Plant Cd2+ and Zn2+ status effects on root and shoot heavy metal accumulation in Thlaspi caerulescens

* In this study we address the impact of changes in plant heavy metal, (i.e. zinc (Zn) and cadmium (Cd)) status on metal accumulation in the Zn/Cd hyperaccumulator, Thlaspi caerulescens. * Thlaspi caerulescens plants were grown hydroponically on both high and low Zn and Cd regimes and whole-shoot and -root metal accumulation, and root (109)Cd(2+) influx were determined. * High-Zn-grown (500 microm Zn) plants were found to be more Cd-tolerant than plants grown in standard Zn conditions (1 microm Zn). Furthermore, shoot Cd accumulation was significantly greater in the high-Zn-grown plants. A positive correlation was also found between shoot Zn accumulation and increased plant Cd status. Radiotracer (109)Cd root flux experiments demonstrated that high-Zn-grown plants maintained significantly higher root Cd(2+) influx than plants grown on 1 microm Zn. It was also found that both nickel (Ni) and copper (Cu) shoot accumulation were stimulated by high plant Zn status, while manganese (Mn) accumulation was not affected. * A speculative model is presented to explain these findings, suggesting that xylem loading may be one of the key sites responsible for the hyperaccumulation of Zn and Cd accumulation in Thlaspi caerulescens.     

Accumulation, detoxification, and genotoxicity of heavy metals in Indian mustard (Brassica juncea L.)

Plants of Indian mustard (Brassica juncea L.) were exposed to different concentrations (15, 30, 60, 120 microM) of (Cd, Cr, Cu, Pb) for 28 and 56 d for accumulation and detoxification studies. Metal accumulation in roots and shoots were analyzed and it was observed that roots accumulated a significant amount of Cd (1980 microg g(-1) dry weight), Cr (1540 microg g(-1) dry weight), Cu (1995 microg g(-1) dry weight), and Pb (2040 microg g(-1) dry weight) after 56 d of exposure, though in shoot this was 1110, 618, 795, and 409 microg g(-1) dry weight of Cd, Cr, Cu, and Pb, respectively. In order to assess detoxification mechanisms, non-protein thiols (NP-SH), glutathione (GSH) and phytochelatins (PCs) were analyzed in plants. An increase in the quantity of NP-SH (9.55), GSH (8.30), and PCs (1.25) micromol g(-1) FW were found at 15 microM of Cd, however, a gradual decline in quantity was observed from 15 microM of Cd onwards, after 56 d of exposure. For genotoxicity in plants, cytogenetic end-points such as mitotic index (MI), micronucleus formation (MN), mitotic aberrations (MA) and chromosome aberrations (CA) were examined in root meristem cells of B. juncea. Exposure of Cd revealed a significant (P < 0.05) inhibition of MI, induction of MA, CA, and MN in the root tips for 24 h. However, cells examined at 24 h post-exposure showed concentration-wise recovery in all the endpoints. The data revealed that Indian mustard could be used as a potential accumulator of Cd, Cr, Cu, and Pb due to a good tolerance mechanisms provided by combined/concerted action of NP-SH, GSH, and PCs. Also, exposure of Cd can cause genotoxic effects in B. juncea L. through chromosomal mutations, MA, and MN formation.     

Aquatic and terrestrial plant species with potential to remove heavy metals from storm-water

Remediation of storm-water polluted with heavy metals should be possible in percolation systems, ponds, or wetlands. The aim of this work was to find plant species for such systems that are efficient in the uptake of Zn, Cu, Cd, and Pb. Plants were collected from percolation and wetland areas and analyzed for heavy metal concentrations. Results showed that submersed and free-floating plants had the capacity to take up high levels of Cu, Zn, and Pb into their shoots. With roots having a concentration factor above 1, the terrestrial plants show efficient stabilization of Cd and Zn and emergent plants show corresponding stabilisation of Zn. In addition, Potamogeton natans, Alisma plantago-aquatica, and Filipendula ulmaria were used in a controlled experiment. The shoots of P. natans and the roots of A. plantago-aquatica were found to accumulate even higher concentrations of Zn, Cu, and Pb than found in the field-harvested plants. Similar results were found for Cd in shoots and Pb in roots of F. ulmaria. Our conclusion is that submersed plant species seem to be the most efficient for removal of heavy metals from storm-water.     

Effects of phytoremediation and application of organic amendment on the mobility of heavy metals in a polluted soil profile

This research aims to assess the effect of the application of biosolids compost and phytoremediation on the mobility of total and biodisponibles (DTPA) fractions of cadmium, copper, lead, and zinc from different horizons of a superficially contaminated soil. Leaching experiment in soil columns was proposed. Treatments contemplated application of compost biosolid and phytoremediation. Two destructive samplings were performed. Total and DTPA trace metals were identified in each horizon. The overall performance of the various elements in its total and DTPA forms show greater concentration in horizon A and fewer gradients between horizons Bt and BC, thus assuming that the high content of clay in horizon Bt (62.9%) limits its movement through the horizons. In the mobile nutrients, a greater mobility was evidenced in DTPA fractions if compared to Total fractions. In the horizon A, the more mobile metals, such as Zn and Cd, evidenced a greater percentage of DTPA/Total fractions in all treatments. The application of compost with or without plant diminished the mobilization of Zn, Cu, and Cd Total, thus limiting a potential leaching to inferior horizons. However, this effect was not observed in the DTPA fraction.     

Characterization of cadmium- and lead- phytochelatin complexes formed in a marine microalga in response to metal exposure

Phytochelatins (PC_n) are thiol-containing peptides with general structure (-Glu-Cys)_n-Gly enzymatically synthesized by plants and algae in response to metal exposure. They are involved in the cellular detoxification mechanism for their capability to form stable metal-phytochelatin complexes. The speciation of Cd and Pb complexes with phytochelatins has been studied in laboratory cultures of the marine diatom Phaeodactylum tricornutum. An approach based on size-exclusion chromatography (SEC) with off-line detection of phytochelatins, by reverse-phase HPLC, and metal ion, by atomic absorption spectrometry, has been used. The formation of Cd- and Pb-PC_n complexes with n-value from 3 to 6 was demonstrated. The metal-PC_n complexes formed with Cd appear to be different from those formed with Pb for the number of molecules of peptide involved in the complex and for the amount of the metal ion bound. The chromatographic behaviour of metal-PC_n complexes is consistent with Pb-PC_n complexes in which only a molecule of peptide binds the metal ion, and with Cd-PC_n complexes containing two or more molecules of peptide. The metal/peptide molar ratio in Cd-PC_n complexes was higher that in Pb-PC_n complexes. The formation of Cd- or Pb-PC₂ complexes was not demonstrated, probably for a dissociation during the cellular extract preparation. The effectiveness of phytochelatins in the detoxification of these two metal ions in this alga is discussed.     

Plant cell responses to heavy metals: molecular and physiological aspects

The effect of lead, cadmium and cooper on protein pattern, free radicals and antioxidant enzymes in root of Lupinus luteus L. were investigated. Heavy metals inhibited growth of lupin roots, which was accompanied by increased synthesis and accumulation of a 16 kDa polypeptide (Przymusiński et al. 1991 Biochem. Physiol. Pflanzen., 187:51–57). This component has been earlier identified as immunologically related to Cu,Zn-superoxide dismutase (Przymusiński et al. 1995 Env.Exp.Bot., 35:485–495). However, more detailed study revealed that this stress-stimulated protein is composed of four to six polypeptides of different electrophoretic mobility. The most abundant polypeptides of the 16kDa region were found to be closely homologous to pathogen related proteins. The number and intensity of these polypeptides was highly variable in roots of individual seedlings, which suggests that they might represent separate allelic forms. Electron paramagnetic spectra revealed that at low lead concentrations the amplitude of the first derivative was similar to the control and distinctly increased at higher metal concentrations. On the other hand, at the lower lead concentrations the activity of antioxidant enzymes increased, whereas at higher metal doses the enzyme activities did not raise further (SOD) or even dropped (CAT, APOX). This implies that the responses of antioxidant system to lead is dose-dependent stimulated by low metal concentrations, whereas at the higher metal level the free radical emission is beyond the quenching capacity of antioxidant enzymes, which in turn might contribute to the reduced root growth. The effect of various heavy metals: Pb²⁺, Cd²⁺ and Cu²⁺ on phytochelatins and antioxidant enzymes depends on the kind of metal ion. Pb²⁺ and Cd²⁺ stimulated the PCs formation whereas Cu²⁺ was not effective. On the other hand, in root exposed to Cu the activity of catalase (CAT) was the highest as was the production of H₂O₂. The strong oxidative effect of Cu²⁺ ions which were not complexed by PCs suggests that these peptides might by involved in the cellular defense system by binding excessive heavy metal ions. On the basis of our results it can be concluded that in lupin roots exposed to heavy metals there is a complex defense system against metal phytotoxicity, which comprises of specific proteins, antioxidant enzymes and phytochelatins.     

Cadmium uptake, translocation and tolerance in the hyperaccumulator Arabidopsis halleri

Arabidopsis halleri is a well-known zinc (Zn) hyperaccumulator, but its status as a cadmium (Cd) hyperaccumulator is less certain. Here, we investigated whether A. halleri can hyperaccumulate Cd and whether Cd is transported via the Zn pathway. Growth and Cd and Zn uptake were determined in hydroponic experiments with different Cd and Zn concentrations. Short-term uptake and root-to-shoot transport were measured with radioactive 109Cd and 65Zn labelling. A. halleri accumulated > 1000 mg Cd kg(-1) in shoot dry weight at external Cd concentrations >or= 5 microm, but the short-term uptake rate of 109Cd was much lower than that of 65Zn. Zinc inhibited short-term 109Cd uptake kinetics and root-to-shoot translocation, as well as long-term Cd accumulation in shoots. Uptake of 109Cd and 65Zn were up-regulated, respectively, by low iron (Fe) or Zn status. A. halleri was much less tolerant to Cd than to Zn. We conclude that A. halleri is able to hyperaccumulate Cd partly, at least, through the Zn pathway, but the mechanisms responsible for cellular Zn tolerance cannot detoxify Cd effectively.     

Use ofCladophora glomerata to monitor heavy metals in rivers

Methods were developed for the use ofCladophora glomerata to monitor heavy metal concentrations in flowing waters. At least under conditions without marked fluctuations in ambient metal concentration, there was no detectable difference in the metal concentrations of young plants between terminal 2-cm lengths of filament and whole plants. In order to establish the relationship between metal concentration in plant and that in water, 60 algal and water samples were analyzed from sites in northern England for Fe, Cu, Zn, Cd and Pb. Other environmental variables were measured at the time in order to assess their influence on metal accumulation. There were highly significant correlations for each of the five metals between concentrations in alga and water. The regression equations relating metal in alga to metal in water permit an unknown environmental metal concentration to be estimated from the algal concentration. Multiple stepwise regression analyses were used to indicate environmental factors which may influence metal accumulation; for instance, Fe appears to have a positive influence on Cu accumulation. In generalCladophora accumulates much less metal than bryophytes, but the slope relating metal in alga to metal in water is steeper, particularly for Pb. This means thatCladophora is especially useful where there is a need for a sensitive indicator of differences between sites or sampling occasions.     

Inhibition of photosynthesis by heavy metals

Inhibition of photosynthesis by heavy metals is well documented. In this review the results are compared between in vitro experiments on isolated systems (chloroplasts, enzymes ­.), experiments on excised leaves and intact plants and algae in vivo. In vitro experiments suggest potential sites of heavy metal interaction with photosynthesis at several levels of organisation, which are not necessarily confirmed in vivo. Analytical data on subcellular heavy metal level are generally missing to discuss their mechanism of action in the intact organism. In the field factors such as soil characteristics and air pollution have to be taken into account for assessing the mechanism of action of heavy metals on photosynthesis in plants, growing in a polluted erea.paper presented at the FESPP meeting in Strasbourg (1984)     

Characterization of cadmium uptake, translocation and storage in near-isogenic lines of durum wheat that differ in grain cadmium concentration

Here we examined several physiological properties of two near-isogenic lines of durum wheat (Triticum turgidum var. durum) that differ in grain cadmium accumulation, to identify the function of a gene locus that confers differential grain Cd concentrations. Time- and concentration-dependent uptake and translocation studies using 109Cd were conducted on nutrient solution-grown seedlings. Root extracts were analysed by inductively coupled plasma emission spectrometry, gel filtration and capillary electrophoresis to determine the interaction between Cd and phytochelatins (PCs) in storage of Cd in roots. The two isolines did not differ in time- or concentration-dependent root Cd uptake, but the low grain-Cd-accumulating isoline showed decreased movement of Cd from roots to shoots. All buffer-soluble Cd extracted from roots of both isolines was in the form of a low-molecular-weight PC-containing complex. The data suggest that PC synthesis is not a limiting factor in the differential storage of Cd in roots, and that movement of Cd through the root and into the transpiration stream may be the cause of differential Cd partitioning in the two isolines.     

Pigment characterisation of suspended matter in association with particulate heavy metal loadings in the German Bight and Elbe Estuary

The pigment fingerprints, determined by HPLC, of suspended matter from different areas of the German Bight and from the Elbe Estuary are presented. These include areas with suspended matter concentrations varying between 10 mg l^–1 in the deeper waters in the middle of the Bight and 150 mg l^–1 in the Elbe mouth. Pigment data allows the identification of three groups of suspended matter (marine, turbidity zone and Elbe) for the numerous different locations. The changes in pigment concentrations in the suspended matter over tidal cycles are considered. The presence of pigments and pigment breakdown productsi.e. lutein and chlorophyllides is used to assess the possible condition of the phytoplankton present in the suspended matter. These pigment characteristics are compared with the loading of the heavy metals cadmium, copper, manganese, iron, zinc and lead in the particulate matter. It has been demonstrated that the pigments of the planktonic material can be used very effectively to identify different types of suspended matter and that the heavy metal loadings of the suspended matter were significantly correlated with chlorophyll-a.     

Enteromorpha as a monitor of heavy metals in estuaries

An account is given of the use of Enteromorpha to monitor zinc, cadmium, mercury and lead pollution in six estuaries and the British North Sea coast. The ranges for each element were: Zn, 19–437 µg g^–1; µg g^–1 Cd, 0.07–4.8 µg g^–1; Hg, 0.02–0.23 µg g^–1. It is suggested that tissue analysis of Enteromorpha is one of the most useful biological techniques available in estuaries for pin-pointing aqueous (as opposed to sediment) metal contamination, and also for providing data suitable for world-wide comparisons. Provisional values are given for concentrations corresponding to moderate and high pollution.Deceased     

Effects of Interaction Between Cadmium and Plumbum on Phytochelatins and Glutathione Production in Wheat （Triticum aestivum L.）

Phytochelatins (PCs) may function as a potential biomarker for metal toxicity. However, less attention has been paid to the effects of metal interactions on the production of PCs and glutathione (GSH), the most prominent cellular thiol. In the present study, the effects of interactions between cadmium (Cd) and plumbum (Pb) on the production of PCs and GSH were monitored over a period of 14 d in wheat (Triticum aestivum L.) tissues. The results showed that combination of Cd and Pb led to synergistic growth inhibition in wheat. Exposure to Cd or Pb increased levels of PCs in a concentration-, tissue-, and time-dependent manner. Cadmium was more effective that Pb in increasing PCs production. Compared with the effects of Cd or Pb alone on the production of PCs, the combination of Cd and Pb acted synergistically, resulting in an enhanced production of PCs. Cadmium also stimulated GSH production in a concentration-, tissue-, and time-dependent manner. However, Pb had no obvious effects on GSH levels. The combination of Pb and Cd antagonized GSH production over the course of the growth period. The results of the present study suggest that metal interactions should be considered in the application of PCs and GSH as potential biomarkers for the evaluation of metal toxicity.