Assessment of Lead Ecotoxicity in Water using the Amphibian Larvae (<Emphasis Type="BoldItalic">Xenopus laevis</Emphasis>) and Preliminary Study of its Immobilization in Meat and Bone Meal Combustion Residues

Lead (Pb) is a major chemical pollutant of the environment. It has been associated with human activities for the last 6000 years. Quite rightly, it remains a public health concern today. The present investigation evaluates the toxic potential of Pb in larvae of the toad Xenopus laevis after 12 days exposure in lab conditions. Acute toxicity, genotoxicity and Pb bioaccumulation were analyzed. The genotoxic effects were analyzed in the circulating blood from the levels of micronucleus induction according to the French standard micronucleus assay (AFNOR 2000 Association française de normalization. Norme NFT 90-325. Qualité de l’Eau. Evaluation de la génotoxicité au moyen de larves d’amphibien (Xenopus laevis, Pleurodeles waltl)). Lead bioaccumulation was analyzed in the liver of larvae at the end of exposure. Moreover, the toxic potential of lead, in aquatic media, was investigated in the presence of meat and bone meal combustion residues (MBMCR) known to be rich in phosphates and a potential immobiliser of lead. Previously, acute toxicity and genotoxicity of MBMCR alone were evaluated using Xenopus larvae. The results obtained in the present study demonstrated: (i) that lead is acutely toxic and genotoxic to amphibian larvae from 1 mg Pb/l and its bioaccumulation is significant in the liver of larvae from the lowest concentration of exposure (1 μg Pb/l), (ii) MBMCR were not acutely toxic nor genotoxic in Xenopus larvae, (iii) lead in presence of MBMCR induced inhibition or reduction of the toxic and genotoxic potential of lead in water at concentrations that do not exceed the capacity of MBMCR of Pb-binding (iv) Pb accumulation in larvae exposed to lead with MBMCR in water was lower than Pb-accumulation in larvae exposed to lead alone except at the concentration of 0.01 mg Pb/l suggesting complex mechanisms of MBMCR interaction in organisms. The results confirm the high toxicity and genotoxicity of lead in the aquatic compartment and suggest the potential utility of MBMCR for use in remediation.     

Pb hyperaccumulation and tolerance in common buckwheat (Fagopyrum esculentum Moench)

Common buckwheat grown in Pb-contaminated soil was found to accumulate a large amount of Pb in its leaves (8,000 mg/kg DW), stem (2,000 mg/kg DW), and roots (3,300 mg/kg DW), without significant damage. This indicates that buckwheat is a newly recognized Pb hyperaccumulator, which is defined as a plant containing over 1,000 mg/kg of Pb in its shoots on a dry-weight basis. Moreover, it was shown that application of the biodegradable chelator methylglycinediacetic acid trisodium salt at concentrations of up to 20 mmol/kg resulted in a more than five times higher concentration of Pb in the shoot without notable growth inhibitation at up to 10 mmol/kg. These results indicate that buckwheat is a potential phytoremediator of Pb-contaminated soils.     

铅对山西省路域优势草本植物生长的影响及铅累积特征

采用温室盆栽试验,研究了不同浓度铅(0、500、1000、1500mg.kg-1)对14种山西省路域优势草本植物生长的影响及其铅吸收积累特征.结果表明:在14种草本植物中,随着铅浓度的增大,反枝苋和高丹草表现出明显的中毒症状,其他12种植物的株高和生物量与对照相比均无显著降低,表现出对铅污染具有一定的耐受性;藜和新麦草植株的地上部铅含量最低,各浓度铅处理下平均值分别为12.70和11.33mg.kg-1,地上部与根的铅含量比(S/R)最低,分别为0.12和0.10,表明二者为低积累植物,可用于铅污染土壤的植被恢复;红叶苋和绿叶苋植株地上部的铅迁移量最高,1500mg.kg-1铅处理下每百株铅迁移总量分别为53.37和45.29mg,可作为修复铅污染土壤的先锋植物.     

Phytoremediation of lead (Pb) and Arsenic (As) by Melastoma malabathricum L. from Contaminated Soil in Separate Exposure

This study was conducted to investigate the uptake of lead (Pb) and arsenic (As) from contaminated soil using Melastoma malabathricum L. species. The cultivated plants were exposed to As and Pb in separate soils for an observation period of 70 days. From the results of the analysis, M. malabathricum accumulated relatively high range of As concentration in its roots, up to a maximum of 2800 mg/kg. The highest accumulation of As in stems and leaves was 570 mg/kg of plant. For Pb treatment, the highest concentration (13,800 mg/kg) was accumulated in the roots of plants. The maximum accumulation in stems was 880 mg/kg while maximum accumulation in leaves was 2,200 mg/kg. Only small amounts of Pb were translocated from roots to above ground plant parts (TF < 1). However, a wider range of TF values (0.01–23) for As treated plants proved that the translocation of As from root to above ground parts was greater. However, the high capacity of roots to take up Pb and As (BF > 1) is indicative this plants is a good bioaccumulator for these metals. Therefore, phytostabilisation is the mechanism at work in M. malabathricum's uptake of Pb, while phytoextraction is the dominant mechanism with As.     

卡拉麦里南部工业区梭梭和琵琶柴重金属空间分布及污染评价

对五彩湾工业区周边21个采样点的梭梭(Hadoxylon)和琵琶柴(Reaummuria soongorica)的Zn、Cu、Cr、Pb、As、Hg 6种重金属元素含量测定,运用统计学方法和地统计插值法分析其茎叶和根部重金属含量变化情况和空间分布特征,并综合评价其污染程度和潜在生态危害性。结果表明:梭梭和琵琶柴植株Hg含量的最大值和均值都超出新疆土壤背景值0.017 mg/kg,除梭梭根部的均值0.060 mg/kg未超出国家土壤背景值0.065 mg/kg,在二者其他部位的最大值和均值都已超出;琵琶柴整株中Cr最大值72.62 mg/kg和Zn最大值97.61 mg/kg均超出新疆土壤背景值49.3 mg/kg和国家土壤背景值61 mg/kg,Pb元素未被检出。插值精度方面,Hg、As的RMSE较小,分别为0.263和0.443,预测模型中Hg的R~2为0.72,Cu的R~2为0.67,能较好地估计预测样点的重金属含量,Zn的R~2为0.31,精度较低;插值结果,琵琶柴中的Zn、Cr、As、Hg含量较高的区域均在工业园区内部及周围,受人为扰动程度较大。梭梭和琵琶柴中Hg元素分别为中度污染和重度污染;Hg元素为中等潜在危害程度高于其他4种元素。     

Cd and Pb Contents in Soil,Plants, and Grasshoppers along a Pollution Gradient in Huludao City,Northeast China

Cd and Pb contents in soil, plants, and two grasshopper species (Locusta migratoria manilensis and Acrida chinensis) were examined to quantify the influence ranges of zinc smelting on heavy metal contamination. Samples were collected simultaneously from Huludao City, a chemical and nonferrous smelting base in Northeast China. Cd and Pb contamination in soil and plants were serious. Cd and Pb contents were 13.32 and 8.83 mg/kg in L. migratoria manilensis and 16.67 and 15.00 mg/kg in A. chinensis, respectively. Correlation analysis indicated the same metal source for Cd and Pb in soil, plants, and grasshoppers. Cd and Pb contents in soil, plants, and grasshoppers were all significantly related to distances far from the zinc smelter in good negative logarithm model. The fitting curves indicated that the influence radius of the smelter on heavy metal contamination was about 4,000 m for soil and plants and about 2,000 m for grasshoppers.     

Toxic Effects of Pb on Anatomy and Hypericin Content in Hypericum perforatum L.

Lead (Pb) is one of the most common heavy metal contaminants in the environment. The present study was therefore undertaken to determine the effects of Pb on structural characteristics and hypericin production in Hypericum perforatum. Mature plants were treated with contaminated soil in seven treatments (75, 150, 300, 600, 800, 1000, and 1500 mg/kg Pb in soil) with three repeats per treatment every 14 days. Maximum observed Pb content in shoot parts was observed in the treatments with 600 and 1500 mg/kg Pb. The Pb concentration in roots was higher than in shoot parts, enhanced with increasing Pb concentration in the soil. In this study, Pb treatment significantly influenced the morphology, anatomy, and hypericin content in the plant. Anatomical characteristics of leaf, stem, and root affected by Pb contamination, as well as scanning electron microscopy (SEM) studies, revealed structural changes in stomata and epicuticular waxes. Under Pb toxicity, anatomical symptoms occurred in leaves, including increase in sizes of epidermal cells, mesophyll tissue, and diameter of stems and roots, as well as amplified vascular bundles and pith area. This, therefore, indicated that metal contamination can change the chemical composition of this plant. Maximum hypericin content was observed in the treatment containing 600 mg/kg Pb in soil, which then decreased.     

镉和铅污染对烤烟根区土壤微生物及烟叶品质的影响

通过盆栽试验,研究了Cd和Pb污染对烤烟根区微生物和烟叶品质的影响.结果表明,随着Cd、Pb处理浓度的增加,烤烟根区4种微生物数量均呈减少趋势;低浓度Cd(≤10 mg·kg^-1)和Pb(≤300 mg·kg^-1)下,烤烟根区4种微生物数量降幅最大;烤烟烟叶中品质指标糖/碱比和氮/碱比升高,化学成分组成趋于不协调,不利于烟叶香味的形成.烟叶K含量与Cd和Pb污染浓度呈负相关,与Cd污染下根区钾细菌数量呈显著(或极显著)正相关,与Pb污染根区钾细菌数量相关不显著.     

Soil Nitrogen Fertilization Effects on Phytoextraction of Cadmium and Lead by Tobacco (Nicotiana tabacum L.)

A greenhouse experiment using 24 plastic pots filled with 6 kg of Pb- and Cd-contaminated soil was carried out. In all 24 pots, soils were heavy metal–contaminated with 10 mg Cd kg^?1 soil and 500 mg of Pb kg^?1 soil by using CdCl and PbNO₃. Two-month-old tobacco (Nicotiana tabacum L.) plants were used to extract these heavy metals. Results showed that tobacco is able to remove Cd and Pb from contaminated soils and concentrate them in its harvestable part, that is, it could be very useful in phytoextraction of these heavy metals. Increasing additions of ammonium nitrate to soil (50, 100, and 150 mg N kg^?1 soil) significantly (p ≤ .05) increased aboveground Cd and Pb accumulation during a 50-day experimental period, whereas increasing additions of urea to soil (50 and 100 mg N kg^?1 soil) did not show these effects at the same significance levels. Increasing additions of ammonium nitrate to soil shows as dry matter increases, both accumulated Cd and accumulated Pb also increase when tobacco plants are growing under Pb- and Cd-contaminated soil conditions. Higher Pb concentrations depress Cd/Pb ratios for concentrations and accumulations, suggesting that Pb negatively affects Cd concentration and/or accumulation.     

Effects of AM Inoculation and Organic Amendment, Alone or in Combination, on Growth, P Nutrition, and Heavy-Metal Uptake of Tobacco in Pb-Cd-Contaminated Soil

As toxic pollutants commonly found in tobacco (Nicotiana tabacum L.) products, lead (Pb) and cadmium (Cd) can enter the human body via smoking and thus pose a potential health risk to smokers. We conducted a greenhouse experiment to study the effects of arbuscular mycorrhizal (AM) inoculation with Glomus intraradices BEG 141 and organic amendment with cattle manure, alone or in combination, on the growth, P nutrition, and heavy-metal uptake by tobacco plants grown in soil to which was added Pb-Cd at 0/0, 350/1, 500/10, and 1,000/100?mg?kg^?1, respectively. In general, AM colonization and plant growth were greatly reduced by Pb-Cd contamination, whereas organic amendment alleviated Pb-Cd stress and showed some beneficial effects on AM symbiosis and some soil parameters. AM inoculation, alone or in combination with organic amendment, increased plant dry weights and improved P nutrition significantly at all Pb-Cd addition levels, and, in most cases, it decreased Pb and Cd concentrations in tobacco plants and DTPA-extractable concentrations in soil. AM inoculation increased total glomalin-related soil protein (GRSP) concentrations in soil to which Pb-Cd was added. The higher soil pH and GRSP contents and the lower DTPA-extractable Pb and Cd concentrations contributed by AM inoculation and/or organic amendment may be contributing factors that lead to higher growth promotion and lower metal toxicity and uptake by plants. Our findings suggest that AM inoculation in combination with organic manure may be a potential method for not only tobacco production but phytostabilization of Pb-Cd-contaminated soil.     

Cross-protection and selectable marker genes in plant transformation

Summary Selectable marker genes play an important role in plant transformation. The level of selection pressure is generally established by generating a kill curve for the selectable marker. In most cases, the lowest concentration which kills all explants is used. This study examined two selectable marker genes, phosphinothricin acetyl transferase (PAT) and hygromycin phosphotransferase (HPT), in transformation of tobacco leaf disks. Experiments to determine the lethal level of the herbicide, glufosinate-ammonium (phosphinothricin) (PPT) using a leaf-disk regeneration assay established that no shoots regenerated at 2 to 4 mg PPT per 1. Likewise with the antibiotic, hygromycin (HYG), no plants regenerated at 50 mg hygromycin per 1. In contrast, after cocultivation of the leaf disks withAgrobacterium tumefaciens containing either the PAT or HPT gene in combination with a Bt gene for insect resistance, plants were successfully regenerated from leaf disks at 2 to 4 mg PPT per 1 and 50 mg hygromycin per 1. However, most plants regenerated at 2 and 3 mg PPT per 1 were found to be nontransformed (95–100% escapes) by i) Southern-blot analysis, ii) herbicide application test, and iii) insect feeding bioassay. On the other hand, plants that regenerated on 50 mg hygromycin per 1 and 4 mg PPT per 1 were transgenic as determined by Southern analysis, leaf assay for PPT or HYG resistance, and death of tobacco budworms feeding on these leaves. This study showed a significant level of cross-protection and/or transient expression of the PAT selectable marker gene allowing escapes (95–100%) at selection levels of 2 and 3 mg PPT per 1 which completely kill controls. On the other hand, the HPT gene at 50 mg is efficient in selecting for T-DNA integration.     

Effects of Zn<Superscript>2+</Superscript> on nodulation and growth of a South American actinorhizal plant, <Emphasis Type="Italic">Discaria americana</Emphasis> (Rhamnaceae)

Discaria americana is a xerophytic shrub which lives in symbiosis with an actinomycete of the genus Frankia. The objective of this paper was to investigate the effects of high soil Zn²⁺ concentrations on growth and nodulation on the association Discaria americana–Frankia with the aim of determining if this association is suitable for improving contaminated soils. Two experiments were performed in 1 dm³ pots containing soil and different Zn additions, from 0 to 2,000 mg Zn²⁺ kg⁻¹ dry soil, with or without N fertilization. Zn additions strongly delayed shoot and root growth, but once growth was initiated, the biomass production of the plants supplied with moderate Zn amounts did not differ from the control plants. Zn reduced the final nodule number, but not the total nodule biomass. At the end of the experiment only the highest Zn treatments showed a lower nodule weight than the control plants, while N addition completely inhibited nodulation. It is concluded than Zn reduces the number of Frankia infections, but once the actinomycete is inside the roots, nodules can continue growing according to plant demand for N, compensating the reduced nodule number with more biomass. On the other hand, there is a toxic effect of Zn itself on plants when present in very high concentrations.     

The Role of Mycorrhizae Associated with Vetiver Grown in Pb-/Zn-Contaminated Soils: Greenhouse Study

The effects of mycorrhizae on growth and uptake of N, P, Zn, and Pb by plants were investigated in a greenhouse trial using vetiver grass (Vetiveria zizanioides) as host. Inoculation of the host plants with arbuscular mycorrhizal fungi (AMF), Glomus mosseae and G. intraradices spores, significantly increased the growth and P uptake. Mycorrhizal colonization increased Pb and Zn uptake by plants under low soil metal concentrations (at 0 and 10 mg/kg of Pb or Zn), whereas under higher concentrations (at 100 and 1,000 mg/kg of Pb or Zn), it decreased Pb and Zn uptake. P concentration in soil was negatively correlated with mycorrhizal colonization as well as Zn or Pb concentrations. The results showed that inoculation of the host plants with AMF protects them from the potential toxicity caused by increased uptake of Pb and Zn, but the degree of protection varied according to the fungus and host plant combination. The potential of arbuscular mycorrhizae in phytoremediation of the Zn‐ or the Pb‐contaminated soils is discussed in this article.     

The response of tomato (Lycopersicon esculentum) to different concentrations of inorganic and organic compounds of arsenic

Tomato plants were cultivated in greenhouse and water solutions of arsenite (As(III)), arsenate (As(V)), methylarsonic acid (MA) and dimethylarsinic acid (DMA) were applied individually into cultivation substrate at two As levels, 5 and 15 mg kg⁻¹ of the substrate. Comparing the availability of arsenic compounds increased in order arsenite = arsenate < MA < DMA where the arsenic contents in plants decreased during vegetation period. Within a single plant, the highest arsenic concentration was found in roots followed in decreasing order by leaves, stems, and fruits regardless of arsenic compound applied. Arsenic toxicity symptoms reflected in suppressed growth of plants and a lower number and size of fruits were most significant with DMA treatment. However, the highest accumulation of arsenic by plants growing in the soil containing DMA was caused by higher mobility of this compound in the soil due to its lower sorption affinity. Our results confirmed substantial role of transformation processes of arsenic compounds in soil in uptake and accumulation of arsenic by plants.     

EDTA-assisted heavy-metal uptake by poplar and sunflower grown at a long-term sewage-sludge farm

Little information is available concerning the efficacy of chelates applied to biosolids (sewage-sludge)-treated soil for heavy-metal removal. The purpose of the experiment was to determine the availability to sunflower (Helianthus annuus L.) and hybrid poplar (Populus deltoides Marsh. × P. nigra L.) seedlings, of non-essential (Cd, Ni, Pb) and essential heavy metals (Cu, Fe, Mn, Zn) in field soil injected with biosolids since 1976 and treated with ethylenediamine-tetraacetic acid (EDTA) in 2001. Sunflower was grown at two densities, 20000 and 60000 plants/ha, and poplar at 10000 plants/ha. The tetrasodium salt of EDTA was applied at rates of 0, 0.5, 1, and 2 g EDTA salt per kg surface (25-cm depth) soil. The EDTA did not affect uptake by poplar of the three non-essential (Cd, Ni, Pb) and four essential (Cu, Fe, Mn, Zn) heavy metals. For sunflower, the 1.0 g/kg rate of chelate addition resulted in maximal removal of the three non-essential heavy metals (Cd, Ni, Pb). Uptake of the essential heavy metals by sunflower was little affected by the EDTA. At the 20000 plants/ha density, leaves of sunflower grown with 1.0 g EDTA Na₄2H₂O per kg soil accumulated more Cd, Ni, and Pb than leaves of sunflower grown without the EDTA salt. At this density, concentrations of Cd in leaves of sunflower without EDTA and with 1.0 g/kg EDTA salt were 2.2 and 6.5 g/g, respectively; for Ni, they were 6.7 and 19.2 g/g, respectively; and for Pb, they were 15.6 and 46.9 g/g, respectively. At the 60000 plants/ha density, stems of sunflower grown with 1.0 g EDTA Na₄2H₂O per kg soil accumulated more Cd, Ni, and Pb than stems of sunflower grown without the EDTA salt. At this density, concentrations of Cd in stems of sunflower without EDTA and with 1.0 g/kg EDTA salt were 0.6 and 4.6 g/g, respectively; for Ni, they were 1.7 and 17.6 g/g, respectively; and for Pb, they were 5.2 and 42.8 g/g, respectively. Removal of the non-essential heavy metals by sunflower was greater at the higher plant density (60000 plants/ha) compared to the lower one (20000 plants/ha).     

Distribution of Cd, Pb, Zn, Mo, and S in juvenile and mature Brassica napus L. var. napus

The study was conducted at three locations in the Savinjska region of Slovenia, where soil is contaminated with heavy metals due to the zinc industry (Cinkarna Celje). In Ponikva the soil to a depth of 30 cm contains 0.8 mg kg(-1) Cd, 32.2 mg kg(-1) Pb, and 86 mg Zn kg(-1), in Medlog 1.4 mg kg(-1) Cd, 37.4 mg kg(-1) Pb, and 115 mg kg(-1) Zn and in Skofja vas 10.9 mg kg(-1) Cd, 239.7 mg kg(-1) Pb, and 1356 mg kg(-1) Zn. The pH at the selected sites was between 7.3 and 7.6. In the beginning of September 2006 two hybrids of Brassica napus L. var. napus, PR45 D01 and PR46 W31 suitable for production of biodiesel obtained from Pioneer Seeds Holding GmbH, were sown. After 96 days juvenile and after 277 days mature plants were collected. Parts of plants (root, shoot and seed) were separated and Cd, Pb, Zn, Mo, and S determined by ultra-trace ICP-MS. We compared the uptake of Cd, Pb, Zn, Mo and S in different parts of juvenile and mature plants of the two different hybrids, TF (translocation factor), BAF (bioaccumulation factor), and PP (phytoextraction potential) were calculated. The mature hybrid PR46 W31 had higher shoot/root ratio and higher PP for metals (Cd, Pb, and Zn) and lower PP for the micronutrient (Mo) and macronutrient (S) on the polluted site. The study demonstrated the potential use of oilseed rape on multiply polluted soils for production of 1st and 2nd generation biofuels. The potential restoration of degraded land could also disburden the use of agricultural land.     

Effects of Cd and Zn on the development of annual xylem rings of young Norway spruce (Picea abies) plants

Three-year-old spruce (Picea abies) saplings were planted and cultivated for 2 years in pots with 3 1 substrate, consisting of a homogenized mixture of sand, peat and forest soil with a high organic content (volume ratio 11.52). This substrate was amended with 10–180 mol Cd [kg soil dry weight (DW)]^–1, 50–7500 mol Zn (kg soil DW)^–1 (determined with 1 M ammonium acetate extracts) or combinations of both elements. Annual xylem growth rings in stems of plants treated with 50 mol Cd (kg soil DW)^–1 or 7500 mol Zn (kg soil DW)^–1 were significantly narrower than in control plants. Growth reductions were more pronounced in the second year of the experiment. The contents of Cd and Zn in stem wood and needles were positively correlated with the substrate concentrations. The Mg contents of the spruce needles were inversely correlated with soil concentrations of Cd and Zn. Root development was impeded at moderate concentrations of Cd (50 mol kg^–1) or Zn (1000 mol kg^–1) in the substrate. The adverse effects of potentially toxic trace elements, like Cd or Zn, on xylem growth of spruce plants are discussed with regard to possible growth reductions in forest trees under field conditions.     

Heavy metal (Pb) accumulation in metallophytes as influenced by the variations in rhizospheric and non-rhizospheric soils physico-chemical characteristics

Activities at root-soil interface determine the solubility and uptake of metals by plants. Metal accumulation in plant species (Imperata cylindrical, Cynodon dactylon, Eleucine indica, Gomphrena celosoides, Sporobolus pyramidalis, Chromolaena odorata and Rhynchospora corymbosa) growing on Pb contaminated site as influenced by variations in physico-chemical characteristics, dissolved organic matter (DOM), Pb fractionation and different functional groups (using Fourier Transmittance Infra-red) of rhizospheric and non-rhizospheric soils was assessed. The electrical conductivity (2660–5520 µs) and Pb concentrations (51390.0–64080.0 mg/kg) were more in non-rhizospheric than rhizospheric soils having 276 µs to 3160 µs EC and 3289.0 to 44850.0 mg/kg Pb. More nutrients, DOM and carbohydrates functional groups (C-O; 1100 -1000 and O-H; 3700–3600) were found in rhizospheric compared to non-rhizospheric soils. The pH was slightly acidic (5.0–5.54) and E. indica with the lowest pH (5.0) accumulated highest Pb concentrations in shoot (8030 mg/kg) and root (16380 mg/kg) while C. odorata with highest values of pH, P, Ca and Mg in rhizospheric soil accumulated the least (root; 331.6 and shoot: 209.0 mg/kg). Pb was more in organic and residual fractions of rhizospheric and non-rhizospheric soils respectively. Reduction in pH, EC coupled with nutrients and DOM availability increased Pb uptake by plants.     

Interspecific differences in Zn,Cd and Pb accumulation by freshwater algae and bryophytes

The relationships between the concentrations of zinc, cadmium and lead in aquatic plants and the concentrations of these metals in the ambient water have been compared for three algae (Lemanea fluviatilis, Cladophora glomerata, Stigeoclonium tenue), one liverwort (Scapania undulata) and three mosses (Amblystegium riparium, Fontinalis antipyretica, Rhynchostegium riparioides). The data to establish these relationships are all based on our own studies, some published already, some here for the first time. They come from a wide range of streams and rivers in Belgium, France, Germany, Ireland, Italy and the U.K. There were significant bivariate positive relationships between concentrations of Zn, Cd and Pb in water and plant for all species except Cd and Pb in Stigeoclonium tenue. When relationships were compared using datasets with total or filtrable metals in water, most differences were slight. However there were marked differences both between species and between metals. Comparison for the seven species of Zn in the plant when aqueous Zn is 0.01 mg l^–1, a concentration at which all seven were found, shows that the four bryophytes had the highest concentrations; however the two green algae had steeper slopes (representing change in concentration in plant in response to change in aqueous concentration). Lemanea fluviatilis had a slope closer to that of the bryophytes, but the concentration was about one order of magnitude lower. All seven species were found at a concentration of 0.01 mg l^–1 Pb, and at this concentration there were almost two orders of magnitude difference between the species which accumulated the most (Scapania undulata) and the one which accumulated the least (Cladophora glomerata). The steepest slope was however shown by C. glomerata.When multiple stepwise regression was applied, the aqueous metal under consideration was the first variable extracted in only nine of the 21 regressions. However one of the other heavy metals (aqueous or accumulated) was extracted first in all but one of the other regressions, presumably because the occurrences of Zn, Cd and Pb were strongly cross-correlated. The principal non-heavy metal factor extracted for Zn and Cd, but not Pb, was aqueous Ca. The relevance of these results to the use of aquatic plants for monitoring heavy metals is discussed.