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.     

Cd、Zn及其交互作用对互花米草中重金属的积累、亚细胞分布及化学形态的影响

通过盆栽试验,研究了Cd、Zn及其交互作用下互花米草中Cd、Zn的含量及积累量,并分析了Cd、Zn在互花米草中的亚细胞分布及化学形态。结果表明：Cd-Zn处理组互花米草地上部及根部Cd含量显著高于Cd处理组;Cd-Zn处理组根部Zn含量显著低于Zn处理组,但地上部差异不显著,说明Zn促进Cd的吸收,Cd抑制Zn的吸收。Cd-Zn处理组互花米草地上部Cd积累量显著高于Cd处理组,但是根部Cd积累量却显著低于Cd处理组;Zn处理组地上部及根部Zn积累量均显著高于对照组及Cd-Zn处理组。Cd单因素胁迫下,Cd主要分布在细胞壁,Cd-Zn交互作用下,Cd在胞液中的分配比例高于其他细胞组分;Zn单因素及Cd-Zn交互作用下,Zn在胞液中的分配比例均较高,总的分配比例呈现以下趋势：胞液>细胞壁>细胞器,说明Zn的添加影响了Cd的亚细胞分布,Cd的出现对Zn在互花米草细胞中的分布影响不明显。Cd和Zn在互花米草叶中主要以氯化钠提取态存在,表明互花米草中Cd和Zn多以果胶酸盐结合态或蛋白质结合态存在。     

Assessing the potential for zinc and cadmium phytoremediation with the hyperaccumulator Thlaspi caerulescens

Thlaspi caerulescens is a Zn and Cd hyperaccumulator, and has been tested for its phytoremediation potential. In this paper we examine the relationships between the concentrations of Zn and Cd in soil and in T. caerulescens shoots, and calculate the rates of Zn and Cd extraction from soil. Using published data from field surveys, field and pot experiments, we show that the concentrations of Zn and Cd in the shoots correlate with the concentrations of Zn and Cd in soils in a log-linear fashion over three orders of magnitude. There is little systematic difference between different populations of T. caerulescens in the relationship between soil and plant Zn concentrations. In contrast, populations from southern France are far superior to those from other regions in Cd accumulation. Bioaccumulation factors (plant to soil concentration ratio) for Zn and Cd decrease log-linearly with soil metal concentration. Model calculations show that phytoremediation using T. caerulescens is feasible when soil is only moderately contaminated with Zn and Cd, and the phytoremediation potential is better for Cd than for Zn if the populations from southern France are used. Recent progress in the understanding of the mechanisms of Zn and Cd uptake by T. caerulescens is also reviewed.     

Zn、Cd及其复合对小麦幼苗吸收Ca,Fe,Mn的影响

研究了溶液培养条件下,Cd,Zn及其复合对小麦幼苗吸收Ca,Fe,Mn的影响,结果表明,小麦幼苗对Zn,Cd的吸收随溶液中Cd^2 ,Zn^2 浓度的升高而增加,Cd,Zn同时存在时与其单独作用时幼苗对它们的吸收不同,Zn影响幼苗对Cd的吸收,Cd对Zn的吸收起抑制作用,Ca,Mn的吸收随溶液中Cd^2 ,Zn2 浓度升高而呈下降趋势,在Cd单独处理组和Zn单独处理组中Fe的吸收随Cd2 ,Zn2 浓度升高而增加,但在Zn Cd处理组中,Fe的吸收则呈下降趋势,其效应试学与作物具体部位有关。     

Foliar Spraying and Seed Soaking of Zinc Fertilizers Decreased Cadmium Accumulation in Cucumbers Grown in Cd-Contaminated Soils

It is important to use proper agronomic management to reduce cadmium (Cd) accumulation in plants, ensuring food safety. To find the most effective agronomic approach, the effect of foliar spraying and seed soaking of zinc (Zn) fertilizers on Cd accumulation in cucumbers (Cucumis sativus L.) grown in two soil Cd levels (2 and 5 mg kg^?1 Cd) with and without an immobilizing amendment (red mud, RM) was investigated in the present study. The results showed that the treatment of foliar Zn or seed Zn significantly decreased the Cd concentration in cucumber shoots by about 12–36% in Cd-contaminated soils without amendment. Combined with RM treatment, the foliar Zn treatment further decreased the Cd concentration in cucumber shoots by up to 48–66% in Cd-contaminated soils. There were significant negative correlations between Cd and Zn concentrations in shoots of cucumbers grown in soils treated with RM and foliar Zn. The results revealed that the cucumber seedlings treated with RM and foliar Zn had a higher capacity for limiting the transfer of Cd to aboveground tissues. The results also suggested that increasing seed Zn concentrations sufficiently might act as an efficient, economic, and practical method for decreasing Cd uptake in crops grown in mildly Cd-contaminated and Zn-deficient soils.     

Cadmium and Zinc Concentrations in Soils and Oil Palm Tissues as Affected by Long-Term Application of Phosphate Rock Fertilizers

The study was conducted to determine Cd and Zn concentrations in soils with three different ages of oil palm trees (<10, >15, >20 years) and plant parts as affected by long-term application of phosphate rock fertilizers. To investigate this issue on Malaysia's oil palm plantations, six soil series that undergo a scheduled fertilizer program were collected. Jawa, Selangor, and Sedu Series were selected from a plantation in the coastal area, whilst Munchong, Rengam, and Segamat Series were collected from a plantation in the inland area. Fronds and fruitlets were also collected along with the soils (paired sampling). To verify the findings of the field study, Cd and Zn adsorption isotherms were prepared. Competitive adsorption between Zn and Cd was also conducted. There was no accumulation of Cd in all the soil series but Zn accumulated in the Selangor and Segamat Series. Fruitlets (edible part) show no increase in Cd and Zn concentrations, but Jawa and Selangor Series show increasing values of Zn concentrations in the fronds. A correlation study reveals that soil pH and clay content were the soil properties that controlled Cd and Zn concentrations in soils. Cadmium in soils and soil solutions most probably contributed to the Cd concentrations in fruitlets, whereas Cd and Zn in soils may influence the uptake of these elements by the oil palm trees. The highest Cd and Zn accumulation in Segamat Series Soil had been shown in the adsorption isotherm study. Cadmium adsorption was decreased in the presence of Zn, leading to no accumulation of Cd in all the soil series.     

The influence of zinc on apical uptake of cadmium in the gills and cadmium influx to the circulatory system in zebrafish (Danio rerio)

Zn (0-16 microM) effects on apical Cd uptake from the water into the branchial epithelium and influx of Cd from the water to the circulatory system in zebrafish (Danio rerio) were studied in three experiments. Apical Cd uptake was decreased by Zn in all three experiments. In fish exposed to 1-600 nM Cd (experiment 1), apical Cd uptake showed saturation kinetics at 2 and 4 microM Zn, and a competitive interaction was indicated. At 16 microMZn, Cd uptake increased linearly. Cadmium influx did not show saturation kinetics, but was inhibited by 16 microM Zn at low Cd exposure concentrations. In fish exposed to 0.1-600 nM Cd (experiment 2), Cd uptake was inhibited by 16 microM Zn, whereas at 30 nM Cd uptake was inhibited by 2 microM Zn. Similarly, 2 microM Zn did not influence Cd uptake in fish exposed to 0.1-2 nM Cd (experiment 3), whereas 2 microM Zn inhibited uptake at 8-30 nM Cd. Zinc also inhibited Cd influx at higher Cd concentrations. However, at lower Cd exposures, a Zn-induced increased influx was indicated. Zinc influences the Cd uptake and influx processes at several sites in the branchial epithelial cells, indicating that influx of Zn2+ and Cd2+ occurs through common pathways.     

紫色土外源锌、镉形态的生物有效性

通过对7种形态Zn,Cd与有效态Zn,Cd(DTPA提取）的相关分析及各形态Zn,Cd与植株Zn,Cd含量的通径分析,评价各形态Zn,Cd的生物有效性及对植株的相对贡献和途径。结果表明,交换态Zn,Cd与有效态Zn,Cd呈极显著正相关（r=0.954,0.953),交换态Zn,Cd与植株Zn,Cd含量的通径系数为1．267、1．168,交换态Zn,Cd有效性高,具有最大的生物活性,对植株Zn,Cd含量的贡献最大。其它形态如碳酸盐结合态锌、氧化锰结合态Zn、Cd通过交换态作用于植株的通径链系数（间接通径系数）分别为0．856、0．592、0．723,上述形态对植株Zn,Cd含量也有影响,具有一定的生物相对有效性,但通过交换态间接作用。交换态不仅自身具有最大的生物有效性,而且还作为其它形态Zn,Cd生物有效性的中介,为紫色土其它Zn,Cd库流向植株的主要通道。     

Relationships between Cadmium, Zinc, Cd-Peptide, and Organic Acid in Tobacco Suspension Cells

Responses of tobacco (Nicotiana tabacum) suspension cells to Cd and Zn were studied in the presence and absence of ligand of Cd-peptide in order to understand the role of this peptide versus other mechanisms in Cd and Zn accumulation and accommodation in plants. With 45 micromolar Cd and 300 micromolar Zn (non-growth-inhibiting levels), metals appeared rapidly within cells, and intracellular Cd and Zn reached medium concentrations after 6 to 10 hours. Cd-peptide was observed in response to Cd after 2 hours, but this form only accounted for ~30% of soluble Cd after 24 hours. Peptide was not observed in cells exposed to 300 micromolar Zn for up to 7 days. Organic acid-to-metal stoichiometry indicated that endogenous organic acid content of cells was more than sufficient to complex absorbed metals and no evidence was found for stimulation of organic acid biosynthesis by Cd or Zn. Metal-complexing potential of organic acids for Cd and Zn versus endogenous cations is discussed as is vacuolar-extravacuolar distribution of metals. The absence of Cd-peptide does not limit Cd-accumulation in the system studied. Results suggest that tobacco suspension cells accommodate the presence of non-growth-inhibiting and growth-inhibiting levels of Cd and Zn by sequestration in the vacuole as complexes with endogenous organic acids and that this may be a principal means for accommodation of Cd as well as Zn in the presence and absence of Cd-peptide.     

Reversal of Cadmium-Induced Thyroid Dysfunction by Selenium, Zinc, or Their Combination in Rat

The aim of this study was to evaluate the potential benefit of combined treatment with zinc (Zn) and selenium (Se) in reversing cadmium (Cd)-induced thyroid dysfunction compared to Se or Zn treatment alone in rats exposed to Cd. For this purpose, 30 adult male Wistar albino rats were equally divided into control and four treated groups receiving either 200 ppm Cd (as CdCl2), 200 ppm Cd + 500 ppm Zn (as ZnCl2), 200 ppm Cd + 0.1 ppm Se (as Na2SeO3), or 200 ppm Cd + 500 ppm Zn + 0.1 ppm Se in their drinking water for 35 days. The results showed that Cd exposure increased significantly the relative thyroid weight (RTW), the thyroid Cd concentration, and the serum thyroid stimulating hormone (TSH) level, whereas the serum thyroxine (T4) level was decreased compared to control rats. The treatment of Cd-exposed rats with Se alone only partially protected from the Cd-induced decrease in serum T4 level. The treatment of Cd-exposed animals with Zn alone partially protected against Cd-induced thyroid dysfunction by maintaining normal RTW and by decreasing Cd concentration in the thyroid. It also partially prevents Cd-induced decrease in serum T4 level. The combined treatment of Cd-exposed animals with Se and Zn induced a more significant decrease in the thyroid Cd concentration than the Zn supplement and a total correction of the RTW. This treatment was also more effective than that with Se or Zn alone in reversing Cd-induced decrease in serum T4 level and Cd-induced increase in serum TSH level. Se and Zn can have a synergistic role against Cd-induced thyroid dysfunction.     

不同有机肥对土壤镉锌生物有效性的影响

在南方典型稻田设置连续4年施用猪粪、鸡粪、稻草的定位试验,监测施用不同有机肥条件下土壤及水稻植株镉(Cd)、锌(Zn)含量的变化,研究有机肥对土壤Cd、Zn活性及其交互作用的影响.结果表明: 施用有机肥(猪粪、鸡粪、稻草)对土壤全Cd、有效态Cd含量及Cd活性皆无显著影响,但有增加土壤Cd全量的趋势,且显著增加土壤全Zn、有效态Zn含量及Zn活性.施用猪粪、鸡粪、稻草皆可降低稻米Cd含量,降Cd效果为猪粪>鸡粪>稻草,猪粪处理水稻稻米、茎、叶Cd含量分别比对照下降37.5%、44.0%、36.4%;鸡粪处理水稻米、茎、叶Cd含量分别比对照下降22.5%、33.8%、22.7%;而稻草处理水稻米Cd含量比对照下降7.5%,但茎、叶Cd含量比对照分别增加8.2%、22.7%;施用猪粪、鸡粪降低稻米Cd含量主要是降低了水稻植株对土壤Cd的富集,而施用稻草则主要是降低了水稻茎Cd向稻米的转运.施用有机肥还增加了水稻茎Zn含量,施用猪粪、鸡粪、稻草的水稻茎Zn含量比单施化肥分别增加53.4%、41.2%、13.9%,但对水稻稻米、叶Zn含量无显著影响.Zn、Cd在土壤、植株茎中皆表现出显著的拮抗作用,土壤及水稻茎Zn含量的增加显著抑制了水稻米、茎、叶对Cd的吸收积累,且随土壤有效态Zn/Cd含量比值的增加,Zn、Cd竞争土壤吸附不是抑制水稻吸收积累Cd的主控因子,而Zn、Cd竞争吸收才是影响水稻吸收积累Cd的主控因子.     

锌对镉胁迫下马蔺生长、镉积累及生理抗性的影响

通过溶液培养研究了不同浓度锌(Zn)对镉(Cd)胁迫下马蔺(Iris lacteavar. chinen-sis)生长、Cd积累及抗氧化酶活性等的影响.结果表明:加入1~100mg.L-1Zn后,Cd胁迫下马蔺地上部Cd含量变化不大,而地下部Cd含量显著增加并呈先增后降的趋势;1mg.L-1Zn处理下马蔺地下部Cd含量最高,比对照显著增加51.4%;Zn浓度高于1mg.L-1后Cd含量均出现不同程度的下降,但仍高于对照.与单独Cd处理(10mg.L-1)相比,添加低浓度Zn(1~10mg.L-1)后,马蔺地上部生物量和叶绿素含量呈增加趋势,丙二醛(MDA)含量降低;在10mg.L-1Zn处理下,叶绿素a、b含量达到峰值,分别增加5.21%和22.27%,MDA含量降低25.46%,表明低浓度Zn缓解了Cd对马蔺的毒害.随Zn浓度的增加,Zn对Cd毒害的缓解作用逐渐降低,当溶液中Zn达到一定浓度(100mg.L-1)时,马蔺毒害加重,其生物量、叶绿素含量均下降,MDA含量显著增加.在试验胁迫浓度范围内超氧化物歧化酶(SOD)、过氧化物酶(POD)活性始终呈增加趋势,表明马蔺在受到Cd胁迫及Cd-Zn共存的条件下SOD和POD起着重要的抗氧化保护作用.     

Effects of land irrigation with partially-treated wastewater on Frankia survival and infectivity

Phytoextraction of Cd by some populations of Thlaspi caerulescens which have the ability to co-hyperaccumulate Cd and Zn requires information about the distribution of both metals within the plant at the organ-level. This work was conducted to determine whether the distribution and solubility of Cd and Zn in Thlaspi caerulescens are affected by the age of plant and organ, and whether Cd and Zn have a common distribution in the plant in soils contaminated by both metals. A series of pot experiments were conducted where a Cd- and Zn-hyperaccumulating population was grown on soils contaminated by Cd and Zn. Temporal changes in metal concentration of roots and of shoots was recorded, along with the water and CaCl₂ solubility of metals in the plant organs. Also, leaves were grouped according to their age and their respective content of Cd and Zn was measured. Both metals were present at higher concentrations in leaves than in roots. The whole-plant content of Zn decreased with time while that of Cd increased or remained unchanged. At harvest, young leaves exhibited higher Cd concentration than older, but the reverse was true for Zn. Both metals were more soluble in dry leaves and senescent leaves than in fresh material, and Zn was more water-soluble than Cd. In conclusion, the distribution of Cd and Zn in the hyperaccumulator T. caerulescensvaried according to the organ and plant age, and Cd and Zn were shown to have a different distribution within the plant.     

锌肥对不同基因型大麦吸收积累镉的影响

对土壤添加不同Zn、Cd条件下两种基因型(Sahara和Clipper)大麦对Zn、Cd的吸收积累研究表明,在本实验条件下土壤添加Zn、Cd对植物地上部生物量没有显著影响,但土壤添加Zn抑制植物根系生长,在土壤不缺Zn情况下添加Zn<20mg·kg^-1时并没有对大麦体内Cd浓度产生显著影响;当土壤Zn添加量达到40mg·kg^-1时,植物体内Cd浓度明显降低,植物吸收Cd的总量随着土壤添加Zn的增加而显著下降,这主要是由于根系生物量的下降所致,两个基因型大麦品种Zn效率存在显著差异,但这一差异对植物吸收Cd的总量没有影响,Zn高效品种Sahara根部Cd浓度显著低于Clipper。     

N、P、K肥对香根草修复土壤镉、锌污染效率的影响

通过盆栽试验研究在30 mg/kg镉(Cd)污染土壤条件下N[CO(NH2)2:100、200、300 mg/kg土]、P(P2O5:50、100、200 mg/kg土)和K(KCl:100、200、300 mg/kg土)处理对香根草修复土壤Cd和锌(Zn)污染效率的影响。结果表明:3种N处理能促进香根草地上部生长,而且显著提高地上部特别是叶的Cd和Zn含量,导致其修复效率成倍显著增加;200 mg/kg K处理显著提高Zn修复效率,但300 mg/kg K和50、200 mg/kg P处理却显著降低Cd、Zn修复效率。因此,为改善香根草对较贫瘠土壤中Cd、Zn污染的修复效率,应对香根草适施N肥,并控制或者不施P、K肥为佳。     

Soil-applied zinc and copper suppress cadmium uptake and improve the performance of cereals and legumes

The present study aimed to evaluate the effect of soil-applied Zn and Cu on absorption and accumulation of Cd applied through irrigation water in legume (chickpea and mung bean) and cereal (wheat and maize) crops. The results revealed that Cd in irrigation water at higher levels (2 and 5 mg L^?1) significantly (p < 0.05) reduced the plant biomass while the soil application of Zn and Cu, singly or combined, favored the biomass production. Plant tissue Cd concentration increased linearly with the increasing application of Cd via irrigation water. While Cd application caused a redistribution of metals in grains, straw, and roots with the highest concentration of Cd, Zn, and Cu occurred in roots followed by straw and grains. Zinc addition to soil alleviated Cd toxicity by decreasing Cd concentration in plant tissues due to a possible antagonistic effect. The addition of Cu to the soil had no consistent effects on Zn and Cd contents across all crops. Inhibitory effects of Cd on the uptake and accumulation of Zn and Cu have also been observed at higher Cd load. Thus, soil-applied Zn and Cu antagonized Cd helping the plant to cope with its toxicity and suppressed the toxic effects of Cd in plant tissues, thus favoring plant growth.     

Subcellular localisation of Cd and Zn in the leaves of a Cd-hyperaccumulating ecotype of<Emphasis Type="Italic"> Thlaspi caerulescens</Emphasis>

Thlaspi caerulescens (Ganges ecotype) is able to accumulate large concentrations of cadmium (Cd) and zinc (Zn) in the leaves without showing any toxicity, suggesting a strong internal detoxification. The distribution of Cd and Zn in the leaves was investigated in the present study. Although the Cd and Zn concentrations in the epidermal tissues were 2-fold higher than those of mesophyll tissues, 65–70% of total leaf Cd and Zn were distributed in the mesophyll tissues, suggesting that mesophyll is a major storage site of the two metals in the leaves. To examine the subcellular localisation of Cd and Zn in mesophyll tissues, protoplasts and vacuoles were isolated from plants exposed to 50 M Cd and Zn hydroponically. Pure protoplasts and vacuoles were obtained based on light-microscopic observation and the activities of marker enzymes of cytosol and vacuoles. Of the total Cd and Zn in the mesophyll tissues, 91% and 77%, respectively, were present in the protoplast, and all Cd and 91% Zn in the protoplast were localised in the vacuoles. Furthermore, about 70% and 86% of total Cd and Zn, respectively, in the leaves were extracted in the cell sap, suggesting that most Cd and Zn in the leaves is present in soluble form. These results indicate that internal detoxification of Cd and Zn in Thlaspi caerulescens leaves is achieved by vacuolar compartmentalisation.     

重金属Cd－Zn对水稻的复合污染和生态效应

重金属Ｃｄ－Ｚｎ对水稻的复合污染和生态效应周启星（浙江农业大学,杭州３１００２９）吴燕玉,熊先哲（中国科学院沈阳应用生态研究所,１１００１５）ＣｏｍｐｏｕｎｄｐｏｌｌｕｔｉｏｎｏｆＣｄａｎｄＺｎａｎｄｉｔｓｅｃｏｌｏｇｉｃａｌｅｆｆｅｃｔｏｎｒｉｃｅ...     

Cadmium accumulation in populations of Thlaspi caerulescens and Thlaspi goesingense

The capacity to accumulate cadmium (Cd) and zinc (Zn) was compared in Thlaspi goesingense and four populations of Thlaspi caerulescens . Two populations of T. caerulescens were grown in hydroponics at five concentrations of Cd. In addition, plants were grown in pots containing compost in which three different concentrations of Cd and two concentrations of Zn were added. A field trial was conducted to compare Zn and Cd uptake by three populations of T. caerulescens on nine selected plots of the Woburn Market Garden Experiment (UK) which had been contaminated to different degrees with heavy metals owing to past applications of sewage sludge. Results show that the four populations of T. caerulescens had the same ability to hyperaccumulate Zn but were significantly different in terms of Cd accumulation. Two populations of T. caerulescens from Southern France accumulated much more Cd than the populations from Prayon (Belgium) and Whitesike (UK). Generally, uptake of Cd was not decreased by increased concentrations of Zn in the substrate. These results indicate that the mechanisms of Cd and Zn hyperaccumulation are not identical in this species. This is the first report of hyperaccumulation of Cd by T. goesingense , but the growth of this species was markedly reduced by the large concentrations of Zn in the substrate. Future work should focus on the differences between Cd and Zn uptake in hyperaccumulator plants at the species and population level.     

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.