μM Pb Research Articles

Interactions between Lead on Nutrients (Ca2+, K+, N), and their Consequences on Growth in Sesuvium portulacastrum and Brassica juncea

Lead is one of the most toxic metals in the environment and causes drastic morphological and physiological deformities in plants. Growth restriction, chlorosis and necrosis are usually accompanied with a large disturbance of the uptake of essential elements. The aim of this work is to study the effects of lead on mineral nutrient acquisition (Ca2+, K+, N), and their consequences on growth in two different species: halophytic (Sesuvium portulacastrum) and glycophytic (Brassica juncea). Seedlings were grown for 21 days in split-root conditions. One half of the root system was immersed in complete nutrient solution supplemented with 400 μM Pb(NO2)3, and the other half was immersed in a Pb2+-free medium, containing all nutrients or deprived of potassium or calcium or nitrogen. Using this approach, we demonstrated that lead interfere mainly with the absorption of Ca2+ by the roots, thus limiting the growth of plants and their ability to accumulate this metal. We propose that the increase of Ca2+ availability in soils could improve the growth of both species in the presence of lead. This would be essential for improving their utility for phytoremediation of this metal in contaminated soils.

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.

Effect of Lead on Lipid Peroxidation, Phospholipids Composition, and Methylation in Erythrocyte of Human

Lead (Pb) is one of the most abundant heavy metals on earth considered as number one environmental persistent toxin and health hazard affecting millions of people in all age groups. After entering bloodstream, 99% of Pb is accumulated in erythrocytes and causes poisoning. Toxic Pb effects on erythrocytes membrane's composition of phosphatidyl serine (PS), phosphatidyl ethanolamine (PE), phosphatidyl choline (PC), and sphingomyelin (SM), and phospholipids transmethylation were determined. Lipid peroxidation in Pb-exposed erythrocytes was evaluated as malondialdehyde (MDA) formation in presence of Fe and vitamin E to understand severity of Pb toxicity and its mitigation. Pb (0.5-5.0 μM) degraded PS (12 to 31%, P < 0.05-0.001) and elevated SM (19-51%, P < 0.05-0.001). Composition of PC and PE were diminished (22%) and elevated (29%), respectively, with higher Pb exposure (5.0 μM, P < 0.001). Pb toxicity suppressed (P < 0.001) transmethylation of phospholipids in membranes (34, 41, and 50%, respectively, with 0.5, 2.5, and 5.0 μM). Pb-induced dose-related MDA production (P < 0.05-0.001) in erythrocytes was obtained, which was accentuated in presence of Fe (P < 0.05-0.001). The vitamin E mitigated (P < 0.05-0.01) the severity of Pb-induced lipid peroxidation. The ratio PS/SM showed maximum change of -27 (P < 0.01), -30 (P < 0.01), and -54% (P < 0.001), respectively at 0.5, 2.5, and 5.0 μM Pb exposures. Ratios PC/SM and PS/PE were at the second, whereas PE/PS at the third order. The study suggests that the mechanisms underlying distortion of compositional phospholipids, inhibition of transmethylation, and exasperated phospholipid peroxidative damage are the active phenomena of Pb toxicity in erythrocytes.

Excluder Strategies in Response to Pb Toxicity in Matricaria Chamomilla

Lead is among the most contaminant metals in ecosystem and has different effects on anatomical and morphological characters of different plant species. Matricaria chamomilla is an important medicinal plant which contains 120 chemical compositions. In this research, the effects of lead on production of apigenin as a significant medicinal substance were studied. Furthermore, structural changes of M. chamomilla were studied in different stages of development by electron and optic microscopes. So the plants in first stage of development, were transplanted to hydroponic culture with five-treatment design of 0, 60, 120, 180, 240 µM Pb. The plants under treatment were harvested in first stage of development, as well as in shooting and flowering stages for measurement of Pb absorption. According to this study, morphological changes were not visually observed during the treatment process in different stages; nevertheless, reduction in total biomass was detected in different stages compared to control plants. The results of SEM study of leaf indicated structural changes in stomata and epicuticular waxes in different stages. Also in anatomical studies we noticed several changes in cross section of leaf, stem and root. The obtained results indicated that in different stages, Pb accumulation in roots and shoots has increased under Pb stress during various stage of development. Moreover, Pb stress has no effect on apigenin content. Therefore, M. chamomilla is an excluder species which is tolerant to Pb stress.

Influence of EDTA and citric acid on lead-induced oxidative stress to Vicia faba roots

In spite of substantial advancement in recent past, the role of metal speciation in assessing biogeochemical behaviour of Pb is still topical. Organic ligands are capable to modify Pb speciation in nutrient/soil solution and in turn its soil–plant transfer and toxicity. In this sense, the main objective of this study was to evaluate the effect of organic ligands on Pb-induced oxidative stress to Vicia faba roots. V. faba seedlings grown to controlled hydroponic system were treated with 5 μM Pb as lead nitrate in the presence and absence of organic ligands viz ethylenediaminetetraacetic acid (EDTA) and citric acid (CA) for 1, 4, 8, 12, and 24 h. The chemical speciation of Pb (percent free and chelated Pb) in nutrient solution in the presence and absence of organic ligands was calculated using Visual Minteq speciation model. The effect of chemical speciation on Pb-induced oxidative stress to V. faba roots was investigated using plant enzymatic antioxidative system [superoxide dismutases (SOD), guaiacol peroxidise (GPX), ascorbate peroxidase (APX), glutathione reductase (GR), and catalase (CAT)]. The antioxidant enzymes activities were determined using ultraviolet spectrophotometer. The activities of SOD, GPX, APX, and GR significantly increased whereas that of CAT decreased in V. faba roots under Pb alone treatment. Lead-induced increase/decrease in antioxidant enzymes activities was not linear but varies with treatment exposure time. EDTA dose dependently inhibited Pb-induced changes in antioxidant enzymes activities. However, CA did not cause any significant change in Pb-induced variation in antioxidant enzymes activities, but delayed or slightly reduced the Pb effect. The present study suggested that physiological responses of V. faba roots to Pb toxicity vary with applied Pb form and duration of exposure. EDTA can inhibit Pb-induced toxicity to V. faba seedlings by forming stable Pb-EDTA complexes due to its high binding strength for Pb. However, CA had no effect on Pb-induced toxicity to V. faba roots due to weak complexation with Pb.

Physiological and morphological responses of Lead or Cadmium exposed Chlorella sorokiniana 211-8K (Chlorophyceae).

The heavy metal pollution in soils and aquatic environments is a serious ecological problem. In the green-microalga Chlorella sorokiniana 211-8K (Chlorophyceae) exposed to ions of lead (Pb) and cadmium (Cd) we studied the metabolic responses to the toxicity of these two heavy metals. Our data indicate that both the pollutants alter the alga cell ultrastructure and its physiological characteristics (growth, photosynthesis, respiration, enzyme activities). The toxic effects of the two metals resulted time-dependent to the exposure. After 24 h of treatment with 250 μM Pb or Cd, photosynthesis was inhibited until to 77 and 86%, however respiration was strongly enhanced up to 300 and 350%, respectively. In the algal cells Pb or Cd exposure induced a reduction in the content of the total chlorophylls and a decrease of the soluble protein levels, significantly compromising the growth, particularly in cultures cadmium-treated. We report data on ultrastructural changes induced by the two heavy metals; they affected overall chloroplast ultrastructure of the alga. Most importantly, the O-acetyl-L-serine(thiol)lyase (OASTL) activity was appreciably increased after only 2 h of Cd exposure, indicating the existence of a link between the metal contamination and cysteine synthesis. Then, Chlorella sorokiniana cells seem to better tolerate high concentrations of Pb while appear to be more sensitive to Cd ions. These results provide some additional information that can lead to better understand consequences of heavy metal poisoning in microalgae.

Are newborn rat-derived neural stem cells more sensitive to lead neurotoxicity?

Lead ion (Pb2+) has been proven to be a neurotoxin due to its neurotoxicity on mammalian nervous system, especially for the developing brains of juveniles. However, many reported studies involved the negative effects of Pb2+ on adult neural cells of humans or other mammals, only few of which have examined the effects of Pb2+ on neural stem cells. The purpose of this study was to reveal the biological effects of Pb2+ from lead acetate [Pb (CH3COO)2] on viability, proliferation and differentiation of neural stem cells derived from the hippocampus of newborn rats aged 7 days and adult rats aged 90 days, respectively. This study was carried out in three parts. In the first part, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT viability assay) was used to detect the effects of Pb2+ on the cell viability of passage 2 hippocampal neural stem cells after 48-hour exposure to 0–200 μM Pb2+. In the second part, 10 μM bromodeoxyuridine was added into the culture medium of passage 2 hippocampal neural stem cells after 48-hour exposure to 0–200 μM Pb2+, followed by immunocytochemical staining with anti-bromodeoxyuridine to demonstrate the effects of Pb2+ on cell proliferation. In the last part, passage 2 hippocampal neural stem cells were allowed to grow in the differentiation medium with 0–200 μM Pb2+. Immunocytochemical staining with anti-microtubule-associated protein 2 (a neuron marker), anti-glial fibrillary acidic protein (an astrocyte marker), and anti-RIP (an oligodendrocyte marker) was performed to detect the differentiation commitment of affected neural stem cells after 6 days. The data showed that Pb2+ inhibited not only the viability and proliferation of rat hippocampal neural stem cells, but also their neuronal and oligodendrocyte differentiation in vitro. Moreover, increased activity of astrocyte differentiation of hippocampal neural stem cells from both newborn and adult rats was observed after exposure to high concentration of lead ion in vitro. These findings suggest that hippocampal neural stem cells of newborn rats were more sensitive than those from adult rats to Pb2+ cytotoxicity.

A metal mixture induces transformation upon antioxidant depletion in a hepatic cell line

Introduction. Metals are ubiquitous soil, air, and water pollutants. A mixture of arsenic cadmium and lead, in particular, has commonly been found in the vicinity of smelter areas. The mixture of As-Cd-Pb has been shown to be carcinogenic, and transforming potential and oxidative stress have been proposed as principal mechanisms involved in this process. The aim of this work was to explore the role of the antioxidant barrier in the establishment of cell transformation upon chronic exposure to a metal mixture containing 2 μM NaAsO2, 2 μM. CdCl2, and 5 μM Pb(C2H3O2)2-3H2O in WRL-68 cells-a non-transformed human hepatic cell line.Material and methods. In this study, we used a WRL-68 cell model of human embryonic hepatic origin treated with antioxidant inhibitors (L-Buthionine-sulfoxamine and aminotriazole) to test the role of the antioxidant barrier in the establishment of cell transformation upon chronic exposure to a metal mixture of As-Cd-Pb (2 μM NaAsO2, 2 μM CdCl2 and 5 μM Pb(C2H3O2)2•3H2O). We evaluated oxidative damage markers, including reactive oxygen species, lipid peroxidation, and genotoxicity, as well as antioxidant response markers, including glutathione concentration, catalase activity, and superoxide dismutase activity, wich promote morphological transformation, which can be quantified by foci formation.Results. As expected, we found an increase in the intracellular concentration of the metals after treatment with the metal mixture. In addition, treatment with the metal mixture in addition to inhibitors resulted in a large increase in the intracellular concentration of cadmium and lead. Our results describe the generation of reactive oxygen species, cytotoxicity, genotoxicity, and oxidative damage to macromolecules that occurred exclusively in cells that were morphologically transformed upon exposure to a metal mixture and antioxidant barrier inhibition.Conclusion. Our results show the importance of the antioxidant barrier role in the protection of cellular integrity and the transformation potential of this metal mixture via free radicals.

Lead at 2.5 and 5.0 μM induced aberrant MH-II surface expression through increased MII exocytosis and increased autophagosome formation in Raw 267.4 cells

Aberrant major histocompatibility complex class II (MHC-II) surface expression on antigen presenting cells (APCs) is associated with dysregulated immune homeostasis. Lead (Pb) is known to increase MHC-II surface expression on murine peritoneal macrophages ex vivo at concentrations exceeding 25μM. Little data exist examining this effect at physiologically relevant concentrations. To address this deficit, we examined the effects of Pb on MHC-II surface expression, secondary T-cell activation markers (CD80, CD86, CD40), cell viability, cellular metabolic activity, and β-hexosaminidase activity in RAW 267.4 macrophage cell lines, with changes in cell ultrastructure evaluated by electron and confocal microscopy. Pb induced an increase in MHC-II, CD86, and lysosome-associated LAMP-1 and LAMP-2 surface mean expression during one doubling cycle (17h), which was mirrored by increased β-hexosaminidase activity. Although cell viability was unaffected, cellular metabolism was inhibited. Electron microscopy revealed evidence of lipid vacuolization, macroautophagy and myelin figure formation in cells cultured with either Pb or LPS. Confocal microscopy with antibodies against LC3B showed a punctate pattern consistent with the presence of mature autophagosomes. Collectively, these data suggest that 2.5–5.0μM Pb increased MHC-II surface expression by inhibiting metabolic activity, inducing autophagy, and increasing MHC-II trafficking in a macrophage cell line.

Alleviation of Lead Toxicity by Silicon is Related to Elevated Photosynthesis, Antioxidant Enzymes Suppressed Lead Uptake and Oxidative Stress in Cotton

Lead (Pb) is known to cause highly toxic direct and indirect effects on plants, animals and humans, like other metals of its group. The mitigating effects of exogenous Silicon (Si) application on Pb stressed cotton plants were investigated in this hydroponic study. The following growth parameters were observed chlorophyll contents, photosynthesis, growth, Pb uptake and accumulation, Malondialdehyde (MDA), Hydrogen Peroxide (H2O2) contents, Electrolyte Leakage (EL), soluble protein and the activities of major antioxidant enzymes (superoxide dismutase, SOD; guaiacol peroxidase, GPX; ascorbate peroxidase, APX and catalase, CAT). Six treatments with three replicates including control (basal nutrients comprising neither lead nor silicon), 1 mM silicon (Si), 50 μM Pb, 100 μM Pb, 50 μM Pb with 1 mM Si and 100 μM Pb with 1 mM Silicon (Si) were studied.

Effects of chronic and acute lead treatments on the biophysical properties of erythrocyte membranes, and a comparison with model membranes

Rat erythrocytes, or erythrocyte membrane ghosts, have been subjected to either chronic (drinking water containing 15 mM lead acetate for 3 months) or acute (10−9–10−2 M lead acetate for 1 h) Pb2+ treatments and subsequent changes in membrane properties have been measured. Pb2+ concentration in chronically treated rat plasma was 1.8 μM, which is one order of magnitude above normal values. Membrane permeability, or hemolysis, was increased in both cases. A comparative study using liposomes, in the form of large unilamellar vesicles, also indicated an increase in membrane permeability. Membrane microviscosity, or acyl chain molecular order, measured as DPH fluorescence polarization, showed an increased order in the acute treatments, at least below 700 μM Pb2+, and a similar increase in chronically treated rats. The correlation between acute and chronic treatments, and between cell and model membranes, suggests that the present observations may be relevant in the pathogenesis of lead intoxication in humans.

Impact of lead and mercuric ions on the interleukin-2-dependent proliferation and survival of T cells

Mercury and lead are widespread in the environment, causing chronic exposure of a large population to low concentrations of these metals. While several studies demonstrated that low levels of both metals affect the immune system, little is known about underlying molecular mechanisms. The objective of this study was to investigate the impact of mercuric (Hg(2+)) and lead (Pb(2+)) ions on T cells. Up to 100 μM Pb(NO(3))(2) had no effect on cellular viability and proliferation. In contrast, HgCl(2) caused a concentration-dependent decline of viable leukocytes and especially of activated T cells. Additionally, Hg(2+) induced reactive oxygen species (ROS) generation accompanied by the loss of mitochondrial transmembrane potential, measured by Dihydrorhodamine and Rhodamine-123, respectively. The antioxidant N-acetylcysteine partially reversed the toxic effects of Hg(2+), pointing to an involvement of ROS. The major cytokine controlling T-cell survival and proliferation is interleukin (IL)-2. Hg(2+) had no effect on the secretion of IL-2, but on IL-2 mediated signal transduction pathways, reducing phosphorylation of the downstream kinases ERK1/2 and AKT. Moreover, Hg(2+) led to an arrest of the cells in the S phase of the cell cycle. Taken together, these data fit a model in which Hg(2+) disrupts mitochondria, and the resulting release of ROS inhibits IL-2-dependent signal transduction, reducing proliferation and survival of T cells.

A Portable and Power-Free Microfluidic Device for Rapid and Sensitive Lead (Pb2+) Detection

A portable and power-free microfluidic device was designed for rapid and sensitive detection of lead (Pb2+). 11-mercaptoundecanoic acid (MUA)-functionalized gold nanoparticles (MUA-AuNPs) aggregated in the presence of Pb2+ for the chelation mechanism. When we performed this analysis on a polydimethylsiloxane (PDMS) microfluidic chip, the aggregations deposited onto the surface of chip and formed dark lines along the laminar flows in the zigzag microchannels. This visual result can be observed by the naked eye through a microscope or just a drop of water as a magnifier. Ten μM Pb2+ was successfully detected.

Differences in Zn and Pb resistance of two ecotypes of the microalga Eustigmatos sp. inhabiting metal loaded calamine mine spoils

Soil-dwelling microalgae as pioneer organisms may play an essential role in degraded post-industrial areas. Zn and Pb resistance of two morphologically identical strains (E120, E5) of the soil microalga Eustigmatos sp. (Eustigmatophyceae) inhabiting two extremely metal polluted calamine mine spoils and Eustigmatos vischeri (Ev) from algal collection was compared. To compare Zn and Pb resistance of algal strains, toxicity parameters (72/96 h-EC50) were determined during their exposure to high Zn (50–800 μM) and Pb (5–80 μM) concentrations. Both Zn- and Pb-EC50 values increased as follows: Ev < E5 < E120. The E120 strain from the mine spoil with the highest metal contents appeared to be two times more Zn and Pb resistant (96 h-EC50 = 416 μM Zn, 39.8 μM Pb) than E5 strain from the less metal polluted site and 4.7–8.8 times more than Ev. Differences in Zn and Pb accumulation as well as in metal-induced membrane lipid peroxidation were also observed. The present study highlights the evolution of algal ecotypes of high but various Zn and Pb resistance as a result of natural exposure to different metal concentrations in their habitats. The algae of high heavy metal resistance could have a practical application in remediation of contaminated soils of anthropogenic origin.

Heavy metal induced ecophysiological function alterations in the euhalophyte Suaeda salsa

Heavy metal accumulation affects the physiological status of plants. Suaeda salsa L. is used to investigate the toxic effects of cadmium (Cd) and lead (Pb) either alone or mixtures under the static test conditions. Cd-Pb mixture exposure can decrease lignin content and weaken the increase. Mitochondrial calcium content significantly reduced at 30 µM Cd and Pb exposure. Cd-Pb mixture exposure can increase calcium content under the same concentration exposure. Soluble sugar levels noted a significant decrease in Cd, Pb and Cd-Pb mixture exposure. The accumulations of Cd, Pb in S. salsa were significantly increased with exposure time. Soluble protein (SP) in S. salsa at 30 µM concentration treatments decreased with exposure time. Heat shock protein 70 (HSP70) was enhanced lightly along with the increase of added Cd-Pb from 30 to 70 &3181;M and then decreased below the controls which present a synergistic effect. Heat shock protein 60 (HSP60) increased slightly with the increase of Cd-Pb from 30 to 110 µM, and then decreased hereafter and significantly inhibited at 150 ƒEM (p<0.05). Moreover, Cd-Pb mixture exposure significantly increased the Rubisco activity under lower concentration and presented antagonistic effect. At the same time, the viability percent decreased as increase Cd-Pb concentration exposure (p0.05), it presents a dose-dependent manner. Mitochondrial cells treated with Cd-Pb exposure obviously reduced the reactive oxygen species (ROS) levels in mitochondrial cells. Key words : Suaeda salsa, heavy metal, ecophysiological function.

Lead induces an osteoarthritis‐like phenotype in articular chondrocytes through disruption of TGF‐β signaling

Lead remains a significant environmental toxin, and we believe we may have identified a novel target of lead toxicity in articular chondrocytes. These cells are responsible for the maintenance of joint matrix, and do so under the regulation of TGF-β signaling. As lead is concentrated in articular cartilage, we hypothesize that it can disrupt normal chondrocyte phenotype through suppression of TGF-β signaling. These experiments examine the effects of lead exposure in vivo and in vitro at biologically relevant levels, from 1 nM to 10 µM on viability, collagen levels, matrix degrading enzyme activity, TGF-β signaling, and articular surface morphology. Our results indicate that viability was unchanged at levels ≤100 µM Pb, but low and high level lead in vivo exposure resulted in fibrillation and degeneration of the articular surface. Lead treatment also decreased levels of type II collagen and increased type X collagen, in vivo and in vitro. Additionally, MMP13 activity increased in a dose-dependent manner. Active caspase 3 and 8 were dose-dependently elevated, and treatment with 10 µM Pb resulted in increases of 30% and 500%, respectively. Increasing lead treatment resulted in a corresponding reduction in TGF-β reporter activity, with a 95% reduction at 10µM. Levels of phosphoSmad2 and 3 were suppressed in vitro and in vivo and lead dose-dependently increased Smurf2. These changes closely parallel those seen in osteoarthritis. Over time this phenotypic shift could compromise maintenance of the joint matrix.

Physiological and biochemical responses induced by lead stress in Spirodela polyrhiza

The effects of increasing lead concentration on the activities of superoxide dismutase (SOD), peroxides (POD) and catalase (CAT), levels of ascorbate (AsA), reduced glutathione (GSH), Pb accumulation and its influence on nutrient elements, polyamines (PAs) content, as well as activities of polyamine oxidase (PAO) and ornithine decarboxylase (ODC), were investigated in Spirodela polyrhiza. POD and CAT activities increased progressively followed by a decline, while SOD activity gradually fell. The effect of Pb application on AsA content was similar to that seen for POD and CAT activities. GSH content initially rose but then declined. A significant enhancement in Pb accumulation was observed, except in the 25 μM Pb treatments. Nutrient elements were also affected. Moreover, Pb stress induced a considerable decrease in total spermidine (Spd), while the levels of total putrescine (Put) and spermine (Spm) initially increased at 25 μM Pb but then declined. Free and perchloric acid soluble conjugated (PS-conjugated) PAs contents changed in a similar way to total PAs. In addition, Pb stress induced a continuous accumulation of perchloric acid insoluble bound (PIS-bound) Spm and an initial accumulation of PIS-bound Put and Spd. The ratio of free (Spd + Spm)/Put significantly declined whereas the ratio of total (Spd + Spm)/Put rose at low Pb concentrations (25 and 50 μM). PAO activity rose gradually with an increase in Pb concentration, reaching peak values at 100 μM, while ODC activity first increased at 25 μM Pb and then declined. The results indicated that the tolerance of S. polyrhiza to Pb stress was enhanced by activating the antioxidant system, preventing the entry of the Pb ion and altering the content of polyamines.

Evaluation of Lead Toxicity in Erica andevalensis as an Alternative Species for Revegetation of Contaminated Soils

Although revegetation using native flora is a low cost way to stabilize soil and restore the landscape contaminated with metals, little is known regarding the Pb-tolerance of many of these species. For this purpose, we evaluated the tolerance of Erica andevalensis to Pb by growing plants in nutrient solutions with increasing concentrations of Pb (up to 100 μM). Plant growth and different physiological parameters were determined to ascertain tolerance to metal stress. Additionally, an electron microscopy study coupled with EDX-analysis was performed to get clues on the Pb uptake and translocation from roots into stem and leaves. The LOEC (the lowest observed effect concentration) of Pb was 40 μM while the IC50 (inhibition concentration) was 80 μM Pb. Chemical analysis revealed a root>stem>leaf accumulation pattern. There was a severe reduction in fresh biomass and chlorophyll concentration at the highest Pb dose. The SEM-EDX study indicated that Pb was mostly located in root epidermal tissues. The blockage of Pb on the root probably avoided its toxic effects by limiting Pb transport to other tissues.

Lead (Pb)-induced biochemical and ultrastructural changes in wheat (Triticum aestivum) roots

The focus of the present study was to explore lead (Pb)-induced metabolic alterations vis-à-vis ultrastructural changes in wheat roots to establish Pb toxicity syndrome at a structural level. Pb (50-500 μM) enhanced malondialdehyde (an indicator of lipid peroxidation) and hydrogen peroxide content, and electrolyte leakage, thereby suggesting reactive oxygen species-induced disruption of membrane integrity and oxidative stress in wheat roots. The activities of superoxide dismutases and catalases enhanced upon Pb exposure, whereas those of ascorbate and guaiacol peroxidases declined. Pb-induced metabolic disruption was manifested in significant alterations in wheat root ultrastructure as analyzed by transmission electron microscopy. Pb caused thinning of cell wall (at 50 μM), formation of amoeboid protrusions and folds and intercellular spaces, and appearance of lesions and nicks/breaks (at ≥ 250 μM Pb). Pb was deposited along the cell walls as dark precipitates. At ≤ 250 μM Pb, the number of mitochondria increased significantly, whereas structural damage in terms of change of shape and disintegration was observed at ≥ 250 μM Pb. Pb reduced the size of nucleoli and induced puff formation (at 250 μM), resulting in complete disintegration/disappearance of nucleolus at 500 μM. The study concludes that Pb inhibited wheat root growth involving an ROS-mediated oxidative damage vis-à-vis the ultrastructural alterations in cell membrane and disruption of mitochondrial and nuclear integrity.

Localization and quantification of Pb and nutrients in Typha latifolia by micro-PIXE

Typha latifolia is a plant species widely used for phytoremediation. Accumulation, localization and distribution of Pb and mineral nutrients were investigated in roots, rhizomes and leaves of Typha latifolia grown at 0, 50, 100 and 250 μM Pb concentrations in a pot experiment under controlled conditions. Bulk elemental concentrations were determined by X-ray fluorescence (XRF) spectroscopy whereas micro-proton-induced X-ray emission (micro-PIXE) was used for element localization in root and rhizome tissues. Gradual increase in bulk Pb concentrations was observed in Typha latifolia roots and rhizomes treated with increasing Pb concentrations, however in rhizomes Pb concentrations were an order of magnitude lower than in roots. In leaves Pb concentrations were around the limit of detection for XRF (~20 μg g(-1)). An increase in concentration of K and Ca in roots, rhizomes and leaves, of iron and zinc in roots and leaves, and of Mn in rhizomes was observed either at 50 and/or 100 μM Pb treatments, whereas for K and Ca in roots, rhizomes and leaves, Fe and Zn in roots and leaves and Mn in rhizomes, or at 250 μM Pb treatment the increase was seen for concentrations of Fe and Zn in rhizomes and Cu in roots. Mn concentrations decreased with Pb treatments in roots and leaves. Element localization using micro-PIXE analysis demonstrated Pb accumulation in epidermal and cortical tissues of treated roots and rhizomes, while in endodermis and vascular tissues Pb was not detected. A displacement of Ca from epidermal to cortical tissues was observed in Pb treated roots and rhizomes, pointing to cell wall immobilization of Pb as one of the tolerance mechanisms in Typha latifolia. High level of colocalization of Pb with P (r = 0.60), S (r = 0.37) and Zn (r = 0.70) was observed in Pb treated roots, while in rhizomes colocalization with the mentioned elements was still positive, but not that prominent. These results indicate that Pb may form complexes with phosphorus and sulfur compounds in roots and rhizomes, which may also represent attraction sites for binding Zn. Because of its large root and rhizome surface area acting as main sites for Pb adsorption, Typha latifolia may represent potentially efficient plant species for phytoremediation of Pb contaminated soils and waters.