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 dissolved organic matter on bioavailability of chlorotoluron to wheat

Chlorotoluron (Chl) is a phenylurea herbicide and is widely used for controlling weeds. While it has brought great benefits to crop production, it has also resulted in contamination to ecosystem. In this study, we investigated accumulation of chlorotoluron (Chl) and biological responses of wheat plants as affected by dissolved organic matter (DOM). Wheat seedlings grown under 10 mg kg⁻¹ Chl for 4 d showed a low level of chlorophyll accumulation and damage to plasma membrane. The growth was inhibited by exposure of chlorotoluron. Treatment with 50 mg DOC kg⁻¹ DOM derived either from sludge (DOM-SL) or straw (DOM-ST) attenuated the chlorotoluron toxicity to plants. Both DOMs decreased activities of catalase, peroxidase and superoxide dismutase in Chl-treated seedlings. However, an increased glutathione S-transferases activity was observed under the same condition. Wheat plants treated with Chl in the presence of DOM accumulated less Chl than those treated with Chl alone. Moreover, in the presence of DOM, bioconcentration factor (BCF) decreased whereas translocation factors increased. Analyses with FT-IR spectra confirmed the regulatory role of DOMs in reducing Chl accumulation in wheat.     

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.     

Effect of exogenous abscisic acid on the level of antioxidants in Atractylodes macrocephala Koidz under lead stress

This study hypothesized that the positive or negative effects of exogenous abscisic acid (ABA) on oxidative stress caused by lead were dose dependent. The effects of different levels of ABA (2.5, 5, and 10 mg L^?1) on lead toxicity in the leaves of Atractylodes macrocephala were studied by investigating plant growth, soluble sugars, proteins, lipid peroxidation, and antioxidative enzymes. Excess Pb inhibited root dry weight, root length, and the number of lateral roots, but increased shoot growth. In addition, lead stress significantly decreased the levels of chlorophyll pigments, protein, and activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and peroxidase (POD). Different levels of ABA significantly increased SOD, CAT, POD, and APX activities, but decreased the level of hydrogen peroxide and malondialdehyde in nonstressed plants. Exogenous application of 2.5 mg L^?1 ABA detoxified the stress-generated damages caused by Pb and also enhanced plant growth, soluble sugars, proteins, and all four antioxidant enzyme activities but reduced Pb uptake of lead-stressed plant compared to lead treatment alone. However, the toxic effects of Pb were further increased by the applications of 5 and 10 mg L^?1 ABA. The levels of antioxidants caused by a low concentration of exogenous ABA might be responsible for minimizing the Pb-induced toxicity in A. macrocephala.     

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.     

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.     

Joint stress of chlorimuron-ethyl and cadmium on wheat Triticum aestivum at biochemical levels

Biochemical responses to joint stress of chlorimuron-ethyl and cadmium (Cd) in wheat Triticum aestivum were examined. The joint action of chlorimuron-ethyl and Cd weakened the inhibition of Cd or chlorimuron-ethyl on the formation of chlorophyll. It was deduced that wheat plants had the capability to protect themselves by increasing the activity of the antioxidant enzyme peroxidase (POD) with the exposure time. The joint effect of chlorimuron-ethyl and Cd on the superoxide dismutase (SOD) activity in leaves was additive, while the joint effect on the SOD activity in roots was determined by the interaction of chlorimuron-ethyl and Cd in wheat. It was also concluded that the change of malondialdehyde (MDA) content in wheat might not be a good biomarker in the oxidative damage by chlorimuron-ethyl, while a decrease in the soluble protein content and POD activity in roots could be considered as a biomarker in the damage of wheat by chlorimuron-ethyl and Cd.     

Involvement of abscisic acid in regulating antioxidative defense systems and IAA-oxidase activity and improving adventitious rooting in mung bean [Vigna radiata (L.) Wilczek] seedlings under cadmium stress

In vitro experiments were conducted to investigate the effects of abscisic acid (ABA) and Cd on antioxidative defense systems and indole-3-acetic acid (IAA) oxidase during adventitious rooting in mung bean [Vigna radiata (L.) Wilczek] seedlings. The exogenous ABA significantly enhanced the number and fresh weight of the adventitious roots. CdCl₂ strongly inhibited adventitious rooting. Pretreatment with 10 μM ABA clearly alleviated the inhibitory effect of Cd on rooting. ABA significantly reduced superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD), and catalase (CAT) activities, as well as the levels of glutathione (GSH) and ascorbic acid (ASA) during adventitious rooting. ABA strongly increased IAA-oxidase activity during the induction (0–12 h) and expression (after 48 h) phases and increased the phenols levels. Cd treatment significantly reduced the activities of SOD, APX, POD, and IAA oxidase, as well as GSH level. Cd strongly increased ASA levels. ABA pretreatment counteracted Cd-induced alterations of certain antioxidants and antioxidative enzymes, e.g., remarkably rescued APX and POD activities, reduced the elevated SOD and CAT activities and ASA levels, and recovered the reduced GSH levels, caused by Cd stress. Thus, the physiological effects of the combination of ABA and Cd treatments were opposite of those obtained with Cd treatment alone, suggesting that ABA involved in the regulation of antioxidative defense systems and the alleviation of wounding- and Cd-induced oxidative stress.     

Influence of cadmium on antioxidant capacity and four microelement concentrations in tomato seedlings (Lycopersicon esculentum)

Tomato (Lycopersicon esculentum) seedlings were grown in four cadmium (Cd) levels of 0-10 microM in a hydroponic system to analyze the antioxidative enzymes, Cd concentration in the plants, and the interaction between Cd and four microelements. The results showed that there was a significant increase in malondialdehyde (MDA) concentration, and superoxide dismutase (SOD) and peroxidase (POD) activities in the plants subjected to 1-10 microM Cd. This indicates that Cd stress induces an oxidative stress response in tomato plants, characterized by an accumulation of MDA and increase in activities of SOD and POD. Root, stem and leaf Cd concentrations increased with its exposure Cd level, and the highest Cd concentration occurred in roots, followed by leaves and stems. A concentration- and tissue-dependent response was found in the four microelement concentrations to Cd stress in the tomato leaves, stems and roots. Regression analysis showed that there was a significantly negative correlation between Cd and Mn, implying the antagonistic effect of Cd on Mn absorption and translocation. The correlation between Cd and Zn, Cu and Fe were inconsistent among leaves, stems and roots.     

Antioxidant responses to simulated acid rain and heavy metal deposition in birch seedlings

This study measured the responses of different anti-oxidants in 2-year-old birch (Betula pendula Roth) seedlings subjected to simulated acid rain (pH 4.0) and heavy metals (Cu/Ni), applied alone or in combination for 2 months. The applied concentrations of pollutants did not significantly affect seedling biomass or total glutathione levels. Acid rain alone increased superoxide dismutase (SOD) activity both in leaves and roots, while heavy metals alone inhibited SOD activity in roots. Both acid rain and heavy metals applied singly increased ascorbate peroxidase (APX) and guaiacol peroxidase (GPX) activities in leaves but decreased activities in roots. In contrast, acid rain and heavy metal treatments increased glutathione reductase (GR) activity in roots but not in leaves. Spraying birch seedlings with a mixture of acid rain and heavy metals increased SOD, APX and GPX activities in leaves and GR activity in roots. However, the effects of mixed pollutants on enzyme activities usually were less than the summed effects of individual pollutants. Enzyme responses also depended on where pollutants were applied: spraying pollutants onto the shoots initiated higher responses in SOD, APX and GPX than did application to the soil surface, while the opposite was true for GR.     

Antioxidative response to Cd in a newly discovered cadmium hyperaccumulator, Arabis paniculata F

A hydroponic experiment was carried out to study the effect of cadmium (Cd) on growth, Cd accumulation, lipid peroxidation, reactive oxygen species (ROS) content and antioxidative enzymes in leaves and roots of Arabis paniculata F., a new Cd hyperaccumuator found in China. The results showed that 22-89 microM Cd in solution enhanced the growth of A. paniculata after three weeks, with 21-27% biomass increase compared to the control. Cd concentrations in shoots and roots increased with increasing Cd supply levels, and reached a maximum of 1662 and 8670 mg kg(-1) Cd dry weight at 178 microM Cd treatment, respectively. In roots, 22-89 microM Cd reduced the content of malondialdehyde (MDA), superoxide (O(2)(-1)) and H(2)O(2) as well as the activities of superoxide dismutase (SOD), guaiacol peroxidase (GPX), ascorbate peroxidase (APX) and glutathione reductase (GR). In leaves, the contents of MDA, O(2)(-1) and H(2)O(2) remained unaffected by 22-89 microM Cd, while 178 microM Cd treatment significantly increased the MDA content, 69.5% higher than that of the control; generally, the activities of SOD, catalase (CAT), GPX and APX showed an increasing pattern with increasing Cd supply levels. Our present work concluded that A. paniculata has a great capability of Cd tolerance and accumulation. Moderate Cd treatment (22-89 microM Cd) alleviated the oxidative stress in roots, while higher level of Cd addition (178 microM) could cause an increasing generation of ROS, which was effectively scavenged by the antioxidative system.     

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.     

Arbuscular mycorrhizal fungi can alleviate the adverse effects of chlorothalonil on Oryza sativa L

A glasshouse pot experiment was conducted to investigate the effect of the fungicide chlorothalonil on the growth of upland rice, in the absence or presence of the arbuscular mycorrhizal fungus (AMF) Glomus mosseae (NM and GM treatments). The plants were grown with three concentrations of chlorothalonil (0, 50 and 100 mg kg(-1) soil). Mycorrhizal colonization decreased significantly with increasing chlorothalonil concentrations. Plant biomass decreases were smaller in GM plants than in non-mycorrhizal (NM) plants. Mycorrhizal dependency was the highest with 50 mg kg(-1) chlorothalonil. Chlorothalonil affected physiological processes in upland rice irrespective of inoculation. Chlorothalonil at 50 and 100 mg kg(-1) increased ascorbate peroxidase (APX) activity and soluble protein concentrations in shoots and roots of NM upland rice. However, values of APX, catalase (CAT) and peroxidase (POD) were reduced more in GM plants than in NM plants. These results showed that chlorothalonil induced oxidative stress in upland rice and it is needed to evaluate the side effects of chlorothalonil on rice and AMF.     

Enantioselectivity of racemic metolachlor and S-metolachlor in maize seedlings

Chiral herbicides may have enantioselective effects on plants. In this study, we assessed and compared the enantioselectivity of the chiral herbicides rac-metolachlor and S-metolachlor to maize seedlings. The superoxide dismutase activity (SOD) activity of roots and stem leaves treated by rac-metolachlor was 1.38 and 1.99 times that of roots and stem leaves treated by S-metolachlor. The peroxidase activity (POD) activity of roots and stem leaves was 1.48 and 2.79 times that of roots and stem leaves treated by S-metolachlor, respectively, while the catalase activity (CAT) activity was 4.77 and 8.37 times greater, respectively. The Hill reaction activity of leaves treated by rac-metolachlor were 1.45, 1.33, and 1.14 times those treated by S-metolachlor with treatments of 18.6, 37.2, and 74.4 μM. The differences observed between treatments of rac- and S-metolachlor were significant. Significant differences in maize seedling morphology were also observed between rac- and S-metolachlor treatments. The degradation rate of S-metolachlor in roots was greater than that of rac-metolachlor. The half-lives of rac- and S-metolachlor were 80.6 and 60.3 h at 18.6 μM; 119.5 and 90 h at 37.2 μM; and 169 and 164.8 h at 74.4 μM, respectively. Using the liquid chromatography-mass spectrometry method, hydroxymetolachlor, deschlorometolachlor and deschlorometolachlor propanol were considered to be possible metabolites. We determined the enantioselective toxicity of rac- and S-metolachlor to maize and speculated on the proposed metabolic pathway of metolachlor in maize roots. These results will help to develop an understanding of the proper application of rac- and S-metolachlor in crops, and give some information for environmental safety evaluation of rac- and S-metolachlor.     

Antioxidant metabolism of coffee cell suspension cultures in response to cadmium

The antioxidant responses of coffee (Coffea arabica L.) cell suspension cultures to cadmium (Cd) were investigated. Cd accumulated very rapidly in the cells and this accumulation was directly correlated with an increase in applied CdCl(2) concentration in the external medium. At 0.05mM CdCl(2), growth was stimulated, but at 0.5mM CdCl(2), the growth rate was reduced. An alteration in activated oxygen metabolism was detected by visual analysis as well as by an increase in lipid peroxidation at the higher CdCl(2) concentration. Catalase (CAT; EC 1.11.1.6), glutathione reductase (GR; EC 1.6.4.2) and superoxide dismutase (SOD; EC 1.15.1.1) activity increased, particularly at the higher concentration of CdCl(2). Ascorbate peroxidase (APX; EC 1.11.1.11) activity was increased at the lower CdCl(2) concentration used, but could not be detected in cells growing in the higher CdCl(2) concentration after 24h of growth, whilst guaiacol peroxidase (GOPX; EC 1.11.1.7) did not show a clear response to Cd treatment. An analysis by non-denaturing PAGE followed by staining for enzyme activity, revealed one CAT isoenzyme, nine SOD isoenzymes and four GR isoenzymes. The SOD isoenzymes were differently affected by CdCl(2) treatment and one GR isoenzyme was shown to specifically respond to CdCl(2). The results suggest that the higher concentrations of CdCl(2) may lead to oxidative stress. The main response appears to be via the induction of SOD and CAT activities for the removal of reactive oxygen species (ROS), and by the induction of GR to ensure the availability of reduced glutathione for the synthesis of Cd-binding peptides, which may also be related to the inhibition of APX activity probably due to glutathione and ascorbate depletion.     

Chromium induced lipid peroxidation in the plants of Pistia stratiotes L.: role of antioxidants and antioxidant enzymes

In the plant, Pistia stratiotes L., the effect of different concentrations of chromium (0, 10, 40, 80 and 160 microM) applied for 48, 96 and 144 h was assessed by measuring changes in the chlorophyll, protein, malondialdehyde (MDA), cysteine, non-protein thiol, ascorbic acid contents and superoxide dismutase (SOD), ascorbate peroxidase (APX) and guiacol peroxidase (GPX) activity of the plants. Both in roots and leaves, an increase in MDA content was observed with increase in metal concentration and exposure periods. In roots, the activity of antioxidant enzymes viz. SOD and APX increased at all the concentrations of Cr at 144 h than their controls. The GPX activity of the treated roots increased with increase in Cr concentration at 48 and 96 h of exposures, however, at 144 h its activity was found declined beyond 10 microM Cr. The level of antioxidants in the roots of the treated plant viz. cysteine and ascorbic acid was also found increased at all the concentrations of Cr at 144 h than their respective controls, however, an increase in the non-protein thiol content was recorded up to 40 microM Cr followed by decrease. The chlorophyll content decreased with increase in Cr concentrations and exposure periods. However, the protein content of both roots and leaves were found decreased with increase in Cr concentrations at all the exposure periods except an increase was recorded at 10 microM Cr at 48 h. In Cr treated plants, the no observed effect concentration (NOEC) and lowest observed effect concentration (LOEC) for leaves chlorophyll and protein contents were 40 and 80 microM Cr, respectively after 48 h exposure while NOEC and LOEC for root protein content were 10 and 40 microM, respectively after 48 h. The analysis of correlation coefficient data revealed that the metal accumulation in the roots of the plant was found positively correlated with antioxidant parameters except SOD after 48 h of exposure, however, negatively correlated with most of all the parameters studied at 144 h in both part of the plant. It may be suggested from the present study that toxic concentrations of Cr cause oxidative damage as evidenced by increased lipid peroxidation and decreased chlorophyll and protein contents. However, the higher levels of enzymatic and non-enzymatic antioxidants suggest the reason for tolerating higher levels of metals.     

Ecotoxicological effects of typical personal care products on seed germination and seedling development of wheat (Triticum aestivum L.)

Biochemical responses of wheat (Triticum aestivum L.) seedlings stressed by two typical personal care products (PCPs) – triclosan (TCS) and galaxolide (HHCB) were experimentally investigated to assess their ecological risks. The results showed that wheat shoot and root elongation was significantly inhibited by 50–250 mg L⁻¹ TCS and HHCB. Wheat roots were sensitive to TCS, while shoots were sensitive to HHCB. The median effect concentration (EC₅₀) of TCS and HHCB based on the inhibition of their sensitive sites were 147.8 and 143.4 mg L⁻¹, respectively. Moreover, the damage of wheat seedlings treated by low concentration of TCS and HHCB during a long period cannot be neglected. After a 21-d exposure, 0.2–3.0 mg L⁻¹ TCS and HHCB treatment caused the damage to the accumulation of chlorophyll (CHL), the synthesis of soluble protein (SP), and the activity of peroxidase (POD) and superoxide dismutases (SOD) in different degree. However, different changing trends of these physiological indexes treated by different PCPs were observed after 7-d to 14-d exposures, especially the activity of POD and SOD. The activity of POD and SOD in wheat leaves and roots decreased with an increase in the concentration of TCS and the exposure time. However, the enzyme activities in wheat leaves treated by 0.2–3.0 mg L⁻¹ HHCB increased after a 14-d exposure, and with the prolongation of exposure time, the enzyme activities significantly decreased. The variations in these physiological indexes of wheat could be considered as good biomarkers of serious stress by TCS and HHCB in the 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.     

Nickel accumulation and its effect on growth,physiological and biochemical parameters in millets and oats

With the boom in industrialization, there is an increase in the level of heavy metals in the soil which drastically affect the growth and development of plants. Nickel is an essential micronutrient for plant growth and development, but elevated level of Ni causes stunted growth, chlorosis, nutrient imbalance, and alterations in the defense mechanism of plants in terms of accumulation of osmolytes or change in enzyme activities like guiacol peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD). Ni-induced toxic response was studied in seedlings of finger millet, pearl millet, and oats in terms of seedling growth, lipid peroxidation, total chlorophyll, proline content, and enzymatic activities. On the basis of germination and growth parameters of the seedling, finger millet was found to be the most tolerant. Nickel accumulation was markedly lower in the shoots as compared to the roots, which was the highest in finger millet and the lowest in shoots of oats. Plants treated with a high concentration of Ni showed significant reduction in chlorophyll and increase in proline content. Considerable difference in level of malondialdehyde (MDA) content and activity of antioxidative enzymes indicates generation of redox imbalance in plants due to Ni-induced stress. Elevated activities of POD and SOD were observed with high concentrations of Ni while CAT activity was found to be reduced. It was observed that finger millet has higher capability to maintain homeostasis by keeping the balance between accumulation and ROS scavenging system than pearl millet and oats. The data provide insight into the physiological and biochemical changes in plants adapted to survive in Ni-rich environment. This study will help in selecting the more suitable crop species to be grown on Ni-rich soils.

     

Cadmium accumulation and its influence on lipid peroxidation and antioxidative system in an aquatic plant, Bacopa monnieri L

Bacopa monnieri L. plants exposed to 10, 50, 100 and 200 microM cadmium (Cd) for 48, 96 and 144 h were analysed with reference to the accumulation of metal and its influence on various enzymatic and non-enzymatic antioxidants, thiobarbituric acid reactive substances (TBARS), photosynthetic pigments and protein content. The accumulation of Cd was found to be increased in a concentration and duration dependent manner with more Cd being accumulated in the root. TBARS content of the treated roots and leaves increased with increase in Cd concentration and exposure periods, indicating the occurrence of oxidative stress. Induction in the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX) and guiacol peroxidase (GPX) was recorded in metal treated roots and leaves of B. monnieri. In contrast, a significant reduction in catalase activity in Cd treated B. monnieri was observed. An increase was also noted in the levels of cysteine and non-protein thiol contents of the roots of B. monnieri followed by a decline. However, in leaves, cysteine and non-protein thiol contents were found to be enhanced at all the Cd concentrations and exposure periods. A significant reduction in the level of ascorbic acid was observed in a concentration and duration dependent manner. The total chlorophyll and protein content of B. monnieri decreased with increase in Cd concentration at all the exposure periods. Results suggest that toxic concentrations of Cd caused oxidative damage as evidenced by increased lipid peroxidation and decreased chlorophyll and protein contents. However, B. monnieri is able to combat metal induced oxidative injury involving a mechanism of activation of various enzymatic and non-enzymatic antioxidants.