Effect of chromium species on phytochemical and physiological parameters in Datura innoxia

In soil, chromium can be found in two main valence states: hexavalent Cr(VI) and trivalent Cr(III). In this study, we investigated the impact of Cr on photosynthetic gas exchange, photosystem II (PSII) activity, Cr translocation and accumulation, proline content and alkaloids production, i.e. scopolamine and hyoscyamine, in Datura innoxia. Cr uptake was influenced by its oxidation state and its concentration in growth medium. The plant roots were determined as being the main organ of Cr accumulation. Cr(VI) was more toxic than Cr(III) as indicated by reduction in plant biomass and net photosynthesis. The stomatal conductance showed a similar trend to that of photosynthetic capacity. Cr(III) and Cr(VI) had a different impact on substomatal CO(2) concentration then Cr toxicity was related to its oxidation states. In plants stressed with a Cr(VI) excess, a down regulation of PSII activity was observed with an impairment of photochemical activity. Indeed, the maximum quantum yield of PSII (F(v)/F(m)), the quantum yield of PSII (PhiPSII) and the efficiency of excitation capture by open centers (F'(v)/F'(m)) decreased. Cr(III) had little effects on PSII primary photochemistry, whatever its form induces an increase of scopolamine content without changes in hyoscyamine content in leaves of D. innoxia. These results provide that chromium contamination can change the secondary metabolites composition of leaves, thereby, impacting the quality, safety and efficacy of natural plant products synthesized by D. innoxia plants.     

Chromium accumulation by the hyperaccumulator plant Leersia hexandra Swartz

Leersia hexandra Swartz (Gramineae), which occurs in Southern China, has been found to be a new chromium hyperaccumulator by means of field survey and pot-culture experiment. The field survey showed that this species had an extraordinary accumulation capacity for chromium. The maximum Cr concentration in the dry leaf matter was 2978 mg kg(-1) on the side of a pond near an electroplating factory. The average concentration of chromium in the leaves was 18.86 times as that in the pond sediment, and 297.41 times as that in the pond water. Under conditions of the nutrient solution culture, it was found that L. hexandra had a high tolerance and accumulation capacity to Cr(III) and Cr(VI). Under 60 mg l(-1) Cr(III) and 10 mg l(-1) Cr(VI) treatment, there was no significant decrease of biomass in the leaves of L. hexandra (p>0.05). The highest bioaccumulation coefficients of the leaves for Cr(III) and Cr(VI) were 486.8 and 72.1, respectively. However, L. hexandra had a higher accumulation capacity for Cr(III) than for Cr(VI). At the Cr(III) concentration of 10 mg l(-1) in the culture solution, the concentration of chromium in leaves was 4868 mg kg(-1), while at the same Cr(VI) concentration, the concentration of chromium in leaves was only 597 mg kg(-1). These results confirmed that L. hexandra is a chromium hyperaccumulator which grows rapidly with a great tolerance to Cr and broad ecological amplitude. This species could provide a new plant resource that explores the mechanism of Cr hyperaccumulation, and has potential for usage in the phytoremediation of Cr-contaminated soil and water.     

Phytotoxicity of cobalt, chromium and copper in cauliflower

Cauliflower (Brassica oleracea L. var. Botrytis cv. Maghi) was grown in refined sand with complete nutrition (control) and at 0.5 mM each of Co, Cr and Cu. In cauliflower, compared to that of excess Cu or Cr, the visible effects of excess Co appeared first and were most pronounced. Excess of each heavy metal restricted the biomass of cauliflower, concentrations of Fe, chlorophylls a and b, protein and activity of catalase in leaves in the order Co>Cu>Cr. The translocation of Cr from roots to tops was minimum and that of Co was maximum when cauliflower was individually supplied with excess Co, Cu or Cr. In cauliflower each heavy metal inhibited the concentration of most of the macro- and micronutrients. The translocation of P, S, Mn, Zn and Cu from roots to tops of cauliflower were affected most significantly by Co and least by Cr. In contrast to excess Cu or Cr, Co significantly decreased the water potential and transpiration rates and increased diffusive resistance and relative water content in leaves of cauliflower.     

Effect of lead on growth and nitrate assimilation of Vigna radiata (L.) Wilczek seedlings in a salt affected environment

The inhibition of seedling growth and nitrate reductase activity in 5 d old Vigna radiata (L.) Wilczek cv. Pusa Baisakhi in the presence of 1.0 mM lead acetate increased drastically, if NaCl (6 and 12 EC) was also present in the nutrient media along with the metal salt. Correspondingly higher endogenous Na+ levels were accumulated in the roots and leaves of seedlings in presence of the two stresses. On the other hand, the levels of endogenous lead get reduced in presence of NaCl in both the roots and leaves. Roots accumulated more Pb2+ and Na+ than the leaves. The two stresses affect more drastically in the additive or even synergistic manner during the early growth phase of the seedlings.     

Uptake studies of environmentally hazardous (51)Cr in Mung beans

Attempt has been made to study the accumulation behaviour of a common plant, Mung bean (Vigna radiata) towards Cr(III) and Cr(VI) to have an insight on the migration and bio-magnification of Cr. For this purpose healthy germinated Mung bean seeds were sown in the sand in the presence of Hoagland's nutrient solution containing measured amount of K(2)(51)Cr(2)O(7) and (51)Cr(NO(3))(3).9H(2)O. Growth rate was also studied in the presence and absence of phosphate salts in the medium. It has been found that the transfer of chromium from soil to plant is significantly low (maximum 5% for both Cr(III) and Cr(VI)). Maximum accumulation of Cr occurs in the root with respect to the total chromium accumulation by the plant. Other parts of the Mung bean plant, e.g. cotyledons, shoot and leaves, show negligible accumulation. Therefore, the chance of direct intake of Cr through food as well as through the grazing animals to human body is less.     

Changes of organic acid exudation and rhizosphere pH in rice plants under chromium stress.

The effect of chromium (Cr) stress on the changes of rhizosphere pH, organic acid exudation, and Cr accumulation in plants was studied using two rice genotypes differing in grain Cr accumulation. The results showed that rhizosphere pH increased with increasing level of Cr in the culture solution and with an extended time of Cr exposure. Among the six organic acids examined in this experiment, oxalic and malic acid contents were relatively higher, and had a significant positive correlation with the rhizosphere pH, indicating that they play an important role in changing rhizosphere pH. The Cr content in roots was significantly higher than that in stems and leaves. Cr accumulation in plants was significantly and positively correlated with rhizosphere pH, and the exudation of oxalic, malic and citric acids, suggesting that an increase in rhizosphere pH, and exudation of oxalic, malic and citric acid enhances Cr accumulation in rice plants.     

Are either sod and catalase or the polyamines involved in the paraquat resistance of Conyza canadensis ?

Abstract

Paraquat/atrazine coresistant (PqAR) and paraquat resistant (PqR) horseweed (Conyza canadensis /L./ Cronq.) plants showed ‐ in the first hour after 0.5 mM paraquat spraying ‐ a decreased catalase activity followed by a slight increase. However, the enzyme activity remained always below the initial value. Sensitive plants showed a significant increase of catalase activity in the first 4 hour after spraying. The transient character of paraquat inhibition, the recovery of photosyn‐thetic activity of the PqAR Conyza plants (characterized by variable fluorescence) after spraying remained unaffected by the Superoxide dismutase (SOD) inhibitor, diethyldithiocarbamate. This indicates that SOD is not involved in the resistance mechanism. Untreated resistant biotypes showed about 2.5 times higher total polyamine and putrescine level than the sensitive one. 100 μM of exogenously added putrescine was observed as having a protecting effect against paraquat in floated leaves of the sensitive biotype only. The resistant leaves were unaffected probably on account of their higher endogenous polyamine level. It is concluded that polyamines may play a role in the paraquat resistance of Conyza canadensis.     

Arsenic accumulation and translocation in the submerged macrophyte Hydrilla verticillata (L.f.) Royle

Worldwide contamination of arsenic in aquatic systems requires the development of a cost-effective, in situ phytoremediation technology. Hydrilla verticillata (L.f.) Royle, a submerged macrophyte widely distributed throughout the world, has the potential to effectively remove heavy metals from water. In order to understand the potential of H. verticillata for As phytofiltration and its impacts on As cycling in the water system, we investigated As accumulation, speciation and translocation in H. verticillata plants. Plant shoots showed a significant accumulation of As, with a maximum of >700 μg g⁻¹ dry weight (DW) after exposure to 20 μM arsenate [As(V)] or arsenite [As(III)] for 4 d, with no significant differences between the As(V) and As(III) treatments (P > 0.05). In addition, results of an in planta transport experiment showed that, after exposure of root and shoot to 2 μM As(V) and As(III) for 4 d, the bioconcentration factor (BCF) in roots for As(V) was almost twofold than that of As(III). Higher As BCFs in roots compared to shoots was also observed. Arsenic accumulated primarily in the cell walls of root cells (>73% of the total As in roots) and in the soluble parts of leaves (>60% of the total As in leaves). Regardless of the form of As supplied [As(III) or As(V)], As(V) was the dominant form in roots and As(III) was the dominant form in leaves. Further, basipetal translocation of As in this plant (?17%) was markedly higher than acropetal translocation (?3%). Because of accumulation of As in the shoot and immobilization of As below ground in roots, H. verticillata is a potential As phytofiltrator for bioremediation.     

Interaction of bioaccumulation of heavy metal chromium with water relation, mineral nutrition and photosynthesis in developed leaves of Lolium perenne L

Contamination by chromium (Cr) is widespread in agricultural soils and industrial sites. This heavy metal represents a risk to human health. In order to gain fundamental insights into the nature of the adaptation to Cr excess, the characterisation of physiological indices, including responses of photosynthetic gas exchange and chlorophyll a fluorescence along with changes in mineral nutrient contents and water status were studied in ray grass (Lolium perenne L.). Increased concentrations of Cr(VI) (0-500 microM Cr) in the Co?c and Lessaint nutrient solution were applied. The growth of Lolium perenne is decreased by chromium and the leaves have lost their pigments. Chromium accumulation was greater in roots than in leaves and reached 2450 and 210 microg g(-1) DW, respectively with 500 microM Cr(VI) in nutrient medium. The physiological parameters were severely reduced by this heavy metal. Cr induced toxicity arising from 100 microM Cr(VI) and resulted in a modification of mineral content in roots and leaves, especially for Ca, Mg and Fe. The chromium stress decreased CO2 assimilation rates mainly due to stomatal closure, which reduced water loss by transpiration without decreasing the cellular available CO2. The fluorescence parameters associated with photosystem II (PSII) activity and the photochemical activity are modified by chromium. Non-radiative energy dissipation mechanisms were triggered during stress since non-photochemical quenching was increased and efficiency of excitation capture by open centers was reduced.     

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.     

Phytotoxic lesions of chromium in maize

Chromium (Cr) is fairly abundant in the earth's crust and ranks fourth among the 29 elements of biological importance. Besides natural sources, Cr enters biotic components of the ecosystem in various ways. Of other major industrial sources, tanning and chrome-plating industries are prominent sources. Cr(VI) form of chromium is highly reactive and influences both plants and animals. Due to Mn present in soil, Cr(III) is oxidized to Cr(VI) which remains in soil for a long time and can affect plant growth and development. Since maize is an important food and fodder plant for human beings and cattle, a study was conducted to investigate the effects of Cr on some metabolic activities of maize (Zea mays L. cv. Ganga 5). Chromium caused visible lesions of interveinal chlorosis. Young leaves showed vein clearing. Also, a papery appearance was observed in leaves. Margins of leaves were curled and the leaves appeared pale at greater Cr exposure. Concentrations of both chlorophyll a and b were reduced by exposure to Cr, the activities of ribonuclease and phenyl phosphatase were greater while the activity of iron-porphyrin enzyme catalase was less and the activity of amylase was also much less in plants exposed to Cr. Chromium also caused retardation of soluble protein. Accumulation of Cr in roots was much at all the levels of chromium supply. Exposure to Cr resulted in reduction in grain production and quality.     

Effects of elevated CO2 concentration on the polyamine levels of field-grown soybean at three O3 regimes

Effects of increased ozone (O3) and carbon dioxide (CO2) on polyamine levels were determined in soybean (Glycine max L. Merr. cv. Clark) grown in open-top field chambers. The chamber treatments consisted of three O3 regimes equal to charcoal filtered (CF), non-filtered (NF), and non-filtered plus 40 nl litre(-1) O3 and CO2 treatments equal to 350, 400 and 500 microl litre(-1) for a total of nine treatments. Leaf samples were taken at three different times during the growing season. Examination of growth and physiological characteristics, such as photosynthesis, stomatal resistance, and shoot weight, revealed that increasing CO2 ameliorated the deleterious effects of increased O3. Results from the initial harvest, at the pre-flowering growth stage (23 days of treatment), showed that increasing O3 at ambient CO2 caused increases in putrescine (Put) and spermidine (Spd) of up to six-fold. These effects were lessened with increased CO2. Elevated CO2 increased polyamines in plants treated with CF air, but had no effect in the presence of ambient or enhanced O3 levels. Leaves harvested during peak flowering (37 days of treatment) showed O3-induced increases in Put and Spd at ambient CO2 concentrations. However, increased CO2 levels inhibited this response by blocking the O3-induced polyamine increase. Leaves harvested during the pod fill stage (57 days of treatment) showed no significant O3 or CO2 effects on polyamine levels. Our results demonstrate that current ambient O3 levels induce the accumulation of Put and Spd early in the growing season and that further increases in O3 could result in even greater polyamine increases. These results are consistent with a possible antiozonant function for polyamines. The ability of increased CO2 to protect soybeans from O3 damage, however, does not appear to involve polyamine accumulation.     

Accumulation of chromium (VI) from aqueous solutions using water lilies (Nymphaea spontanea)

This study describes an investigation using tropical water lilies (Nymphaea spontanea) to remove hexavalent chromium from aqueous solutions and electroplating waste. The results show that water lilies are capable of accumulating substantial amount of Cr(VI), up to 2.119 mg g(-1) from a 10 mg l(-1) solution. The roots of the plant accumulated the highest amount of Cr(VI) followed by leaves and petioles, indicating that roots play an important role in the bioremediation process. The maturity of the plant exerts a great effect on the removal and accumulation of Cr(VI). Plants of 9 weeks old accumulated the most Cr(VI) followed by those of 6 and 3 weeks old. The results also show that removal of Cr(VI) by water lilies is more efficient when the metal is present singly than in the presence of Cu(II) or in waste solution. This may be largely associated with more pronounced phytotoxicity effect on the biochemical changes in the plants and saturation of binding sites. Significant toxicity effect on the plant was evident as shown in the reduction of chlorophyll, protein and sugar contents in plants exposed to Cr(VI) in this investigation.     

Mercury in salt marshes ecosystems: Halimione portulacoides as biomonitor

Mercury concentrations were quantified in Halimione portulacoides (roots, stems and leaves) as well as in sediments from eight Portuguese estuarine systems, covering seventeen salt marshes with distinct degrees of mercury contamination. The concentration of mercury in the sediments ranged from 0.03 to 17.0 microg g(-1). The results show that the accumulation of mercury differed according to the organ of the plant examined and the concentration of mercury in the sediments. Higher mercury concentrations were found in the roots (up to 12.9 microg g(-1)) followed by the leaves (up to 0.12 microg g(-1)), while the stems had the lowest concentrations (up to 0.056 microg g(-1)). A linear model explained the relation between the concentrations of mercury in the different plant organs: roots and stems (R(adj)(2)=0.75), stems and leaves (R(adj)(2)=0.85) and roots and leaves (R(adj)(2)=0.78). However, the results show that the variation of mercury concentration in the roots versus mercury concentration in the sediments was best fitted by a sigmoidal model (R(adj)(2)=0.89). Mercury accumulation in the roots can be described in three steps: at a low range of mercury concentrations in the sediments (from 0.03 up to 2 microg g(-1)), the accumulation of mercury in roots is also low reaching a maximum concentration of 1.3 microg g(-1); the highest rates of mercury accumulation in the roots occur in a second step, until the concentrations of mercury in the sediments reach approximately 4.5 microg g(-1); after reaching this maximum value, the rate of mercury accumulation in the roots slows down leading to a plateau in the concentration of mercury in the roots of about 9.4 microg g(-1), which corresponds to a mercury concentration in the sediments of about 11 microg g(-1). A linear model explained also the accumulation of mercury in leaves versus the mercury concentration in the sediments (R(adj)(2)=0.88). Differences in responses of roots and leaves are explained by the dynamics of the plant organs: old roots are mineralised in situ close to new roots, while leaves are renewed. Previous studies have already shown that H. portulacoides is a bioindicator for mercury and the results from this work sustain that H. portulacoides may also be used as a biomonitor for mercury contamination in salt marshes. Nevertheless, caution should be taken in the application of the models, concerning the life cycle of the species and the spatial variability of the systems.     

Response of antioxidants in sunflower (Helianthus annuus L.) grown on different amendments of tannery sludge: its metal accumulation potential

The interaction of metals present in tannery waste and their tolerance in the plants of sunflower (Helianthus annuus L.) was studied in the present paper under field conditions. Effects of 100% tannery sludge and various amendments of tannery sludge (10%, 25%, 35%, 50%, 75%) along with one set of control were studied on the physiological and biochemical parameters of the plant along with their metal accumulation potential after 30, 60 and 90 d after sowing. The plants of H. annuus were found effective in the accumulation of metals (Cr, Fe, Zn and Mn) in roots, shoots and leaves, however, the level of toxic metal, Cr was found below detection limit in the seeds of the plant. The oil was extracted from the seeds of the plant and the level of oil content was increased up to 35% tannery sludge as compared to control followed by decrease at higher tannery sludge ratio. An increase in the chlorophyll, protein, cysteine, non-protein thiol and sugar contents was observed at the lower amendment of tannery sludge at initial exposure periods followed by decrease than their respective controls. Malondialdehyde content in the roots and leaves was increased beyond 50% sludge amendments at all the exposure periods as compared to control. However, proline and ascorbic acid contents of the roots and leaves of the plant increased at all the exposure periods and sludge amendments, compared to their respective controls.     

Species-dependent chromium accumulation, lipid peroxidation, and glutathione levels in germinating kiwifruit pollen under Cr(III) and Cr(VI) stress

The accumulation of chromium by germinating kiwifruit pollen appears to be significantly affected by Cr species, Cr concentration and calcium availability. Cr(III) accumulation always occurred in a linear manner while Cr(VI) uptake followed a logarithmic model. In the absence of exogenous calcium, Cr(III) accumulation was much higher than that of Cr(VI). It was observed that, as the Cr(III) concentration increased, there was a significant decrease in the endogenous calcium content of pollen, ultimately leading to complete calcium depletion after 90 min of incubation at 150 microM Cr(III). This loss of calcium could be responsible for the strong inhibition of tube emergence and growth following exposure of pollen to Cr(III). Indeed, when exogenous calcium was added to the kiwifruit pollen culture medium, significant growth recovery and reduced Cr(III) uptake occurred; the opposite was true in Cr(VI)-treatments. A significant rise in lipid peroxide production occurs in the presence of both Cr species; the effect was more pronounced following Cr(VI) exposure. Finally, glutathione pool dynamics appears to be differentially affected by chromium species and concentrations. In conclusion, results of the present study have provided important information regarding the different activity profiles of Cr(III) and Cr(VI) in relation to kiwifruit pollen performance, and have also demonstrated differences in some biochemical responses of pollen to metal stress.     

Clonal differences in survival capacity, copper and zinc accumulation, and correlation with leaf polyamine levels in poplar: A large-scale field trial on heavily polluted soil

Three ex situ collections of poplar clones from natural populations of Populus alba and P. nigra growing in northern Italy were assessed for their genetic dissimilarity (GD) by means of amplified fragment length polymorphism (AFLP). The high GD evidenced within populations was exploited for screening 168 clones in a field trial on heavy metal-polluted soil. After one growth season, clonal differences in plant survival and growth were observed. On the basis of performance, six clones were singled out, and used to evaluate copper and zinc accumulation in different organs. Clonal differences in metal concentrations were most evident for leaves and stems; one clone of P. alba (AL35) had a distinctly higher concentration of both metals in the roots. Leaf polyamine (putrescine, spermidine, spermine) profiles correlated with tissue metal concentrations, depending on the clone, plant organ and metal. In particular, the high metal-accumulating clone AL35 exhibited a dramatically higher concentration of free and conjugated putrescine. Overall, the results indicate that, given the high GD of Populus even within populations, it is possible to identify genotypes best suited for soil clean-up, and useful also for investigating physiological markers associated with high metal accumulation/tolerance     

Assessment of macro and microelement accumulation capability of two aquatic plants

The concentrations of four macroelements (C, N, P, S) and eight trace metals (Cd, Cr, Cu, Fe, Mn, Ni, Pb, Zn) were measured in the leaves and roots of the emergent plant, Phragmites communis Trin., and in the shoots and roots of the submersed Najas marina L., taken from Lake Averno (Naples, Italy). Phragmites communis leaves showed higher concentrations of carbon, nitrogen and phosphorus than roots, while the roots exhibited significantly higher concentrations of sulphur and trace metals. Najas marina roots also showed higher concentrations of sulphur and trace metals than shoots, but these differences were less marked than in Phragmites communis except for sulphur. Sulphur was the only macronutrient to show the highest concentrations in the roots. Phragmites communis roots had higher values of Cr, Cu, Fe, Mn and Ni than Najas marina roots. By contrast, Cd, Cr, Fe, Ni, Pb and Zn concentrations were higher in Najas marina shoots than in Phragmites communis leaves. Phragmites communis, available through the year, showing high capability to accumulate trace metals in the roots, appears a good monitor of lake contamination, better than Najas marina.     

Excess lead alters growth, metabolism and translocation of certain nutrients in radish

To elucidate the deleterious effects of excess lead on radish (Raphanus sativus) cv. Jaunpuri plants were grown in refined sand in complete nutrient solution for 30 days. On the 31st day lead nitrate was superimposed at 0.1 and 0.5mM to radish for 65 days. A set of plants in complete nutrient solution was maintained as control for the same period without lead. Excess Pb at 0.5mM showed growth depression with interveinal chlorosis on young leaves at apex. Excess Pb reduced the fresh and dry weight pronouncedly at d 65. Lead accumulation reduced the concentration of chlorophyll, iron, sulphur (in tops), Hill reaction activity and catalase activity whereas increased the concentration of phosphorus, sulphur (in roots) and activity of peroxidase, acid phosphatase and ribonuclease in leaves of radish.     

Impact of nano-CuO stress on rice (Oryza sativa L.) seedlings

Indiscriminate release of metal oxide nanoparticles (NPs) into the environment due to anthropogenic activities has become a serious threat to the ecological system including plants. The present study assesses the toxicity of nano-CuO on rice (Oryza sativa cv. Swarna) seedlings. Three different levels of stress (0.5 mM, 1.0 mM and 1.5 mM suspensions of copper II oxide, <50 nm particle size) were imposed and seedling growth performance was studied along control at 7 and 14 d of experiment. Modulation of ascorbate–glutathione cycle, membrane damage, in vivo ROS detection, foliar H₂O₂ and proline accumulation under nano-CuO stress were investigated in detail to get an overview of nano-stress response of rice. Seed germination percentage was significantly reduced under stress. Higher uptake of Evans blue by nano-CuO stressed roots over control indicates loss of root cells viability. Presence of dark blue and deep brown spots on leaves evident after histochemical staining with NBT and DAB respectively indicate severe oxidative burst under nano-copper stress. APX activity was found to be significantly increased in 1.0 and 1.5 mM CuO treatments. Nevertheless, elevated APX activity might be insufficient to scavenge all H₂O₂ produced in excess under nano-CuO stress. That may be the reason why stressed leaves accumulated significantly higher H₂O₂ instead of having enhanced APX activity. In addition, increased GR activity coupled with isolated increase in GSH/GSSG ratio does not seem to prevent cells from oxidative damages, as evident from higher MDA level in leaves of nano-CuO stressed seedlings over control. Enhanced proline accumulation also does not give much protection against nano-CuO stress. Decline in carotenoids level might be another determining factor of meager performance of rice seedlings in combating nano-CuO stress induced oxidative damages.