Effects of Cd2+ on K+, Ca2+ and N uptake in two halophytes Sesuvium portulacastrum and Mesembryanthemum crystallinum: consequences on growth

One of the limits of Cd2+-phytoextraction is the high toxicity of this metal to plants. Growth restriction, chlorosis and necrosis are usually accompanied with a large disturbance of the uptake of essential elements. This work aims to study the effects of cadmium (Cd2+) on potassium (K+), calcium (Ca2+) and nitrogen (N) acquisition, and their consequences on growth in two halophytes species: Sesuvium portulacastrum and Mesembryanthemum crystallinum. Seedlings were grown for 30 days in split-root conditions. One half of the root system was immersed in complete nutrient solution (Basal medium (B)) supplemented with 100 microM Cd2+, and the other half was immersed in a Cd2+-free medium, containing all nutrients (B/Cd plants) or deprived of potassium ((B-K)/Cd) or calcium ((B-Ca)/Cd) or nitrogen ((B-N)/Cd). Using this approach, we demonstrated that K+ and Ca2+ uptake was impaired in roots exposed to Cd2+. Concerning N, we noticed no indication of uptake inhibition by Cd2+. However, restriction of K+ uptake by roots was compensated by an increase in the K+-use efficiency, so that growth was not inhibited. Calcium uptake was strongly limited by Cd2. This inhibition was accompanied by a reduction in growth of ((B-Ca)/Cd) plants. Thus, we conclude that Cd2+ limits growth of both halophytes through restriction imposed on Ca2+ uptake. We suggest that the increase of Ca2+ availability in soils could improve the growth of both species in the presence of Cd2+. This would be essential for improving their utility for extraction of this metal by from salty contaminated soils.     

Uptake and localisation of lead in the root system of Brassica juncea

The uptake and distribution of Pb sequestered by hydroponically grown (14days growth) Brassica juncea (3days exposure; Pb activities 3.2, 32 and 217microM) was investigated. Lead uptake was restricted largely to root tissue. Examination using scanning transmission electron microscopy-energy dispersive spectroscopy revealed substantial and predominantly intracellular uptake at the root tip. Endocytosis of Pb at the plasma membrane was not observed. A membrane transport protein may therefore be involved. In contrast, endocytosis of Pb into a subset of vacuoles was observed, resulting in the formation of dense Pb aggregates. Sparse and predominantly extracellular uptake occurred at some distance from the root tip. X-ray photoelectron spectroscopy confirmed that the Pb concentration was greater in root tips. Heavy metal rhizofiltration using B. juncea might therefore be improved by breeding plants with profusely branching roots. Uptake enhancement using genetic engineering techniques would benefit from investigation of plasma membrane transport mechanisms.     

Oxidative stress in duckweed (Lemna minor L.) caused by short-term cadmium exposure

The mechanisms of plant defence against cadmium toxicity have been studied by short-term exposure of Lemna minor L. (common duckweed) to concentrations of CdCl2 ranging from 0 to 500microM. High accumulation of cadmium was observed (12,320+/-2155microgg(-1) at 500microM CdCl2), which caused a gradual decrease of plant growth, increased lipid peroxidation, and weakened the entire antioxidative defence. Total glutathione concentration decreased significantly; however, the concentration of oxidized glutathione remained stable. The responses of four antioxidant enzymes showed that catalase was the most inhibited after CdCl2 exposure, ascorbate peroxidase and guaiacol peroxidase moderately, and glutathione reductase least. The total antioxidative potential revealed an induced antioxidative network at 0.1microM CdCl2 (137+/-13.2% of the control) and its reduction to only 47.4+/-4.0% of the control at higher cadmium concentrations. The possible application of the examined biomarkers in ecotoxicological research is discussed.     

Abolishing cadmium toxicity in Chlorella vulgaris by ascorbic acid, calcium, glucose and reduced glutathione

Addition of ascorbic acid, calcium (Ca2+), glucose and reduced glutathione (GSH) to the cadmium (Cd2+)-amended Kuhl medium significantly accelerated the growth and chlorophyll content of Chlorella vulgaris. This enhancement was found to be dose dependent. Their relative effectiveness against Cd2+ on growth and chlorophyll content has been arranged as GSH > Ca2+ > glucose > ascorbic acid. However, Ca2+ offered more protection against Cd2+ toxicity on protein contents, followed by ascorbic acid, GSH and glucose. The different concentrations of all amelioratives completely alleviated the toxicity of Cd2+ on total carbohydrates to exceed the control level. Photosynthetic oxygen evolution showed more response to Ca2+ and glucose in alleviating Cd2+ toxicity. On the other hand, at certain concentrations, GSH, Ca2+, ascorbic acid and glucose stimulated respiratory oxygen uptake over the control level. The uptake of Cd2+ dropped significantly in cultures supplemented with 20 microM ascorbic acid, glucose and GSH, while Ca2+ (30 microM) significantly lowered the Cd2+ uptake by 58% indicating that Ca2+ had a more antagonistic effect against Cd2+.     

Removal of chromium from abrasive blast media by leaching and electrochemical precipitation

Water is effective in leaching out Cr6+ from a mixture of paint powders and abrasive blast media. However, acids such as HNO3, HCl, and H2SO4 significantly enhance the leaching procedure. Cr ions in the leaching solutions are successfully removed by electrochemical precipitation. The consumable Fe electrodes generate ferrous ions to cause the reduction of Cr6+ to Cr3+. Cr3+ ions along with Fe2+ and Fe3+ are then removed mainly by precipitation as Cr(OH)3, Fe(OH)2, and Fe(OH)3 near the cathode where OH- ions are generated by water electrolysis. The electrochemical process is capable of discharging low levels of Cr6+, less than 1 mg/L, without pH adjustment.     

Competitive adsorption characteristics of Co2+, Ni2+, and Cr3+ by IRN-77 cation exchange resin in synthesized wastewater

Adsorption properties of Co2+, Ni2+, and Cr3+ on an Amberlite IRN-77 cation exchange resin were investigated in batch systems. Levels of adsorption rapidly approached an equilibrium state within 1 h. The adsorption characteristics of each metal onto the resin were accurately represented by Langmuir isotherms. Co2+ and Ni2+, which have an equivalent electrovalence, displayed similar levels of adsorption onto the resin when they coexisted in the solution. However, when Cr3+ was added to the solution it competitively replaced Co2+ and Ni2+ ions that had been previously adsorbed onto the resin, resulting in the desorption of these metals into the solution. The result was likely due to a higher adsorption affinity of Cr3+ relative to Co2+ and Ni2+. This implies that interactively competitive adsorption of multi-cations onto the resin should be thoroughly considered when contemplating the efficient operation of an ion exchange process in the treatment of industrial wastewater.     

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.     

Metallothioneins induction and antioxidative response in aquatic worms Tubifex tubifex (Oligochaeta, Tubificidae) exposed to copper

Metallothioneins (MTs), are low molecular weight proteins, mainly implicated in metal ion detoxification. Increase in MT contents is considered as a specific biomarker of metal exposure. Recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth, and antioxidative defences. Tubifex tubifex were exposed to different copper concentrations (50, 100, and 200 microgl(-1)) for 7 and 15 days. MT levels in exposed worms increased significantly (p<0.05) after 7 and 15 days of exposure to different concentrations of copper (maximum +208% for 100 microgl(-1) after 7 days of exposure). Also important perturbation in metal-metallothionein content occurred, along with an increase in total soluble protein content in all treated worms after 7 and 15 days (max. +88.49%). Catalase activities (CAT) in Cu treated-worms were significantly increased, and demonstrated a development of antioxidative defenses. Additionally a reduction of gulathione-S-transferase (GST) was observed in all treated worms after 7 days of exposure to Cu (max. -44.42%). The high induction of MTs observed during T. tubifex exposure to Cu make them potentially useful biomarkers to monitor metal pollution.     

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.     

Trichoderma atroviride F6 improves phytoextraction efficiency of mustard (Brassica juncea (L.) Coss. var. foliosa Bailey) in Cd, Ni contaminated soils

Trichoderma atroviride F6, isolated from decaying feather and resistant to 100 mg l(-1) Cd2+ and 250 mg l(-1) Ni2+, was applied for rhizoremediation of Cd, Ni and Cd-Ni combination contaminated soils through association with Brassica juncea (L.) Coss. var. foliosa. The strain significantly alleviated the cellular toxicity of cadmium and nickel to plants. Inoculation of B. juncea (L.) Coss. var. foliosa with T. atroviride F6 resulted a 110%, 40% and 170% increase in fresh weight in Cd, Ni and Cd-Ni contaminated soils, respectively (P<0.05). The translocation factors and metal bioconcentration factors calculated for the inoculated plant were increased compared to the noninoculated plants. The results indicated that the efficiency of phytoextraction for B. juncea (L.) Coss. var. foliosa enhanced after inoculating with T. atroviride F6. The fungal treated plants grown in Cd-Ni combination contaminated soils showed higher phytoextraction efficiency than those in Cd or Ni contaminated soils. Thus, it is suggested that the fungus T. atroviride F6 endowed with organic-degrading capabilities could be exploited for fungi-assisted phytoremediation of mixed organic-metal contaminated soils.     

Field intercomparison of precipitation measurements performed within the framework of the Pan European Intensive Monitoring Program of EU/ICP Forest

A 6-month field intercomparison study on precipitation measurements was performed at Schagerbrug near the west coast in the Netherlands. Twenty bulk sampling systems and two wet-only samplers were evaluated on accuracy, sampling strategy and performance under field conditions. Bulk precipitation fluxes of NO3-, NH4+, H+ and Kjeldahl-N generally could be determined with a greater accuracy than bulk precipitation fluxes of SO4(2-), Na+, Cl-, Mg2+, Ca2+, Alkalinity and H+. Bulk precipitation fluxes of K+ generally had the lowest accuracy. Only 20% of the sampling systems differed less than 10% from the best estimate, whereas most systems (60%) differed more than 20% from the best estimate. The inaccuracy induced by the sampling system appeared to be much larger than that resulting from the analysis of the samples by different laboratories as determined by ring-tests.     

Reduction of chromate (CrO4(2-)) by an enrichment consortium and an isolate of marine sulfate-reducing bacteria

An enrichment consortium and an isolate (isolate TKW) of sulfate-reducing bacteria (SRB) have been obtained from metal-contaminated marine sediments of Tokwawan, Hong Kong SAR. These bacteria are capable of reducing highly toxic and soluble hexavalent chromium (Cr6+) enzymatically into less toxic and insoluble trivalent chromium (Cr3+) under anaerobic conditions. The enrichment consortium almost completely (98.5%) reduced 0.6 mM Cr6+ in 168 h and the rate of reduction was 0.5 g (Cr6+) g(protein)(-1)h(-1). In comparison, with Cr6+ as the sole electron acceptor (as a surrogate for SO4(2-)), isolate TKW reduced 94.5% of the initially added Cr6+ (0.36 mM) in 288 h, with the rate of 0.26 g (Cr6+) g(protein)(-1)h(-1). Adsorption by these bacteria was not the major mechanism contributing to the transformation or removal of Cr6+. The biomass and Cr3+ in the cultures increased simultaneously with the reduction of Cr6+. These indigenous SRB might have potential application in bioremediation of metal contaminated sediments.     

The response of Lemna trisulca L. to cadmium

Lemna trisulca was grown, using aseptic culture techniques in a filter-sterilized medium, a portion of which was replaced regularly during experiments. L. trisulca responded to the addition of 0.64 microM Cd with a reduction in multiplication rate (MR) 2 +/- 1 days after exposure. The internal Cd content reached 1000 +/- 140 microg Cd/g (dry wt) within 2 days exposure to 0.64 microM Cd. The final yield was reduced by an average of c.8% for each day of exposure to 0.64 microM Cd in a 14 day experiment. This implies that an equilibration period should be used for short-term bioassay tests before the effect of a toxicant is determined. Pretreating L. trisulca with 0.08 or 0.32 microM Cd for 6 weeks had no significant effect on MR or Cd uptake when plants were subsequently exposed to a range of Cd concentrations or grown in a control medium. This suggests that L. trisulca does not become acclimated to elevated Cd concentrations. The MR of L. trisulca fluctuated over a period of almost 600 days and the doubling time ranged from 1.6 to 2.4 days. This produced more than a fivefold difference in final yield in experiments of 14 days duration. The reduction in MR in response to 0.32 microM Cd during this same 600 days period averaged 24% with a coefficient of variation of 38%, and varied with the MR of control cultures. Fluctuations in the intrinsic growth rate and the effect of a toxicant on L. trisulca could potentially confound the assessment of toxicity and must be carefully considered when designing test protocols for aquatic plants.     

24-Epibrassinolide protects against the stress generated by salinity and nickel in Brassica juncea

The plants of Brassica juncea (L) were grown in the presence of NaCl and/or NiCl2 and were sprayed with 1muM of 24-epibrassinolide (EBL) at 15 days after sowing (DAS) and were sampled at 30 DAS. The plants exposed to NaCl and/or NiCl2 exhibited a significant decline in growth, the level of pigments and photosynthetic parameters. However, the follow up treatment with EBL detoxified the stress generated by NaCl and/or NiCl2 and significantly improved the above parameters. The NaCl and/or NiCl2 increased electrolyte leakage and lipid peroxidation, and decreased the membrane stability index (MSI) and relative water content. However, the EBL treatment in absence of the stress improved the MSI and relative water content but could not influence electrolyte leakage and lipid peroxidation. The antioxidative enzymes and the level of proline exhibited a significant increase in response to EBL as well as to NaCl and/or NiCl2 stress.     

The ability of Azolla caroliniana to remove heavy metals (Hg(II), Cr(III), Cr(VI)) from municipal waste water

The aim of this paper was to investigate the capacity of a small water fern, Azolla caroliniana Willd. (Azollaceae), to purify waters polluted by Hg and Cr. Many plants are capable of accumulating heavy metals (called hyperaccumulators) and one of them is the water fern A. caroliniana. During 12 days of the experiment the fern was grown on the nutrient solution containing Hg2+, Cr3+ and CrO4(2-) ions, each in a concentration 0.1, 0.5 and 1.0 mg dm(-3). The presence of these ions caused a 20-31% inhibition of A. caroliniana growth, the highest in the presence of Hg(II) ions, in comparison to the control. After day 12 of the experiment, metal contents the solution decreased to 0-0.25 mg dm(-3), and this decrease comprised between 74 (Cr3+ 1.0 mg dm(-3) treatment) and 100% (CrO4(2-) 0.1 mg dm(-3) treatment). The fern took a lesser quantity of the metals from 0.1 mg dm(-3) treatments compared to 0.5 and 1.0 mg dm(-3) treatments. In the A. caroliniana tissues the concentration of heavy metals under investigation ranged from 71 to 964 mg kg(-1) dm; the highest level being found for Cr(III) containing nutrient solution.     

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

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

利用芦竹修复重金属污染湿地的研究

研究了芦竹(Arundodonaxlinn)对Cu2+、Ni2+、Cr6+重金属污染湿地的响应和对该污染湿地的影响。实验结果表明,芦竹对这3种重金属离子有一定的耐受性,并有不同程度的吸收,Cu2+、Ni2+和Cr6+去除率分别为63.8%,42.3%和34.4%。芦竹在100mg/kg浓度的Cu2+、Ni2+污染湿地中生长正常,在低浓度(55mg/kg)Cr6+污染中能存活,但生长速度较慢。在100mg/kg浓度Cr6+污染湿地中,出现急性中毒现象,半月后致死。在重金属污染环境中,芦竹普遍出现失绿现象,但除高浓度Cr6+(100mg/kg)以外,都能正常存活,表现出较强的适应性。     

Influence of sodium nitrilotriacetate (NTA) and citric acid on phenolic and organic acids in Brassica juncea grown in excess of cadmium

Brassica juncea cv. 426308 was grown in soils containing 150 mg Cd(2+)kg(-1) soil. After 38 days, the soil was amended with two rates of citric acid or NTA (10 and 20 mmol kg(-1) soil). Control soil was not amended with chelates. Plants were harvested during growth, immediately before and seven days after chelate addition. Shoot composition of organic and phenolic acids and shoot Cd(2+) concentration were determined. Cadmium concentration remained constant during the growth and increased following NTA and citric acid amendments depending on chelate type and concentration. The highest increments in Cd(2+) were measured after the addition of NTA. Compared to the control, 10 and 20 NTA-treated plants showed two- and three-fold increases, respectively. At 150 mg Cd(2+)kg(-1) soil the amount of organic and phenolic acids in the leaves of B. juncea was always higher than that detected in the control. A direct correlation between organic acid concentration and cadmium content was detected both during growth and after chelate addition. On the contrary, the amount of phenols seemed to be correlated with the metal content only in non-amended and NTA-treated plants. The 10 and 20 citric acid additions caused 45% and 90% increases in shoot phenolic acids although cadmium content rose to a smaller extent. The inhibition of citrate synthase and the entrance of phosphoenolpyruvate in shikimate pathway leading to the formation of aromatic compounds might come into play. The increase in phenylalanine ammonialyase activity following citric acid amendments suggested this metabolic response.