Chemically enhanced phytoextraction of lead-contaminated soils

The effects of the combined application of soil fungicide (benomyl) and ethylenediaminetetraacetic acid (EDTA) on lead (Pb) phytoextraction by ryegrass (Lolium perenne) were examined. Twenty-five pots of Pb-contaminated soil (200 mg Pb kg(-1)) were seeded with ryegrass and randomly arranged into the following treatments: (1) Control, (2) benomyl, (3) EDTA, (4) benomyl and EDTA (B+E), and (5) benomyl followed by an application of EDTA 14 days later (B .. . E). Chemicals were applied when plants had reached maximum growth. Plants were analyzed for foliage Pb concentration using inductively coupled argon plasma (ICAP) spectrometry. The synergistic effects of the combined benomyl and EDTA application (treatments 4 and 5) were made evident by the significantly (p < 0.05) highest foliage Pb concentrations. However, the foliage dry biomass was significantly lowest for plants in treatments 4 and 5. The bioaccumulation factor (BF) and phytoextraction ratio (PR) were highest for plants in treatment 5 followed by plants in treatment 4.     

Augmented Phytoextraction of Lead (Pb2+)-Polluted Soils: A Comparative Study of the Effectiveness of Plant Growth Regulators,EDTA, and Plant Growth–Promoting Rhizobacteria

Abstract

The role of gibberellic acid (GA3), indole-3-acetic acid (IAA), plant growth–promoting bacteria (Rhizobium and Azotobacter), and a synthetic chelator (EDTA; ethylenediaminetetraacetic acid) in lead (Pb) phytoextraction was evaluated using Parthenium hysterophorus (dicot, unpalatable noncrop) and Zea mays (monocot food/forage crop) plants at the flowering stage. Various plant parts were analyzed by atomic absorption/flame spectrophotometer for their Pb content. Both plant growth regulators and both growth-promoting bacteria significantly increased the plant growth in Pb-polluted soils, whereas EDTA significantly decreased growth and biomass of both plants. EDTA increased the Pb uptake (μg g^?1 dry biomass), but the total plant Pb accumulation was decreased. GA3 and IAA significantly increased both uptake and translocation, and the maximum total Pb in the entire plant of Parthenium was found with GA3 foliar spray, whereas in Z. mays the total Pb was maximum in the plant treated with GA3 in combination with EDTA, followed by the GA3 foliar spray treatment. Overall, the GA3 foliar application showed superior response compared with all other treatments. Further research is recommended to observe the role of endogenous GA3 levels in correlation with metal phytoextraction in different plants.     

EDTA-Facilitated Phytoremediation of Soil with Heavy Metals from Sewage Sludge

The objective of this research was to determine the effect of the chelate EDTA (ethylenediaminetetraacetic acid), which is used in phytoremediation, on plant availability of heavy metals in liquid sewage sludge applied to soil. Sunflower (Helianthus annuus L.) was grown under greenhouse conditions in a commercial potting soil; the tetrasodium salt of EDTA (EDTA Na₄) was added at a rate of 1 g kg^-1 to half the pots. Immediately after seeds were planted, half of the pots with each soil (with or without EDTA) were irrigated with 60 ml sludge, and half were irrigated with 60 ml tap water. For the subsequent five irrigations, plants in soil with EDTA received either sludge or tap water containing 0.5 g EDTA Na₄ per 1000 ml, and plants in soil without EDTA received sludge or tap water without EDTA. Of the four heavy metals whose extractable concentrations in the soil were measured (Cu, Fe, Mn, and Zn), only Zn had a higher concentration in sludge-treated soil with EDTA compared to sludge-treated soil without EDTA. The concentrations of Fe, Cu, and Mn were similar in sludge-treated soil with and without EDTA. Of the three heavy metals whose total concentrations in the soil were measured (Cd, Pb, Cr), Pb (<10 mg kg^-1) and Cd (< 1 mg kg^-1) were below detection limits, and Cr was unaffected by treatment. The concentration of all measured elements in plants (Cd, Cu, Fe, Zn, Pb) was higher than the concentrations measured in the soil. With no EDTA, sludge-treated plants had a higher concentration of the five heavy elements than plants grown without sludge. Cadmium was lower in sludge-treated plants with EDTA than plants with EDTA and no sludge. After treatment with EDTA, the concentrations of Cu, Fe, and Zn were similar in plants with and without sludge. Lead was higher in plants with EDTA than plants without EDTA, showing that EDTA can facilitate phytoremediation of soil with Pb from sewage sludge.     

Effects of cadmium and lead on ferric chelate reductase activities in sugar beet roots

The effects of the heavy metals Cd and Pb on the activity of the enzyme ferric chelate reductase (FC-R, E.C. 1.6.99.13) have been studied in excised sugar beet root tips. The activity of this enzyme is markedly increased by iron deficiency. Metals were used as chloride salts or chelated with EDTA, and chemical speciation was carried out to predict the metal chemical species in equilibrium both in the ferric reductase assay and in the nutrient solutions. Three different heavy metal treatments were used. First, effects of Cd and Pb on the functioning of the FC-R were assessed in Fe-deficient plants, by including metals in the enzyme assay medium only. Results indicate that 50 μM CdCl₂ or Cd-EDTA did not affect FC-R activities even when assay time was as long as 2 h, whereas Pb slightly decreased enzyme activity only at concentrations of 2 mM. Second, short-time Cd and Pb pre-treatments (30–60 min) were imposed on intact Fe-deficient plants before carrying out the assay of FC-R activity. These short-term treatments induced significant decreases in the FC-R activities previously induced by Fe deficiency. With Cd, effects were more pronounced at higher concentrations, and they were stronger when Cd was in the free ion form than when present in the form of Cd-EDTA chelate. Third, prolonged Cd and Pb treatments were imposed on plants grown on 45 μM Fe-EDTA to assess the long-term effects of heavy metals on the induction of the FC-R enzyme. These long-term heavy metal treatments caused a significant increase in the root FC-R activities, indicating that Cd and Pb induce a deficiency in Fe in sugar beet that in turn elicits FC-R activity. The increases, however, are not as large as those found in total absence of Fe.     

Comparison of EDTA- and Citric Acid-Enhanced Phytoextraction of Heavy Metals in Artificially Metal Contaminated Soil by Typha Angustifolia

A pot experiment was conducted to study the performance of EDTA and citric acid (CA) addition in improving phytoextraction of Cd, Cu, Pb, and Cr from artificially contaminated soil by T. angustifolia. T. angustifolia showed the remarkable resistance to heavy metal toxicity with no visual toxic symptom including chlorosis and necrosis when exposed to metal stress. EDTA-addition significantly reduced plant height and biomass, compared with the control, and stunted plant growth, while 2.5 and 5 mM CA addition induced significant increases in root dry weight. EDTA, and 5 and 10 mM CA significantly increased shoot Cd, Pb, and Cr concentrations compared with the control, with EDTA being more effective. At final harvest, the highest shoot Cd, Cr, and Pb concentrations were recorded in the treatment of 5 mM EDTA addition, while maximal root Pb concentration was found at the 2.5 mM CA treatment. However, shoot Cd accumulation in the 10 mM CA treatment was 36.9% higher than that in 2.5 mM EDTA, and similar with that in 10 mM EDTA. Shoot Pb accumulation was lower in 10 mM CA than that in EDTA treatments. Further, root Cd, Cu, and Pb accumulation of CA treatments and shoot Cr accumulation in 5 or 10 mM CA treatments were markedly higher than that of control and EDTA treatments. The results also showed that EDTA dramatically increased the dissolution of Cu, Cr, Pb, and Cd in soil, while CA addition had less effect on water-soluble Cu, Cr, and Cd, and no effect on Pb levels. It is suggested that CA can be a good chelator candidate for T. angustifolia used for environmentally safe phytoextraction of Cd and Cr in soils.     

The role of root damage in the chelate-enhanced accumulation of lead by Indian mustard plants

In the present study, increasing ethylenediaminetetraacetic acid (EDTA) concentration from 0 to 0.5 mmol L(-1) resulted in progressive increases in root elongation and in shoot and root dry matter (DM) of Indian mustard seedlings (Brassica juncea. L.) exposed at 0.5 mmol L(-1) of lead (Pb). The highest concentration of Pb in the shoots of Indian mustard reached 1140 mg kg(-1) dry weight (DW) in the treatment with 0.5 mmol L(-1) of Pb + 0.25 mmol L(-1) of EDTA. A significantly positive correlation was found between the concentrations of Pb and EDTA in the shoots of mustard. Roots were pretreated with an MC (methanol:trichloromethane) solution, 0.1 mol L(-1) of HCl, and 65 degrees C hot water. The plants were then exposed to 0.5 mmol L(-1) of Pb + 3 mmol L(-1) of EDTA in solution for 2 d. The pretreatments with MC, HCl, and hot water all increased the concentration of Pb in shoots by 14-, 7-, and 15-fold, respectively, compared with the shoots that had not been pretreated. Therefore, some physiological damage to roots would be useful to enhance the uptake of metal by plants and to minimize the application of doses of chelates in the practical operation of chelate-assisted phytoremediation.     

Effect of arbuscular mycorrhizal fungi on phytoextraction by corn (Zea mays) of lead-contaminated soil

The role of arbuscular mycorrhizal fungi (AMF) in lead (Pb) uptake by corn (Zea mays) grown in soil supplemented with Pb was examined. Plants were subjected to four Pb levels: 0 (control); 10 (low); 100 (medium); and 500 mg L(-1) (high). At each Pb level, plants were grown in soil without and with fungicide (benomyl) (20 mg kg(-1)) to suppress AMF activity. Benomyl significantly reduced AMF colonization at high. medium, and zero Pb exposures. Benomyl application resulted in significantly lower concentrations of phosphorus in leaves at low and medium Pb exposures. The benomyl-treated plants had higher Pb and manganese concentrations in leaves than plants not treated with benomyl. In addition, benomyl-treated plants had generally lower concentrations of zinc and copper in leaves than plants not treated with benomyl. These results suggest that the role of AMF in heavy metal uptake is metal specific. Based on this work, the use of benomyl on soils contaminated with Pb can be recommended in phytoextraction.     

Lead uptake,toxicity and accumulation in <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> plants

The effects of lead were investigated in bean plants (Phaseolus vulgaris L. cv. Zlota Saxa) grown hydroponically in nutrient solution and exposed to Pb(NO₃)₂ (0.1, 0.5, 1 mM) with or without equimolar concentrations of chelator ethylenediaminetetraacetic acid (EDTA). The roots treated only with Pb(NO₃)₂ accumulated up to 25 g(Pb) kg⁻¹(d.m.), during 4-d exposure. However, in bean plants exposed to 0.5 mM Pb + 0.5 mM EDTA or 1 mM Pb + 1 mM EDTA 2.5 times less Pb was determined. In bean plants treated only with Pb, less than 6 % of total lead accumulated was transported to the aboveground parts, while in the case of plants grown with Pb + EDTA, around 50 % of total Pb was transported to the shoots.     

Mechanisms of microbially enhanced Fe acquisition in red clover (Trifolium pratense L.)

Soil microorganisms may play an important role in plant Fe uptake from soils with low Fe bioavailability, but there is little direct experimental evidence to date. We grew red clover, an Fe-efficient leguminous plant, in a calcareous soil to investigate the role of soil microbial activity in plant Fe uptake. Compared with plants grown in non-sterlie (NS) grown plants, growth and Fe content of the sterile(s) grown plants was significantly inhibited, but was improved by foliar application of Fe EDTA, indicating that soil microbial activity should play an important role in plant Fe acquisition. When soil solution was incubated with phenolic root exudates from Fe-deficient red clover, a few microbial species thrived while growth of the rest was inhibited, suggesting that the Fe-deficient (-Fe) root exudates selectively influenced the rhizosphere's microbial community. Eighty six per cent of the phenolic-tolerant microbes could produce siderophore [the Fe(III) chelator] under -Fe conditions, and 71% could secrete auxin-like compounds. Interestingly, the synthetic and microbial auxins (MAs) significantly enhanced the Ferric reduction system, suggesting that MAs, in addition to siderophores, are important to plant Fe uptake. Finally, plant growth and Fe uptake in sterilized soil were significantly increased by rhizobia inoculation. Root Fe-EDTA reductase activity in the -Fe plant was significantly enhanced by rhizobia infection, and the rhizobia could produce auxin but not siderophore under Fe-limiting conditions, suggesting that the contribution of nodulating rhizobia to plant Fe uptake can be at least partially attributed to stimulation of turbo reductase activity through nodule formation and auxin production in the rhizosphere. Based on these observations, we propose as a model that root exudates from -Fe plants selectively influence the rhizosphere microbial community, and the microbes in turn favour plant Fe acquisition by producing siderophores and auxins.     

不同pH值下丛枝菌根真菌对枳生长及铁吸收的影响

摘要：【目的】本文对营养液不同pH值下丛枝菌根(arbuscular mycorrhiza)真菌地表球囊霉（Glomus versiforme）对枳[Poncirus trifoliata]实生苗生长及植株铁营养状况的影响进行了初步研究。【方法】采用盆栽砂培试验,分别施浇pH 5.0、6.0、7.0和8.0的霍格兰营养液（含50 μM Fe-EDTA）;常规方法测定植株生长指标;曲利苯蓝染色法测定菌根侵染率;分光光度法测定叶绿素含量和根系三价铁螯合物还原酶活性;原子吸收分光光度法测定叶片钾和活性铁含量;钒     

Accumulation and distribution of iron, cadmium, lead and nickel in cucumber plants grown in hydroponics containing two different chelated iron supplies

Cucumber plants grown in hydroponics containing 10 μM Cd(II), Ni(II) and Pb(II), and iron supplied as Fe(III) EDTA or Fe(III) citrate in identical concentrations, were investigated by total-reflection X-ray fluorescence spectrometry with special emphasis on the determination of iron accumulation and distribution within the different plant compartments (root, stem, cotyledon and leaves). The extent of Cd, Ni and Pb accumulation and distribution were also determined. Generally, iron and heavy-metal contaminant accumulation was higher when Fe(III) citrate was used. The accumulation of nickel and lead was higher by about 20% and 100%, respectively, if the iron supply was Fe(III) citrate. The accumulation of Cd was similar. In the case of Fe(III) citrate, the total amounts of Fe taken up were similar in the control and heavy-metal-treated plants (27-31 μmol/plant). Further, the amounts of iron transported from the root towards the shoot of the control, lead- and nickel-contaminated plants were independent of the iron(III) form. Although Fe mobility could be characterized as being low, its distribution within the shoot was not significantly affected by the heavy metals investigated.     

Iron-Stress Induced Redox Activity in Tomato (Lycopersicum esculentum Mill.) Is Localized on the Plasma Membrane

Tomato plants (Lycopersicum esculentum Mill.) were grown for 21-days in a complete hydroponic nutrient solution including Fe³⁺-ethylenediamine-di(o-hydroxyphenylacetate) and subsequently switched to nutrient solution withholding Fe for 8 days to induce Fe stress. The roots of Fe-stressed plants reduced chelated Fe at rates sevenfold higher than roots of plants grown under Fe-sufficient conditions. The response in intact Fe-deficient roots was localized to root hairs, which developed on secondary roots during the period of Fe stress. Plasma membranes (PM) isolated by aqueous two-phase partitioning from tomato roots grown under Fe stress exhibited a 94% increase in rates of NADH-dependent Fe³⁺-citrate reduction compared to PM isolated from roots of Fe-sufficient plants. Optimal detection of the reductase activity required the presence of detergent indicating structural latency. In contrast, NADPH-dependent Fe³⁺-citrate reduction was not significantly different in root PM isolated from Fe-deficient versus Fe-sufficient plants and proceeded at substantially lower rates than NADH-dependent reduction. Mg²⁺-ATPase activity was increased 22% in PM from roots of Fe-deficient plants compared to PM isolated from roots of Fe-sufficient plants. The results localized the increase in Fe reductase activity in roots grown under Fe stress to the PM.     

EFFECT OF INDOLE-3-ACETIC ACID,KINETIN, AND ETHYLENEDIAMINETETRAACETIC ACID ON PLANT GROWTH AND UPTAKE AND TRANSLOCATION OF LEAD,MICRONUTRIENTS, AND MACRONUTRIENTS IN ALFALFA PLANTS

Alfalfa plants germinated and grown for 15 d in soil containing 80 mg Pb kg^?1 were treated with ethylenediaminetetraacetic acid (EDTA) at 0.8 mM and indole-3-acetic acid-kinetin (IAA-KN) at 100 μM. Fifteen days after the treatment application, the concentration of lead (Pb), macronutrients, and micronutrients was determined using inductively coupled plasma/optical emission spectroscopy. The chlorophyll content and plant growth were also measured. Roots of plants exposed to Pb alone, Pb–EDTA, and Pb–EDTA-IAA-KN had 160, 140, and 150 mg Pb kg^?1 DW, respectively. Pb was not detected in the stems of plants exposed to Pb alone; however, stems of plants treated with EDTA and EDTA–IAA-KN had 78 and 142 mg Pb kg^?1 DW, respectively. While the Pb concentration in leaves of plants treated with EDTA and EDTA–IAA-KN was 92 and 127 mg kg^?1 DW, respectively. In addition, EDTA and EDTA–IAA-KN significantly increased the translocation of zinc and manganese to leaves. The x-ray absorption spectroscopic studies demonstrated that Pb(II) was transported from roots to leaves without a change in the oxidation state.     

Biofertilizers and/or some micronutrients role on wheat plants grown on newly reclaimed soil

Pot experiment was conducted to study the effect of biofertilizers (inoculation with different bacterial isolates), foliar spraying with some micronutrients (Mn, Zn, Fe and Mn+Zn+Fe) and their interaction on growth, physiological parameters and nutrients content of wheat plants grown on reclaimed soil. Pot experiment was conducted in the greenhouse of National Research center, The experimental design was split plot with four replicates. Four biofertilizer treatments (un‐inoculated, Bacillus polymyxa, Azotobacter chroococcum or Azosprillium barasilense) were used and randomly distributed in the main pots. The foliar treatments with micronutrients were randomly distributed in the sub plots. The growth parameters (plant height, leaf area, roots, shoots and whole plant dry weights and number of tillers & leaves per plant); some physiological parameters (soluble sugar %, protein %, polysaccharide %, chl. A+b μg cm^?1 leaf per plant, carotenoids μg g^?1, IAA mg kg^?1 and psll mol DCPIP reduced per mg chl. per h) and nutrient contents (N, P, K, Mg, Mn, Zn and Cu) of wheat plants were significantly increased by inoculating wheat grains with different bacteria as compared with un‐inoculated plants (control). The highest values of all the mentioned parameters were obtained by using Azospirillum brasilense followed by Azotobacter chroococcum and Bacillus polymyxa in decreasing order. Foliar spraying treatments significantly increased the growth parameters, physiological parameters as well as nutrients content of wheat plants as compared with control. Highest values were obtained by using (Mn+Fe+Zn) treatment followed by Zn, Fe and Mn in decreasing order. Micronutrients in wheat plants differed as the foliar treatments were differed, so application of any micronutrient individually significantly increased its content and enhanced the content of other micronutrients in wheat. Interaction between the used biofertilizers and foliar spraying with micronutrients significantly affected all the studied parameters of wheat plants, the highest were obtained by inoculating wheat grains with Azospirillum brasilense and spraying the plants with (Mn+Fe+Zn) treatment, while the lowest values were attained by un‐inoculated grains (control) and spraying the wheat plants with tap water (control). Effective microorganisms in combination with micronutrients could be recommended to farmers to lead higher wheat yield.     

Investigation of Arbuscular mycorrhizal Fungus and EDTA Efficiencies on Lead Phytoremediation by Sunflower in a Calcareous Soil

Lead (Pb) contamination of soils is a widespread problem. Mycorrhizal inoculation and synthetic chelators such as ethylenediaminetetraacetic acid (EDTA) may be useful for improving phytoremediation efficiency in Pb-contaminated soils. A greenhouse experiment was performed to study the influence of inoculation with arbuscular mycorrhizal fungus (AMF), Glomus mosseae, and addition of EDTA on phytoremediation of Pb by sunflowers (Helianthus annuus) in a calcareous soil. The experiment was a completely randomized design in a factorial arrangement with five levels of Pb, two levels of mycorrhizal treatments, and two levels of EDTA. Inoculation increased root colonization as Pb levels increased, but the addition of EDTA decreased it. Shoot and root dry matter yields increased by inoculation; however, they decreased with EDTA and Pb levels in co-application treatments. Pb concentration in shoots was significantly higher than that in roots, indicating a translocation factor greater than 1. Inoculation or addition of EDTA significantly increased Pb in roots and its translocation to shoots. The uptake index (UI) value increased in co-application of EDTA and AMF and the individual application of them; it is, therefore, concluded that both AMF and EDTA are effective in phytoremediation of Pb by sunflowers in the studied soil.     

Effects of soil treatments and cultivars on 'early dying' disease of potatoes caused by Globodera rostochiensis and Verticilliurn dahliae

Plots in a field infested with Globodera rostochiensis and Verticillium dahliae were treated in 1971 with methyl bromide, dazomet, aldicarb, benomyl or aldicarb + benomyl and potatoes (cv. Pentland Dell) were grown in four consecutive years. In 1971, all treatments delayed the development of haulm symptoms, decreased soil populations of G. rostochiensis and increased yields. In the second year yields were increased by all treatments except dazomet whereas in the third and fourth years only benomyl or aldicarb + benomyl increased yields. Soil populations of V. dahliae were decreased by methyl bromide in 1971 but not by other treatments or in later years. In 1976, Pentland Dell and Pentland Crown were grown on an infested field in plots fumigated with methyl bromide. Fumigation slightly delayed shoot emergence but increased plant height, ground cover and the size and persistence of the leaves; development of haulm symptoms was delayed and yield increased. Fumigating with methyl bromide at five times the normal rate was more beneficial to Pentland Dell than to Pentland Crown. In a comparison of 10 cultivars in 1975, symptoms developed sooner and yields were smaller on a field infested with G. rostochiensis and V. dahliae than on a non-infested field and on both sites Maris Peer plants died soonest and yielded least. Pentland Crown yielded most on the non-infested field and Maris Piper on the infested field.     

GA3与EDTA强化黑麦草修复Pb污染土壤及其解毒机制

采用盆栽土培方法研究了生长调节剂GA₃和重金属螯合剂EDTA强化黑麦草修复Pb外源浓度分别为250和500 mg·kg^-1的污染土壤以及黑麦草Pb解毒机制.结果表明： 细胞壁沉积和液泡区室化在黑麦草地上部分Pb的解毒中起重要作用.EDTA单独使用提高了植物体内Pb的浓度和Pb在细胞可溶组分和细胞器组分中的比例,增强了Pb对植物的毒害作用,从而使植物生物量显著下降(P<0.05).叶面喷洒低浓度GA₃(1或10 μmol·L^-1)显著促进黑麦草对Pb的富集,但Pb在细胞器组分中的比例降低,缓解了Pb对植物细胞的伤害,从而促进了黑麦草的生长(P<0.05),其中1 μmol·L^-1 GA₃作用最显著.100 μmol·L^-1 GA₃降低了黑麦草体内Pb浓度,但Pb在细胞可溶组分和细胞器组分中的比例提高,使黑麦草的生物量明显低于对照.低浓度GA₃可在一定程度上缓解Pb和/或EDTA对黑麦草的毒害,生物量表现为低浓度GA₃ 单独>低浓度GA₃+EDTA>EDTA单独使用.低浓度GA₃和EDTA联用时的协同作用强化了黑麦草对Pb的富集,其中,在外源Pb浓度为500 mg·kg^-1处理组中,EDTA+1 μmol·L^-1 GA₃使黑麦草地上部分Pb浓度和提取效率分别达到1250.6 mg·kg^-1和1.1%.因此,1 μmol·L^-1 GA₃与EDTA联用在强化Pb污染土壤植物修复中具有很好的应用潜力.     

Chelator-enhanced lead accumulation in Agropyron elongatum cv Szarvasi-1 in hydroponic culture

Hydroponic culture was applied to compare the efficiency of K2EDTA and citrate in mobilizing Pb for accumulation in Agropyron elongatum cv. Szarvasi-1 and their effects on some physiological characteristics of the plants. The plants were grown in nutrient solutions containing 0, 10, and 100 microM Pb(NO3)2 combined with chelating agents added to the nutrient solutions after 21 days of growth, in 3 concentrations (0, 100, and 500 microM). The effects were measured after eight days. The energy grass proved to be greatly resistant to the treatments, as was reflected in the slight inhibition of growth, the resistance of the photosynthetic electron transport chain and the chlorophyll composition and the lack of change in the malone-dialdehyde content. Fundamental differences can be identified between the effects of EDTA and citrate. Citrate had only a little effect on the physiological parameters, which may be due to the strongly increasing lead accumulation with increasing concentration of Pb in the nutrient solution. Additionally, citrate ensured a higher biomass yield with higher shoot Pb accumulation compared to EDTA in almost all treatments. Concerning biomass reduction, 10 microM Pb applied together with K2EDTA has the most deleterious effects on energy grass. The effects correlated with the concentration of EDTA.     

The effect of EDTA and EDDS on lead uptake and localization in hydroponically grown Pisum sativum L.

Pisum sativum plants were treated for 3 days with an aqueous solution of 100 μM Pb(NO₃)₂ or with a mixture of lead nitrate and ethylenediaminetetraacetic acid (EDTA) or [S,S]-ethylenediaminedisuccinic acid (EDDS) at equimolar concentrations. Lead decline from the incubation media and its accumulation and localization at the morphological and ultrastructural levels as well as plant growth parameters (root growth, root and shoot dry weight) were estimated after 1 and 3 days of treatment. The tested chelators, especially EDTA, significantly diminished Pb uptake by plants as compared to the lead nitrate-treated material. Simultaneously, EDTA significantly enhanced Pb translocation from roots to shoots. In the presence of both chelates, plant growth parameters remained considerably higher than in the case of uncomplexed Pb. Considerable differences between the tested chelators were visible in Pb localization both at the morphological and ultrastructural level. In Pb+EDTA-treated roots, lead was mainly located in the apical parts, while in Pb+EDDS-exposed material Pb was evenly distributed along the whole root length. Transmission electron microscopy and EDS analysis revealed that in meristematic cells of the roots incubated in Pb+EDTA, large electron-dense lead deposits were located in vacuoles and small granules were rarely noticed in cell walls or cytoplasm, while after Pb+EDDS treatment metal deposits were restricted to the border between plasmalemma and cell wall. Such results imply different ways of transport of those complexed Pb forms.     

Solubility and accumulation of metals in Chinese brake fern, vetiver and rostrate sesbania using chelating agents

Greenhouse experiments were conducted to study the effects of chelating agents on the growth and metal accumulation of Chinese brake fern (Pteris vittata L.), vetiver (Vetiveria zizanioides L.), and rostrate sesbania (Sesbania rostrata L.) in soil contaminated with arsenic (As), Cu, Pb, and Zn. Among the five chelating agents used [ethylenediaminetriacetic acid (EDTA), hydroxyethylenediaminetriacetic acid (HEDTA), nitrilotriacetic acid (NTA), oxalic acid (OA), and phytic acid (PA)], OA was the best to mobilize As, EDTA to mobilize Cu and Pb, and HEDTA to mobilize Zn from soil, respectively. The biomass of vetiver was the highest, followed by rostrate sesbania. All chelating agents inhibited the growth of Chinese brake fern and rostrate sesbania, but HEDTA significantly increased the aboveground biomass of vetiver. Dry weights of both Chinese brake fern and rostrate sesbania decreased with increasing EDTA concentrations amended in the soil, especially in treatments with high EDTA concentrations. EDTA and HEDTA enhanced Cu, Zn, and Pb, but lowered As accumulation in all three plant species, except for As in vetiver, while OA significantly enhanced As accumulation in the aboveground part of vetiver. Concentrations of Cu, Zn, and Pb in the aboveground parts of plants increased significantly with the increase of EDTA concentrations and treatment time. In addition to As, Chinese brake fern also accumulated the highest Cu, Pb, and Zn in its aboveground parts among the three plant species grown in metal-contaminated soil with EDTA/HEDTA treatments. This species, therefore, can be used to simultaneously clean up As, Cu, Pb, and Zn from contaminated soils with the aid of EDTA or HEDTA.