Physiological and proteomic alterations in rice (Oryza sativa L.) seedlings under hexavalent chromium stress

Main Conclusion

Rice plants employ two strategies to cope with Cr toxicity: immobilizing Cr ions into cell walls to reduce its translocation and activating antioxidant defense to mitigate Cr-induced oxidative stress. The investigation aimed at understanding the physiological and proteomic responses of rice seedlings to hexavalent chromium (Cr⁶⁺) stress was conducted using two rice genotypes, which differ in Cr tolerance and accumulation. Cr toxicity (200 µM) heavily increased the accumulation of H₂O₂ and \({\text{O}}_{2}^{{ \cdot-}}\) , enhanced lipid peroxidation, decreased cell viability and consequently inhibited rice plant growth. Proteomic analyses suggest that the response of rice proteome to Cr stress is genotype- and Cr dosage-dependent and tissue specific. Sixty-four proteins, which show more than fourfold difference under either two Cr levels, have been successfully identified. They are involved in a range of cellular processes, including cell wall synthesis, energy production, primary metabolism, electron transport and detoxification. Two proteins related to cell wall structure, NAD-dependent epimerase/dehydratase and reversibly glycosylated polypeptide were greatly up-regulated by Cr stress. Their enhancements coupled with callose accumulation by Cr suggest that cell wall is an important barrier for rice plants to resist Cr stress. Some enzymes involved in antioxidant defense, such as ferredoxin-NADP reductase, NADP-isocitrate dehydrogenase, glyoxalase I (Gly I) and glutamine synthetase 1 (GS1) have also been identified in response to Cr stress. However, they were only detected in Cr-tolerant genotype, indicating the genotypic difference in the capacity of activating the defense system to fight against Cr-induced oxidative stress. Overall, two strategies in coping with Cr stress in rice plants can be hypothesized: (i) immobilizing Cr ions into cell walls to reduce its translocation and (ii) activating antioxidant defense to mitigate Cr-induced oxidative stress.     

Comparative study of alleviating effects of GSH, Se and Zn under combined contamination of cadmium and chromium in rice (Oryza sativa)

A hydroponic experiment was conducted to study the ameliorative effects of separate or combined application of exogenous glutathione (GSH), selenium (Se) and zinc (Zn) upon 20 μM cadmium (Cd) plus 20 μM chromium (Cr) heavy metal stress (HM) in rice seedlings. The results showed that HM caused a marked reduction in seedling height, chlorophyll content (SPAD) and biomass, and activities of catalase (CAT) and ascorbate peroxidase (APX) in leaves and H⁺-ATPase in roots/leaves, but elevated superoxide dismutase (SOD) and guaiacol peroxidase (POD) activities in leaves with elevated malondialdehyde (MDA) accumulation both in leaves and roots over the control. The best mitigation effect was recorded in HM+GSH+Zn and HM+GSH (addition of GSH+Zn and GSH to HM solution), which greatly alleviated HM-induced growth inhibition and oxidative stress. Compared with HM alone, HM+GSH and HM+GSH+Zn markedly reduced Cr uptake and translocation but not affected Cd concentration; improved H⁺-ATPase activity and Fe, Zn, Mn uptake and translocation, and repressed MDA accumulation. Meanwhile exogenous GSH and GSH+Zn counteracted HM-induced response of antioxidant enzymes, via suppressing HM-induced dramatic increase of root/leaf SOD and leaf POD activities, and elevating stress-depressed leaf APX and leaf/root CAT activities.     

Glutathione-Mediated Alleviation of Chromium Toxicity in Rice Plants

A hydroponic experiment was conducted to determine the possible effect of exogenous glutathione (GSH) in alleviating chromium (Cr) stress through examining plant growth, chlorophyll contents, antioxidant enzyme activity, and lipid peroxidation in rice seedlings exposed to Cr toxicity. The results showed that plant growth and chlorophyll content were dramatically reduced when rice plants were exposed to 100 μM Cr. Addition of GSH in the culture solution obviously alleviated the reduction of plant growth and chlorophyll content. The activities of some antioxidant enzymes, including superoxide dismutase, catalase (CAT) and glutathione reductase in leaves, and CAT and glutathione peroxidase in roots showed obvious increase under Cr stress. Addition of GSH reduced malondialdehyde accumulation and increased the activities of these antioxidant enzymes in both leaves and roots, suggesting that GSH may enhance antioxidant capacity in Cr-stressed plants. Furthermore, exogenous GSH caused significant decrease of Cr uptake and root-to-shoot transport in the Cr-stressed rice plants. It can be assumed that GSH is involved in Cr compartmentalization in root cells.     

Role of ethylene in alleviation of cadmium-induced photosynthetic capacity inhibition by sulphur in mustard

Sulphur (S) assimilation leads to the formation of glutathione (GSH) and alleviation of cadmium (Cd) stress. GSH is synthesized from its immediate metabolite cysteine, which also serves as a metabolite for ethylene formation through S‐adenosyl methionine. To assess the role of ethylene in S‐induced alleviation of Cd stress on photosynthesis, the effects of S or ethephon (ethylene source) on GSH and ethylene were examined in mustard (Brassica juncea L. cv. Varuna). Sufficient‐S at 100 mg S kg^?1 soil alleviated Cd‐induced photosynthetic inhibition more than excess‐S (200 mg S kg^?1 soil) via ethylene by increased GSH. Under Cd stress, plants were less sensitive to ethylene, despite high ethylene evolution, and showed photosynthetic inhibition. Ethylene sensitivity of plants increased with ethephon or sufficient‐S, triggering the induction of an antioxidant system, and leading to increased photosynthesis even under Cd stress. The effects of ethephon and S under Cd stress were similar. The effects of S were reversed by ethylene biosynthesis inhibitor, aminoethoxyvinylglycine (AVG), suggesting that ethylene plays an important role in S‐induced alleviation of Cd stress on photosynthesis.     

In vitro Interactions of Thallium with Components of the Glutathione-dependent Antioxidant Defence System

We investigated the hypothesis that thallium (Tl) interactions with the glutathione-dependent antioxidant defence system could contribute to the oxidative stress associated with Tl toxicity. Working in vitro with reduced glutathione (GSH), glutathione reductase (GR) or glutathione peroxidase (GPx) in solution, we studied the effects of Tl⁺ and Tl³⁺ (1-25 μM) on: (a) the amount of free GSH, investigating whether the metal binds to GSH and/or oxidizes it; (b) the activity of the enzyme GR, that catalyzes GSH regeneration; and (c) the enzyme GPx, that reduces hydroperoxide at expense of GSH oxidation. We found that, while Tl⁺ had no effect on GSH concentration, Tl³⁺ oxidized it. Both cations inhibited the reduction of GSSG by GR and the diaphorase activity of this enzyme. In addition, Tl³⁺per se oxidized NADPH, the cofactor of GR. The effects of Tl on GPx activity depended on the metal charge: Tl⁺ inhibited GPx when cumene hydroperoxide (CuOOH) was the substrate, while Tl³⁺-mediated GPx inhibition occurred with both substrates. The present results show that Tl interacts with all the components of GSH/GSSG antioxidant defence system. Alterations of this protective pathway could be partially responsible for the oxidative stress associated with Tl toxicity.     

γ‐Aminobutyric acid addition alleviates ammonium toxicity by limiting ammonium accumulation in rice (Oryza sativa) seedlings

Excessive use of nitrogen (N) fertilizer has increased ammonium (NH₄⁺) accumulation in many paddy soils to levels that reduce rice vegetative biomass and yield. Based on studies of NH₄⁺ toxicity in rice (Oryza sativa, Nanjing 44) seedlings cultured in agar medium, we found that NH₄⁺ concentrations above 0.75 mM inhibited the growth of rice and caused NH₄⁺ accumulation in both shoots and roots. Use of excessive NH₄⁺ also induced rhizosphere acidification and inhibited the absorption of K, Ca, Mg, Fe and Zn in rice seedlings. Under excessive NH₄⁺ conditions, exogenous γ‐aminobutyric acid (GABA) treatment limited NH₄⁺ accumulation in rice seedlings, reduced NH₄⁺ toxicity symptoms and promoted plant growth. GABA addition also reduced rhizosphere acidification and alleviated the inhibition of Ca, Mg, Fe and Zn absorption caused by excessive NH₄⁺. Furthermore, we found that the activity of glutamine synthetase/NADH‐glutamate synthase (GS; EC 6.3.1.2/NADH‐GOGAT; EC1.4.1.14) in root increased gradually as the NH₄⁺ concentration increased. However, when the concentration of NH₄⁺ is more than 3 mM, GABA treatment inhibited NH₄⁺‐induced increases in GS/NADH‐GOGAT activity. The inhibition of ammonium assimilation may restore the elongation of seminal rice roots repressed by high NH₄⁺. These results suggest that mitigation of ammonium accumulation and assimilation is essential for GABA‐dependent alleviation of ammonium toxicity in rice seedlings.     

Kinetics,biochemical and factorial analysis of chromium uptake in a multi-ion system by Tradescantia pallida (Rose) D. R. Hunt

Discharge of wastewater from electroplating and leather industries is a major concern for the environment due to the presence of toxic Cr⁶⁺ and other ions, such as sulfate, nitrate, phosphate, etc. This study evaluated the potential of Tradescantia pallida, a plant species known for its Cr bioaccumulation, for the simultaneous removal of Cr⁶⁺, SO₄^2?, NO₃^?, and PO₄^3?. The effect of different co-ions on Cr⁶⁺ removal by T. pallida was examined following the Plackett-Burman design of experiments carried out under batch hydroponics conditions. The results revealed a maximum removal of 84% Cr⁶⁺, 87% SO₄^2?, 94% NO₃^? and 100% PO₄^3? without any phytotoxic effect on the plant for an initial Cr⁶⁺ concentration in the range 5–20 mg L^?1. SO₄^2? and NO₃^? enhanced Cr uptake at a high initial Cr concentration (20 mg L^?1), whereas PO₄^3? did not affect Cr uptake both at high and low initial Cr concentrations. The Cr⁶⁺ removal kinetics in the presence of different ions was well described by the pseudo-second-order kinetic model which revealed that both biosorption and bioaccumulation of the metal played an important role in Cr⁶⁺ removal. Increase in the total carbohydrate and protein content of the plant following Cr⁶⁺ and co-ions exposure indicated a good tolerance of the plant toward Cr⁶⁺ toxicity. Furthermore, enhancement in the lipid peroxidation and catalase activity in T. pallida upon Cr⁶⁺ exposure revealed a maximum stress-induced condition in the plant. Overall, this study demonstrated a very good potential of the plant T. pallida for Cr⁶⁺ removal from wastewater even in the presence of co-ions.     

Experimental Research Showing the Reduction of Naloxone-Place Aversion by Oral Zinc Administration in Rats

Selenium (Se) can play a protective role against heavy metal toxicity. This experiment aims to evaluate the effect of Se supplementation at different doses on the chicken brains. Oxidative stress was induced in the chicken brains by chromium(VI). A total of 105 Hyland brown male chickens were randomly divided into seven groups, including the control group, poisoned group [6%LD₅₀ K₂Cr₂O₇ body weight (B.W.)], and detoxification groups K₂Cr₂O₇ (6%LD₅₀) + Se (0.31, 0.63, 1.25, 2.50, and 5.00 Na₂SeO₃ mg/kg B.W.) orally in water for 42 days. The chickens were detected by the activities of mitochondrial membrane potential, 2′-benzoyloxycinnamaldehyde, superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), and Ca²⁺-ATPase. Cr(VI) administration caused histopathological damage. In addition, changes in oxidative stress indicators were observed in the chicken’s brains. Se supplement increased the levels of GSH, mitochondrial membrane potential (MMP), and Ca²⁺-ATPase and reduced MDA activity in the detoxification groups. However, the high-dose Se supplementation groups of 2.50 and 5.00 mg/kg reduced the activities of GSH, MMP, and Ca²⁺-ATPase; increased the brain–body ratio; and increased SOD activity. In conclusion, Cr(VI) exposure caused oxidative stress. Se exerted a remission effect on toxic responses in the chicken brains. However, a high Se concentration was synergistic to the toxic effect of Cr(VI).     

Cytokinin-like growth regulators mitigate toxic action of zinc and nickel ions on maize seedlings

Maize (Zea mays L.) seedlings grown in water culture in the presence of zinc and nickel ions were used with an effort to alleviate heavy metal toxicity by treating seeds with thidiazuron and kinetin (synthetic growth regulators with cytokinin-like activity). Heavy metals were shown to decrease germinability of seeds, suppress seedling growth, alter membrane permeability, and inhibit the activity of ascorbate peroxidase. Synthetic cytokinin-like agents alleviated deteriorative effects of heavy metals; the extent of alleviation depended on toxicant species and its concentration. The toxic effect of Zn²⁺ was effectively relieved by kinetin, whereas the Ni²⁺ toxicity was preferentially alleviated by thidiazuron.     

Effect of Chelate-Assisted Hexavalent Chromium on Physiological Changes,Biochemical Alterations,and Chromium Bioavailability in Crop Plants—An In Vitro Phytoremediation Approach

This study revealed heavy metal–induced physiological and biochemical alterations in crop seedlings by supplementing chelating agents in the nutrient solution. Hexavalent chromium (Cr⁺⁶) induces several toxic effects in hydroponically grown rice, wheat, and green gram seedlings. A noticeable decrease was observed in root length, shoot length, biomass content, and chlorophyll biosynthesis of the seedlings grown in the nutrient solutions supplemented with Cr⁺⁶ at 100 μM. The seedling growth was stimulated with supplement of chelating agents such as EDTA, DTPA, and EDDHA. An increase in proline content was noticed with the application of Cr⁺⁶ (100 μM) in nutrient solutions. Stimulated activities of antioxidant enzymes such as catalase and peroxidase were noticed with increasing concentrations of chromium. Cr bioaccumulation was significantly high in roots of seedlings treated with Cr⁺⁶ at 100 μM in nutrient solution. Shoot translocation of Cr as depicted by transportation index (Ti) values for different crops were enhanced with the application of chelating agents. The total accumulation rate (TAR) for Cr was enhanced with the supplementation of DTPA in rice and wheat, whereas the application of EDDHA was found effective for increasing the accumulation rate of Cr in green gram seedlings. This study demonstates the role of chelating agents in lessening the toxic effects of Cr⁺⁶. The chelating agents supplemented with Cr⁺⁶ in the culture medium enhanced the Cr bioavailability in plants.     

Comparative Studies of Tri- and Hexavalent Chromium Cytotoxicity and Their Effects on Oxidative State of Saccharomyces cerevisiae Cells

Chromium is a significant mutagen and carcinogen in environment. We compared the effects of tri- and hexavalent chromium on cytotoxicity and oxidative stress in yeast. Cell growth was inhibited by Cr³⁺ or Cr⁶⁺, and Cr⁶⁺ significantly increased the lethal rate compared with Cr³⁺. Both Cr³⁺ and Cr⁶⁺ can enter into the yeast cells. The percent of propidium iodide permeable cells treated with Cr³⁺ is almost five times as that treated with the same concentration of Cr⁶⁺. Levels of TBARS, O₂ ^?, and carbonyl protein were significantly increased in both Cr⁶⁺- and Cr³⁺-treated cells in a concentration- and time-dependent manner. Moreover, the accumulation of these stress markers in Cr⁶⁺-treated cells was over the Cr³⁺-treated ones. The decreased GSH level and increased activity of GPx were observed after 300 μM Cr⁶⁺-exposure compared with the untreated control, whereas there was no other change of GSH content in cells treated with Cr³⁺ even at very high concentration. Exposure to both Cr³⁺ and Cr⁶⁺ resulted in the decrease of activities of SOD and catalase. Furthermore, the effect of Cr⁶⁺ is stronger than that of Cr³⁺. Null mutation sensitivity assay demonstrated that the gsh1 mutant was sensitive to Cr⁶⁺ other than Cr³⁺, the apn1 mutant is more sensitive to Cr⁶⁺ than Cr³⁺, and the rad1 mutant is sensitive to both Cr⁶⁺ and Cr³⁺. Therefore, Cr³⁺ can be concluded to inhibit cell growth probably due to the damage of plasma membrane integrality in yeast. Although both tri- and hexavalent chromium can induce cytotoxicity and oxidative stress, the action mode of Cr³⁺ is different from that of Cr⁶⁺, and serious membrane damage caused by Cr³⁺ is not the direct consequence of the increase of lipid peroxidation.     

Ecophysiological responses of water hyacinth exposed to Cr3+ and Cr6+

Due to its wide industrial use, chromium (Cr) is considered a serious environmental pollutant of aquatic bodies. In order to investigate the ecophysiological responses of water hyacinth [Eichhornia crassipes (Mart.) Solms] to Cr treatment, plants were exposed to 1 and 10 mM Cr₂O₃ (Cr³⁺) and K₂Cr₂O₇ (Cr⁶⁺) concentrations for two or 4 days in a hydroponic system. Plants exposed to the higher concentration of Cr⁶⁺ for 4 days did not survive, whereas a 2 days treatment with 1 mM Cr³⁺ apparently stimulated growth. Analysis of Cr uptake indicated that most of the Cr accumulated in the roots, but some was also translocated and accumulated in the leaves. However, in plants exposed to Cr⁶⁺ (1 mM), a higher translocation of Cr from roots to shoots was observed. It is possible that the conversion from Cr⁶⁺ to Cr³⁺, which immobilizes Cr in roots, was not total due to the presence of Cr⁶⁺, causing deleterious effects on gas exchange, chlorophyll a fluorescence and photosynthetic pigment contents. Chlorophyll a was more sensitive to Cr than chlorophyll b. Cr³⁺ was shown to be less toxic than Cr⁶⁺ and, in some cases even increased photosynthesis and chlorophyll content. This result indicated that the F_v/F₀ ratio was more effective than the F_v/F_m ratio in monitoring the development of stress by Cr⁶⁺. There was a linear relationship between qP and F_v/F_m. No statistical differences were observed in NPQ and chlorophyll a/b ratio, but there was a tendency to decrease these values with Cr exposure. This suggests that there were alterations in thylakoid stacking, which might explain the data obtained for gas exchanges and other chlorophyll a fluorescence parameters.     

Preliminary studies on the involvement of glutathione metabolism and redox status against zinc toxicity in radish seedlings by 28-Homobrassinolide

The effect of exogenous application of 28-Homobrassinolide (HBR) on radish (Raphanus sativus L.) seedlings under zinc (Zn²⁺) stress on glutathione (GSH) production, consumption and changes in redox status was investigated. Zinc toxicity resulted in oxidative burst as evidenced by increased accumulation of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) content. These stress indices were significantly decreased by HBR supplementation. Under Zn²⁺ stress, GSH pool was decreased, while the contribution of oxidized glutathione (GSSG) to total GSH increased (GSSH/GSH ratio), this translated into significant reduction of GSH redox homeostasis. In addition, an increase of phytochelatins (PCs) was observed. In radish seedlings under Zn²⁺ stress, the activities of gamma-glutamylcysteine synthetase (γ-ECS), glutathione synthetase (GS), glutathione peroxidase (GPX), glutathione-S-transferase (GST) and cysteine (Cys) levels increased but the activity of glutathione reductase (GR) decreased. However, application of HBR increased the GSH pool and maintained their redox ratio by increasing the enzyme activities of GSH biosynthesis (γ-ECS and GS) and GSH metabolism (GR, GPX and GST). The results of present study are novel in being the first to demonstrate that exogenous application of HBR modulates the GSH synthesis, metabolism and redox homeostasis to confer resistance against Zn²⁺ induced oxidative stress.     

Development of a Multi-Species Biotic Ligand Model Predicting the Toxicity of Trivalent Chromium to Barley Root Elongation in Solution Culture

Little knowledge is available about the influence of cation competition and metal speciation on trivalent chromium (Cr(III)) toxicity. In the present study, the effects of pH and selected cations on the toxicity of trivalent chromium (Cr(III)) to barley (Hordeum vulgare) root elongation were investigated to develop an appropriate biotic ligand model (BLM). Results showed that the toxicity of Cr(III) decreased with increasing activity of Ca²⁺ and Mg²⁺ but not with K⁺ and Na⁺. The effect of pH on Cr(III) toxicity to barley root elongation could be explained by H⁺ competition with Cr³⁺ bound to a biotic ligand (BL) as well as by the concomitant toxicity of CrOH²⁺ in solution culture. Stability constants were obtained for the binding of Cr³⁺, CrOH²⁺, Ca²⁺, Mg²⁺ and H⁺ with binding ligand: log K_CrBL 7.34, log K_CrOHBL 5.35, log K_CaBL 2.64, log K_MgBL 2.98, and log K_HBL 4.74. On the basis of those estimated parameters, a BLM was successfully developed to predict Cr(III) toxicity to barley root elongation as a function of solution characteristics.     

Chromium interference in iron nutrition and water relations of cabbage

Cabbage (Brassica oleracea var. capitata cv. Snowball), known to be responsive to potentially toxic elements, was investigated for chromium (Cr³⁺) effect on iron metabolism and water relations. After 6 weeks growth in sand culture, a set of plants was supplied with 500 μM Cr³⁺ (CrCl₃), superimposed over the full nutrient solution (control). Exposure to excess Cr³⁺ led to increased accumulation of Cr, more in roots than in leaves, and to the development of toxicity symptoms. In decreasing chlorophyll concentration and the activities of heme enzymes, catalase and peroxidase, the excess Cr³⁺ effect resembled Fe deficiency. These changes, associated with decrease in Fe accumulation in Cr³⁺ treated plants, indicate that by reducing absorption of Fe, Cr³⁺ impairs the Fe requiring steps of chlorophyll and heme biosynthesis. In spite of lower water saturation deficit, the leaves of Cr³⁺ treated plants showed decrease in leaf water potential, associated with increase in diffusive resistance and lowering of transpiration rate, indicating development of water stress. Enhanced accumulation of proline in Cr³⁺ treated plants also suggested this. Observed changes in water stress parameters in Cr³⁺ stressed plants indicate that plant exposure to excess supply of Cr³⁺ reduces the physiological availability of water.     

Complexation of Al(III) with reduced glutathione in acidic aqueous solutions

The complexation of reduced glutathione (GSH) in its free and Al(III)-bound species in acidic aqueous solutions was characterized by means of multi-analytical techniques: pH-potentiometry, multinuclear (¹H, ¹³C and ²⁷Al) and two-dimensional nuclear Overhauser enhancement NMR spectroscopy (¹H, ¹H-NOESY), electrospray mass spectroscopy (ESI-MS), and ab initio electronic structure calculations. The following results were found. In the 25 °C 0.1 M KCl and 37 °C 0.15 M NaCl ionic medium systems, Al³⁺ coordinates with the important biomolecule GSH through carboxylate groups to form various mononuclear 1:1 (AlHL, AlH₂L and AlH₋₁L), 1:2 (AlL₂) complexes, and dinuclear (Al₂H₅L₂) species, where H₄L⁺ denotes totally protonated GSH. Besides the monodentate complexes through carboxylate groups, the amino groups and the peptide bond imino and carbonyl groups may also be involved in binding with Al³⁺ in the bidentate and tridentate complexes. The present data reinforce that the glycine carboxylate group of GSH has a higher microscopic complex formation constant than γ-glutamyl carboxylate. Compared with simple amino acids, the tripeptide GSH displays a greater affinity for the Al³⁺ ion and thus may interfere with aluminum’s biological role more significantly.     

Hydroxyl radical formation and lipid peroxidation enhancement by chromium

Chromium VI compounds have been shown to be carcinogenic in occupationally exposed humans, and to be genotoxic, mutagenic, and carcinogenic in a variety of experimental systems. In contrast, most chromium III compounds are relatively nontoxic, noncarcinogenic, and nonmutagenic. Reduction of Cr⁶⁺ leads to reactive intermediates, such as Cr⁵⁺, Cr⁴⁺, or other radical species. The molecular mechanism for the intracellular Cr⁶⁺ reduction has been the focus of recent studies, but the details are still not understood. Our study was initiated to compare the effect of Cr⁶⁺-hydroxyl radical formation and Cr⁶⁺-induced lipid peroxidation vs those of Cr³⁺. Electron spin responance measurements provide evidence for the formation of long-lived Cr⁵⁺ intermediates in the reduction of Cr⁶⁺ by glutathione reductase in the presence of NADPH and for the hydroxyl radical formation during the glutathione reductase catalyzed reduction of Cr⁶⁺. Hydrogen peroxide suppresses Cr⁵⁺ and enhances the formation of hydroxyl radical. Thus, Cr⁵⁺ intermediates catalyze generation of hydroxyl radicals from hydrogen peroxide through a Fenton-like reaction. Comparative effects of Cr⁶⁺ and Cr³⁺ on the development of lipid peroxidation were studied by using rat heart homogenate. Heart homogenate was incubated with different concentrations of Cr⁶⁺ compounds at 22°C for 60 min. Lipid peroxidation was determined as thiobarbituric acid reacting materiels (TBA-RM). The results confirm that Cr⁶⁺ induces lipid peroxidation in the rat heart homogenate. These observations might suggest a possible causative role of lipid peroxidation in Cr⁶⁺ toxicity. This enhancement of lipid peroxidation is modified by the addition of some metal chelators and antioxidants. Thus, strategies for combating Cr⁶⁺ toxicity should take into account the role of the hydroxy radicals, and hence, steps for blocking its chain propagation and preventing the formation of lipid peroxides.     

Removal of Cr6+ from groundwater using zero-valence iron in the laboratory

Abstract

In this paper, we describe a series of laboratory experiments which quantify the rate of Cr⁶⁺ reduction by Fe⁰. The main goal of these experiments was to determine the removal efficiency of Cr⁶⁺ by iron. The results indicate that Fe⁰ reduces Cr⁶⁺ to Cr³⁺ under alkaline and slightly acidic conditions. The removal efficiency rises with an increase of the initial concentration of Cr⁶⁺ (1 mg/L to 10 mg/L) when the quantity of Fe⁰ is stable. The removal efficiency increases as the quantity of Fe⁰ is raised when other conditions are constant. The removal efficiency would not be affected by other inorganic ions unless they were present at very high concentrations. When the initial concentration Cr⁶⁺ is 10mg/L and pH is 6.5–7.7, the final concentration of Cr⁶⁺ in effluent is less than 0.05 mg/L and the total Fe is less than 0.3 mg/L in effluent.     

Oxalic acid enhances Cr tolerance in the accumulating plant Leersia hexandra Swartz

This study examined the relationship between oxalic acid and Cr tolerance in an accumulating plant Leersia hexandra Swartz. The plants grown in hydroponics were exposed to Cr at 0, 5, 30, and 60 mg/L (without oxalate), and 0, 40, and 80 mg/L concentrations of Cr (with 70 mg/L oxalate or without oxalate). The results showed that more than 50% of Cr in shoots was found in HCl-extracted fraction (chromium oxalate) when the plants were exposed to Cr. Cr supply significantly increased oxalate concentration in shoots of L. hexandra (p < 0.05), but did not increase oxalate concentration in roots. Under 80 mg/L Cr stress, electrolyte leakages from roots and shoots with oxalate treatment were both significantly lower than those without oxalate treatment (p < 0.05), indicating exogenous oxalate supply alleviated Cr-induced membrane damage. Oxalate added to growth solution ameliorated reduction of biomass and inhibition of root growth induced by Cr, which demonstrated that application of oxalate helped L. hexandra tolerate Cr stress. However, oxalate supply did not affect the Cr concentrations both in roots and shoots of L. hexandra. These results suggest that oxalic acid may act as an important chelator and takes part in detoxifying chromium in internal process of L. hexandra.     

Effect of fertilization on exudation,dehydrogenase activity,iron-reducing populations and Fe++ formation in the rhizosphere of rice (Oryza sativa L.) in relation to iron toxicity

Summary To explain the mechanism of iron toxicity, greenhouse and growth chamber (¹⁴CO₂ atmosphere) experiments were carried out. In pot experiments (with a typical iron-toxic soil and a fertile clay) we studied the effect of N, P, K and Ca+Mg fertilization (alone or in combination) on dehydrogenase activity, Fe⁺⁺ formation, and the populations of iron-reducing bacteria in the rhizosphere of rice IR22 and IR42. Fe uptake by the plants was measured at regular intervals. Dehydrogenase activity, the number of N₂-fixing iron-reducing bacteria, and the formation and uptake of Fe⁺⁺ decreased with increased supply of K, Ca, and Mg. This effect was clearer with IR22 (susceptible to iron toxicity) than with IR42 (releatively tolerant). Increased exudation and Fe uptake by IR36 at low nutrient and high Fe supply were recorded in a growth chamber experiment. Nutritional conditions, exudation rate (a measure of metabolic root leakage), the iron-reducing activity of the rhizosphere, and Fe⁺⁺ uptake by wetland rice appear to be clearly related. Iron toxicity is considered a physiological disorder caused by multiple nutritional soil stress rather than by a low pH and high Fe supply per sé.