Cytochrome c Oxydase Subunit I Gene is Up-regulated by Cadmium in Freshwater and Marine Bivalves

Inhibition of the mitochondrial electron transfer chain and induction of reactive oxygen species (ROS) production are one of the roots of cadmium (Cd) toxicity. To appreciate the impact of Cd on mitochondria, we focused on the expression of CoxI gene which encodes the subunit I of the Cytochrome c oxidase (complex IV of the respiratory chain). CoxI gene expression was studied by real-time quantitative PCR in three species: two freshwater bivalves (Corbicula fluminea and Dreissena polymorpha) and one marine bivalve (diploid or triploid Crassostrea gigas). Bivalves were exposed for 10 or 14 days to 0.13 μM Cd²⁺ and 15.3 μM Zn²⁺ in controlled laboratory conditions. We demonstrate that in the three mollusk species CoxI gene was up-regulated by Cd. Zinc (Zn), which is known to have antioxidant properties, had no effect on CoxI gene expression. In the presence of Cd and Zn, CoxI gene inducibility was lower than after a single Cd exposure, in each species; result that could not be fully explained by a decreased Cd accumulation. CoxI gene induction by Cd was 4.8-fold higher in triploid oysters than in diploid ones, indicating a possible influence of triploidy on animal responses to Cd contamination.     

Metallothionein responses in the Asiatic clam (Corbicula fluminea) after exposure to trivalent arsenic

The main objective of this work was to evaluate arsenic effects on metallothionein (MT) induction by exposing a freshwater Asiatic clam (Corbicula fluminea) to different concentrations of this metalloid. The presence of MT-like proteins was detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis and compared with a standard rabbit MT. In addition, the polarographic response showed good correspondence between standard MT and MT-like curves from C. fluminea, allowing MT quantification. The results show that clams exposed to different concentrations of arsenic are able to induce significant levels of MTs. Although variability was found in MT induction, significant differences in MT levels were found after 28 days of exposure in all treatments in comparison with the controls, suggesting that exposure to arsenic induced MT-like proteins in C. fluminea.     

Changes in phytochelatins and their biosynthetic intermediates in red spruce (<Emphasis Type="Italic">Picea rubens </Emphasis>Sarg.) cell suspension cultures under cadmium and zinc stress

Cell suspension cultures of red spruce (Picea rubens Sarg.) were selected to study the effects of cadmium (Cd) and zinc (Zn) on phytochelatins (PCs) and related metabolites after 24 h exposure. The PC₂ and its precursor, γ-glutamylcysteine (γ-EC) increased two to fourfold with Cd concentrations ranging from 12.5 to 200 μM as compared to the control. However, Zn-treated cells showed a less than twofold increase in γ-EC and PC₂ levels as compared to the control even at the highest concentration of 800 μM. In addition, unidentified higher chain PCs were also found in both the Cd and Zn treated cells and they increased significantly with increasing concentrations of Cd and Zn. The cellular ratio of PC₂ : Cd or Zn content clearly indicated that Cd (with ratios ranging from 0.131 to 0.546) is a more effective inducer of PC₂ synthesis/accumulation than Zn (with ratios ranging from 0.032 to 0.102) in red spruce cells. A marginal decrease in glutathione (GSH) was observed in both Cd and Zn treated cells. However, the GSH precursor, cysteine, declined twofold with all Cd concentrations while the decrease with Zn was 1.5–2-fold only at the higher treatment concentrations of Zn as compared to control. In addition, changes in other free amino acids, polyamines, and inorganic ions were also studied. These results suggest that PCs and their biosynthetic intermediates play a significant role in red spruce cells protecting against Cd and Zn toxicity.     

Effect of zinc and cadmium on physiological and production characteristics in <Emphasis Type="Italic">Matricaria recutita</Emphasis>

Effects of zinc (12–180 μM) alone and in mixtures with 12 μM Cd on metal accumulation, dry masses of roots and shoots, root respiration rate, variable to maximum fluorescence ratio (F_V/F_M), and content of photosynthetic pigments were studied in hydroponically cultivated chamomile (Matricaria recutita) plants. The content of Zn in roots and shoots increased with the increasing external Zn concentration and its accumulation in the roots was higher than that in the shoots. While at lower Zn concentrations (12 and 60 μM) the presence of 12 μM Cd decreased Zn accumulation in the roots, treatment with 120 and 180 μM Zn together with 12 μM Cd caused enhancement of Zn content in the root. Presence of Zn (12–120 μM) decreased Cd accumulation in roots. On the other hand, Cd content in the shoots of plants treated with Zn + Cd exceeded that in the plants treated only with 12 μM Cd. Only higher Zn concentrations (120 and 180 μM) and Zn + Cd mixtures negatively influenced dry mass, chlorophyll (Chl) and carotenoid content, F_V/F_M and root respiration rate. Chl b was reduced to a higher extent than Chl a.     

Effects of Metal Combinations on the Production of Phytochelatins and Glutathione by the Marine Diatom Phaeodactylum tricornutum

Copper, Cd and Zn can be found at elevated concentrations in contaminated estuarine and coastal waters and have potential toxic effects on phytoplankton species. In this study, the effects of these metals on the intracellular production of the polypeptides phytochelatin and glutathione by the marine diatom Phaeodactylum tricornutum were examined in laboratory cultures. Single additions of Cu and Cd (0.4 μM Cu² and 0.45 μM Cd²⁺) to the culture medium induced the production of short-chained phytochelatins ((γ-Glu-Cys)_n-Gly where n = 2–5), whereas a single addition of Zn (2.2 μM Zn²⁺) did not stimulate phytochelatin production. Combination of Zn with Cu resulted in a similar phytochelatin production compared with a single Cu addition. The simultaneous exposure to Zn and Cd led to an antagonistic effect on phytochelatin production, which was probably caused by metal competition for cellular binding sites. Glutathione concentrations were affected only upon exposure to Cd (85% increase) or the combination of Cd with Zn (65% decrease), relative to the control experiment. Ratios of phytochelatins to glutathione indicated a pronounced metal stress in response to exposures to Cu or Cd combined with Zn. This study indicates that variabilities in phytochelatin and glutathione production in the field can be explained in part by metal competition for cellular binding sites.     

Urinary levels of metallothioneins and metals in subjects from a semiindustrialized area in Tarragona Province of Spain

The monitoring of heavy metals is important if adverse effects on health are to be avoided. In humans, metallothionein (MT) has been used as a biomonitor for the assessment of cadmium (Cd). In the present study, subjects drawn from the population of Tarragona Province (NE Spain) were investigated. Urinary MT, zinc (Zn), and copper (Cu) concentrations, corrected for creatine concentrations, were determined in 625 samples from healthy subjects aged between 10 and 65 yr. Mean values of MT and Cu in females were higher than those in males, with levels of 29.5 (23.8) vs. 22.7 (24.9) μg MT/creatinine (p<0.001) and 4.8 (6.1) vs 3.4 (4.9) μg Cu/g creatinine (p<0.001). No differences between males and females were observed with respect to urinary Zn: 78.0 (66.4) vs 73.0 (85.5) μg/g creatinine, respectively (p=0.332). Significantly higher MT, Zn, and Cu values were observed in the females aged 15–19 yr and, in the age group of 50–54 yr, only in the Zn and Cu values, when compared with those in males. Significant positive correlations of MT vs Zn and Cu as well as correlations of Zn vs Cu levels were observed in both genders. The present findings confirm the proposed role of MT as a biomonitor of mineral status.     

Organic Acids Accumulation and Antioxidant Enzyme Activities in Thlaspi caerulescens under Zn and Cd Stress

Growth, organic acid and phytochelatin accumulation, as well as the activity of several antioxidative enzymes, i.e. superoxide dismutase (SOD), ascorbate peroxidase (APX) guaiacol peroxidase (POX) and catalase (CAT) were investigated under Zn and Cd stress in hydroponically growing plants of Thlaspi caerulescens population from Plombières, Belgium. Tissue Zn and Cd concentration increased (the highest concentration of both was in roots) as the concentration of these metals increased in the nutrient solution. Increasing Zn concentration enhanced plant growth, while with Cd it declined compared to the control. Both metals stimulated malate accumulation in shoots, Zn also caused citrate to increase. Zn did not induce phytochelatin (PC) accumulation. In plants exposed to Cd, PC concentration increased with increasing Cd concentration, but decreased with time of exposure. Under Zn stress SOD activity increased, but APX activity was higher at 500 and 1000 μM Zn and CAT activity only at 500 μM Zn in comparison with the control. CAT activity decreased in Cd- and Zn-stressed plants. The results suggest that relative to other populations, a T. caerulescens population from Plombières, when grown in hydroponics, was characterized by low Zn and Cd uptake and their translocation to shoots and tolerance to both metals. The accumulation of malate and citrate, but not PC accumulation was responsible for Zn tolerance. Cd tolerance seems to be due to neither PC production nor accumulation of organic acids.     

Arabidopsis thaliana and Thlaspi caerulescens respond comparably to low zinc supply

The main objective of this research was to study the response of Arabidopsis thaliana L. and Thlaspi caerulescens J. & C. Presl to different Zn supplies. The A. thaliana plants were exposed to Zn-deficiency (0 and 0.05 μM Zn) and compared to the plants grown on media containing standard Zn (2 μM). T. caerulescens plants were also exposed to Zn-deficiency (0.05 μM Zn), but as this is a Zn hyperaccumulator species, also to high Zn (1,000 μM Zn). Plants were compared to plants grown on standard Zn media (100 μM Zn). Both A. thaliana and T. caerulescens were found to be heavily affected by Zn deficiency, showing similar retarded growth and reduced reproduction phenotypes, and even less reduction in biomass production in T. caerulescens than in A. thaliana. T. caerulescens plants were similarly affected when grown on high Zn concentrations, with comparable effects on reproductive tissues as seen on low Zn supply.     

Metallothionein as Potential Biomarker of Cadmium Exposure in Persian Sturgeon (<Emphasis Type="Italic">Acipenser persicus</Emphasis>)

Metallothionein (MT) concentration in gills, liver, and kidney tissues of Persian sturgeon (Acipenser persicus) were determined following exposure to sublethal levels of waterborne cadmium (Cd) (50, 400, and 1,000 μg l⁻¹) after 1, 2, 4, and 14 days. The increases of MT from background levels were 4.6-, 3-, and 2.8-fold for kidney, liver, and gills, respectively. The results showed that MT level change in the kidney is time and concentration dependent. Also, cortisol measurement revealed elevation at the day 1 of exposure and followed by MT increase in the liver. Cd concentrations in the cytosol of experimental tissues were measured, and the results indicated that Cd levels in the cytosol of liver, kidney, and gills increased 240.71-, 32.05-, and 40.16-fold, respectively, 14 days after exposure to 1,000 μg l⁻¹ Cd. The accumulation of Cd in cytosol of tissues is in the order of liver > gills > kidney. Pearson correlation coefficients showed that the MT content in kidney is correlated with Cd concentration, the value of which is more than in liver and gills. Thus, kidney can be considered as a tissue indicator in A. persicus for waterborne Cd contamination.     

Cadmium accumulation and interactions with zinc, copper, and manganese, analysed by ICP-MS in a long-term Caco-2 TC7 cell model

The influence of long-term exposure to cadmium (Cd) on essential minerals was investigated using a Caco-2 TC7 cells and a multi-analytical tool: microwave digestion and inductively coupled plasma mass spectrometry. Intracellular levels, effects on cadmium accumulation, distribution, and reference concentration ranges of the following elements were determined: Na, Mg, Ca, Cr, Fe, Mn, Co, Ni, Cu, Zn, Mo, and Cd. Results showed that Caco-2 TC7 cells incubated long-term with cadmium concentrations ranging from 0 to 10 μmol Cd/l for 5 weeks exhibited a significant increase in cadmium accumulation. Furthermore, this accumulation was more marked in cells exposed long-term to cadmium compared with controls, and that this exposure resulted in a significant accumulation of copper and zinc but not of the other elements measured. Interactions of Cd with three elements: zinc, copper, and manganese were particularly studied. Exposed to 30 μmol/l of the element, manganese showed the highest inhibition and copper the lowest on cadmium intracellular accumulation but Zn, Cu, and Mn behave differently in terms of their mutual competition with Cd. Indeed, increasing cadmium in the culture medium resulted in a gradual and significant increase in the accumulation of zinc. There was a significant decrease in manganese from 5 μmol Cd/l exposure, and no variation was observed with copper.     

Characterization of zinc transport by divalent metal transporters of the ZIP family from the model legume <Emphasis Type="Italic">Medicago truncatula</Emphasis>

To understand how plants from the Fabaceae family maintain zinc (Zn) homeostasis, we have characterized the kinetics of three Zn transporting proteins from the ZIP family of divalent metal transporters in the model legume Medicago truncatula. Of six ZIP’s studied, MtZIP1, MtZIP5 and MtZIP6 were the only members from this family determined to transport Zn and were further characterized. MtZIP1 has a low affinity for Zn with a K_m of 1 μM as compared to MtZIP5 and MtZIP6 that have a higher affinity for Zn with K_m of 0.4 μM and 0.3 μM, respectively. Zn transport by MtZIP1 was more sensitive to inhibition by copper (Cu) concentrations than MtZIP5 and MtZIP6, because 3 μM Cu inhibited Zn transport by 80% in MtZIP1 while 5 μM Cu was required to achieve the same inhibition of Zn transport in MtZIP5 and MtZIP6. Cadmium (Cd) had a greater effect on the ability of MtZIP1 to transport Zn than MtZIP5 and MtZIP6, because at a concentration of 3 μM Cd, the Zn transport by MtZIP1 was inhibited 55% and the transport of Zn by MtZIP5 and MtZIP6 was inhibited by 20–30%. However, only MtZIP6 transported Cd at higher rates than those observed in the control plasmid pFL61, demonstrating a low affinity for Cd based on a K_m of 57 μM. These results suggest that Medicago truncatula has both high and low affinity Zn transporters to maintain Zn homeostasis and that these transporters may function in different compartments within the plant.     

The Response of Metallothionein and Malondialdehyde after Exclusive and Combined Cd/Zn Exposure in the Crab Sinopotamon henanense

The purpose of this paper is to show the interactions of Cd and Zn in the freshwater crab Sinopotamon henanense through metallothionein (MT) and malondialdehyde (MDA) level measurements. Laboratory acclimated S.henanense were exposed to Cd (50 µg/L, 100 µg/L, 500 µg/L ), and Zn (100 µg/L, 1000 µg/L) alone and in combined treatments (100 µg/L Zn+50 µg/L Cd, 100 µg/L Zn+100 µg/L Cd, 100 µg/L Zn+500 µg/L Cd, 1000 µg/L Zn+50 µg/L Cd, 1000 µg/L Zn+100 µg/L Cd, 1000 µg/L Zn+500 µg/L Cd) for 7, 14, 21, 28, 35 days. The results demonstrated that the MDA contents increased with exposure time and dose and showed time- and dose-dependence in both gills and hepatopancreas of S.henanense after single Cd exposure, while the changes of MDA levels were not significant with single Zn exposure. The MDA levels decreased when the crabs were exposed to metal mixtures compared to Cd exposure alone, indicating that Zn mediated the cellular toxicity of Cd. MT contents increased after single Cd exposure and also showed a time- and dose-dependence, in a tissue-specific way. Zn showed a limited ability of MT induction both in gills and hepatopancreas of S.henanense. The MT contents represented not a simple addition of single metal exposures but were enhanced at a higher concentration of Zn combined with different Cd concentrations compared to single metal exposure. Whether MT can be used as a biomarker for complex field conditions need to be considered cautiously since different induction patterns of MT were found among single Zn, Cd and combined groups. It is suggested that several biomarkers together as a suite should be used in the monitoring of heavy metal pollution in the aquatic environment.     

Variation in response to heavy metals during vegetative growth in <Emphasis Type="Italic">Dorycnium pentaphyllum</Emphasis> Scop.

Seeds, young plants and adult plants of the perennial Mediterranean leguminous shrub Dorycnium pentaphyllum Scop. were exposed to Cd (1–100 μM) or Zn (10–10,000 μM) on nutrient solution. This species is resistant to Cd and Zn at different phenological stages. The lowest doses of Zn and Cd improved seed germination and young seedling growth, while only the highest doses of both heavy metals inhibited germination and decreased growth. High doses of Cd reduced seed imbibition and young seedling water content, while Zn did not. Osmotic adjustment was more efficient in Zn-treated young plants than in Cd-treated ones, while chlorophyll concentrations decreased in the former but not in the latter. Those differences were not observed anymore in adult plants. Exclusion processes were more efficient at the adult stage than at the young seedling stage and were more marked in response to Zn than to Cd. It is concluded that D. pentaphyllum could be used for phytostabilization of heavy metal-contaminated areas. The physiological strategies of tolerance, however, differ according to the age of the plants and the nature of the metal.     

Chl-<Emphasis Type="Italic">a</Emphasis> fluorescence parameters as biomarkers of metal toxicity in fluvial biofilms: an experimental study

A study was carried out to evaluate the sensitivity of different chlorophyll-a (chl-a) fluorescence parameters measured in freshwater biofilms as metal pollution biomarkers of short- and long-term metal exposures at environmentally realistic concentrations. A microcosm experiment was performed using indoor channels. Mature biofilms were exposed from hours to weeks to three different treatments: No-Metal, Zn (400 μg l⁻¹); and Zn plus Cd (400 μg l⁻¹ and 20 μg l⁻¹, respectively). Metal concentration was based on a real case study: the Riou-Mort River (France). Biofilms exposed to Zn bioaccumulated similar Zn contents per dry weight to those exposed to the mixture (Zn plus Cd) causing a similar inhibition of the effective quantum yield (\Upphi_\textM^￠)(\Upphi_{\text{M}}^{\prime}) during the first hours of exposure. A reduction of the algal biomass, a shift in the community composition (a high reduction of diatoms), a reduction of the maximal quantum yield (Φ_M) and a strong reduction of non-photochemical quenching (NPQ) were observed from day 14 until the end of the experiment (35 days). The results indicate that the effects of the metal mixture present in the Riou-Mort on biofilms could be attributed to Zn toxicity. The use of a set of chl-a fluorescence measurements, including photochemical and NPQ parameters, are recommended as a reliable biomarker tool box to evaluate both short- and long-term effects of metals on biofilms containing oxygenic photoautotrophs, suggesting its use in field applications.     

Effects of temperature and temperature acclimation on survival of zebra mussels (Dreissena polymorpha) and Asian clams (Corbicula fluminea) under extreme hypoxia

Following acclimation to 5°, 15° or 25°C for 14days, samples of 30 Dreissena polymorpha (zebra mussels) andCorbicula fluminea (Asian clams) were held in either aerated(control) or extremely hypoxic N₂ gassed water (PO2 < 3%of full air saturation). Mortality was negligible in all aeratedcontrols. Mean hypoxia tolerance in D. polymorpha ranged from3–4 days at 25°C to 38–42 days at 5°C. Hypoxiatolerance time of zebra mussels increased significantly withdeclining test temperature (P < 0.001) and increasing acclimationtemperature (P < 0.001). Larger zebra mussels were more tolerantthan smaller individuals. Asian clams were 2–7 times moretolerant of hypoxia than zebra mussels, surviving a mean of11.8 and 35.1 days at 25°C and 15°C, respectively, andwithout mortality for 84 days at 5°C, and were not influencedby temperature acclimation. At 25°C, larger specimens ofAsian clams were less tolerant of hypoxia than smaller individuals.Both species are amongst the least hypoxia tolerant freshwaterbivalve molluscs, reflecting their prevalence in well-oxygenatedshallow water habitats. Prolonged exposure to extreme hypoxiamay provide an efficacious control strategy, particularly forD. polymorpha (Received 12 January 1998; accepted 30 September 1998)     

Metallothionein induction in freshly isolated rat hepatocytes

The control of metallothionein (MT) synthesis was investigated in freshly prepared rat hepatocytes in experiments of short-term duration. Viability and metabolic function were maintained in incubations of 6-h duration. MT synthesis was measurable in hepatocytes from fed rats at Zn concentrations down to 1 μM. Zn and dexamethasone induced concentration-dependent increases in the synthesis of MT with maximal increases above the 5-h control of 3.2- and 2.5-fold, respectively. Zn induction of MT was first measurable at 2 h and was inhibited by actinomycin C. Although initial (0 h) MT concentrations in hepatocytes from fasted rats were double those from fed rats, after 6-h incubation in the presence of 50 μM Zn, the fasted rat hepatocytes showed only half the MT concentrations of the fed rat hepatocytes. Glucagon and interleukin-6 (IL-6) were less effective inducers and increased MT synthesis by 28 and 17%, respectively. IL-6 (100 U/mL) was found to have an additive effect on MT synthesis above that of Zn alone (1–50 μM) or Zn plus dexamethasone (1 μM). A supernatant from LPS-stimulated macrophages increased MT synthesis by 40%. The basal MT synthesis was not increased by either tumor necrosis factor-α (TNF-α) or interleukin-1 (IL-1). All incubations were carried out in the presence of RPMI 1640 medium with Hepes (20 mM), bicarbonate (24 mM), and fatty acid-free albumin (FAFA; 0.5% w/v). MT synthesis was also seen using Krebs bicarbonate buffer with glucose (10 mM), Hepes (20 mM), and FAFA (0.5% w/v), and although the level of MT synthesis was less than in RPMI, the increases in concentrations of MT at 5 h were 225, 139, 36 and 20% for Zn, dexamethasone, glucagon, and control, respectively. It is concluded that MT synthesis occurs in freshly prepared hepatocytes and that these cells are responsive to some of the established inducers of MT. This system enables the study of MT synthesis in individual rats in various metabolic and pathological states.     

Sensitivity to Cd or Zn of host and symbiont of ectomycorrhizal Pinus sylvestris L. (Scots pine) seedlings

Pinus sylvestris seedlings infected with either the ectomycorrhizal (ECM) fungus Paxillus involutus or Suillus variegatus were exposed to a range of Cd or Zn concentrations. This was done to investigate the relationship between the sensitivity of ECM fungi and their host plants over a wide range of concentrations. P. involutus ameliorated the toxicity of Cd and Zn to P. sylvestris with respect to root length, despite significant inhibition of ECM infection levels by Cd (Cd EC₅₀ [effective concentration which inhibits ECM infection by 50%] values were: P. involutus 3.7 μg g^-1 Cd; S. variegatus 2.3 μg g^-1 Cd). ECM infection by P. involutus also decreased Cd and Zn transport to the plant shoots at potentially toxic concentrations and also influenced the proportion of Zn transported to the roots and shoots, with a higher proportion retained in the roots of the seedlings. ECM infection did increase host biomass production, but this was not affected by the presence of Cd or Zn. Root and shoot biomass production by P. sylvestris, in both the presence and absence of ECM fungi, was unaffected by Cd and Zn at all concentrations tested. This revised version was published online in August 2006 with corrections to the Cover Date.     

Oxidative Stress Induced by Cadmium in the C6 Cell Line: Role of Copper and Zinc

In this report, we have investigated the role of copper (Cu) and zinc (Zn) in oxidative stress induced by cadmium (Cd) in C6 cells. Cells were exposed to 20 μM Cd, 500 μM Cu, and 450 μM Zn for 24 h. Then, toxic effects, cellular metals levels, oxidative stress parameters, cell death, as well as DNA damage were evaluated. Cd induced an increase in cellular Cd, Cu, and Zn levels. This results not only in the inhibition of GSH-Px, GRase, CAT, and SOD activities but also in ROS overproduction, oxidative damage, and apoptotic cell death not related to Cu and Zn mechanisms. The thiol groups and GSH levels decreased, whereas the lipid peroxidation and DNA damage increased. The toxicity of Zn results from the imbalance between the inhibition of antioxidant activities and the induction of MT synthesis. The increase in Cu and Zn levels could be explained by the disruption of specific transporter activities, Cd interference with signaling pathways, and metal displacement. Our results suggest that the alteration of Cu and Zn homeostasis is involved in the oxidative stress induced by Cd.     

Physiological behavior of Scenedesmus sp. during exposure to elevated levels of Cu and Zn and after withdrawal of metal stress

Summary. A 48 h exposure of Scenedesmus sp. to sublethal concentrations of Cu (2.5 and 10 μM) and Zn (5 and 25 μM) caused a concentration-dependent inhibition of growth, photosynthesis, respiration, NO₃ ⁻ uptake, and nitrate reductase (EC 1.6.6.1) activity, and a reduction in protein, carbohydrate, and photosynthetic-pigment levels with a concomitant increase in intracellular levels of the test metals. After exposure, algal cells were transferred to the basal medium without the excess level of test metals, to study the recovery of various processes. The growth of the test algae had not recovered up to 12 h after transfer to the basal medium, but some physiological parameters such as photosynthesis and respiration recovered within 6 h. The quicker recovery of photosynthesis and respiration might be used as acclimatory responses as they prepare a background for the recovery of other parameters, including growth, of the test alga by generating energy, forming photosynthate, and establishing the usual catabolism to attain normal conditions. Most of the processes recovered completely or almost completely after being stressed with 2.5 μM Cu or 5 μM Zn. However, the maintenance of a relatively high level of Cu and Zn in the cells previously exposed to 10 μM Cu and 25 μM Zn slowed down the recovery of different processes, which did not fully recover even at the end of the experiment after 96 h. The present study demonstrates that a chain of metabolic events, beginning with respiration and photosynthesis and continuing with assimilation and uptake of nutrients and subsequent restoration of other metabolic processes, is involved in the recovery of the algae from Cu and Zn stress. Each studied parameter seems to play an important role in balancing the cellular homeostasis during recovery from metal stress. Correspondence and reprints: Department of Bioscience and Biotechnology, Banasthali Vidyapith, Banasthali 304 022, Rajasthan, India.     

Cadmium effects on growth and antioxidant enzymes activities in Miscanthus sinensis

Plants of Miscanthus sinensis (cv. Giganteus) were grown in hydroponics for three months in nutrient solution with 0, 2.2, 4.4 and 6.6 μM CdNO₃. Growth parameters, catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX) and superoxide dismutase (SOD) activities were analysed in leaves and roots collected after 1-and 3-month exposure. Dry biomass of all miscanthus organs was affected by Cd concentration both after 1-and 3-month exposure. No visible symptoms of Cd toxicity were observed in shoots and rhizomes of plants grown in presence of Cd. In contrast, roots became shorter and thicker and the whole root system more dense and compact already after one month of treatment with 6.6 μM Cd. The lower Cd concentration increased the enzymes activities after 3 months in leaves and only after 1-month in roots, while a decrease in activity was observed at higher Cd concentrations.