EDTA-enhanced phytoremediation of lead-contaminated soil by the halophyte Sesuvium portulacastrum

The low bioavailability of Pb and low number of Pb-tolerant plant species represent an important limitation for Pb phytoextraction. It was recently suggested that halophyte plant species may be a promising material for this purpose, especially in polluted salt areas while Pb mobility may be improved by synthetic chelating agents. This study aims to evaluate Pb extraction by the halophyte Sesuvium portulacastrum in relation to the impact of EDTA application. Seedling were cultivated during 60 days on Pb artificially contaminated soil (200, 400, and 800 ppm Pb) in the presence or in the absence of EDTA (3 g kg^?1 soil). Results showed that upon to 400 ppm, Pb had no impact on plant growth. However, exogenous Pb induce a decrease in shoot K⁺ while it increased shoot Mg²⁺ and had no impact on shoot Ca²⁺ concentrations. Lead concentration in the shoots increased with increasing external Pb doses reaching 1,390 ppm in the presence of 800 ppm lead in soil. EDTA addition had no effect on plant growth but strongly increased Pb accumulation in the shoot which increased from 1,390 ppm in the absence of EDTA to 3,772 ppm in EDTA-amended plants exposed to 800 ppm exogenous Pb. Both Pb absorption and translocation from roots to shoots were significantly enhanced by EDTA application, leading to an increase in the total amounts of extracted Pb per plant. These data suggest that S. portulacastrum is very promising species for decontamination of Pb²⁺-contaminated soil and that its phytoextraction potential was significantly enhanced by addition of EDTA to the polluted soil.     

The effect of EDDS and citrate on the uptake of lead in hydroponically grown Matthiola flavida

Root and shoot lead concentrations and the impact of chelating agents on these were investigated in two populations of the novel metallophyte Matthiola flavida. Plants were exposed in hydroponics to Pb(NO₃)₂, supplied alone, or in combination with citric acid, or EDDS. When supplied at concentrations expected to bind about 95% of the Pb in a solution containing 1-μM Pb (1000 μM citrate or 3.1 μM EDDS, respectively), the root and shoot Pb concentrations were dramatically lowered, in comparison with a 1-μM free ionic Pb control exposure. A 1-mM EDDS + 1-μM Pb treatment decreased the plants’ Pb concentrations further, even to undetectable levels in one population. At 100 μM Pb in a 1-mM EDDS-amended solution the Pb concentration increased strongly in shoots, but barely in roots, in comparison with the 1-μM Pb + 1-mM EDDS treatment, without causing toxicity symptoms. Further increments of the Pb concentration in the 1-mM EDDS-amended solution, i.e. to 800 and 990 μM, caused Pb hyperaccumulation, both in roots and in shoots, associated with a complete arrest of root growth and foliar necrosis. M. flavida seemed to be devoid of constitutive mechanisms for uptake of Pb-citrate or Pb-EDDS complexes. Hyperaccumulation of Pb-EDDS occurred only at high exposure levels. Pb-EDDS was toxic, but is much less so than free Pb. Free EDDS did not seem to be toxic at the concentrations tested.     

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.     

Effect of transpiration on Pb uptake by lettuce and on water soluble low molecular weight organic acids in rhizosphere

The effect of transpiration (high and low) on Pb uptake by leaf lettuce and on water soluble low molecular weight organic acids (LMWOAs) in rhizosphere has been studied. After two weeks of growth the plants were cultured in greenhouse for more four weeks and two days. Pb(NO(3))(2) solutions of different concentrations (100, 200, and 300 mg l(-1) of Pb) were then added to the quartz sand pots of different plants and studies were initiated. Blank experiments (without treating the quartz sand pots with Pb(NO(3))(2) solutions) were also run in parallel. No significant differences in the growth of the plants with the concentrations of added Pb(NO(3))(2) solutions were observed by both low and high transpirations at the end of the 0, 3rd, and 10th days of studies. The total evaporation of the volatiles during 10 days did not depend on the concentration of Pb(2+) but with high transpiration the rate of evaporation was significantly higher than with low transpiration. Uptake of Pb by shoots and roots of the plants was found to be proportional to the concentration of various Pb(NO(3))(2) solutions added and more accumulation was observed in roots than in shoots at the end of 3rd and 10th days. High transpiration created more Pb uptake than low transpiration did. One volatile acid, propionic acid and nine non-volatile acids, lactic, glycolic, oxalic, succinic, fumaric, oxalacetic, D-tartaric, trans-aconitic, and citric acids in rhizosphere quartz sands were identified and quantified by gas chromatography (GC) analysis. D-Tartaric and citric acids were major among the non-volatile acids. The amount of LMWOAs in rhizosphere quartz sands increased with the higher amount of Pb uptake and also with the duration of studies. The total quantities of the LMWOAs in the rhizosphere quartz sands were significantly higher under high transpiration with 300 mg l(-1) Pb solution addition at the end of 10th day. The present study shows prominent correlation between transpiration and uptake of heavy metal and interesting correlation between Pb contaminated level and quantity of water soluble LMWOAs in rhizosphere quartz sands. The latter thus deserves of further studies.     

The role of citric acid on the phytoremediation of heavy metal contaminated soil

Adsorption and hydroponics experiments were conducted to study the role of citric acid on the phytoremediation of heavy metal contaminated soil. The results show that addition of citric acid decreased the adsorption of both lead and cadmium, such an effect was bigger for cadmium than for lead. The decrease in the adsorption of Pb and Cd was mainly due to a decrease of pH in the presence of citric acid. The presence of citric acid could alleviate the toxicity of Pb and Cd to radish, and stimulate their transportation from root to shoot. The studies of heavy metal forms using sequential extraction demonstrated that lead was mainly existed as FHAC (a lower bioavailable form) in the root, while F(HCl) was the dominant form in the leaf. The addition of citric acid to the soil changed the concentration and relative abundance of all the forms. The detoxifying effect of citric acid to Pb in shoots might result from the transformation of higher toxic forms into lower toxic forms. Cadmium was mainly present as F(NaCl), therefore, it had higher toxicity than lead. The addition of citric acid increased the abundance of F(H2O) + F(NaCl), indicating that citric acid treatment could transform cadmium into more transportable forms.     

Classification and identification of metal-accumulating plant species by cluster analysis

Identification and classification of metal-accumulating plant species is essential for phytoextraction. Cluster analysis is used for classifying individuals based on measured characteristics. In this study, classification of plant species for metal accumulation was conducted using cluster analysis based on a practical survey. Forty plant samples belonging to 21 species were collected from an ancient silver-mining site. Five groups such as hyperaccumulator, potential hyperaccumulator, accumulator, potential accumulator, and normal accumulating plant were graded. For Cd accumulation, the ancient silver-mining ecotype of Sedum alfredii was treated as a Cd hyperaccumulator, and the others were normal Cd-accumulating plants. For Zn accumulation, S. alfredii was considered as a potential Zn hyperaccumulator, Conyza canadensis and Artemisia lavandulaefolia were Zn accumulators, and the others were normal Zn-accumulating plants. For Pb accumulation, S. alfredii and Elatostema lineolatum were potential Pb hyperaccumulators, Rubus hunanensis, Ajuga decumbens, and Erigeron annuus were Pb accumulators, C. canadensis and A. lavandulaefolia were potential Pb accumulators, and the others were normal Pb-accumulating plants. Plant species with the potential for phytoextraction were identified such as S. alfredii for Cd and Zn, C. canadensis and A. lavandulaefolia for Zn and Pb, and E. lineolatum, R. hunanensis, A. decumbens, and E. annuus for Pb. Cluster analysis is effective in the classification of plant species for metal accumulation and identification of potential species for phytoextraction.     

Lead accumulation and association with Fe on Typha latifolia root from an urban brownfield site

Synchrotron X-ray microfluorescence and X-ray absorption near-edge microstructure spectroscopy techniques were applied to Typha latifolia (cattail) root sections and rhizosphere soils collected from a brownfield site in New Jersey to investigate lead (Pb) accumulation in T. latifolia roots and the role of iron (Fe) plaque in controlling Pb uptake. We found that Pb and Fe spatial distribution patterns in the root tissues are similar with both metals present at high concentrations mainly in the epidermis and at low concentrations in the vascular tissue (xylem and phloem), and the major Pb and Fe species in T. latifolia root are Pb(II) and Fe(III) regardless of concentration levels. The sequestration of Pb by T. latifolia roots suggests a potential low-cost remediation method (phytostabilization) to manage Pb-contaminated sediments for brownfield remediation while performing wetland rehabilitation.     

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.     

Effects of EDTA and low molecular weight organic acids on soil solution properties of a heavy metal polluted soil

A pot experiment was conducted to study the effects of EDTA and low molecular weight organic acids (LMWOA) on the pH, total organic carbon (TOC) and heavy metals in the soil solution in the rhizosphere of Brassica juncea grown in a paddy soil contaminated with Cu, Zn, Pb and Cd. The results show that EDTA and LMWOA have no effect on the soil solution pH. EDTA addition significantly increased the TOC concentrations in the soil solution. The TOC concentrations in treatments with EDTA were significantly higher than those in treatments with LMWOA. Adding 3 mmol kg(-1) EDTA to the soil markedly increased the total concentrations of Cu, Zn, Pb and Cd in the soil solution. Compared to EDTA, LMWOA had a very small effect on the metal concentrations. Total concentrations in the soil solution followed the sequence: EDTA > citric acid (CA) approximately oxalic acid (OA) approximately malic acid (MA) for Cu and Pb; EDTA > MA > CA approximately OA for Zn; and EDTA > MA > CA > OA for Cd. The labile concentrations of Cu, Zn, Pb and Cd showed similar trends to the total concentrations.     

Nitrate facilitates cadmium uptake, transport and accumulation in the hyperaccumulator Sedum plumbizincicola

The aims of this study are to investigate whether and how the nitrogen form (nitrate (NO₃ ^–) versus ammonium (NH₄ ⁺)) influences cadmium (Cd) uptake and translocation and subsequent Cd phytoextraction by the hyperaccumulator species Sedum plumbizincicola. Plants were grown hydroponically with N supplied as either NO₃ ^– or NH₄ ⁺. Short-term (36 h) Cd uptake and translocation were determined innovatively and quantitatively using a positron-emitting ¹⁰⁷Cd tracer and positron-emitting tracer imaging system. The results show that the rates of Cd uptake by roots and transport to the shoots in the NO₃ ^– treatment were more rapid than in the NH₄ ⁺ treatment. After uptake for 36 h, 5.6 (0.056 μM) and 29.0 % (0.290 μM) of total Cd in the solution was non-absorbable in the NO₃ ^– and NH₄ ⁺ treatments, respectively. The local velocity of Cd transport was approximately 1.5-fold higher in roots (3.30 cm h^?1) and 3.7-fold higher in shoots (10.10 cm h^?1) of NO₃ ^–- than NH₄ ⁺-fed plants. Autoradiographic analysis of ¹⁰⁹Cd reveals that NO₃ ^– nutrition enhanced Cd transportation from the main stem to branches and young leaves. Moreover, NO₃ ^– treatment increased Cd, Ca and K concentrations but inhibited Fe and P in the xylem sap. In a 21-day hydroponic culture, shoot biomass and Cd concentration were 1.51 and 2.63 times higher in NO₃ ^–- than in NH₄ ⁺-fed plants. We conclude that compared with NH₄ ⁺, NO₃ ^– promoted the major steps in the transport route followed by Cd from solution to shoots in S. plumbizincicola, namely its uptake by roots, xylem loading, root-to-shoot translocation in the xylem and uploading to the leaves. S. plumbizincicola prefers NO₃ ^– nutrition to NH₄ ⁺ for Cd phytoextraction.     

Changes of organic acid exudation and rhizosphere pH in rice plants under chromium stress.

The effect of chromium (Cr) stress on the changes of rhizosphere pH, organic acid exudation, and Cr accumulation in plants was studied using two rice genotypes differing in grain Cr accumulation. The results showed that rhizosphere pH increased with increasing level of Cr in the culture solution and with an extended time of Cr exposure. Among the six organic acids examined in this experiment, oxalic and malic acid contents were relatively higher, and had a significant positive correlation with the rhizosphere pH, indicating that they play an important role in changing rhizosphere pH. The Cr content in roots was significantly higher than that in stems and leaves. Cr accumulation in plants was significantly and positively correlated with rhizosphere pH, and the exudation of oxalic, malic and citric acids, suggesting that an increase in rhizosphere pH, and exudation of oxalic, malic and citric acid enhances Cr accumulation in rice plants.     

Analysis of phytochelatin complexes in the lead tolerant vetiver grass [Vetiveria zizanioides (L.)] using liquid chromatography and mass spectrometry

Ethylenediamene tetraacetic acid (EDTA) has been used to mobilize soil lead (Pb) and enhance plant uptake for phytoremediation. Chelant bound Pb is considered less toxic compared to free Pb ions and hence might induce less stress on plants. Characterization of possible Pb complexes with phytochelatins (PC_n, metal-binding peptides) and EDTA in plant tissues will enhance our understanding of Pb tolerance mechanisms. In a previous study, we showed that vetiver grass (Vetiveria zizanioides L.) can accumulate up to 19,800 and 3350 mg Pb kg⁻¹ dry weight in root and shoot tissues, respectively; in a hydroponics set-up. Following the basic incubation study, a greenhouse experiment was conducted to elucidate the efficiency of vetiver grass (with or without EDTA) in remediating Pb-contaminated soils from actual residential sites where Pb-based paints were used. The levels of total thiols, PC_n, and catalase (an antioxidant enzyme) were measured in vetiver root and shoot following chelant-assisted phytostabilization. In the presence of 15 mM kg ⁻¹ EDTA, vetiver accumulated 4460 and 480 mg Pb kg⁻¹ dry root and shoot tissue, respectively; that are 15- and 24-fold higher compared to those in untreated controls. Despite higher Pb concentrations in the plant tissues, the amount of total thiols and catalase activity in EDTA treated vetiver tissues was comparable to chelant unamended controls, indicating lowered Pb toxicity by chelation with EDTA. The identification of glutathione (referred as PC₁) (m/z 308.2), along with chelated complexes like Pb-EDTA (m/z 498.8) and PC₁-Pb-EDTA (m/z 805.3) in vetiver root tissue using electrospray tandem mass spectrometry (ES-MS) highlights the possible role of such species towards Pb tolerance in vetiver grass.     

The impact of EDTA on lead distribution and speciation in the accumulator Sedum alfredii by synchrotron X-ray investigation

The in vivo localization and speciation of lead (Pb) in tissues of the accumulator Sedum alfredii grown in EDTA-Pb and Pb(NO₃)₂ was studied by synchrotron X-ray investigation. The presence of EDTA-Pb in solution resulted in a significant reduction of Pb accumulation in S. alfredii. Lead was preferentially localized in the vascular bundles regardless of treatments but the intensities of Pb were lower in the plants treated with EDTA. Lead was predominantly presented as a Pb-cell wall complex in the plants regardless of its supply form. However, a relatively high proportion of Pb was observed as Pb-EDTA complex when the plant was treated with EDTA-Pb, but as a mixture of Pb₃(PO₄)₂, Pb-malic, and Pb-GSH when cultured with ionic Pb. These results suggest that EDTA does not increase the internal mobility of Pb, although the soluble Pb-EDTA complex could be transported and accumulated within the plants of S. alfredii.     

Ca(2+) -dependent plant response to Pb(2+) is regulated by LCT1

Tobacco plants transformed with TaLCT1 were cultured on Knop's medium with modified calcium concentrations (0.01-3 mM) in the presence of Pb(2+), and in soil contaminated by lead. A 4-5 microM Pb(2+) administered in the presence of 1 mM Ca(2+) inhibited the root growth of transgenic plants to much lesser degree than of control plants, whereas in the presence of 3mM Ca(2+) no differences were found between the studied lines. The reduction of Pb(2+) toxicity in the presence of 1 mM Ca(2+) was not accompanied by a change in the lead tissue concentration. However, when Ca(2+) level in the medium was lowered to 0.01 mM, several fold higher root/shoot Pb ratio in transgenic plants was observed, twofold increase in the total amount of metal accumulated, and lower concentration of Pb in the xylem sap. Results suggest the involvement of TaLCT1 in the regulation of Ca-dependent Pb-detoxification, and under conditions of low calcium in lead uptake and distribution.     

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.     

Determination of malic acid and other C4 dicarboxylic acids in atmospheric aerosol samples

An ion chromatographic method was developed which is able to separate five unsubstituted and hydroxy C4 dicarboxylic acids, succinic, malic, tartaric, maleic and fumaric acid, besides the other unsubstituted C2-C5 dicarboxylic acids, oxalic, malonic and glutaric acids, as well as inorganic ions in samples extracted from atmospheric particulate matter. By the application of this method it was found for both rural and urban sites and for various types of air masses that in the summer-time malic acid is the most prominent C4 diacid (64 ng m(-3) by average), exceeding succinic acid concentration (28 ng m(-3) by average) considerably. In winter-time considerably less, a factor of 4-15, C4 acids occurred and succinic acid was more concentrated than malic acid. Tartaric, fumaric and maleic acids were less concentrated (5.1, 5.0 and 4.5 ng m(-3) by average, respectively). Tartaric acid was observed for the first time in ambient air. The results indicate that in particular anthropogenic sources are important for the precursors of succinic, maleic and fumaric acids. Biogenic sources seem to influence the occurrence of malic acid significantly.     

Growth and metal accumulation in vetiver and two Sesbania species on lead/zinc mine tailings

The lead (Pb)/zinc (Zn) tailings contained high concentrations of heavy metals (total Pb, Zn, Cu and Cd concentrations 4164, 4377, 35 and 32 mg kg(-1), respectively), and low contents of major nutrient elements (N, P, and K) and organic matter. A field trial was conducted to compare growth performance, metal accumulation of Vetiver (Vetiveria zizanioides) and two legume species (Sesbania rostrata and Sesbania sesban) grown on the tailings amended with domestic refuse and/or fertilizer. It was revealed that domestic refuse alone and the combination of domestic refuse and artificial fertilizer significantly improved the survival rates and growth of V. zizanioides and two Sesbania species, especially the combination. However, artificial fertilizer alone did not improve both the survival rate and growth performance of the plants grown on tailings. Roots of these species accumulated similar levels of heavy metals, but the shoots of two Sesbania species accumulated higher (3-4 folds) concentrations of Pb, Zn, Cu and Cd than shoots of V. zizanioides. Most of the heavy metals in V. zizanioides were accumulated in roots, and the translocation of metals from roots to shoots was restricted. Intercropping of V. zizanioides and S. rostrata did not show any beneficial effect on individual plant species, in terms of height, biomass, survival rate, and metal accumulation, possibly due to the rather short experimental period of 5 months.     

Accumulation of Cd,Cu and Zn in shoots of maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) exposed to 0.8 or 20 nM Cd during vegetative growth and the relation with xylem sap composition

This work focuses on the exposure of maize plants to nanomolar concentrations of Cd, which is relevant for agricultural soils cropped with food and feed plants. Maize plants were cultivated in nutrient solution at 0.8 or 20 nM Cd during the vegetative growth stages. No significant hormesis or toxic effects of Cd were observed on maize growth, but a decrease in the allocation of Cd to shoots between the 0.8 and 20 nM Cd exposures revealed that the plants already responded to these low concentrations of Cd according to a shoot Cd excluder strategy. The Cd, Cu and Zn concentrations in shoots decreased with time as the result of an early decrease in the root/shoot ratio and of a decrease in the coefficient of allocation to aboveground for Zn and Cd at 20 nM. As a consequence, shoots of young plants were richer in micronutrients Cu and Zn but also in toxic Cd. The rate of delivery of Cd, Cu and Zn from xylem sap was successfully used to predict the time course of concentrations of Cd, Cu and Zn in the shoot. However, it overestimated the actual concentrations of Cd in the shoot, presumably because the reallocation of this trace element from shoots back to roots was not taken into account.     

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.