Lead Uptake, Toxicity, and Detoxification in Plants - Archive ouverte
Roberto Wells | Download | Embed to your website
- Jul 12, 2012
- Page(s): 26
- Size: 393.27 kB
Lead Uptake, Toxicity, and Detoxification in Plants Bertrand Pourrut, Muhammad Shahid, Camille Dumat, Peter Winterton, Eric Pinelli To cite this version: Bertrand Pourrut, Muhammad Shahid, Camille Dumat, Peter Winterton, Eric Pinelli. Lead Uptake, Toxicity, and Detoxification in Plants. Reviews of Environmental Contamination and Toxicology, Springer Verlag, 2011, vol. 213, pp. 113-136. �10.1007/978-1-4419-9860-6_4�. �hal-00717188� HAL Id: hal-00717188 https://hal.archives-ouvertes.fr/hal-00717188 Submitted on 12 Jul 2012 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés.
Open Archive Toulouse Archive Ouverte (OATAO) OATAO is an o pen ac cess re pository that c ollects th e work of To ulouse re searchers a nd makes it freely available over the web where possible. This is an author-deposited ver sion p ublished in: http://oatao.univ-toulouse.fr/ Eprints ID: 5674 To link to this article: DOI:10.1007/978-1-4419-9860-6_4 http://dx.doi.org/10.1007/978-1-4419-9860-6_4 To c ite t his vers ion: Pourrut, Bertrand and Shahid, Muhammad and Dumat, Camille and Winterton, Peter and Pinelli, Eric Lead Uptake, Toxicity, and Detoxification i n Plants. (2011) Revi ews o f Environmental Contami nation and Toxicology, vol. 213 . pp. 113-136. ISSN 0179-5953 Any correspondence concerning this service should be sent to the repository administrator: [emailprotected]
Lead Uptake, Toxicity, and Detoxification in Plants Bertrand Pourrut, Muhammad Shahid, Camille Dumat, Peter Winterton, and Eric Pinelli Contents 1 Introduction 2 Retention, Mobility, and Bioavailability of Lead in Soil 3 Lead Behavior in Plants 3.1 Lead Uptake by Plants 3.2 Lead Accumulation in Plants 4 General Effects of Lead on Plants 4.1 Effects on Germination and Growth 4.2 Effects on Proteins 4.3 Water Status Effects 4.4 Mineral Nutrition Effects 4.5 Photosynthesis Effects 4.6 Respiration Effects 4.7 Genotoxicity 4.8 Oxidative Stress and Lipid Peroxidation 5 Mechanisms of Lead Tolerance 5.1 Passive Mechanisms 5.2 Inducible Mechanisms 5.3 Antioxidant Enzymes 6 Conclusions and Perspectives 7 Summary References Bertrand Pourrut and Muhammad Shahid – equivalent first authors. E. Pinelli (B) EcoLab (Laboratoire d’écologie fonctionnelle), INP-ENSAT, 31326 Castanet-Tolosan, France; EcoLab (Laboratoire d’écologie fonctionnelle), UMR 5245 CNRS-INP-UPS, 31326 Castanet-Tolosan, France e-mail: [emailprotected]
1 Introduction Plants are the target of a wide range of pollutants that vary in concentration, speci- ation, and toxicity. Such pollutants mainly enter the plant system through the soil (Arshad et al. 2008) or via the atmosphere (Uzu et al. 2010). Among common pol- lutants that affect plants, lead is one of the most toxic and frequently encountered (Cecchi et al. 2008; Grover et al. 2010; Shahid et al. 2011). Lead continues to be used widely in many industrial processes and occurs as a contaminant in all envi- ronmental compartments (soils, water, the atmosphere, and living organisms). The prominence of environmental lead contamination results both from its persistence (Islam et al. 2008; Andra et al. 2009; Punamiya et al. 2010) and from its present and past numerous sources. These sources have included smelting, combustion of leaded gasoline, or applications of lead-contaminated media (sewage sludge and fertilizers) to land (Piotrowska et al. 2009; Gupta et al. 2009; Sammut et al. 2010; Grover et al. 2010). In 2009, production of recoverable lead from mining operations was 1690, 516, and 400 thousand metric tons by China, Australia, and the USA, respectively (USGS 2009). Despite a long history of its beneficial use by humankind, lead has no known bio- logical function in living organisms (Maestri et al. 2010) and is now recognized as a chemical of great concern in the new European REACH regulations (EC 1907/2006; Registration, Evaluation, Authorization, and Restriction of Chemicals). Moreover, lead was reported as being the second most hazardous substance, after arsenic, based on the frequency of occurrence, toxicity, and the potential for human exposure by the Agency for Toxic Substances and Disease Registry (ATSDR 2003). The trans- fer of lead from polluted soils to plants was therefore widely studied, especially in the context of food quality, use in phytoremediation, or in biotesting (Arshad et al. 2008; Uzu et al. 2009). Lead is known to induce a broad range of toxic effects to living organism, includ- ing those that are morphological, physiological, and biochemical in origin. This metal impairs plant growth, root elongation, seed germination, seedling develop- ment, transpiration, chlorophyll production, lamellar organization in the chloroplast, and cell division (Sharma and Dubey 2005; Krzesłowska et al. 2009; Gupta et al. 2009, 2010; Maestri et al. 2010). However, the extent of these effects varies and depends on the lead concentration tested, the duration of exposure, the intensity of plant stress, the stage of plant development, and the particular organs studied. Plants have developed various methods for responding to toxic metal exposures. They have internal detoxification mechanisms to deal with metal toxicity that includes selective metal uptake, excretion, complexation by specific ligands, and compartmentaliza- tion (Gupta et al. 2009; Krzesłowska et al. 2010; Maestri et al. 2010; Sing et al. 2010; Jiang and Liu 2010). The various responses of plants to lead exposure are often used as tools (bioindi- cators) in the context of environmental quality assessment. To develop tools that are relevant for ecotoxicological studies, it is essential to understand the mechanisms involved in plant uptake, transfer, and toxicity. This is especially true in selected research areas, such as choice of plant species, when polluted soils are under study
(i.e., reduced transfer when studying vegetables or increased transfer when phytoex- traction is desired). For example, legumes are considered more suitable to grow on contaminated soil than Umbelliferae, Liliaceae, Compositae, and Chenopodiaceae, because they take up reduced amounts of lead (Alexander et al. 2006). The reduced lead uptake by vegetables minimizes the threat of lead introduction to the food chain. In contrast, phytoextraction requires plants that can sequester excessive amounts of lead in their biomass without incurring damage to basic metabolic functions (Arshad et al. 2008; Zaier et al. 2010). Pelargonium (Arshad et al. 2008) and Brassica napus (Zaier et al. 2010) are characterized as Pb hyperaccumulators, and they can extract huge amounts of lead from contaminated soil without showing morpho- phytotoxicity symptoms. Indeed, these plants have efficient natural detoxification mechanism to alleviate lead toxicity. In this review, we propose to trace the rela- tionship that exists between lead uptake, accumulation, translocation, and toxicity in plants. 2 Retention, Mobility, and Bioavailability of Lead in Soil Lead occurs naturally in the earth’s crust (Arias et al. 2010) and its natural levels remain below 50 mg kg–1 (Pais and Jones 2000). But, anthropogenic activities often modify the amount and nature of lead species present in soil. In soils, lead may occur as a free metal ion, complexed with inorganic constituents (e.g., HCO3 – , CO3 2– , SO4 2– , and Cl– ), or may exist as organic ligands (e.g., amino acids, fulvic acids, and humic acids); alternatively lead may be adsorbed onto particle surfaces (e.g., Fe-oxides, biological material, organic matter, and clay particles) (Uzu et al. 2009; Tabelin and Igarashi 2009; Sammut et al. 2010; Vega et al. 2010). Anthropogenic- sourced lead generally accumulates primarily in the surface layer of soil, and its concentration decreases with depth (Cecchi et al. 2008). Because of its strong bind- ing with organic and/or colloidal materials, it is believed that only small amounts of the lead in soil are soluble, and thereby available for plant uptake (Kopittke et al. 2008; Punamiya et al. 2010). However, lead behavior in soil, in the context of species, solubility, mobility, and bioavailability, is largely controlled by complex interactions governed by many biogeochemical factors (Punamiya et al. 2010). These factors include pH (Kopittke et al. 2008; Lawal et al. 2010; Vega et al. 2010), redox conditions (Tabelin and Igarashi 2009), cation-exchange capacity (Vega et al. 2010), soil mineralogy (Dumat et al. 2006), biological and microbial conditions (Arias et al. 2010), amount of lead present (Bi et al. 2010; Cenkci et al. 2010; Lawal et al. 2010), organic and inor- ganic ligand levels (Padmavathiamma and Li 2010; Sammut et al. 2010; Shahid et al. 2011), competing cation levels (Kopittke et al. 2008; Komjarova and Blust 2009), and plant species involved (Kovalchuk et al. 2005; Bi et al. 2010; Liu et al. 2010). Such factors may act individually or in combination with each other and may alter the soil behavior of the lead present, as well as the rate of uptake by plants.
Lead bioavailability is strongly influenced by its speciation and, in particular, by the concentration of free lead ions present (Dumat et al. 2006; Uzu et al. 2009; Sammut et al. 2010; Shahid et al. 2011). This is because the most significant plant uptake route for many cationic metals (and especially for the free metal ion) is via the soil solution in dissolved form (Punamiya et al. 2010). Moreover, the free lead ion concentration in soils depends on the adsorption/desorption processes in which it participates (Vega et al. 2010). 3 Lead Behavior in Plants 3.1 Lead Uptake by Plants With the exception of the special conditions that exist for plants cultivated near metal recycling industries (Uzu et al. 2010), the main pathway by which plants accumulate metals is through root uptake from soils (Sharma and Dubey 2005; Uzu et al. 2009). Part of the lead present in the soil solution is adsorbed onto the roots, and then becomes bound to carboxyl groups of mucilage uronic acid, or directly to the polysaccharides of the rhizoderm cell surface (Seregin and Ivanov 2001). Lead adsorption onto roots has been documented to occur in several plant species: Vigna unguiculata (Kopittke et al. 2007), Festuca rubra (Ginn et al. 2008), Brassica juncea (Meyers et al. 2008), Lactuca sativa (Uzu et al. 2009), and Funaria hygro- metrica (Krzesłowska et al. 2009, 2010). Once adsorbed onto the rhizoderm roots surface, lead may enter the roots passively and follow translocating water streams. However, lead absorption is not uniform along plant roots as a lead concentration gradient from root apex can be observed (Tung and Temple 1996; Seregin et al. 2004). Indeed, the highest lead concentrations can be found in root apices, where root cells are young and have thin cell walls (with the exception of root cap cells) that facilitate root uptake (Tung and Temple 1996; Seregin et al. 2004). Moreover, the apical area is the area where rhizodermic pH is the lowest, which increases solubility of lead in the soil solution. At the molecular level, the mechanism by which lead enters roots is still unknown. Lead may enter the roots through several pathways, and a particular pathway is through ionic channels. Although, lead uptake is a non-selective phe- nomenon, it nonetheless depends on the functioning of an H+ /ATPase pump to maintain a strong negative membrane potential in rhizoderm cells (Hirsch et al. 1998; Wang et al. 2007). Inhibition of lead absorption by calcium is well-known (Garland and Wilkins 1981; Kim et al. 2002) and is associated with competition between these two cations for calcium channels (Huang and Cunningam 1996). Several authors have demonstrated that Ca2+ -permeable channels are the main path- way by which lead enters roots (Wang et al. 2007; Pourrut et al. 2008). The use of transgenic plants has shown that lead can penetrate into roots through alternative non-selective pathways, such as cyclic nucleotide-gated ion channels (Arazi et al. 1999; Kohler et al. 1999) or via low-affinity cation transporters (Wojas et al. 2007).
Reduced uptake and translocation of lead to aerial plant parts of vegetables is considered to be beneficial in preventing lead from entering the food chain. However, reduced uptake and translocation of lead to aerial plant parts, when plants are used to remediate polluted soils, is a major problem. Indeed, soil remediation requires plants (hyperaccumulators) that can take high lead levels up and translocate it to aerial plant parts with no or minimal toxicity. The amount of lead that moves from soil to penetrate into plants can be measured by what is called “the transfer factor”; this factor is defined as the ratio that exists between the concentration of lead in the plant vs. the concentration of lead in the soil (Arshad et al. 2008; Bi et al. 2010; Liu et al. 2010). This transfer factor will be different for different plant species and will change as soil physical and chemical properties are altered (Arshad et al. 2008; Bi et al. 2010; Liu et al. 2010). Generally, plants having a transfer factor greater than 1 are categorized as hyperaccumulators, whereas those with transfer factor less than 1 are termed as non-accumulators of lead (Arshad et al. 2008). 3.2 Lead Accumulation in Plants Once lead has penetrated into the root system, it may accumulate there or may be translocated to aerial plant parts. For most plant species, the majority of absorbed lead (approximately 95% or more) is accumulated in the roots, and only a small fraction is translocated to aerial plant parts, as has been reported in Vicia faba, Pisum sativum, and Phaseolus vulgaris (Piechalak et al. 2002; Małecka et al. 2008; Shahid et al. 2011), V. unguiculata (Kopittke et al. 2007), Nicotiana tabacum, (Gichner et al. 2008), Lathyrus sativus (Brunet et al. 2009), Zea mays (Gupta et al. 2009), Avicennia marina (Yan et al. 2010), non-accumulating Sedum alfredii (Gupta et al. 2010), and Allium sativum (Jiang and Liu 2010). Although many metals display the translocation restriction phenomenon mentioned above, this phenomenon is not common to all heavy metals. Notwithstanding, this phenomenon in plants is both specific and very intense for lead. When entering the root, lead mainly moves by apoplast and follows water streams until it reaches the endodermis (Tanton and Crowdy 1971; Lane and Martin 1977). There are several reasons why the transport of lead from roots to aerial plant parts is limited. These reasons include immobilization by negatively charged pectins within the cell wall (Islam et al. 2007; Kopittke et al. 2007; Arias et al. 2010), precipita- tion of insoluble lead salts in intercellular spaces (Kopittke et al. 2007; Islam et al. 2007; Meyers et al. 2008; Małecka et al. 2008), accumulation in plasma membranes (Seregin et al. 2004; Islam et al. 2007; Jiang and Liu 2010), or sequestration in the vacuoles of rhizodermal and cortical cells (Seregin et al. 2004; Kopittke et al. 2007). However, these reasons are not sufficient to explain the low rate of lead translo- cation from root to shoot. The endoderm, which acts as a physical barrier, plays an important role in this phenomenon. Indeed, following apoplastic transport, lead is blocked in the endodermis by the Casparian strip and must follow symplastic
transport. In endodermis cells, the major part of lead is sequestered or excreted by plant detoxification systems (c.f. Section 5.2). Several hyperaccumulator plant species, such as Brassica pekinensis and Pelargonium, are capable of translocating higher concentrations of lead to aerial plant parts, without incurring damage to their basic metabolic functions (Xiong et al. 2006; Liu et al. 2008; Arshad et al. 2008). A specific hyperaccumulator species can accumulate more than 1000 ppm lead (Maestri et al. 2010). Indeed, these plants exude substances from roots that dissolve metals in soil (Arshad et al. 2008) that increases uptake and translocation (by employing certain metal cation transporters/genes). Moreover, they can tolerate higher concentrations of lead ions because they have various detoxification mechanisms, which may include selective metal uptake, excretion, complexation by specific ligands, and compartmentaliza- tion. In addition, translocation of lead to aerial plant parts increases in the presence of organic chelators like ethylenediaminetetraacetate (EDTA) (Liu et al. 2008; Zaier et al. 2010; Barrutia et al. 2010) or certain species of micro-organisms (Arias et al. 2010; Punamiya et al. 2010). Recently, Liu et al. (2010) reported that, in 30 B. pekinensis cultivars, increased soil lead levels also increased the percent translo- cation to aerial plant parts. High concentrations of lead are known to destroy the physical barrier formed by the Casparian strip. Transportation of metals from plant roots to shoots requires movement through the xylem (Verbruggen et al. 2009) and, when it occurs, is probably driven by tran- spiration (Liao et al. 2006). Arias et al. (2010) demonstrated high lead deposition in xylem and phloem cells of mesquite plants by using X-ray mapping. After pen- etrating into the central cylinder of the stem, lead can again be transported via the apoplastic pathway. The lead is then translocated to leaf areas via vascular flow (Sharma and Dubey 2005; Krzesłowska et al. 2010). While passing through the xylem, lead can form complexes with amino or organic acids (Roelfsema and Hedrich 2005; Vadas and Ahner 2009; Maestri et al. 2010). However, lead may also be transferred in inorganic form, as is cadmium. To express the degree of lead translocation, some authors have used a translocation factor (lead in aerial parts/lead in roots) (Arshad et al. 2008; Uzu et al. 2009; Liu et al. 2010). When this factor is used, the numeric value is normally rather low, which indicates that lead has been sequestered in the roots (Uzu et al. 2009; Liu et al. 2010). 4 General Effects of Lead on Plants 4.1 Effects on Germination and Growth When plants are exposed to lead, even at micromolar levels, adverse effects on germination and growth can occur (Kopittke et al. 2007). Germination is strongly inhibited by very low concentrations of Pb2+ (Tomulescu et al. 2004; Islam et al. 2007). Lead-induced inhibition of seed germination has been reported in Hordeum
vulgare, Elsholtzia argyi, Spartina alterniflora, Pinus halepensis, Oryza sativa, and Z. mays (Tomulescu et al. 2004; Islam et al. 2007; Sengar et al. 2009). At higher concentrations, lead may speed up germination and simultaneously induce adverse affects on the length of radical and hypocotyl in E. argyi (Islam et al. 2007). Inhibition of germination may result from the interference of lead with protease and amylase enzymes (Sengar et al. 2009). Lead exposure in plants also strongly limits the development and sprouting of seedlings (Dey et al. 2007; Gichner et al. 2008; Gopal and Rizvi 2008). At low concentrations, lead inhibits the growth of roots and aerial plant parts (Islam et al. 2007; Kopittke et al. 2007). This inhibition is stronger for the root, which may be correlated to its higher lead content (Liu et al. 2008). Lead toxicity may also cause swollen, bent, short and stubby roots that show an increased number of sec- ondary roots per unit root length (Kopittke et al. 2007). Recently, Jiang and Liu (2010) reported mitochondrial swelling, loss of cristae, vacuolization of endoplas- mic reticulum and dictyosomes, injured plasma membrane and deep colored nuclei, after 48–72 h of lead exposure to A. sativum roots. Arias et al. (2010) reported significantly inhibited root elongation in Mesquite (Prosopis sp.). Plant biomass can also be restricted by high doses of lead exposure (Gopal and Rizvi 2008; Gichner et al. 2008; Islam et al. 2008; Piotrowska et al. 2009; Sing et al. 2010). Under severe lead toxicity stress, plants displayed obvious symptoms of growth inhibition, with fewer, smaller, and more brittle leaves having dark purplish abaxial surfaces (Islam et al. 2007; Gupta et al. 2009). Plant growth retardation from lead exposure may be attributed to nutrient metabolic disturbances (Kopittke et al. 2007; Gopal and Rizvi 2008) and disturbed photosynthesis (Islam et al. 2008). In most cases, the toxic effect of lead on plant growth is time and dose dependent (Dey et al. 2007; Gupta et al. 2009, 2010). However, the effect of low concentrations is not clearly established, and the observed growth inhibition is not necessarily correlated to a reduction in biomass (Kosobrukhov et al. 2004; Yan et al. 2010). Moreover, the effect of lead toxicity varies with plant species, i.e., hyperaccumulators naturally tolerate more lead toxicity than do sensitive plants (Arshad et al. 2008). 4.2 Effects on Proteins Similar to what occurs with other heavy metals, lead interacts with cytoplasmic proteins. The effect of lead on the total concentration of protein is unclear, although high concentrations may decrease the protein pool (Chatterjee et al. 2004; Mishra et al. 2006; Garcia et al. 2006; Piotrowska et al. 2009). This quantitative decrease in total protein content is the result of several lead effects: acute oxidative stress of reactive oxygen species (ROS) (Piotrowska et al. 2009; Gupta et al. 2009), modification in gene expression (Kovalchuk et al. 2005), increased ribonuclease activity (Gopal and Rizvi 2008), protein utilization by plants for the purposes of lead detoxification (Gupta et al. 2009), and diminution of free amino acid content
(Gupta et al. 2009) that is correlated with a disturbance in nitrogen metabolism (Chatterjee et al. 2004). However, certain amino acids, like proline, increase under lead stress (Qureshi et al. 2007). Such proteins play a major role in the tolerance of the plant to lead. In contrast, low concentrations of lead increase total protein con- tent (Mishra et al. 2006). This protein accumulation may defend the plant against lead stress (Gupta et al. 2010), particularly for proteins involved in cell redox main- tenance. If true, then such proteins act in a way similar to how ascorbate functions or similar to how metals are sequestered by glutathione (GSH) or phytochelatins (PCs) (Brunet et al. 2009; Liu et al. 2009; Yadav 2010; Jiang and Liu 2010). In addition to a quantitative change, lead can affect the qualitative composition of cell proteins. The protein profile of root cells in bean seedlings was modified after lead expo- sure (Beltagi 2005). Such modification can be correlated to the change that occurs in the transcriptome profile of several enzymes including isocitrate lyase, cysteine proteinase SAG12, serine hydroxymethyltransferase, and arginine decarboxylase (Kovalchuk et al. 2005). 4.3 Water Status Effects The disruption of plant water status after lead treatment has been addressed in many studies (Brunet et al. 2009). Results of such exposures show a decrease in transpira- tion, as well as reduction of the moisture content (Barcelo and Poschenrieder 1990; Patra et al. 2004). Reduced transpiration may result from reduced leaf surface area for transpiration that is caused by decreased leaf growth (Elibieta and Miroslawa 2005). However, some plant species that have high stomatal density are capable of coping with such effects (Kosobrukhov et al. 2004; Elibieta and Miroslawa 2005). Lead reduces plant cell wall plasticity, and thereby influences the cell turgor pres- sure. The decrease in concentrations of molecules that control cell turgor, such as sugars and amino acids, further accentuates the phenomenon of lead influence on turgor pressure (Barcelo and Poschenrieder 1990). The change in turgor pressure, particularly in the guard cells, interferes with stomatal opening and closing. To maintain cell turgor pressure, plants synthesize high concentrations of osmolytes, particularly proline under lead stress conditions (Qureshi et al. 2007). Stomatal opening/closing is controlled by abscisic acid (ABA), a phytohor- mone (Roelfsema and Hedrich 2005). The presence of Pb2+ ions causes a large accumulation of ABA in roots and aerial plant parts (Parys et al. 1998; Atici et al. 2005; Cenkci et al. 2010), leading to stomatal closure (Mohan and Hosetti 1997). Stomatal closure strongly limits gas exchange with the atmosphere, and water losses by transpiration (Parys et al. 1998). According to Elibieta and Miroslawa (2005), the foliar respiration of plants is also reduced by lead expo- sure, because the deposition of a cuticle layer, for example, on Glycine max leaf surfaces, is affected. Moreover, a CO2 /O2 imbalance in plants from lead-induced oxidative phosphorylation and respiratory disorders may also disrupt plant water status.
4.4 Mineral Nutrition Effects Results from multiple studies demonstrate that nutrient uptake by plants is signifi- cantly affected by the presence of lead (Chatterjee et al. 2004; Sharma and Dubey 2005; Gopal and Rizvi 2008). Although data are insufficient to allow a definitive conclusion to be drawn, it is known that lead affects plant mineral uptake. It is also known that lead exposure decreases the concentration of divalent cations (Zn2+ , Mn2+ , Mg2+ , Ca2+ , and Fe2+ ) in leaves of Z. mays (Seregin et al. 2004), O. sativa (Chatterjee et al. 2004), Brassica oleracea (Sinha et al. 2006), Medicago sativa (Lopez et al. 2007), V. unguiculata (Kopittke et al. 2007), and Raphanus sativus (Gopal and Rizvi 2008). But, it is not possible to conclude if this decrease results from blockage of root absorption, a decrease in translocation from roots to aerial plant parts, or a change in distribution of these elements in the plant. The effect of lead on mineral accumulation in aerial plant parts, in most cases, follows a common trend. In roots, the trend varies according to plant species or the intensity of the imposed stress (Lopez et al. 2007; Kopittke et al. 2007; Gopal and Rizvi 2008). The decreased absorption of nutrient in the presence of lead may result from competition (e.g., those with atomic size similar to lead) or changes in physiological plant activities. According to Sharma and Dubey (2005), the strong interaction of K+ ions with lead could result from their similar radii (Pb2+ : 1.29 Å and K+ : 1.33 Å): these two ions may compete for entry into the plant through the same potassium channels. Similarly, lead effects on K+ -ATPase and -SH groups of cell membrane proteins cause an efflux of K+ from roots. However, lead does not cause nitrogen efflux. The general reduction in the concentration of inorganic nitrogen in all plant parts could be induced by the reduced activity of nitrate reductase, the rate-limiting enzyme in the nitrate assimilation process (Xiong et al. 2006; Sengar et al. 2009). Xiong et al. (2006) reported that lead exposure (4 and 8 mmol kg–1 ) significantly decreased shoot nitrate content (70 and 80%), nitrate reductase activity (100 and 50%), and free amino acid content (81 and 82%) in B. pekinensis. 4.5 Photosynthesis Effects Photosynthesis inhibition is a well-known symptom of lead toxicity (Xiong et al. 2006; Hu et al. 2007; Liu et al. 2008; Piotrowska et al. 2009; Sing et al. 2010; Cenkci et al. 2010). This inhibition is believed to result from the following indirect effects of lead rather than from a direct effect: • distorted chloroplast ultrastructure from the affinity lead has for protein N and S ligands (Elibieta and Miroslawa 2005; Islam et al. 2007), • decreased ferredoxin NADP+ reductase and delta-aminolevulinic acid dehy- dratase (ALAD) activity at the origin of chlorophyll synthesis inhibition (Gupta et al. 2009; Cenkci et al. 2010),
• inhibition of plastoquinone and carotenoid synthesis (Kosobrukhov et al. 2004; Chen et al. 2007; Liu et al. 2008; Cenkci et al. 2010), • obstruction of the electron transport system (Qufei et al. 2009), • inadequate concentration of carbon dioxide via stomatal closure (Romanowska et al. 2002, 2005, 2006), • impaired uptake of essential elements such as Mn and Fe (Chatterjee et al. 2004; Gopal and Rizvi 2008) and substitution of divalent cations by lead (Gupta et al. 2009; Cenkci et al. 2010), • inhibition of Calvin cycle enzymatic catalysis (Mishra et al. 2006; Liu et al. 2008), and • increased chlorophyllase activity (Liu et al. 2008). However, these different effects vary by plant species. Generally, chlorophyll b is more sensitive than chlorophyll a (Xiong et al. 2006). The mechanism of chlorophyll breakdown into phytol, magnesium and the primary cleavage product of the porphyrin ring occur in four consecutive steps. This reaction is catalyzed by chlorophyllase, Mg-dechelatase, pheophorbide a oxygenase, and red chlorophyll catabolite reductase. Loss of the typical chlorophyll green color occurs only after cleavage of the porphyrin ring (Harpaz-Saad et al. 2007). The decrease observed in photosynthetic activity is often a more sensitive measure than is pigment content. 4.6 Respiration Effects When exposed to lead, photosynthetic plants usually experience harmful effects on respiration and adenosine triphosphate (ATP) content. Unlike the photosynthetic activity, the effect of lead on respiratory activity has been little studied (Seregin and Ivanov 2001). All the studies carried out on respiratory activity deal with leaves, whereas the effect of the Pb2+ ions on the respiratory activity of roots remains unknown. Lead is reported to affect the activity of ribulose-bisphosphate carboxy- lase in C3 plants that control CO2 assimilation, without affecting oxygenase activity (Assche and Clijsters 1990). Therefore, it is quite possible that photosynthesis is significantly reduced without any effect on photorespiration being induced, thus increasing the relative ratio of photorespiration to photosynthesis. Parys et al. (1998) reported that the CO2 concentration of P. sativum in leaves increased significantly after exposure to lead nitrate, most probably from the reduced photosynthetic and increased respiration activity. Romanowska et al. (2002) stressed that Pb2+ -induced increases in respiration resulted only from dark (mitochondrial) respiration, while photorespiration was unaffected. The stimulation of dark respiration by lead was observed in leaves or protoplasts of P. sativum and H. vulgare (Romanowska et al. 2002, 2005, 2006). Moreover, the stimulation of respiration was well correlated with increased production of ATP in mitochondria, resulting in the high energy demands of the plant to combat lead effects being met.
It has been also shown that divalent cations (e.g., Pb, Zn, Cd, Co, and Ni) can bind to mitochondrial membranes, disrupting the electron transport that could lead to decoupling of phosphorylation (Romanowska et al. 2002, 2006). An increase in the respiratory rate of 20–50% was observed by Romanowska et al. (2002) in the detached leaves of C3 plants (P. sativum and H. vulgare) and C4 plants (Z. mays), when they were exposed to 5 mM Pb(NO3 )2 for 24 h. Glycine, succinate, and malate substrates were more fully oxidized in mitochondria, isolated from lead- treated P. sativum leaves, than in mitochondria from control leaves (Romanowska et al. 2002). Lead caused an increase in ATP content as well as an increase in the ATP/ADP ratio in P. sativum and H. vulgare leaves (Romanowska et al. 2005, 2006). Rapid fractionation of H. vulgare protoplasts, incubated under conditions of low and high CO2 , indicated that the increased ATP/ADP ratio in lead-treated leaves mainly resulted from the production of mitochondrial ATP. The activity of NAD+ - malate dehydrogenase in protoplasts of barley leaves treated with lead was threefold higher than that in protoplasts from control leaves (Romanowska et al. 2005). Lead also significantly inhibited Hill reaction activity in spinach chloroplasts, in addition to photophosphorylation; moreover, lead had a more conspicuous effect on cyclic photophosphorylation than on noncyclic photophosphorylation (Romanowska et al. 2008). Recently, Qufei and Fashui (2009) reported that the accumulation of Pb2+ in photosystem II resulted in damage to its secondary structure and induced decreased absorbance of visible light and inhibited energy transfer among amino acids. Moreover, Jiang and Liu (2010) reported mitochondrial swelling, loss of cristae, and vacuolization of endoplasmic reticulum and dictyosomes during a 48–72 h lead exposure in A. sativum. 4.7 Genotoxicity The antimitotic effect of lead is one of its best known toxic effects on plants (Patra et al. 2004; Shahid et al. 2011). Indeed, Hammett (1928) demonstrated long ago that lead induces a dose-dependent decrease in mitotic activity in root cells of Allium cepa, which was later described in detail by Wierzbicka (1999) and Patra et al. (2004). In V. faba roots, lead shortened the mitotic stage and prolonged inter- phase, thus lengthening the cell cycle (Patra et al. 2004). The first step by which lead induces plant toxicity is the binding of the Pb2+ ion to cell membranes and to the cell wall. This produces rigidity in these components and reduces cell divi- sion. The second step is the disruption of microtubules that are essential for mitosis. Lead exposure induces disturbances in the G2 and M stages of cell division that leads to the production of abnormal cells at the c-mitosis (colchicine-mitosis) stage. This phenomenon is thought to be accentuated by direct or indirect interactions of lead with the proteins involved in the cell cycle, such as cyclins. Cyclin activity is indirectly dependent on the concentration of GSH. The spindle activity disturbances caused by lead may be transient in some cases, returning the mitotic index to initial levels.
Unlike antimitotic mechanisms, the mechanisms by which lead causes genotox- icity are complex and not yet well understood. At a low concentration, lead did not induce a significant effect on mitosis, but did induce aberrations (chromosomal bridges during anaphase), loss of acentric fragments during meiosis, chromosomal fragmentation, and micronucleus formation (National Toxicology Program 2003; Patra et al. 2004; Cecchi et al. 2008; Marcato et al. 2009; Grover et al. 2010; Barbosa et al. 2010; Shahid et al. 2011). The induction of chromosomal aberra- tions by lead can be explained, in part, by its action of disrupting the microtubule network. Results of in vitro studies have demonstrated that lead creates breaks in single and double strands of DNA and thereby affects horizontal DNA–DNA or DNA–protein links (Rucińska et al. 2004; Gichner et al. 2008; Shahid et al. 2011). Lead may enter the nucleus (Małecka et al. 2008) and bind directly to the DNA or indirectly to protein. After binding to DNA, lead disrupts DNA repair and repli- cation mechanisms. Lead does not induce direct genotoxic effects until it becomes attached to naked DNA (Valverde et al. 2001). Lead can also affect replication by replacing the zinc in the Zn-finger pattern of the enzymes that intervene in DNA repair (Gastaldo et al. 2007). Recently, Cenksi et al. (2010) used a random ampli- fied polymorphic DNA (RAPD) assay that amplifies random DNA fragments of genomic DNA, and they reported that genomic template stability was significantly affected by lead exposure in Brassica rapa. 4.8 Oxidative Stress and Lipid Peroxidation ROS are produced during normal cell metabolism in the chloroplast, either as by- products of the reduction of molecular oxygen (O2 ) or because of excitation in the presence of highly energized pigments. These ROS, such as superoxide radicals (O2 •– ), hydroxyl radicals (•OH), and hydrogen peroxide (H2 O2 ), are also generated following exposure to certain environmental agents. The production of ROS in the cells of aerobic organisms, defined as oxidative stress, is a well-known feature of the toxicity of heavy metals, including lead (Pourrut et al. 2008; Liu et al. 2008; Grover et al. 2010; Yadav 2010; Singh et al. 2010). However, the degree to which this feature is important is dependent on the metal type, specific form of the metal, plant species, exposure time, etc. When ROS forms exhaust cellular antioxidant reserves, they can rapidly attack and oxidize all types of biomolecules, such as nucleic acids, proteins, and lipids (Reddy et al. 2005; Clemens 2006; Hu et al. 2007; Wang et al. 2007; Yadav 2010). Such attacks lead to irreparable metabolic dysfunction and cell death. Lead causes marked changes in the lipid composition of different cell mem- branes (Liu et al. 2008; Piotrowska et al. 2009; Grover et al. 2010; Yan et al. 2010; Singh et al. 2010). The polyunsaturated fatty acids and their esters that are present in lipids show high susceptibility to ROS (Dey et al. 2007; Gupta et al. 2009). Indeed, ROS removes hydrogen from unsaturated fatty acids and forms lipid
radicals and reactive aldehydes, ultimately causing distortion of the lipid bilayer (Mishra et al. 2006). Lead-induced changes in lipid composition and potassium ion leakage were reported in Z. mays (Malkowski et al. 2002). Lead ions are known to induce lipid peroxidation, decrease the level of saturated fatty acids, and increase the unsaturated fatty acid content of membranes in several plant species (Singh et al. 2010). The oxidation of bis-allylic hydrogens on polyunsaturated fatty acids by ROS involves three distinct stages: initiation (formation of the lipid radical), progression (formation of lipid peroxyl radical by reaction between lipid radical and oxygen), and termination (formation of non-radical products after bimolecular interaction of lipid peroxyl radicals) (see details in the reviews of Gurer and Ercal 2000 and Bhattacharjee 2005). These lipid membrane changes cause the formation of abnor- mal cellular structures, such as alterations in the cell membrane (Dey et al. 2007; Islam et al. 2008; Gupta et al. 2009), organelles (e.g., mitochondria), peroxisomes (Małecka et al. 2008; Liu et al. 2008), or chloroplasts (Choudhury and Panda 2004; Elibieta and Miroslawa 2005; Hu et al. 2007). 5 Mechanisms of Lead Tolerance Plants respond to noxious effects of lead in various ways, such as selective metal uptake, metal binding to the root surface, binding to the cell wall, and induc- tion of antioxidants. There are several types of antioxidants to which plants may respond: non-protein thiol (NP-SH), cysteine, glutathione, ascorbic acid, proline, and antioxidant enzymes, such as superoxide dismutase (SOD), ascorbate peroxi- dase (APX), guaiacol peroxidase (GPX), catalase (CAT), and glutathione reductase (GR). However, the response varies with plant species, metal concentration, and exposure conditions. 5.1 Passive Mechanisms Even when small amounts of lead penetrate root cell membranes, it interacts with cellular components and increases the thickness of cell walls (Krzesłowska et al. 2009, 2010). Pectin is a component of plant cell walls. Lead complexation with pectin carboxyl groups is regarded as the most important interaction by which plant cells can resist lead toxicity (Meyers et al. 2008; Jiang and Liu 2010). Krzesłowska et al. (2009) observed that binding of lead to JIM5-P (within the cell wall and its resultant thickening) acted as a physical barrier that restricted lead access to the plasma membrane in F. hygrometrica protonemata. However, later, these authors stated that lead bound to JIM5-P within the cell can be taken up or remobilized by endocytosis, together with this pectin epitope (Krzesłowska et al. 2010).
5.2 Inducible Mechanisms Recently, several authors have reported the presence of transporter proteins among plant cells that play an important role in metal detoxification, by allowing the excretion of metal ions into extracellular spaces (Meyers et al. 2008; Vadas and Ahner 2009; Maestri et al. 2010). The human divalent metal transporter 1 (DMT1), expressed in yeast, has been shown to transport lead via a pH-dependent process (Bressler et al. 2004) in plants. Simultaneously, several ATP-binding cassette (ABC) carriers, such as AtATM3 or AtADPR12 at ATP-binding sites in Arabidopsis, were involved in resistance to lead (Kim et al. 2006; Cao et al. 2008). Although, suspected to act against lead, this detoxification mechanism has not yet been clearly con- firmed. Transcriptome analysis has shown that the gene expression of these carriers is stimulated by lead (Liu et al. 2009). Cellular sequestration is considered to be an important aspect of plant metal homeostasis and plant detoxification of heavy metals (Maestri et al. 2010). The lead, that could be bound by certain organic molecules (Piechalak et al. 2002; Vadas and Ahner 2009), is sequestered in several plant cell compartments: vacuoles (Małecka et al. 2008; Meyers et al. 2008), dictyosome vesicles (Malone et al. 1974), endo- plasmic reticulum vesicles (Wierzbicka et al. 2007), or plasmatubules (Wierzbicka 1998). Cysteine and glutathione (GSH) are known to be non-enzymatic antioxidants in plants. An increase in cysteine content, in response to lead toxicity, has been demonstrated in Arabidopsis thaliana (Liu et al. 2009). Glutathione protects plants from lead stress by quenching lead-induced ROS (Verbruggen et al. 2009; Liu et al. 2009). Moreover, as the substrate for phytochelatin (PC) biosynthesis, the glutathione-related proteins play an important role in heavy metal detoxification and homeostasis (Liu et al. 2009). Lead treatment can induce different GSH genes, including glutathione-synthetase, -peroxidase, and -reductase, and glutamylcysteine synthetase. Glutathione can also enhance accumulation of proline in stressed plants, a role that is associated with reducing damage to membranes and proteins (Liu et al. 2009). Gupta et al. (2010) reported the role of GSH in lead detoxification in S. alfredii, although this was accomplished without any induction of PC. This suggests that GSH may play an important role in detoxifying lead, under stress conditions where PCs are absent. PCs and metallothioneins (MTs) are the best characterized metal-binding ligands in plant cells. These ligands belong to different classes of cysteine-rich heavy metal- binding protein molecules. PCs, the most frequently cited metal protective proteins in plants, are low molecular weight, metal-binding proteins that can form mercaptide bonds with various metals (Maestri et al. 2010) and play an important role in their detoxification in plants (Brunet et al. 2009; Liu et al. 2009; Gupta et al. 2010; Yadav 2010; Jiang and Liu 2010). These thiols are biologically active compounds, whose function is to prevent oxidative stress in plant cells (Verbruggen et al. 2009; Gupta et al. 2010). Their general structure is (γ-glutamyl-cys)n Gly where n = 2–11, and they are synthesized by the action of γ-glutamylcysteine dipeptidyl transpeptidase (phytochelatin synthase; PCS) from GSH (Yadav 2010).
Lead is known to stimulate the production of PC and activate PCS (Mishra et al. 2006; Clemens 2006; Andra et al. 2009; Vadas and Ahner 2009; Sing et al. 2010). It has been proposed that in vivo, phytochelatins are involved in the cellular detoxi- fication and accumulation of several metals, including lead, because of their ability to form stable metal–PC complexes (Clemens 2006; Yadav 2010). Phytochelatin sequesters soluble lead in the cytoplasm before transporting it to vacuoles and chloroplasts (Piechalak et al. 2002; Małecka et al. 2008; Jiang and Liu 2010), thus reducing the deleterious effect of Pb2+ in the cells. The mechanism regulating the passage of the lead–PC complex through the tonoplast is, however, not yet known. Gisbert et al. (2003) reported significantly increased uptake and tolerance to lead and Cd following the induction and over-expression of a wheat gene encoding for phytochelatin synthase (TaPCS1), in Nicotiana glauca. 5.3 Antioxidant Enzymes To cope with the increased production of ROS and to avoid oxidative damage, plants have a system of antioxidant enzymes that scavenge the ROS that are present in different cell compartments (Brunet et al. 2009; Singh et al. 2010; Gupta et al. 2010). Lead-induced toxicity may inhibit the activity of these enzymes or may induce their synthesis (Table 1). However, lead-induced inhibition or induction of antioxidant enzymes is dependent on metal type, specific form of the metal, plant species type, and the duration/intensity of the treatment (Islam et al. 2008; Gupta et al. 2009; Singh et al. 2010). Generally, lead inhibits enzymatic activities and, when this occurs, the values of the inactivation constant (Ki ) range between 10–5 and 2 × 10–4 M (i.e., 50% of enzymatic activities are inhibited in this concentration range) (Seregin and Ivanov 2001). Enzyme inhibition results from the affinity lead has for -SH groups on the enzyme (Sharma and Dubey 2005; Gupta et al. 2009). This is true for more than 100 enzymes, including ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO) and nitrate reductase. Inactivation results from a link at either the catalytic site or elsewhere on the protein and produces an altered tertiary structure. Lead can also produce the same effect by binding to protein-COOH groups (Gupta et al. 2009, 2010). Lead also interacts with metalloid enzymes. Indeed, lead can disrupt plant absorption of minerals that contain zinc, iron, manganese, etc., which are essential for these enzymes. Lead and other divalent cations also can substitute for these metals, and thereby inactivate enzymes, as occurs with ALAD (Gupta et al. 2009; Cenkci et al. 2010). The effect lead has on ROS constitutes another mechanism by which lead exposure affects protein behavior (Gupta et al. 2009, 2010). Lead exposure is also known to stimulate the activities of certain enzymes (Table 1), but the mechanisms of action are, as yet, unclear. It has been proposed that lead activates certain enzymes by modulating gene expression or by restricting the activity of enzyme inhibitors (Seregin and Ivanov 2001). Indeed, antioxidant enzymes scavenge ROS, when they are produced in excess as a consequence of
Table 1 Lead-induced activation (↑) and reduction (↓) of enzymatic activities in different plant species Plant species ↑ Enzyme activity ↓ Enzyme activity References Najas indica SOD, GPX, APX, Sing et al. (2010) CAT, GR Sedum alfredii SOD, APX Gupta et al. (2010) Zea mays SOD, CAT, AsA Gupta et al. (2009) Lathyrus sativus APX, GR, GST Brunet et al. (2009) Wolffia arrhiza CAT, APX Piotrowska et al. (2009) Raphanus sativus POD, ribonuclease CAT Gopal and Rizvi (2008) Elsholtzia argyi CAT SOD, GPX Islam et al. (2008) Kandelia candel SOD, POD, CAT Zhang et al. (2007) Bruguiera SOD, POD, CAT Zhang et al. (2007) gymnorrhiza Cassia angustifolia SOD, APX, GR, Qureshi et al. (2007) CAT Zea mays SOD, POD, CAT Wang et al. (2007) Triticum aestivum SOD, POX CAT Dey et al. (2007) Potamogeton crispus POD, SOD, CAT SOD, CAT Hu et al. (2007) Ceratophyllum SOD, GPX, APX, SOD, GPX, APX, Mishra et al. (2006) demersum CAT, GR CAT, GR Helianthus annuus GR CAT Garcia et al. (2006) Macrotyloma SOD, CAT, POD, Reddy et al. (2005) uniflorum GR, GST Cicer arietinum SOD, CAT, POD, Reddy et al. (2005) GR, GST Taxithelium APX, GPOX, CAT, Choudhury and nepalense Panda (2004) Oryza sativa SOD, GPX, APX, GR, CAT Verma and Dubey CAT, GR (2003) SOD, superoxide dismutase; APX, ascorbate peroxidase; GPX, guaiacol peroxidase; CAT, cata- lase; GR, glutathione reductase; AsA, ascorbic acid; GST, GSH S-transferase; GSH, glutathione; POD, peroxidase metal toxicity. Superoxide dismutase, a metallo-enzyme present in various cell com- partments, is considered to be the first defense against oxidative stress (Mishra et al. 2006). It catalyses the dismutation of two superoxide radicals to H2 O2 and oxygen and thus maintains superoxide radicals at steady-state levels (Islam et al. 2008; Gupta et al. 2009). H2 O2 is a very strong oxidant and requires quick removal to avoid oxidative toxicity; removal is achieved by the action of APX in the ascorbate-glutathione cycle or by GPX and CAT in the cytoplasm and in other cell compartments (Mishra et al. 2006). The role of GSH and glutathione reductase in the H2 O2 -scavenging mechanism in plant cells (Piechalak et al. 2002) is well estab- lished in the Halliwell–Asada enzyme pathway. Moreover, antioxidant enzymes may be activated from the increased concentration of their substrates, instead of direct interaction with lead (Islam et al. 2008).
6 Conclusions and Perspectives Lead is a major inorganic global pollutant and numerous studies have revealed its biogeochemical behavior and impact on the biosphere. Based on these studies, as cited in this review, it is concluded as follows: (1) Lead has been in use since antiquity, because of its many useful proper- ties. The continued use of lead in many industrial processes has increased its concentration to toxic levels in all environmental compartments. (2) Lead forms stable complexes with different compounds in soil and tends to be stored in the soil. The fate and behavior of lead in soil is affected by its form, solubility, mobility, and bioavailability and is controlled by many biogeochemi- cal parameters, such as soil pH, redox conditions, cation-exchange capacity, soil mineralogy, biological and microbial conditions, amount and nature of organic and inorganic ligands present, and competing cations. (3) Lead enters plants mainly through the roots via the apoplast pathway or calcium ion channels. Lead can also enter plants in small amounts through leaves. Once in the roots, lead tends to sequester in root cells. Only a limited amount of lead is translocated from roots to shoot tissues, because there are natural plant barriers in the root endodermis (e.g., Casparian strips). (4) Lead has no biological function and induces various noxious effects inside plants. Excessive lead accumulation in plant tissue is toxic to most plants, leading to a decrease in seed germination, root elongation, decreased biomass, inhibition of chlorophyll biosynthesis, mineral nutrition and enzymatic reac- tions, as well as a number of other physiological effects. The intensity of these effects varies depending on the duration of exposure, stage of plant devel- opment, studied organ, and the concentration of lead used in the exposure assessment. (5) Lead-induced production of ROS is the major cause of its toxicity. These free radicals disrupt the redox status of cells, causing oxidative stress and DNA dam- age through oxidation, and lead to irreparable metabolic dysfunction and cell death. (6) Plants defend against lead toxicity through several avoidance or detoxification mechanisms. Plants resist lead entry into their cells via exclusion or they bind lead to their cell walls or other ligands. Plants combat lead-induced increased production of ROS by activating various antioxidant enzymes. (7) The efficiency of detoxification mechanisms determines the final tolerance or sensitivity of plants to metal-induced stress. Plants that have efficient detoxi- fication mechanisms are generally characterized as being hyperaccumulators. Such plants are useful in soil bioremediation for many metal types. Conversely, plants that do not efficiently cope with pollutants are sensitive to metal toxicity and are often used in risk assessment studies. In this review, we raise several questions that need attention if our understanding of the biogeochemical behavior of lead in different environmental compartments is
to be advanced. Lead is known to interfere directly or indirectly with the genetic material to induce ROS and modify (increase or decrease) the activities of certain enzymes in plants. These responses of plants to lead toxicity are often used as tools for risk assessment. However, the mechanisms of action underlying the noxious effects of lead in plants are still unknown. Moreover, most field work performed on the effects of lead on plants is based almost exclusively on the total metal content in polluted soil, even though this is of little significance from an environmental point of view. Indeed, the potential effects of lead and other toxic elements in the environment depend on insights into their physico-chemical distribution, i.e., speciation. Therefore, if environmental scientists are to become better at predicting what the toxicity or environmental impact of lead may be, then additional research into the form of lead present is essential. 7 Summary Lead has gained considerable attention as a persistent toxic pollutant of concern, partly because it has been prominent in the debate concerning the growing anthro- pogenic pressure on the environment. The purpose of this review is to describe how plants take lead up and to link such uptake to the ecotoxicity of lead in plants. Moreover, we address the mechanisms by which plants or plant systems detoxify lead. Lead has many interesting physico-chemical properties that make it a very useful heavy metal. Indeed, lead has been used by people since the dawn of civilization. Industrialization, urbanization, mining, and many other anthropogenic activities have resulted in the redistribution of lead from the earth’s crust to the soil and to the environment. Lead forms various complexes with soil components, and only a small fraction of the lead present as these complexes in the soil solution are phytoavailable. Despite its lack of essential function in plants, lead is absorbed by them mainly through the roots from soil solution and thereby may enter the food chain. The absorption of lead by roots occurs via the apoplastic pathway or via Ca2+ -permeable channels. The behavior of lead in soil, and uptake by plants, is controlled by its speciation and by the soil pH, soil particle size, cation-exchange capacity, root surface area, root exudation, and degree of mycorrhizal transpiration. After uptake, lead primarily accumulates in root cells, because of the blockage by Casparian strips within the endodermis. Lead is also trapped by the negative charges that exist on roots’ cell walls. Excessive lead accumulation in plant tissue impairs various morphological, phys- iological, and biochemical functions in plants, either directly or indirectly, and induces a range of deleterious effects. It causes phytotoxicity by changing cell mem- brane permeability, by reacting with active groups of different enzymes involved in plant metabolism and by reacting with the phosphate groups of ADP or ATP, and by replacing essential ions. Lead toxicity causes inhibition of ATP production,
lipid peroxidation, and DNA damage by over production of ROS. In addition, lead strongly inhibits seed germination, root elongation, seedling development, plant growth, transpiration, chlorophyll production, and water and protein content. The negative effects that lead has on plant vegetative growth mainly result from the fol- lowing factors: distortion of chloroplast ultrastructure, obstructed electron transport, inhibition of Calvin cycle enzymes, impaired uptake of essential elements, such as Mg and Fe, and induced deficiency of CO2 resulting from stomatal closure. Under lead stress, plants possess several defense strategies to cope with lead toxicity. Such strategies include reduced uptake into the cell; sequestration of lead into vacuoles by the formation of complexes; binding of lead by phytochelatins, glutathione, and amino acids; and synthesis of osmolytes. In addition, activa- tion of various antioxidants to combat increased production of lead-induced ROS constitutes a secondary defense system. References Alexander PD, Alloway BJ, Dourado AM (2006) Genotypic variations in the accumulation of Cd, Cu, Pb and Zn exhibited by six commonly grown vegetables. Environ Pollut 144: 736–745 Andra SS, Datta R, Sarkar D, Sarkar D, Saminathan SK, Mullens CP, Bach SB (2009) Analysis of phytochelatin complexes in the lead tolerant vetiver grass [Vetiveria zizanioides (L.)] using liquid chromatography and mass spectrometry. Environ Pollut 157(7):2173–2183 Arazi T, Sunkar R, Kaplan B, Fromm H (1999) A tobacco plasma membrane calmodulin-binding transporter confers Ni2+ tolerance and Pb2+ hypersensitivity in transgenic plants. Plant J 20:171–182 Arias JA, Peralta-Videa JR, Ellzey JT, Ren M, Viveros MN, Gardea-Torresdey JL (2010) Effects of Glomus deserticola inoculation on Prosopis: enhancing chromium and lead uptake and translocation as confirmed by X-ray mapping, ICP-OES and TEM techniques. Environ Exp Bot 68(2):139–148 Arshad M, Silvestre J, Pinelli E, Kallerhoff J, Kaemmerer M, Tarigo A, Shahid M, Guiresse M, Pradere P, Dumat C (2008) A field study of lead phytoextraction by various scented Pelargonium cultivars. Chemosphere 71(11):2187–2192 Assche F, Clijsters H (1990) Effects of metals on enzyme activity in plants. Plant Cell Environ 13(3):195–206 Atici Ö, Ağar G, Battal P (2005) Changes in phytohormone contents in chickpea seeds germinating under lead or zinc stress. Biol Plantarum 49(2):215–222 ATSDR (2003) Agency for Toxic Substances and Disease Registry. http://www.atsdr.cdc.gov/ Barbosa J, Cabral T, Ferreira D, Agnez-Lima L, Batistuzzo de Medeiros S (2010) Genotoxicity assessment in aquatic environment impacted by the presence of heavy metals. Ecotoxicol Environ Saf 73(3):320–325 Barceló J, Poschenrieder C (1990) Plant water relations as affected by heavy metal stress: a review. J Plant Nutr 13(1):1–37 Barrutia O, Garbisu C, Hernández-Allica J, García-Plazaola JI, Becerril JM (2010) Differences in EDTA-assisted metal phytoextraction between metallicolous and non-metallicolous accessions of Rumex acetosa L. Environ Pollut 158(5):1710–1715 Beltagi MS (2005) Phytotoxicity of lead (Pb) to SDS-PAGE protein profile in root nodules of faba bean (Vicia faba L.) plants. Pak J Biol Sci 8(5):687–690 Bhattacharjee S (2005) Reactive oxygen species and oxidative burst: roles in stress, senescence and signal transduction in plants. Curr Sci 89(7):1113–1121
Bi X, Ren L, Gong M, He Y, Wang L, Ma Z (2010) Transfer of cadmium and lead from soil to mangoes in an uncontaminated area, Hainan Island, China. Geoderma 155(1–2):115–120 Bressler JP, Olivi L, Cheong JH, Kim Y, Bannona D (2004) Divalent metal transporter 1 in lead and cadmium transport. Ann N Y Acad Sci 1012:142–152 Brunet J, Varrault G, Zuily-Fodil Y, Repellin A (2009) Accumulation of lead in the roots of grass pea (Lathyrus sativus L.) plants triggers systemic variation in gene expression in the shoots. Chemosphere 77(8):1113–1120 Cao X, Ma LQ, Singh SP, Zhou Q (2008) Phosphate-induced lead immobilization from different lead minerals in soils under varying pH conditions. Environ Pollut 152(1):184–192 Cecchi M, Dumat C, Alric A, Felix-Faure B, Pradere P, Guiresse M (2008) Multi-metal contamina- tion of a calcic cambisol by fallout from a lead-recycling plant. Geoderma 144(1–2):287–298 Cenkci S, Cigerci IH, Yildiz M, Özay C, Bozdag A, Terzi H (2010) Lead contamination reduces chlorophyll biosynthesis and genomic template stability in Brassica rapa L. Environ Exp Bot 67(3):467–473 Chatterjee C, Dube BK, Sinha P, Srivastava P (2004) Detrimental effects of lead phytotoxicity on growth, yield, and metabolism of rice. Commun Soil Sci Plant Anal 35(1–2):255–265 Chen J, Zhu C, Li L, Sun Z, Pan X (2007) Effects of exogenous salicylic acid on growth and H2 O2 - metabolizing enzymes in rice seedlings under lead stress. J Environ Sci (China) 19(1):44–49 Choudhury S, Panda S (2004) Toxic effects, oxidative stress and ultrastructural changes in moss Taxithelium Nepalense (Schwaegr.) Broth. under chromium and lead phytotoxicity. Water Air Soil Pollut 167(1):73–90 Clemens S (2006) Evolution and function of phytochelatin synthases. J Plant Physiol 163(3): 319–332 Dey SK, Dey J, Patra S, Pothal D (2007) Changes in the antioxidative enzyme activities and lipid peroxidation in wheat seedlings exposed to cadmium and lead stress. Braz J Plant Physiol 19(1):53–60 Dumat C, Quenea K, Bermond A, Toinen S, Benedetti MF (2006) Study of the trace metal ion influence on the turnover of soil organic matter in cultivated contaminated soils. Environ Pollut 142(3):521–529 Elzbieta W, Miroslawa C (2005) Lead-induced histological and ultrastructural changes in the leaves of soybean (Glycine max (L.) Merr.). Soil Sci Plant Nutr 51(2):203–212 Garcia JS, Gratão PL, Azevedo RA, Arruda MAZ (2006) Metal contamination effects on sunflower (Helianthus annuus L.) growth and protein expression in leaves during development. J Agric Food Chem 54(22):8623–8630 Garland C, Wilkins D (1981) Effect of calcium on the uptake and toxicity of lead in Hordeum vulgare L. and Festuca ovina L. New Phytol 87(3):581–593 Gastaldo J, Viau M, Bencokova Z, Joubert A, Charvet A, Balosso J, Foray M (2007) Lead contam- ination results in late and slowly repairable DNA double-strand breaks and impacts upon the ATM-dependent signaling pathways. Toxicol Lett 173(3):201–214 Gichner T, Znidar I, Száková J (2008) Evaluation of DNA damage and mutagenicity induced by lead in tobacco plants. Mutat Res Genet Toxicol Environ Mutagen 652(2):186–190 Ginn BR, Szymanowski JS, Fein JB (2008) Metal and proton binding onto the roots of Fescue rubra. Chem Geol 253(3–4):130–135 Gisbert C, Ros R, De Haro A, Walker DJ, Pilar Bernal M, Serrano R, Navarro-Aviñó J (2003) A plant genetically modified that accumulates Pb is especially promising for phytoremediation. Biochem Biophys Res Commun 303(2):440–445 Gopal R, Rizvi AH (2008) Excess lead alters growth, metabolism and translocation of certain nutrients in radish. Chemosphere 70(9):1539–1544 Grover P, Rekhadevi P, Danadevi K, Vuyyuri S, Mahboob M, Rahman M (2010) Genotoxicity evaluation in workers occupationally exposed to lead. Int J Hyg Environ Health 213(2):99–106 Gupta D, Huang H, Yang X, Razafindrabe B, Inouhe M (2010) The detoxification of lead in Sedum alfredii H. is not related to phytochelatins but the glutathione. J Hazard Mater 177(1–3): 437–444
Gupta D, Nicoloso F, Schetinger M, Rossato L, Pereira L, Castro G, Srivastava S, Tripathi R (2009) Antioxidant defense mechanism in hydroponically grown Zea mays seedlings under moderate lead stress. J Hazard Mater 172(1):479–484 Gurer H, Ercal N (2000) Can antioxidants be beneficial in the treatment of lead poisoning? Free Radic Biol Med 29(10):927–945 Hammett FS (1928) Studies in the biology of metals. Protoplasma 5(1):535–542 Harpaz-Saad S, Azoulay T, Arazi T, Ben-Yaakov E, Mett A, Shiboleth YM, Hortensteiner S, Gidoni D, Gal-On A, Goldschmidt EE, Eyal Y (2007) Chlorophyllase is a rate-limiting enzyme in chlorophyll catabolism and is posttranslationally regulated. Plant Cell 19(3):1007–1022 Hirsch RE, Lewis BD, Spalding EP, Sussman MR (1998) A role for the AKT1 potassium channel in plant nutrition. Science 280(5365):918–921 Hu J, Shi G, Xu Q, Wang X, Yuan Q, Du K (2007) Effects of Pb2+ on the active oxygen- scavenging enzyme activities and ultrastructure in Potamogeton crispus leaves. Russ J Plant Physl 54(3):414–419 Huang JW, Cunningham SD (1996) Lead phytoextraction: species variation in lead uptake and translocation. New Phytol 134:75–84 Islam E, Liu D, Li T, Yang X, Jin X, Mahmood Q, Tian S, Li J (2008) Effect of Pb toxicity on leaf growth, physiology and ultrastructure in the two ecotypes of Elsholtzia argyi. J Hazard Mater 154(1–3):914–926 Islam E, Yang X, Li T, Liu D, Jin X, Meng F (2007) Effect of Pb toxicity on root morphol- ogy, physiology and ultrastructure in the two ecotypes of Elsholtzia argyi. J Hazard Mater 147(3):806–816 Jiang W, Liu D (2010) Pb-induced cellular defense system in the root meristematic cells of Allium sativum L. BMC Plant Biol 10:40–40 Kim D, Bovet L, Kushnir S, Noh EW, Martinoia E, Lee Y (2006) AtATM3 is involved in heavy metal resistance in Arabidopsis. Plant Physiol 140(3):922–932 Kim YY, Yang YY, Lee Y (2002) Pb and Cd uptake in rice roots. Physiol Plantarum 116:368–372 Kohler C, Merkle T, Neuhaus G (1999) Characterisation of a novel gene family of putative cyclic nucleotide- and calmodulin-regulated ion channels in Arabidopsis thaliana. Plant J 18(1): 97–104 Komjarova I, Blust R (2009) Effect of Na, Ca and pH on simultaneous uptake of Cd, Cu, Ni, Pb, and Zn in the water flea Daphnia magna measured using stable isotopes. Aquat Toxicol 94(2):81–86 Kopittke PM, Asher CJ, Kopittke RA, Menzies NW (2007) Toxic effects of Pb2+ on growth of cowpea (Vigna unguiculata). Environ Pollut 150(2):280–287 Kopittke PM, Asher CJ, Kopittke RA, Menzies NW (2008) Prediction of Pb speciation in concentrated and dilute nutrient solutions. Environ Pollut 153(3):548–554 Kosobrukhov A, Knyazeva I, Mudrik V (2004) Plantago major plants responses to increase content of lead in soil: growth and photosynthesis. Plant Growth Regul 42(2):145–151 Kovalchuk I, Titov V, Hohn B, Kovalchuk O (2005) Transcriptome profiling reveals similarities and differences in plant responses to cadmium and lead. Mutat Res: Fundam Mol Mech Mutagen 570(2):149–161 Krzeslowska M, Lenartowska M, Mellerowicz EJ, Samardakiewicz S, Wozny A (2009) Pectinous cell wall thickenings formation–a response of moss protonemata cells to lead. Environ Exp Bot 65(1):119–131 Krzesłowska M, Lenartowska M, Samardakiewicz S, Bilski H, Woźny A (2010) Lead deposited in the cell wall of Funaria hygrometrica protonemata is not stable–a remobilization can occur. Environ Pollut 158(1):325–338 Lane SD, Martin ES (1977) A histochemical investigation of lead uptake in Raphanus sativus. New Phytol 79(2):281–286 Lawal O, Sanni A, Ajayi I, Rabiu O (2010) Equilibrium, thermodynamic and kinetic studies for the biosorption of aqueous lead(II) ions onto the seed husk of Calophyllum inophyllum. J Hazard Mater 177(1–3):829–835
Liao Y, Chien SC, Wang M, Shen Y, Hung P, Das B (2006) Effect of transpiration on Pb uptake by lettuce and on water soluble low molecular weight organic acids in rhizosphere. Chemosphere 65(2):343–351 Liu D, Li T, Jin X, Yang X, Islam E, Mahmood Q (2008) Lead induced changes in the growth and antioxidant metabolism of the lead accumulating and non-accumulating ecotypes of Sedum alfredii. J Integr Plant Biol 50(2):129–140 Liu T, Liu S, Guan H, Ma L, Chen Z, Gu H (2009) Transcriptional profiling of Arabidopsis seedlings in response to heavy metal lead (Pb). Environ Exp Bot 67(2):377–386 Liu X, Peng K, Wang A, Lian C, Shen Z (2010) Cadmium accumulation and distribution in populations of Phytolacca americana L. and the role of transpiration. Chemosphere 78(9):1136–1141 López ML, Peralta-Videa JR, Benitez T, Duarte-Gardea M, Gardea-Torresdey JL (2007) Effects of lead, EDTA, and IAA on nutrient uptake by alfalfa plants. J Plant Nutr 30(8): 1247–1261 Maestri E, Marmiroli M, Visioli G, Marmiroli N (2010) Metal tolerance and hyperaccumulation: costs and trade-offs between traits and environment. Environ Exp Bot 68(1):1–13 Malone C, Koeppe DE, Miller RJ (1974) Localization of lead accumulated by corn plants. Plant Physiol 53(3):388–394 Małecka A, Piechalak A, Morkunas I, Tomaszewska B (2008) Accumulation of lead in root cells of Pisum sativum. Acta Physiol Plant 30(5):629–637 Małkowski E, Kita A, Galas W, Karcz W, Kuperberg JM (2002) Lead distribution in corn seedlings (Zea mays L.) and its effect on growth and the concentrations of potassium and calcium. Plant Growth Regul 37(1):69–76 Marcato-Romain C, Guiresse M, Cecchi M, Cotelle S, Pinelli E (2009) New direct contact approach to evaluate soil genotoxicity using the Vicia faba micronucleus test. Chemosphere 77(3):345–350 Meyers DER, Auchterlonie GJ, Webb RI, Wood B (2008) Uptake and localisation of lead in the root system of Brassica juncea. Environ Pollut 153(2):323–332 Mishra S, Srivastava S, Tripathi R, Kumar R, Seth C, Gupta D (2006) Lead detoxification by coontail (Ceratophyllum demersum L.) involves induction of phytochelatins and antioxidant system in response to its accumulation. Chemosphere 65(6):1027–1039 Mohan BS, Hosetti BB (1997) Potential phytotoxicity of lead and cadmium to Lemna minor grown in sewage stabilization ponds. Environ Pollut 98(2):233–238 National Toxicology Program (2003) Report on carcinogens: background document for lead and lead compounds. Department of Health and Human Services, Research Triangle Park, NC Padmavathiamma PK, Li LY (2010) Phytoavailability and fractionation of lead and manganese in a contaminated soil after application of three amendments. Bioresour Technol 101(14): 5667–5676 Pais I, Jones JB (2000) The handbook of trace elements. Saint Lucie Press, Boca Raton, FL, p 223 Parys E, Romanowska E, Siedlecka M, Poskuta J (1998) The effect of lead on photosynthesis and respiration in detached leaves and in mesophyll protoplasts of Pisum sativum. Acta Physiol Plant 20(3):313–322 Patra M, Bhowmik N, Bandopadhyay B, Sharma A (2004) Comparison of mercury, lead and arsenic with respect to genotoxic effects on plant systems and the development of genetic tolerance. Environ Exp Bot 52(3):199–223 Piechalak A, Tomaszewska B, Baralkiewicz D, Malecka A (2002) Accumulation and detoxification of lead ions in legumes. Phytochemistry 60(2):153–162 Piotrowska A, Bajguz A, Godlewska-Zylkiewicz B, Czerpak R, Kaminska M (2009) Jasmonic acid as modulator of lead toxicity in aquatic plant Wolffia arrhiza (Lemnaceae). Environ Exp Bot 66(3):507–513 Pourrut B, Perchet G, Silvestre J, Cecchi M, Guiresse M, Pinelli E (2008) Potential role of NADPH-oxidase in early steps of lead-induced oxidative burst in Vicia faba roots. J Plant Physiol 165(6):571–579
Punamiya P, Datta R, Sarkar D, Barber S, Patel M, Das P (2010) Symbiotic role of glomus mosseae in phytoextraction of lead in vetiver grass [Chrysopogon zizanioides (L.)]. J Hazard Mater 177(1–3):465–474 Qufei L, Fashui H (2009) Effects of Pb2+ on the Structure and Function of Photosystem II of Spirodela polyrrhiza. Biol Trace Elem Res 129(1):251–260 Qureshi M, Abdin M, Qadir S, Iqbal M (2007) Lead-induced oxidative stress and metabolic alterations in Cassia angustifolia Vahl. Biol Plantarum 51(1):121–128 Reddy AM, Kumar SG, Jyothsnakumari G, Thimmanaik S, Sudhakar C (2005) Lead induced changes in antioxidant metabolism of horsegram (Macrotyloma uniflorum (Lam.) Verdc.) and bengalgram (Cicer arietinum L.). Chemosphere 60(1):97–104 Roelfsema MRG, Hedrich R (2005) In the light of stomatal opening: new insights into ‘the Watergate’. New Phytol 167(3):665–691 Romanowska E, Igamberdiev AU, Parys E, Gardeström P (2002) Stimulation of respiration by Pb2+ in detached leaves and mitochondria of C3 and C4 plants. Physiol Plant 116(2): 148–154 Romanowska E, Pokorska B, Siedlecka M (2005) The effects of oligomycin on content of adeny- lates in mesophyll protoplasts, chloroplasts and mitochondria from Pb2+ treated pea and barley leaves. Acta Physiol Plant 27(1):29–36 Romanowska E, Wróblewska B, Drozak A, Siedlecka M (2006) High light intensity protects photosynthetic apparatus of pea plants against exposure to lead. Plant Physiol Biochem 44(5–6):387–394 Romanowska E, Wróblewska B, Drożak A, Zienkiewicz M, Siedlecka M (2008) Effect of Pb ions on superoxide dismutase and catalase activities in leaves of pea plants grown in high and low irradiance. Biol Plantarum 52(1):80–86 Rucińska R, Sobkowiak R, Gwóźdź EA (2004) Genotoxicity of lead in lupin root cells as evaluated by the comet assay. Cell Mol Biol Lett 9(3):519–528 Sammut M, Noack Y, Rose J, Hazemann J, Proux O, Depoux Ziebel M, Fiani E (2010) Speciation of Cd and Pb in dust emitted from sinter plant. Chemosphere 78(4):445–450 Sengar RS, Gautam M, Sengar RS, Sengar RS, Garg SK, Sengar K, Chaudhary R (2009) Lead stress effects on physiobiochemical activities of higher plants. Rev Environ Contam Toxicol 196:1–21 Seregin IV, Ivanov VB (2001) Physiological aspects of cadmium and lead toxic effects on higher plants. Russ J Plant Physiol 48(4):523–544 Seregin IV, Shpigun LK, Ivanov VB (2004) Distribution and toxic effects of cadmium and lead on maize roots. Russ J Plant Physiol 51(4):525–533 Shahid M, Pinelli E, Pourrut B, Silvestre J, Dumat C (2011) Lead-induced genotoxicity to Vicia faba L. roots in relation with metal cell uptake and initial speciation. Ecotoxicol Environ Saf 74(1):78–84 Sharma P, Dubey RS (2005) Lead toxicity in plants. Braz J Plant Physiol 17(1):35–52 Singh R, Tripathi RD, Dwivedi S, Kumar A, Trivedi PK, Chakrabarty D (2010) Lead bioaccu- mulation potential of an aquatic macrophyte Najas indica are related to antioxidant system. Bioresour Technol 101:3025–3032 Sinha P, Dube B, Srivastava P, Chatterjee C (2006) Alteration in uptake and translocation of essential nutrients in cabbage by excess lead. Chemosphere 65(4):651–656 Tabelin C, Igarashi T (2009) Mechanisms of arsenic and lead release from hydrothermally altered rock. J Hazard Mater 169(1–3):980–990 Tanton TW, Crowdy SH (1971) The distribution of lead chelate in the transpiration stream of higher plants. Pestic Sci 2(5):211–213 Tomulescu IM, Radoviciu EM, Merca VV, Tuduce AD (2004) Effect of copper, zinc and lead and their combinations on the germination capacity of two cereals. J Agric Sci 15 Tung G, Temple PJ (1996) Uptake and localization of lead in corn (Zea mays L.) seedlings, a study by histochemical and electron microscopy. Sci Total Environ 188(2–3):71–85 U.S. Geological Survey (2009) http://minerals.usgs.gov/minerals/pubs/commodity/lead/
Uzu G, Sobanska S, Aliouane Y, Pradere P, Dumat C (2009) Study of lead phytoavailability for atmospheric industrial micronic and sub-micronic particles in relation with lead speciation. Environ Pollut 157(4):1178–1185 Uzu G, Sobanska S, Sarret G, Munoz M, Dumat C (2010) Foliar lead uptake by lettuce exposed to atmospheric fallouts. Environ Sci Technol 44:1036–1042 Vadas TM, Ahner BA (2009) Cysteine- and glutathione-mediated uptake of lead and cadmium into Zea mays and Brassica napus roots. Environ Pollut 157(8–9):2558–2563 Valverde M, Trejo C, Rojas E (2001) Is the capacity of lead acetate and cadmium chloride to induce genotoxic damage due to direct DNA-metal interaction? Mutagenesis 16(3):265–270 Vega F, Andrade M, Covelo E (2010) Influence of soil properties on the sorption and retention of cadmium, copper and lead, separately and together, by 20 soil horizons: comparison of linear regression and tree regression analyses. J Hazard Mater 174(1–3):522–533 Verbruggen N, Hermans C, Schat H (2009) Molecular mechanisms of metal hyperaccumulation in plants. New Phytol 181:759–776 Verma S, Dubey RS (2003) Lead toxicity induces lipid peroxidation and alters the activities of antioxidant enzymes in growing rice plants. Plant Sci 164:645–655 Wang H, Shan X, Wen B, Owens G, Fang J, Zhang S (2007) Effect of indole-3-acetic acid on lead accumulation in maize (Zea mays L.) seedlings and the relevant antioxidant response. Environ Exp Bot 61(3):246–253 Wierzbicka M (1998) Lead in the apoplast of Allium cepa L. root tips–ultrastructural studies. Plant Sci 133(1):105–119 Wierzbicka M (1999) Comparison of lead tolerance in Allium cepa with other plant species. Environ Pollut 104(1):41–52 Wierzbicka MH, Przedpełska E, Ruzik R, Ouerdane L, Połeć-Pawlak K, Jarosz M, Szpunar J, Szakiel A (2007) Comparison of the toxicity and distribution of cadmium and lead in plant cells. Protoplasma 231(1):99–111 Wojas S, Ruszczynska A, Bulska E, Wojciechowski M, Antosiewicz DM (2007) Ca2+ -dependent plant response to Pb2+ is regulated by LCT1. Environ Pollut 147(3):584–592 Xiong Z, Zhao F, Li M (2006) Lead toxicity in Brassica pekinensis Rupr.: effect on nitrate assimilation and growth. Environ Toxicol 21(2):147–153 Yadav S (2010) Heavy metals toxicity in plants: an overview on the role of glutathione and phytochelatins in heavy metal stress tolerance of plants. S Afr J Bot 76(2):167–179 Yan ZZ, Ke L, Tam NFY (2010) Lead stress in seedlings of Avicennia marina, a common mangrove species in South China, with and without cotyledons. Aquat Bot 92(2):112–118 Zaier H, Ghnaya T, Ben Rejeb K, Lakhdar A, Rejeb S, Jemal F (2010) Effects of EDTA on phy- toextraction of heavy metals (Zn, Mn and Pb) from sludge-amended soil with Brassica napus. Bioresour Technol 101(11):3978–3983 Zhang F, Wang Y, Lou Z, Dong J (2007) Effect of heavy metal stress on antioxidative enzymes and lipid peroxidation in leaves and roots of two mangrove plant seedlings (Kandelia candel and Bruguiera gymnorrhiza). Chemosphere 67(1):44–50