Changes in proline accumulation and antioxidative enzyme activities in Groenlandia densa under cadmium stress

In this study an experiment was carried out to study the process of stress adaptation in Groenlandia densa (opposite-leaved pondweed) grown under cadmium stress (0–20 mg L^?1 Cd). The results showed that Cd concentrations in plants increased with increasing Cd supply levels and reached a maximum of 0.43 mg kg^?1 DW at 0.5 mg L^?1 Cd concentrations. The level of photosynthetic pigments and soluble proteins decreased only upon exposure to high Cd concentrations. At the same time, the level of malondialdehyde (MDA) increased with increasing Cd concentration. These results suggested an alleviation of stress that was presumably the result of by antioxidants such as superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and glutathione S-transferase (GST) as well as ascorbate peroxidase (APX), which increased linearly with increasing Cd levels. Cellular antioxidants levels showed a decline suggesting a defensive mechanism to protect against oxidative stress caused by Cd. In addition, the proline content in G. densa increased with increasing cadmium levels. These findings suggest that G. densa is equipped with an efficient antioxidant mechanism against Cd-induced oxidative stress which protects the plant's photosynthetic machinery from damage.Our present work concluded that G. densa has a high level of Cd tolerance and accumulation. We also found that moderate Cd treatment (0.05–5 mg L^?1 Cd) alleviated oxidative stress in plants, while the addition of higher amounts of Cd (10–20 mg L^?1) could cause an increasing generation of ROS, which was effectively scavenged by the antioxidative system.     

Parietin in the tolerant lichen Xanthoria parietina (L.) Th. Fr. increases protection of Trebouxia photobionts from cadmium excess

Secondary metabolites of lichens can be involved in production of chelates with heavy metals. We hypothesized that parietin plays important role in protection of photobiont cells in Xanthoria parietina from an excess of cadmium ions. Two types of X. parietina lichen thalli, natural with presence of secondary metabolite parietin (p+) as well as without parietin (p−) were exposed to different doses of cadmium (up to 300 μmol g⁻¹ dw). Based on determination of the total and intracellular Cd-accumulation, ergosterol and thiobarbituric acid reactive substances (TBARS) content did not show statistically significant differences in the response of both types of thalli (p+ and p−). However, a stronger toxic effect of the highest Cd-dose on photosynthetic pigment content and chlorophyll a fluorescence was observed in the parietin-depleted thalli. The protective role of parietin against Cd excess was better supported and concluded from the differences observed in the production of non-protein thiol compounds (cysteine, glutathione and phytochelatins) involved in Cd detoxification. In the p+ thalli Cys content was stable but GSH content slightly decreased in the studied Cd range, while in the p− thalli these compounds were completely absent at high Cd doses. At Cd doses higher than 37.5 μmol Cd g⁻¹ dw, toxic to both types of X. parietina thalli, Cys and GSH contents were significantly higher in p+ than in p− thalli. Also, the photobiont partner in the p+ thalli was better protected of the metal exposition, and able to produce phytochelatins (PCs) over the whole range of metal, while in the p− thalli the production was completely inhibited at 75 μmol Cd g⁻¹ dw and higher concentrations, together with the inhibition of cysteine (Cys) and reduced glutathione (GSH) production. The obtained results indicate that the parietin layer is a natural barrier decreasing Cd access to algal cells in X. parietina. Comparison of PCs production appeared to be the most sensitive marker for estimation of Cd availability to photobiont in the symbiotic system.     

Long-term effects of exogenous silicon on cadmium translocation and toxicity in rice (Oryza sativa L.)

Most nutrient solution studies on the interactions between silicon (Si) and cadmium (Cd) are short term. Here we reported a long-term experiment in which rice (Oryza sativa L.) was cultured for 105 days and harvested at four different growth stages to measure biomass accumulation and Cd uptake and distribution in shoots and roots. Exogenous Si increased shoot biomass by 61–238% and root biomass by 48–173% when the culture solution was free of Cd. When 2 μmol L^?1 Cd was added, Si supply increased shoot and root biomass by 125–171% and by 100–106% compared to the zero-Si treatment. Increasing the Cd concentration to 4 μmol L^?1 decreased the beneficial effects of Si on root and shoot biomass. Silicon supply decreased shoot Cd concentrations by 30–50% and Cd distribution ratio in shoot by 25.3–46%, compared to the treatment without Si supply. Additionally, lower Si supply or more serious Cd stress would lead to roots with bigger biomass and higher Si concentration. Energy-dispersive X-ray microanalysis showed that both Si and Cd accumulated synchronously in the border and middle of phytoliths of the shoots. We conclude that Si enhances plant growth and decreases Cd accumulation in shoots and thereby helps to lower the potential risks of food contamination.     

Cadmium toxicity in tomato (Lycopersicon esculentum) plants grown in hydroponics

The effects of Cd have been investigated in tomato (Lycopersicon esculentum) plants grown in a controlled environment in hydroponics, using Cd concentrations of 10 and 100 μM. Cadmium treatment led to major effects in shoots and roots of tomato. Plant growth was reduced in both Cd treatments, leaves showed chlorosis symptoms when grown at 10 μM Cd and necrotic spots when grown at 100 μM Cd, and root browning was observed in both treatments. An increase in the activity of phosphoenolpyruvate carboxylase, involved in anaplerotic fixation of CO₂ into organic acids, was measured in root extracts of Cd-exposed plants. Also, significant increases in the activities of several enzymes from the Krebs cycle were measured in root extracts of tomato plants grown with Cd. In leaf extracts, significant increases in citrate synthase, isocitrate dehydrogenase and malate dehydrogenase activities were also found at 100 μM Cd, whereas fumarase activity decreased. These data suggest that at low Cd supply (10 μM) tomato plants accumulate Cd in roots and this mechanism may be associated to an increased activity in the PEPC–MDH–CS metabolic pathway involved in citric acid synthesis in roots. Also, at low Cd supply some symptoms associated with a moderate Fe deficiency could be observed, whereas at high Cd supply (100 μM) effects on growth overrule any nutrient interaction caused by excess Cd. Cadmium excess also caused alterations on photosynthetic rates, photosynthetic pigment concentrations and chlorophyll fluorescence, as well as in nutrient homeostasis.     

Cadmium distribution and its effects on molybdate-containing hydroxylases in Phragmites australis

The influence of cadmium (Cd) on physiological and biochemical parameters was studied to elucidate the mechanism of Cd resistance in Phragmites australis. Cadmium concentrations in roots, stems and leaves increased with exogenous Cd concentration, but Cd content in roots was much higher than in shoots. X-ray microanalysis was used to reveal compartments in which Cd accumulated in root cortex. Cadmium concentrations followed a gradient with the sequence: intercellular space > cell wall > vacuole > cytoplasm, indicating that most Cd was immobilized in the apoplast or sequestered into the vacuolar lumen. Sequential extraction of various Cd chelates revealed that more than half of extractable Cd was bound to proteins, whereas 26% was bound to organic acids. Cd-binding protein fractions were found in the roots after gel filtration chromatography, among which a polypeptide with an apparent molecular mass of 14 kDa bound Cd most avidly. One newly synthesized polypeptide of low molecular mass (1 kDa) appeared under Cd pollution, whereas a prominent fraction of 72 kDa disappeared. Four aldehyde oxidase (AO) isoenzyme activities increased significantly in roots under Cd pollution. Cd stress also enhanced xanthine dehydrogenase (XDH) activities in roots. Two AO polypeptides of different molecular sizes were detected in the roots by Western blot assay. The abundance of the 160 kDa subunit correlated with Cd stress, but the amount of the 90 kDa polypeptide did not change under Cd treatment. Enhanced abscisic acid (ABA) contents were observed in roots of P. australis exposed to Cd. The involvement of Cd distribution in plant tissues and subcellular compartments and of AO and XDH enzymatic activities in the acclimation mechanism of P. australis to Cd pollution is discussed herein.     

Silicon mitigates cadmium inhibitory effects in young maize plants

The influence of silicon on the growth of maize plants cultivated in hydroponics in the presence of cadmium (5 μM) was investigated. Four different treatments were used: Control (C), Cadmium (Cd), Silicon (Si) and Cadmium plus Silicon (Cd + Si). The Si concentration was 35 mM. Thirteen-day-old plants were harvested. Growth parameters (length of primary seminal root, leaf area of first and second fully developed leaves, fresh and dry weight of below- and above-ground parts of the plants), and Cd concentration and total amount of Cd in the below- and above-ground parts were determined. In roots, the development of the endodermal barrier was observed by fluorescent staining with Fluorol yellow 088.Inhibitory effects of Cd on plant growth were observed. Silicon treatment in the absence of Cd had positive effects on most of observed growth parameters compared with the control. Moreover, Si in the Cd + Si treatment improved all growth parameters compared with the cadmium treatment. Silicon increased the cell-wall extensibility both in Si and Cd + Si treatments when compared with the control. Alleviation of the Cd-inhibitory effect on maize plants by Si was not due to exclusion of Cd from the plant; in contrast, Cd concentration in below- and above-ground plant parts and the total amount of Cd per plant were significantly higher in the Cd + Si plants than in the Cd treatment. The increased Cd content in Cd + Si plants was correlated with the development of the endodermis; during the second stage of endodermal development, suberin lamellae were formed at a greater distance from the root apex in the Cd + Si than in the Cd treatment. Silicon itself did not influence the development of suberin lamellae in the maize roots compared with the control.     

Physiological mechanism of hypertolerance of cadmium in Kentucky bluegrass and tall fescue: Chemical forms and tissue distribution

Kentucky bluegrass (Poa pratensis) and tall fescue (Festuca arundinacea) are hypertolerant grasses to soil cadmium contamination. Little information is available on their tolerance mechanism. A sand culture and a hydroponic culture experiment were designed to investigate the Cd chemical form changes and its translocation in different tissues. The results showed that Kentucky bluegrass and tall fescue can tolerate 50–200 mg kg⁻¹ of soil Cd stresses and accumulate as high as 4275 and 2559 mg Cd kg⁻¹ DW, respectively, in their shoots without the loss of shoot biomass. Their Cd hypertolerance was correlated with an increase of the undissolved Cd phosphates in the leaves in both grass species, as determined by sequential solvent extraction procedures. The superior Cd tolerance of tall fescue to Kentucky bluegrass was associated with less Cd translocation into the stele of roots and less Cd transported to leaves. The pectate- and protein-integrated Cd forms may be involved in the symplastic translocation of Cd from cortex into stele, and this may lead the higher Cd concentrations in the stele of roots and then above ground leaves via long-distance transport in Kentucky bluegrass.     

Thiol metabolism play significant role during cadmium detoxification by Ceratophyllum demersum L.

In the present study, the level of thiols and activity of related enzymes were investigated in coontail (Ceratophyllum demersum L.) plants to analyze their role in combating the stress caused upon exposure to cadmium (Cd; 0–10 μM) for a duration up to 7 d. Plants showed the maximum accumulation of 1293 μg Cd g^?1 dw after 7 d at 10 μM. Significant increases in the level of total non-protein thiols (NP-SH) including phytochelatins (PCs) as well as upstream metabolites of the PC biosynthetic pathway, cysteine and glutathione (GSH) were observed. In addition, significant increases in the activities of cysteine synthase (CS), glutathione-S-transferase (GST), glutathione reductase (GR), as well as in vitro activation of phytochelatin synthase (PCS), were noticed in response to Cd. In conclusion, under Cd stress, plants adapted to a new metabolic equilibrium of thiols through coordinated synthesis and consumption to combat Cd toxicity and to accumulate it.     

Subcellular distribution and chemical forms of cadmium in Bechmeria nivea (L.) Gaud.

Bechmeria nivea (L.) Gaud. (Ramie) is a promising species for Cd phytoextraction with large biomass and fast growth rate. Nevertheless, little information is available on its tolerance mechanisms towards Cd. Determination of Cd distribution and chemical speciation in ramie is essential for understanding the mechanisms involved in Cd accumulation, transportation and detoxification. In the present study, ramie plants were grown in hydroponics with increasing Cd concentrations (0, 1, 3, 7 mg l^?1). The subcellular distribution and chemical forms of Cd in different tissues were determined after 20 days exposure to this metal. To assess the effect of Cd uptake on plant performance, nitrate reductase activity in leaves and root activity were analyzed during the entire experimental period. Increased Cd level in the medium caused a proportional increase in Cd uptake, and the highest Cd concentration occurred in roots, followed by stems and leaves. Subcellular fractionation of Cd-containing tissues indicated that about 48.2–61.9% of the element was localized in cell walls and 30.2–38.1% in soluble fraction, and the lowest in cellular organelles. Cd taken up by ramie rapidly equilibrated among different chemical forms. Results showed that the greatest amount of Cd was found in the extraction of 1 M NaCl and 2% HAC, and the least in residues in all test tissues. In roots, the subdominant amount of Cd was extracted by d-H₂O and 80% ethanol, followed by 0.6 M HCl. While in stems and leaves, the amount of 0.6 M HCl-extractable Cd was comparable with that extracted by 80% ethanol or d-H₂O. 1 mg l^?1 Cd stimulated nitrate reductase activity in leaves and root activity, while a concentration-dependent inhibitory effect was observed with increasing Cd concentration, particularly at 7 mg l^?1 Cd. It could be suggested that the protective mechanisms evolved by ramie play an important role in Cd detoxification at relatively low Cd concentrations (below 3 mg l^?1 Cd) but become restricted to maintain internal homeostasis with higher Cd stress.     

Cadmium,lead and mercury concentrations and their influence on morphological parameters in blood donors from different age groups from southern Poland

Due to industrial development, environmental contamination with metals increases which leads to higher human exposure via air, water and food. In order to evaluate the level of the present exposition, the concentrations of metals can be measured in such biological materials as human blood. In this study, we assessed the concentrations of cadmium (Cd), mercury (Hg) and lead (Pb) in blood samples from male blood donors from southern Poland (Europe) born in 1994 (n = 30) and between 1947 and 1955 (n = 30). Higher levels of Pb were seen in the group of older men (4.48 vs 2.48 μg/L), whereas the Hg levels were lower (1.78 vs 4.28 μg/L). Cd concentrations did not differ between age groups (0.56 μg/L). The levels of Cd and Pb in older donors were significantly correlated (Spearman R 0.5135). We also observed a positive correlation between the number of red blood cells (RBC) and Hg concentrations in the older group (Spearman R 0.4271). Additionally, we noted numerous correlations among morphological parameters. Based on our results, we can state that metals influence the blood morphology and their concentrations in blood vary among age groups.     

Blood lead and cadmium levels in a six hospital employee population. PESA study, 2009

IntroductionExposure to lead and cadmium is a public health problem due to the broad exposure to these toxic substances among the general population. The objective of this study is to determine blood lead and cadmium concentrations in a working population drawn from six university hospitals in Madrid, Getafe, Cartagena, Santiago de Compostela, Santander and Palma de Mallorca (Spain) and to identify associated factors.Materials and methods951 individuals participated in the study and were administered the standardized PESA^® questionnaire regarding exposure to lead and cadmium. The blood lead and cadmium concentrations were measured by electrothermal atomization atomic absorption spectrometry with Zeeman background correction in Perkin-Elmer spectrometers, guaranteeing the transferability of the results.ResultsThe median overall blood lead concentration was: 1.6 μg/dL (IQR: 0.9–2.7) and that of cadmium was: 0.21 μg/L (IQR: 0.10–0.50). There were significant differences in lead levels between men (2 μg/dL) and women (1.5 μg/dL), postmenopausal (2.6 μg/dL) and premenopausal women (1.1 μg/dL), and between participants who cooked in earthenware (2.1 μg/dL) and those who did not (1.5 μg/dL). The median of cadmium in women (0.24 μg/L) was higher than in men (0.11 μg/L) and was also higher in subjects who smoked (0.70 μg/L) than in non-smokers (0.13 μg/L).ConclusionsA reduction in blood lead and cadmium levels was observed with respect to previous studies carried out in Spain. Nevertheless, the results suggest there are certain factors which increase risk such as age, gender, menopause, age of housing, cooking in lead-glazed earthenware and exposure to cigarette smoke.     

Trace elements in bipolar disorder

IntroductionTrace elements may play an important role in bipolar disorders. The objective of this study is to determine serum copper and zinc, blood lead and cadmium and urine lead, cadmium and thallium concentrations in patients diagnosed with bipolar disorders and to compare these levels with those of a healthy control group.Materials and methodsA total of 25 patients diagnosed with bipolar disorder and 29 healthy subjects participated in this study. Serum copper and zinc concentrations were measured using flame atomic absorption spectrometry; the blood lead and cadmium concentrations were measured by electrothermal atomization atomic absorption spectrometry with Zeeman background correction; urine lead, cadmium and thallium concentrations were measured by inductively coupled plasma mass spectrometry.ResultsMedian blood and urine lead and cadmium levels were significantly higher among the bipolar patients than among the control group: Blood lead (μg/dL): patient median: 3.00 (IQR: 1.40–4.20); control median (μg/dL): 2.20 (IQR: 0.90–3.00) p = 0.040. Blood cadmium (μg/L): patient median: 0.39 (IQR: 0.10–1.15); control median: 0.10 (IQR: 0.10–0.17) p < 0.001. The median of cadmium (μg/L) in patients who smoked (1.20 IQR: 0.44–2.30) was higher than that in non-smokers (0.12 IQR: 0.10–0.34) p < 0.001. There was a statistically significant increase (p = 0.001) in zinc levels among patients in the manic phase (mean 111.28, SD: 33.36 μg/dL) with respect to the control group (mean 86.07, SD: 12.39 μg/dL).ConclusionsThe results suggest that there could be higher levels of some toxic trace elements in the group of patients with bipolar disorder than in the healthy control group.     

Genotypic variation in safflower (Carthamus spp.) cadmium accumulation and tolerance affected by temperature and cadmium levels

Soil pollution is a world-wide problem, with heavy metals being a major part of the concern. To investigate the effect of temperature on cadmium (Cd) uptake and translocation, as well as Cd tolerance in wild and cultivated species of safflower, a hydroponic experiment was conducted under controlled conditions. The responses of four wild genotypes (Isfahan, Arak, Azari, and Shiraz) and four cultivated genotypes (AC-Sterling, 2811, Saffire, and C111) of safflower to nine levels of CdCl₂ (0, 0.5, 1, 5, 10, 20, 50, 100, and 500 μM) in solution were examined under two temperatures (18 and 23 °C). Cadmium sensitivity was determined using the Weibull model on the total dry weight of the plants. Cadmium uptake and translocation were analyzed on 1 μM Cd treated plants. Results revealed that safflower genotypes differed in terms of uptake, translocation, and tolerance to Cd, with AC-Sterling and Arak indicating the most and the least tolerance to Cd, respectively. Relative Cd uptake and Cd concentration in roots and shoots increased with an increase in temperature in all genotypes, with the exception of AC-Sterling. Net accumulation of Cd via root increased with an increase in temperature for the wild Azari and the cultivated 2811, Saffire, and C111, though it decreased for the rest of genotypes. Cadmium translocation to shoots significantly increased with increased temperature in all genotypes. Cadmium translocation from roots to shoots in cultivated genotypes was significantly greater than in wild genotypes. Root Cd concentration in wild genotypes was significantly greater than in cultivated genotypes. It seems that wild and cultivated species of safflower differ in their response to Cd. Furthermore, temperature may affect the plant's tolerance to Cd, probably through accompanying changes in Cd uptake and translocation from root to shoot.     

Response of miscanthus to toxic cadmium applications during the period of maximum growth

Plants of miscanthus were grown in a Cd-free solution up to 1 month before heading and then were exposed to 0, 0.75, 1.5, 2.25 and 3 mg l⁻¹ cadmium for 36 days. All cadmium levels were toxic to miscanthus. Growth response was not dose-dependent and two toxicity thresholds were identified: one between 0 and 0.75 mg l⁻¹ Cd, the other between 2.25 and 3 mg l⁻¹ Cd. The former caused a biomass decrease by about 50%, whereas the latter completely inhibited growth and disrupted the mechanisms that restricted Cd translocation to the shoot. Growth of the aerial part was affected by cadmium more than that of the hypogeal one. Cadmium did not change the N concentration of different plant parts, but markedly reduced the N uptake of the plant, the N net uptake rate (NUR) and the N net translocation rate (NTR) from the rhizome to the aerial part. These two indexes equalled zero when plants ceased to grow. Otherwise, the Cd-NUR increased with Cd supply and the Cd-NTR from rhizome to aerial part showed the highest increment when plants did not grow at all. This suggests different uptake pathways for the two elements, active for nitrogen and passive for cadmium. The Cd concentration and the Cd content markedly increased with all Cd levels, following the order roots ≫ rhizome > culms > leaves. The Cd concentration and the Cd content of aerial organs increased with Cd supply, but increments were highest between 2.25 and 3 mg l⁻¹ Cd. The highest Cd concentrations were recorded in plants grown with 3 mg l⁻¹ Cd and were 41 and 122 mg kg⁻¹, respectively, for the aerial and the hypogeal plant parts. The hypogeal plant part retained most of the cadmium taken up from solution, accounting for approximately 87% of total plant cadmium with the three lower Cd levels, and for 73% with the highest one. The maximum Cd content of the entire plant was achieved with the two higher Cd levels and was approximately 4.7 mg, while the Cd content of the aerial part was highest with 3 mg l⁻¹ Cd (1.2 mg Cd per plant) and that of the hypogeal one with 2.25 mg l⁻¹ Cd (4 mg Cd per plant). The highest aerial content achieved in this experiment was 10-fold that obtained in a previous research when small-sized plants were exposed to the same Cd level.     

Thiol-peptide level and proteomic changes in response to cadmium toxicity in Oryza sativa L. roots

In the present study, rice seedlings were exposed to a range of Cd concentrations (0.1 μM, 1 μM, 10 μM, 100 μM and 1 mM) for 15 days and a combination of different molecular approaches were used to evidence Cd effects and to assess the plants’ ability to counteract metal toxicity. At a macroscopical level, only the highest Cd concentration (1 mM) caused a complete plant growth inhibition, whereas the lowest concentrations seemed to stimulate growth. At genome level, the amplified fragment length polymorphism (AFLP) technique was applied to detect DNA sequence changes in root cells, showing that all the Cd concentrations induced significant DNA polymorphisms in a dose-dependent manner. Data also evidenced the absence of preferential mutation sites.Plant responses were analysed by measuring the levels of gluthatione (GSH) and phytochelatins (PCs), the thiol-peptides involved in heavy metal tolerance mechanisms. Results showed a progressive increase of GSH up to 10 μM of Cd treatment, whereas a significant induction only of PC₃ was detected in roots of plants exposed to 100 μM of Cd. As suggested by the proteome analysis of root tissues, this last concentration strongly induced the expression of regulatory proteins and some metabolic enzymes. Furthermore, the treatment with 10 μM of Cd induced changes in metabolic enzymes, but it mainly activated defence mechanisms by the induction of transporters and proteins involved in the degradation of oxidatively modified proteins.     

Comparative analysis of the contribution of phytochelatins to cadmium and arsenic tolerance in soybean and white lupin

The biosynthesis of phytochelatins (PCs) plays a crucial role in the detoxification and homeostasis of heavy metals and metalloids in plants. However, in an increasing number of plant species metal(loid) tolerance is not well correlated with the accumulation of PCs: tolerant ecotypes frequently contain lower levels of PCs than non-tolerant ecotypes. In this study we have compared the responses of soybean (Glycine max L. cv. Resnik) and white lupin (Lupinus albus L. cv. Marta) to cadmium and arsenate in order to assess the role of homophytochelatins (hPCs) in the tolerance of soybean to these toxic elements. Soybean plants treated with Cd and As showed a high contribution of homo-glutathione (hGSH) to the pool of thiols in shoots in comparison to white lupin. Higher levels of hPCs in Cd-treated soybeans compared to PCs in lupins did not prevent growth inhibition. In contrast, the role of hPCs in the detoxification mechanism to arsenate in soybean seems to be clearer, showing higher thiol concentrations and lower growth reductions than those present in lupin plants.     

The impact of heavy metals on the activity of some enzymes along the barley root

Barley seedlings 48 h after the onset of germination on filter paper treated for 24 h by 1 mM cadmium (Cd), 3 mM nickel (Ni) or 0.5 mM mercury (Hg) showed similar approximately 45% root growth inhibition. Although root growth inhibition was similar, loss of cell viability evaluated, as Evans blue uptake was distinct among Cd, Ni and Hg treated roots. While Cd and Hg caused cell death along the whole barley root (0–8 mm), Ni induced significant loss of cell viability only in root cells 6–8 mm distance from the root tip. Our results suggest that different metabolic processes are activated in different parts of barley root in relation to distance from the root tip during heavy metal (HM) treatment. Some of them are characteristic for several HMs such as inhibition of ascorbic acid oxidase or glutathione-S-transferase stimulation, while others are specific for individual HMs, e.g. activation of acid phosphatase and lipoxygenase by Cd and Hg, or inhibition of ascorbate peroxidase by Ni and Hg treatment.     

Role of iron plaque in Cd uptake by and translocation within rice (Oryza sativa L.) seedlings grown in solution culture

A hydroponics culture experiment was conducted to investigate the effect of iron plaque on Cd uptake by and translocation within rice seedlings grown under controlled growth chamber conditions. Rice seedlings were pre-cultivated for 43 days and then transferred to nutrient solution containing six levels of Fe (0, 10, 30, 50, 80 and 100 mg L⁻¹) for 6 days to induce different amounts of iron plaque on the root surfaces. Seedlings were then exposed to solution containing three levels of Cd (0, 0.1 and 1.0 mg L⁻¹) for 4 days. In order to differentiate the uptake capability of Cd by roots with or without iron plaque, root tips (white root part without iron plaque) and middle root parts (with iron plaque) of pre-cultivated seedlings treated with 0, 30 and 50 mg L⁻¹ Fe were exposed to ¹⁰⁹Cd for 24 h. Reddish iron plaque gradually became visible on the surface of rice roots but the visual symptoms of the iron plaque on the roots differed among treatments. In general, the reddish color of the iron plaque became darker with increasing Fe supply, and the iron plaque was more homogeneously distributed all along the roots. The Fe concentrations increased significantly with increasing Fe supply regardless of Cd additions. The Cd concentrations in dithionite–citrate–bicarbonate (DCB)-extracts and in shoots and roots were significantly affected by Cd and Fe supply in the nutrient solution. The Cd concentrations increased significantly with increasing Cd supply in the solution and were undetectable when no Cd was added. The Cd concentrations in DCB-extracts with Fe supplied tended to be higher than that at Fe₀ at Cd_0.1, and at Cd_1.0, DCB-Cd with Fe supplied was significantly lower. Cd concentrations in roots and shoots decreased with increasing Fe supply at both Cd additions. The proportion of Cd in DCB-extracts was significantly lower than in roots or shoots. Compared to the control seedlings without Fe supply, the radioactivity of ¹⁰⁹Cd in shoots of seedlings treated with Fe decreased when root tips were exposed to ¹⁰⁹Cd and did not change significantly when middle parts of roots were exposed. Our results suggest that root tissue rather than iron plaque on the root surface is a barrier to Cd uptake and translocation within rice plants, and the uptake and translocation of Cd appear to be related to Fe nutritional levels in the plants.     

Cadmium tolerance in Thlaspi caerulescens: I. Growth parameters,metal accumulation and phytochelatin synthesis in response to cadmium

A population of the metallophyte, Thlaspi caerulescens, originating from a Cd–Pb–Zn old mining and smelter site at Plombières (Belgium) was studied. T. caerulescens was cultivated hydroponically to investigate Cd uptake and tolerance. Cd was added to Hoagland’s medium at concentration range from 5 to 500 μM. The plants could tolerate 500 μM Cd in the solution showing only minor visible symptoms of toxicity but with a 32% decrease in fresh weight. After 14 days at 500 μM, Cd content in roots and shoots was 707 and 602 mg kg⁻¹ of dry weight (d.w.), respectively. Application of Cd to hydroponically cultivated T. caerulescens induced the accumulation of PCs in plant roots and shoots. Buthionine sulfoximine (BSO) application almost completely reduced (by 98–100%) the accumulation of PCs without simultaneous increase in plants sensitivity to Cd. These results suggest a minor if any role of PCs in tolerance to Cd of the studied population of T. caerulescens in hydroponics. On the other hand, no PC accumulation was detected either in T. caerulescens plants growing in their natural environment at Plombierès or in plants growing in their native soil in a greenhouse. These results suggest that naturally selected tolerance in T. caerulescens population from Plombières is not associated with enhanced PCs synthesis.     

Changes induced by two levels of cadmium toxicity in the 2-DE protein profile of tomato roots

Tomato is an important crop from nutritional and economical points of view, and it is grown in greenhouses, where special substrates and the use of recycled water imply an increased risk of Cd accumulation. We investigated tomato root responses to low (10 µM) and high (100 µM) Cd concentrations at the root proteome level. Root extract proteome maps were obtained by 2-DE, and an average of 121, 145 and 93 spots were detected in the 0, 10 and 100 µM Cd treatments, respectively. The low Cd treatment (10 µM) resulted in significant and higher than 2-fold changes in the relative amounts of 36 polypeptides, with 27 of them identified by mass spectrometry, whereas the 100 µM Cd treatment resulted in changes in the relative amounts of 41 polypeptides, with 33 of them being identified. The 2-DE based proteomic approach allowed assessing the main metabolic pathways affected by Cd toxicity. Our results suggests that the 10 µM Cd treatment elicits proteomic responses similar to those observed in Fe deficiency, including activation of the glycolytic pathway, TCA cycle and respiration, whereas the 100 µM Cd treatment responses are more likely due to true Cd toxicity, with a general shutdown of carbon metabolism and increases in stress related and detoxification proteins.