Effects of low-molecular-weight organic acids and residence time on desorption of Cu, Cd, and Pb from soils

Effects of low-molecular-weight organic acids (LMWOAs) and residence time on desorption of Cu, Cd, and Pb from two typical Chinese soils were studied. Citric, malic, and acetic acids were chosen as representatives of LMWOAs commonly present in soils. CaCl(2) and NaNO(3) were used in desorption as they were main soil background electrolytes for comparison. Desorption of Cu, Cd, and Pb from both soils followed the descending order: citric acid>malic acid>acetic acid>CaCl(2)>NaNO(3), which was consistent with the order of stability of Cu-, Cd-, and Pb-LMWOAs complexes from large to small and ion exchange ability of Ca(2+) and Na(+). Desorption of metals by inorganic salts decreased with increasing desorption solution pH. Whereas desorption of metals by LMWOAs showed different trend in response to pH change due to their different complexing abilities. Malic and acetic acids released less metals at low pH 3.1 compared with citric acid at pH 7, indicating that pH was not the dominant factor governing the release of metals. In addition, all LMWOAs desorbed more metals than inorganic salts, CaCl(2) and NaNO(3). Therefore, organic ligands played a dominant role in desorption of heavy metals. More metals were released from Jiangxi soil than from Heilongjiang soil due to lower soil pH, CEC, organic matter content and manganese oxide of Jiangxi soil. Generally, desorption of metals decreased with increasing residence time of metals in soils.     

Dissolution of mimetite Pb5(AsO4)3Cl in low-molecular-weight organic acids and EDTA

Due to its relatively low solubility, mimetite Pb₅(AsO₄)₃Cl may control Pb and As(V) solution levels at a low value in contaminated soils. The time-dependent dissolution of mimetite by low-molecular-weight organic acids (LMWOAs) such as acetic, lactic, citric, and ethylene diamine tetra-acetic acid (EDTA) was determined. At pH 3.5, the presence of citric acid or EDTA significantly increases the solubility of mimetite while acetic or lactic acids show little effect. The effect of all organic anions on the dissolution of mimetite increased with the increase in solution pH. The rate of mimetite dissolution depended on the kind and concentration of organic solvents in the sequence r_EDTA > r_lactate > r_acetate > r_citrate. Soluble Pb and As(V) released in LMWOAs and EDTA were higher than the WHO guideline value for these elements in drinking water (10 μg As(V) L⁻¹, 10 μg Pb L⁻¹). This suggests that soil organic acids in rhizosphere can potentially liberate Pb and As(V) from mimetite in contaminated soils.     

The short-term effect of cadmium on low molecular weight organic acid and amino acid exudation from mangrove (Kandelia obovata (S., L.) Yong) roots

The aim of this study was to evaluate short-term concentration and time effects of cadmium on Kandelia obovata (S., L.) Yong root exudation, thereby evaluating and predicting the ecophysiological effects of mangrove to heavy metals at the root level. Mature K. obovata propagules were cultivated in a sandy medium for 3 months, and then six concentrations of Cd (0, 2.5, 5, 10, 20, and 40 mg L^?1) were applied. After exposure time of 24 h and 7 days, respectively, the root exudates of K. obovata were collected and low molecular weight organic acids (LMWOAs) and amino acids of which were analyzed. In addition, we measured glutathione, soluble protein content, and Cd concentration in the plant. We found 10 and 15 types of LMWOAs and amino acids in root exudates of K. obovata with total concentrations ranging from 29.54 to 43.08 mg g^?1 dry weight (DW) roots and from 737.35 to 1,452.46 ng g^?1 DW roots, respectively. Both of them varied in quality and quantity under different Cd treatment strengths and exposure times. Oxalic, acetic, l-malic, tartaric acid, tyrosine, methionine, cysteine, isoleucine, and arginine were dominant. Both LMWOAs and amino acids excreted from K. obovata roots play a key role in Cd toxicity resistance. The responsiveness of amino acids was less than that of LMWOAs. We suggest that the ecological effect of root-excreted free amino acids in the rhizosphere is mainly based on the role of nutrients, supplemented with detoxification to heavy metals.     

Effects of lead and chelators on growth, photosynthetic activity and Pb uptake in Sesbania drummondii grown in soil

Effects of lead (Pb) and chelators, such as EDTA, HEDTA, DTPA, NTA and citric acid, were studied to evaluate the growth potential of Sesbania drummondii in soils contaminated with high concentrations of Pb. S. drummondii seedlings were grown in soil containing 7.5 g Pb(NO(3))(2) and 0-10 mmol chelators/kg soil for a period of 2 and 4 weeks and assessed for growth profile (length of root and shoot), chlorophyll a fluorescence kinetics (F(v)/F(m) and F(v)/F(o)) and Pb accumulations in root and shoot. Growth of plants in the presence of Pb+chelators was significantly higher (P<0.05) than the controls grown in the presence of Pb alone. F(v)/F(m) and F(v)/F(o) values of treated seedlings remained unaffected, indicating normal photosynthetic efficiency and strength of plants in the presence of chelators. On application of chelators, while root uptake of Pb increased four-five folds, shoot accumulations increased up to 40-folds as compared to controls (Pb only) depending on the type of chelator used. Shoot accumulations of Pb varied from 0.1 to 0.42% (dry weight) depending on the concentration of chelators used.     

Influence of metal ion on sorption of p-nitrophenol onto sediment in the presence of cetylpyridinium chloride

Heavy metals and surfactants have a significant effect on the sorption of organic contaminants. In this study, batch equilibrium experiments were carried out to investigate the influence of Pb(NO(3))(2) on the sorption of p-nitrophenol (PNP) onto sediments in the presence of cationic surfactant cetylpyridinium chloride (CPC). Results indicated that in the complex system containing PNP, Pb(NO(3))(2) and CPC, the sorption of PNP decreased with increasing concentration of Pb(NO(3))(2) due primarily to competing for adsorption sites. Likewise, partitioning of PNP in adsorbed surfactant layers and micelles decreased with increasing level of Pb(NO(3))(2). Moreover, the influence of different metal ions (Pb(2+), Cd(2+), Zn(2+)) was examined and results indicated that the presence of heavy metals inhibited the sorption of PNP in the order: Pb(2+)>Cd(2+)>Zn(2+). The competitive effect of the heavy metals was in agreement with the hydration energy and hydrated radius. The results are believed to provide a useful insight into describing the transport and fate of PNP in natural environments.     

Evaluation of metal mobility, plant availability and immobilization by chemical agents in a limed-silty soil

Metal-contaminated soils in the vicinity of industrial sites become of ever-increasing concern. Diagnostic criteria and ecological technologies for soil remediation should be calibrated for various soil conditions; actually, our knowledge of calcareous soil is poor. Silty soils near smelters at Evin (Pas de Calais, France) have been contaminated by non-ferrous metal fallout and regularly limed using foams. Therefore, the mobility, bioavailability, and potential phytotoxicity of Cd, Pb and Zn, were investigated using single soil extractions (i.e. water, 0.1 n Ca(NO(3))(2), and EDTA pH 7), and vegetation experiments, in parallel with a biological test based on (iso)-enzymes in leaves and roots, before and following soil treatment with chemical agents, i.e. Thomas basic slags (TBS), hydrous manganese oxide (HMO), steel shots (ST) and beringite. No visible toxicity symptoms developed on the above-ground parts of ryegrass, tobacco and bean plants grown in potted soil under controlled environmental conditions. Cd, Zn and Pb uptake resulted in high concentrations in the above-ground plant parts, but the enzyme capacities in leaves and roots, and the peroxidase pattern indicated that these metal concentrations were not phytotoxic for beans as test plants. The addition of chemical agents to the soil did not increase biomass production, but treatment with either HMO, ST or beringite markedly decreased the mobility of Cd, Zn and Pb. These agents were proven to be effective in mitigating the Cd uptake by plants. HMO and ST decreased either Pb or Zn uptake by ryegrass. TBS was effective in lowering Pb uptake by the same species. Beringite decreased Cd uptake by beans. If fallout could be restricted, the metal content of food crops in this area should be lowered by soil treatment. However, the differences in Cd uptake between plant species were not suppressed, regardless of the type of agents applied to the soil.     

Sulfur (S)-induced enhancement of iron plaque formation in the rhizosphere reduces arsenic accumulation in rice (Oryza sativa L.) seedlings

The effects of two sulfur (S) sources (SO(4)(2-), S(0)), and three rates of S application (0, 30, 120 mgS/kg) on the formation of iron plaque in the rhizosphere, and on the root surface of rice, and As (arsenic) uptake into rice (Oryza sativa L.) were studied in a combined soil-sand culture experiment. Significant differences in As uptake into rice between +S and -S treatments were observed in relation to S sources, and rates of S application. Concentrations of As in rice shoots decreased with increasing rates of S application. The mechanism could be ascribed to sulfur, induced the formation of iron plaque, since concentrations of Fe in iron plaque on quartz sands in the rhizosphere, and on the root surface of rice increased with increasing rates of S application. The results suggest that sulfur fertilization may be important for the development approaches to reducing As accumulation in rice.     

The effect of nitrogen form on rhizosphere soil pH and zinc phytoextraction by Thlaspi caerulescens

The phytoextraction of Zn may be improved by applying N fertilizers to increase the biomass and Zn content of shoots. Rhizosphere-pH change from uptake of different N forms will affect Zn phyto-availability in the rhizosphere and Zn phytoextraction. This glasshouse study examined the effect of N form on Zn phytoextraction by Thlaspi caerulescens (Prayon). The plants were grown in a Zn-contaminated soil (total Zn 250 mg kg-1 soil; pHwater 5.7) and supplied with (NH4)2SO4, Ca(NO3)2 or urea [(NH2)2CO]. The form was maintained by applying the nitrification inhibitor dicyandiamide. A biodegradable chelator ethylenediaminedisuccinic acid (EDDS) was included for comparison. The addition of N doubled the shoot biomass. The highest shoot Zn content occurred in the Ca(NO3)2 treatment and was associated with the highest rhizosphere pH. The lowest shoot dry weight occurred in the EDDS treatment. The Zn concentration in the shoots increased as the rhizosphere pH increased. A significant correlation occurred between Ca and Zn concentrations in the shoots. This study demonstrated that Ca(NO3)2 is a more effective treatment than , urea or EDDS for enhancing Zn phytoextraction in a mildly acidic soil.     

Relationship between the extractable metals from soils and metals taken up by maize roots and shoots

Two extraction procedures, i.e. a single extraction procedure using low-molecular-weight-organic-acids (LMWOAs) as extractant and a sequential extraction procedure recommended by the European Community Bureau of Reference (BCR), were performed to extract metal fractions from wet rhizosphere soil. And the extracted soil solutions were further fractionated as colloidal and truly dissolved fractions. Heavy metals in maize roots were experimentally defined as metals adsorbed on cell wall and metals taken up by cross-membrane by washing with CaCl(2). The correlation coefficients between extractable metals from soil and taken up by maize roots and shoots were compared between two extraction methods, and a good correlation was obtained if LMWOAs were used. In contrast, the correlation coefficients were poor when the BCR method was used.     

Evaluation of plant availability of soil trace metals by chemical fractionation and multiple regression analysis

Soil samples with a range of chemical and physical properties were collected from 10 different rural regions of China. Trace metals (Ni, Co, Cu, and Pb) in the soils were partitioned by a sequential extraction procedure into Mg(NO(3))(2) extractable (F1), CH(3)COONa extractable (F2), NH(2)OH.HCl extractable (F3), HNO(3)?H(2)O(2) extractable (F4), and residual (F5) fractions. Chemical fractionation showed that F1 fraction of the metals was less than 1% and residue was the dominant form for Cu and Ni in all samples, and for Co in most of the samples. Significant interrelationships of the fractions varied considerably with the different metals. Winter wheat (Triticum aestivum L.) and alfalfa (Medicago sativa L.) had been grown on the soils in a pot-culture experiment under greenhouse conditions for 40 days. Metal availability to the plants was evaluated by simple and multiple regression analysis. The Mg(NO(3))(2) extractable Co (F1) was significantly correlated with Co concentrations in different parts of wheat and in the whole of alfalfa. For the other metals, the independent variables of the multiple regression models, chosen by stepwise selection, were given as: F1 and F2 + F3 + F4 for Ni; F1, F2 + F3 and F4 for Cu; and F3 + F4 for Pb. The results of this study demonstrate that the sequential extraction procedure, in conjunction with multiple regression models using a combination of correlated fractions as an independent variable, may be useful for the prediction of plant absorption of trace metals in soils.     

The effects of exogenous plant growth regulators in the phytoextraction of heavy metals

The term "assisted phytoextraction" usually refers to the process of applying a chemical additive to contaminated soil in order to increase the metal uptake by crop plants. In this study three commercially available plant growth regulators (PGRs) based on cytokinins (CKs) were used to boost the assisted phytoextraction of Pb and Zn in contaminated soil collected from a former manufactured gas-plant site. The effects of EDTA treatment in soil and PGR treatment in leaves of Helianthus annuus were investigated in terms of dry weight biomass, Pb and Zn accumulation in the upper parts of the plants, Pb and Zn phytoextraction efficiency and transpiration rate. Metal solubility in soil and its subsequent accumulation in shoots were markedly enhanced by EDTA. The combined effects of EDTA and cytokine resulted in an increase in the Pb and Zn phytoextraction efficiency (up to 890% and 330%, respectively, compared to untreated plants) and up to a 50% increase in foliar transpiration. Our results indicate that exogenous PGRs based on CKs can positively assist the phytoextraction increasing the biomass production, the metal accumulation in shoots and the plant transpiration. The observed increase in biomass could be related to its action in stimulation of cell division and shoot initiation. On the other hand, the increase in metal accumulation in upper parts of plant could be related to both the role of PGRs in the enhancement of plant resistance to stress (as toxic metals) and the increase in transpiration rate, i.e. flux of water-soluble soil components and contaminants by the regulation of stomatal opening.     

Phytoextraction of metals from a multiply contaminated soil by Indian mustard

The effects of nitrilotriacetate (NTA) and citric acid applications on metal extractability from a multiply metal-contaminated soil, as well as on their uptake and accumulation by Indian mustard (Brassica juncea) were investigated. Desorption of metals from the soil increased with chelate concentration, NTA being more effective than citric acid in solubilising the metals. Plants were grown in a sandy soil collected from a contaminated field site and polluted by Cd, Cr, Cu, Pb and Zn. After 43 days of plant growth, pots were amended with NTA or citric acid at 5 mmol kg-1 soil. Control pots were not treated with any chelate. Harvest of plants was performed 1 week after chelate addition. Soil water-, NH4NO3- and DTPA-extractable Cd, Cu, Pb and Zn fractions were enhanced only in the presence of NTA. In comparison to unamended plants, Indian mustard shoot dry weights suffered significant reductions following NTA application. NTA treatment increased shoot metal concentrations by a factor of 2-3, whereas citric acid did not induce any difference compared to the control. Chromium was detected in the above-ground tissues only after NTA amendment. Due to differences in dry matter yield, a significant enhancement of metal uptake was observed in NTA-treated plants for Cu and Zn.     

Influence of organic acids on the transport of heavy metals in soil

Vegetation historically has been an important part of reclamation of sites contaminated with metals, whether the objective was to stabilize the metals or remove them through phytoremediation. Understanding the impact of organic acids typically found in the rhizosphere would contribute to our knowledge of the impact of plants in contaminated environments. Heavy metal transport in soils in the presence of simple organic acids was assessed in two laboratory studies. In the first study, thin layer chromatography (TLC) was used to investigate Zn, Cd, and Pb movement in a sandy loam soil as affected by soluble organic acids in the rhizosphere. Many of these organic acids enhanced heavy metal movement. For organic acid concentrations of 10mM, citric acid had the highest R(f) values (frontal distance moved by metal divided by frontal distance moved by the solution) for Zn, followed by malic, tartaric, fumaric, and glutaric acids. Citric acid also has the highest R(f) value for Cd movement followed by fumaric acid. Citric acid and tartaric acid enhanced Pb transport to the greatest degree. For most organic acids studied, R(f) values followed the trend Zn>Cd>Pb. Citric acid (10mM) increased R(f) values of Zn and Cd by approximately three times relative to water. In the second study, small soil columns were used to test the impact of simple organic acids on Zn, Cd, and Pb leaching in soils. Citric acid greatly enhanced Zn and Cd movement in soils but had little influence on Pb movement. The Zn and Cd in the effluents from columns treated with 10mM citric acid attained influent metal concentrations by the end of the experiment, but effluent metal concentrations were much less than influent concentrations for citrate <10mM. Exchangeable Zn in the soil columns was about 40% of total Zn, and approximately 80% total Cd was in exchangeable form. Nearly all of the Pb retained by the soil columns was exchangeable.     

Zinc and copper uptake by plants under two transpiration rates. Part II. Buckwheat (Fagopyrum esculentum L.)

To evaluate the environmental risks of irrigating crops with treated wastewater, a study was undertaken to quantify heavy metal uptake by 4-week old buckwheat (Fagopyrum esculentum L.) plants during 18 days of irrigation with 8 different Cu and Zn solutions under two transpiration rates (TR). At 4 weeks, potted buckwheat plants were transferred into one of the two growth chambers, offering either a high or low vapour pressure deficit (VDP) for, respectively, a high or low TR. Triplicate pots received one of the 8 irrigation treatments containing one of two Zn levels (0 and 25 mg/L) combined with one of four Cu levels (0, 5, 10 and 15 mg/L). Daily TR were measured by weighing the evapo-transpired water lost from the planted pot, less was the evaporation loss measured from triplicate non-planted pots. After 0, 6, 12 and 18 days of treatment, the stems and leaves of three randomly selected plants were harvested and after 18 days, the roots were harvested to determine Cu and Zn uptake. The treatments did not affect TR in terms of dry plant mass, indicating the absence of toxic effects. Irrigating with Zn, without Cu, increased dry biomass production, whereas the lowest biomass occurred with 15 and 30 mg/L of Cu with and without 25 mg/L of Zn, respectively, because higher applications of heavy metal significantly reduced soil pH. Plant Cu and Zn uptake increased with TR. With higher levels of Cu, Zn uptake by buckwheat was significantly reduced, while Zn had a slight but non-significant impact on Cu uptake. Previously and in a study exposing wheat plants to the same conditions, Cu significantly increased Zn uptake, while Zn had a slight but insignificant negative effect on Cu uptake. The buckwheat roots contained the greatest levels of Cu and Zn, indicating their role in moderating heavy metal uptake. Also, both Cu and Zn had a synergetic effect on each other in terms of root levels, and a similar observation was made in the earlier similar experiment using wheat plants. Irrigating a buckwheat crop with treated wastewater, with more natural Cu and Zn levels of 0.08 mg/L, could be quite beneficial without endangering the quality of the crop and acidifying the soil pH. The most concentrated experimental solutions contained 300 times more Cu and Zn, to obtain measurable differences.     

Characterization of heavy metal-resistant endophytic bacteria from rape (Brassica napus) roots and their potential in promoting the growth and lead accumulation of rape

Two lead (Pb)-resistant endophytic bacteria were isolated from rape roots grown in heavy metal-contaminated soils and characterized. A pot experiment was conducted for investigating the capability of the two isolates to promote the growth and Pb uptake of rape from Pb-amended soil. The two isolates were identified as Pseudomonas fluorescens G10 and Microbacterium sp. G16 based on the 16S rDNA gene sequence analysis. Strains G10 and G16 exhibited different multiple heavy metal and antibiotic resistance characteristics and increased water-soluble Pb in solution and in Pb-added soil. Root elongation assays demonstrated increases in root elongation of inoculated rape seedlings compared to the control plants. Strain G16 produced indole acetic acid, siderophores and 1-aminocyclopropane-1-carboxylate deaminase. Increases in biomass production and total Pb uptake in the bacteria-inoculated plants were obtained compared to the control. The two strains could colonize the root interior and rhizosphere soil of rape after root inoculation.     

Zinc and copper uptake by plants under two transpiration rates. Part I. Wheat (Triticum aestivum L.)

To evaluate the environmental risk of irrigating crops with treated wastewater, an experiment was conducted using two growth chambers, each offering a different vapour pressure deficit (VPD) for high and low transpiration rates (TR), respectively. One of the two sets of 24 pots planted with 6 week old wheat (Triticum aestivum L.), was placed in each growth chamber, and irrigated in triplicates for 20 days with 8 Zn and Cu solutions (0 and 25 mg Zn/L combined with 0, 5, 15 and 30 mg Cu/L). Water losses from planted and non-planted pots served to measure evapo-transpiration and evaporation, respectively. Pots were monitored for Cu and Zn uptake by collecting three plants (shoot and grain)/pots after 0, 10 and 20 days, and roots in each pot after 20 days, and analyzing these plant parts for dry mass, and Cu and Zn levels. Transpiration rate was not affected by any Cu/Zn treatment, but Cu and Zn uptake increase with the time, irrigation solution level and higher TR, with the roots retaining most Cu and Zn, compared to the shoot followed by the grain. For the shoot and grain, Cu had a significant synergetic effect on Zn uptake, when Zn had slight but insignificant antagonistic effects on Cu uptake. For the roots, Cu and Zn had significant synergetic effect on each other. Regression equations obtained from the data indicate that Cu and Zn levels normally found in treated wastewater (0.08 mg/L) are 300 times lower than those used for the most concentrated experimental solutions (30 and 25 mg/L, respectively) and may, on a long term basis, be beneficial rather than toxic to wheat plants and do not acidify soil pH.     

Soil solution Zn and pH dynamics in non-rhizosphere soil and in the rhizosphere of Thlaspi caerulescens grown in a Zn/Cd-contaminated soil

Temporal changes in soil solution properties and metal speciation were studied in non-rhizosphere soil and in the rhizosphere of the hyperaccumulator Thlaspi caerulescens J. & C. Presl (population from Prayon, Belgium) grown in a Zn- and Cd-contaminated soil. This paper focuses on soil solution Zn and pH dynamics during phytoextraction. The concentration of Zn in both non-rhizosphere and rhizosphere soil solutions decreased from 23 mg/l at the beginning to 2 mg/l at the end of the experiment (84 days after transplanting of seedlings), mainly due to chemical sorption. There was no significant difference in overall Zn concentration between the planted and the unplanted soil solutions (P > 0.05). Soil solution pH decreased initially and then increased slightly in both planted and unplanted soil zones. From 60 to 84 days after transplanting, the pH of the rhizosphere soil solution was higher than that of non-rhizosphere soil solution (P<0.05). Zn uptake by the hyperaccumulator plants was 8.8 mg per pot (each containing 1 kg oven-dry soil) on average. The data indicate that the potential of T. caerulescens to remove Zn from contaminated soil may not be related to acidification of the rhizosphere.     

Survival, Pb-uptake and behaviour of three species of earthworm in Pb treated soils determined using an OECD-style toxicity test and a soil avoidance test

Mature (clitellate) Eisenia andrei Bouché (ultra epigeic), Lumbricus rubellus Hoffmeister (epigeic), and Aporrectodea caliginosa (Savigny) (endogeic) earthworms were placed in soils treated with Pb(NO(3))(2) to have concentrations in the range 1,000 to 10,000 mg Pb kg(-1). After 28 days LC50(-95%confidence limit)(+95%confidence limit) values were E. andrei 5824(-361)(+898) mg Pb kg(-1), L. rubellus 2867(-193)(+145) mg Pb kg(-1) and A. caliginosa2747(-304)(+239) mg Pb kg(-1) and EC50s for weight change were E. andrei2841(-68)(+150) mg Pb kg(-1), L. rubellus1303(-201)(+240) mg Pb kg(-1) and A. caliginosa1208(-206)(+212) mg Pb kg(-1). At any given soil Pb concentration, Pb tissue concentrations after 28 days were the same for all three earthworm species. In a soil avoidance test there was no difference between the behaviour of the different species. The lower sensitivity to Pb exhibited by E. andrei is most likely due to physiological adaptations associated with the modes of life of the earthworms, and could have serious implications for the use of this earthworm as the species of choice in standard toxicological testing.     

The role of endogenous nitric oxide in melatonin-improved tolerance to lead toxicity in maize plants

Melatonin (MT) and nitric oxide (NO) are known as scavengers of free radicals and an antioxidant against biotic and abiotic stresses in plant defense systems. However, whether NO interplays role in MT-induced antioxidant defense remains to be determined in the plants exposed to lead (Pb) toxicity. So, two experiments were designed to evaluate the role of NO in MT-mediated tolerance of maize plants to Pb stress. In the initial experiment, prior to starting different treatments, a solution of 0.05- or 0.10-mM MT was sprayed every other day for a period of 10 days to the leaves of maize plants exposed to Pb stress (0.1-mM PbCl₂). Pb toxicity significantly caused reduction in plant biomass (both fresh and dry), PSII maximum efficiency (F_v/F_m), total chlorophyll, leaf potassium (K), calcium (Ca), and leaf water potential, but it resulted in increased levels of proline, hydrogen peroxide (H₂O₂), malondialdehyde (MDA), electron leakage (EL), leaf Pb, and endogenous NO. An addition experiment was set up to further understand whether NO played role in mitigation of Pb toxicity in maize plants by MT using scavengers of NO and cPTIO combined with the MT treatments. MT-induced tolerance to Pb toxicity was totally eliminated by cPTIO by reversing endogenous NO. The present results clearly indicated that MT mediated the endogenous NO to improve tolerance of maize plants to Pb toxicity. This evidence was also supported by the increases of H₂O₂ and MDA and reduces some antioxidant enzyme activities tested as well as the plant growth inhibition and increased leaf Pb content by application of MT combined with cPTIO.

     

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.