid peroxidation, antioxidative response and the accumulation of lead in roots and shoots of both plants were evaluated after 12 days of lead stress. Lead (Pb) treated plants showed increased levels of lipid peroxidation as evidenced from the increased malondialdehyde content coupled with the increase in the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), glutathione reductase (GR), glutathione S-transferase (GST) compared to control (untreated) plants. Lead stress caused significant changes in the activity of antioxidative enzymes. The effect of lead was found to be concentration dependent. Higher concentration of lead (800 ppm) resulted 2- to 3-fold increase in SOD, catalase and peroxidase activities, 3- to 5-fold increase in GR activity and 3- to 4-fold increase in GST activity in roots and leaves of both horsegram and bengalgram plants. Lead stress caused a significant increase in the rate of peroxidation as showed in the levels of malondialdehyde content in roots and leaves of both plant species. Horsegram registered lower Pb accumulation than bengalgram, however localization of Pb was greater in roots than leaves in both plants. In general, lipid peroxide levels and antioxidative enzyme activities were higher in horsegram than bengalgram and also more in roots than leaves which best concordance with the lead contents of both the plants and organs. These results suggest that Pb toxicity causes oxidative stress in plants and the antioxidative enzymes SOD, CAT, POD, GR, GST could play a pivotal role against oxidative injury.