Solubilization of toxic metal minerals and metal tolerance of mycorrhizal fungi

This work investigates the ability of ericoid mycorrhizal (ErM) and ectomycorrhizal (EcM) fungi to solubilize different toxic metal (Cd, Cu, Pb, Zn)-containing minerals. Minerals were incorporated into solidified agar media and solubilization assessed by measuring clearing of the agar after fungal growth. Measurement of radial growth and biomass dry weight provided indications of metal tolerance: accumulated metal in the biomass was measured by atomic absorption spectrophotometry. Metal tolerance and solubilizing ability varied widely between different mineral and fungal species, and strains derived from sites of differing degrees of metal pollution. Zinc phosphate exhibited the least toxicity and was the easiest to solubilize by the majority of tested fungal isolates. Solubilization of toxic metal minerals was connected with both the pH of the medium and growth and tolerance of fungi and it seems that acidification of the medium was the main mechanism of mineral dissolution for most of the mycorrhizal fungi studied. A very strong lethal effect was observed for ectomycorrhizal isolates (>60% of strains) in the presence of Pb phosphate, carbonate, sulphide and tetraoxide. In contrast, ericoid mycorrhizal isolates were able to grow on Pb-mineral-amended media. A significant proportion of ericoid mycorrhizal cultures (70-90%) solubilized Cd and Cu phosphates and cuprite. None of the ericoid mycorrhizal and ectomycorrhizal fungi were able to produce a clear zone in Pb mineral-containing agar. However, many fungi were able to accumulate mobilized Pb in their mycelia. Differences in toxic metal mineral tolerance, mineral solubilization and metal uptake between populations isolated from metal-polluted and uncontaminated sites were related to the toxic metal which was the main pollutant in the original contaminated environment. In general, metal-tolerant fungi grew and solubilized toxic metal minerals better than non-tolerant isolates.     

土壤重金属污染的植物修复与金属超富集植物及其遗传工程研究

本文综述了土壤重金属污染的植物修复、金属超富集植物及其遗传工程的最新研究进展及存在问题,并提出加紧筛选和发现野生高生物量的金属超富集植物,在现有高生物量作物种质资源中筛选金属超富集作物,应用遗传工程技术把野生植物的超富集基因转移到现有高生物量植物(作物)中,寻找综合、可持续的植物修复手段等对策。     

Relative impact of soil, metal source and metal concentration on bacterial community structure and community tolerance

In a study to assess the sustainable use of sewage sludge application to land, the long-term effect of Zn and Cu contaminated sludge additions on the structure of the bacterial communities (using T-RFLP analysis) and their tolerance to additional metal exposure through pollution-induced community tolerance (PICT) assays was assessed. This used two soils that received metal-rich sludge cake (SC), liquid sludge (LS) or metal salts (MS) additions more than 10 years previously. Soil type had the predominant influence on bacterial community structure and PICT. The source of the metal contamination also had a large influence on community structure and PICT, greater than the effects due to metal concentrations. Nevertheless, in both Zn and Cu contaminated soils, PICT was observed and decreased in the order MS > LS > SC. Within a metal source and site, there was evidence of increased PICT with increasing Zn or Cu contamination, however few differences were significant as a result of high variability between sample replicates. These results highlight the importance of considering soil physico-chemical properties and the source of metal contamination as well as total metal concentrations when considering the long-term effects of metals on soil microbial communities. Further, the matrix that a metal is associated with prior to addition may play an important factor in determining levels of toxicity. This could have consequences for the interpretation and use of data from metal spiking experiments when considering metal limits for sludge application to land.     

Fulvic acids: structure and metal binding

Equilibrium calculations have been carried out at 25°C and an ionic strength of 0.04 to determine the predominant metal binding sites on a well-characterized fulvic acid for magnesium(ll), calcium(ll), manganese(ll), iron(lll), copper(ll) and zinc(II). Statistical estimates of the concentration of 14 sites are obtained from a random molecular model of fulvic acid. Protonation and metal formation constants are estimated from those of simple model ligands and corrected to the above conditions. Results indicate the importance of phthalate sites for all metals except iron(III), salicylate sites for iron(lll) and acetylacetonate sites for copper(ll). Iron (III) ions are found to form ML₂ type complexes extensively. The affinity of the metal ions for fulvic acid as a whole is found to decrease in the order iron(III) > copper(II) > zinc(II) > manganese(Il) > calcium(II) > magnesium (II).     

Humus forms and metal pollution in soil

Smelters in northern France are a serious source of soil pollution by heavy metals. We have studied a poplar plantation downwind of an active zinc smelter. Three humus profiles were sampled at increasing distance from the smelter, and the thickness of topsoil horizons was measured along a transect. We analysed the vertical distribution of humus components and plant debris to assess the impact of heavy metal pollution on the humus forms and on soil faunal activity. We compared horizons within a profile, humus profiles between them, and traced the recent history of the site. Near the smelter, poplar trees are stunted or dead and the humus form is a mor, with a well‐developed holorganic OM horizon. Here faunal activity is inhibited, so there is little faecal deposition and humification of plant litter. At the distant site poplar grows well and faunal activity is intense, so there are skeletonized leaves and many organo–mineral earthworm and millipede faecal pellets. The humus form is a mull, with a well‐developed hemorganic A horizon. The passage from mor to mull along the transect was abrupt, mor turning to mull at 250 m from the smelter, though there was a progressive decrease in heavy metal deposition. This indicates that there was a threshold (estimated to be 20 000 mg Zn kg^?1) in the resilience of the soil foodweb.     

Phytochelatins as biomarkers for heavy metal toxicity in maize: Single metal effects of copper and cadmium

Contamination of soils with heavy metals becomes more and more a problem in many countries all over the world. In areas where metal contaminated soils are used for food crop production, metals relatively mobile within the plant, such as cadmium (Cd) and zinc (Zn) can easily come into the food chain with great risks for human health. Since bioavailability of heavy metals in soils varies with soil and plant characteristics, e.g., mineralogical and organic matter properties of the soil and plant metal susceptibility, prediction of heavy metal uptake by plants by the common soil and plant chemical analysis techniques is often unreliable. Recently, the use of biomarkers has been suggested to be a suitable technique complementing chemical soil analysis. Therefore, the usefulness of the biomarker phytochelatin (PC), a non‐protein thiol, specifically induced in plants suffering from heavy metal stress, was tested. Maize (Zea mays L.) plants were exposed to excess copper (Cu) or Cd in nutrient solution systems and metal and PC concentrations were monitored in plant shoot and root. Results clearly illustrated that very soon after plant exposure to the metal, PC induction started, especially in plant roots. Phytochelatin seems to be a useful early warning system for heavy metal stress in plants.     

植物重金属转运蛋白及其在植物修复中的应用

植物修复技术已成为解决土壤重金属污染的一个绿色环保方法。利用基因工程和分子生物学技术,已经鉴定出一系列与重金属转运相关的蛋白及基因,包括ZIP家族、ABC载体、有机汞裂解酶基因merB、Hg离子还原酶基因merA和金属S蛋白基因MT等。本文着重从细胞、亚细胞水平上综述了一系列重金属转运蛋白的相关分子生物学研究进展及其在植物修复上的应用。     

公路旁土壤中水分和重金属的迁移

Roads with very high traffic loads in regions where soils are low in both pH and sorption capacity might be a source of percolation water loaded with heavy metals. Looking at some ＂worst case＂ scenarios, this study focused on the input of traffic related pollutants and on Pb, Cd, Cu, Zn, Ni and Cr concentrations in the soil matrix and soil solution, respectively. The analysis also included pH and electrical conductivity and at some sites DOC. The investigations were carried out on sandy soils with more or less low pH values at four motorway sites in Germany. The average of daily traffic was about 50 000 up to 90 000 vehicles. Soil pore water was collected in two soil depths and at four distances from the road. The pH in general decreased with increasing distance from the roadside. The elevated pH near the roadside was presumably caused by deposition of dust and weathering residues of the road asphalt, as well as by infiltration of salt that was used during winter time. At these road sites, increased heavy metal concentrations in the soil matrix as well as in the soil solution were found. However, the concentrations seldom exceeded reference values of the German Soil Protection Act. The soil solution concentrations tended to increase from the road edge to 10 m distance, whereas the concentration in the soil matrix decreased. Elevated DOC concentrations corresponded with elevated Cu concentrations but did not substantially change this tendency. High soil water percolation rates were found near the roads. Thus, even low metal concentrations of percolation water could yield high metal loads in a narrow area beside the road.     

Heavy metal effects on soil populations and heavy metal tolerance of Rhizobium meliloti,nodulation, and growth of alfalfa

Effects of heavy metals on rhizobia and the symbiotic association with leguminous hosts are currently unclear. To investigate this problem, we examined Rhizobium meliloti (microsymbiont) and alfalfa (Medicago sativa) (macrosymbiont) collected from soils contaminated with varying concentrations of heavy metals (varying distances from a Zn smelter operating 90 yr.). Soil populations of R. meliloti were not correlated with metal concentrations in soil. The lowest rhizobial population was found in the soil with the highest extractable metal concentrations, but the highest populations were found in soil which was moderately contaminated. A greenhouse study in which alfalfa was grown in the same soils showed no significant trend for nodulation or nitrogenase activity of roots. Highest nodule number and nitrogenase activity were observed in those soils which had the lowest population of R. meliloti. When the heavy metal Minimum Inhibitory Concentration (MIC) of individual isolates was examined, no correlation was found between the MIC and soil metal concentration (total, or water or 0.01 M Ca(NO₃)₂ extractable).These results indicate that even in highly contaminated soils, metal activity was not high enough to exert an antagonistic influence on the soil rhizobial population or the symbiotic association between alfalfa and R. meliloti.     

Hydrolysis of naptalam and structurally related amides: inhibition by dissolved metal ions and metal (hydr)oxide surfaces.

In metal ion-free solutions, the secondary amide naptalam hydrolyzes more rapidly as the pH is decreased; intramolecular nucleophilic attack by a carboxylate side group is very likely involved. Millimolar levels of dissolved Cu(II) and Zn(II) inhibit hydrolysis between pH 3.6 and pH 6.5. Metal ion-naptalam complex formation is important since addition of the competitive ligand citrate lessens the inhibitory effect. The metal (hydr)oxide surfaces Al(2)O(3) and FeOOH inhibit naptalam hydrolysis to a lesser degree; inhibition is proportional to the extent of naptalam adsorbed. Secondary amides (propanil, salicylanilide, and N-1-naphthylacetamide) and tertiary amides (N-methyl-N-1-naphthylacetamide, furalaxyl, and N, N-diethylsalicylamide) that lack carboxylate side groups do not hydrolyze within 45 days of reaction, even when millimolar Cu(II) concentrations are present. Tertiary amides possessing carboxylate side groups (N,N-diethyl-3,6-difluorophthalamic acid and N, N-dimethylsuccinamic acid) do hydrolyze but are insensitive to the presence or absence of Cu(II). The inhibitory effect is believed to occur via metal coordination of (1) the carbonyl group of naptalam, which induces deprotonation of the amide group and makes the substrate less reactive toward nucleophilic attack; (2) the free carboxylate group of naptalam, which blocks intramolecular nucleophilic attack; or (3) a combination of the two.     

The metal — metal interactions in biological systems. Part I. Escherichlia coli

The effect of nine metal ions: Co(II), Cr(III), Mg, Ni(II), Pb(II), Se(IV), Ti(III), V(IV) and Zn and following pairs of them: Cr-Mg, Cr-Se, Cr-Co, Cr-Zn, Cr-Ni, Ni-Mg, Ni-Se, Ni-Co, Ni-Zn, Pb-Mg, Pb-Se, Pb-Co, Pb-Zn, Ti-Mg, Ti-Se, Ti-Co, Ti-Zn, V-Mg, V-Se, V-Co and V-Zn was tested on the growth of Escherichia coli. Each test lasted 4 days. Magnesium is the most potent protective agent in all pairs of metals. The observed effects are not dependent only on concentration but also on time.     

Heavy metal deposition in Scandinavia

Maps showing the regional differences in the atmospheric deposition of Pb, Cd, Hg, Cr, Ni, Cu, Zn, and Fe have been prepared for Finland, Norway and Sweden from data obtained by means of moss analysis, a method previously shown to be a sensitive and reliable tool in measuring heavy metal deposition. In all elements the minima were recorded in northern Norway, the maxima in south-central or south-western Sweden and southernmost Norway. The greatest regional differences were measured in Pb and Cd.     

Heavy metal content and toxicity of mine and quarry soils

Purpose

Soils formed in metallic mines and serpentinite quarries, among other unfavourable features, have high levels of heavy metals. They can release into the environment causing surface and subsurface water contamination, uptake by plants, their accumulation in the food chain and adverse effects on living organisms. In this work, we studied the magnitude of the soils’ toxic effects not only on spontaneous plants but also on two species with phytoremediation potential.

Materials and methods

Several soils from two different exploitations were selected: a lead and zinc mine and a serpentinite quarry. Soils were characterized, and the pseudo-total and extractable contents of Co, Cr and Ni in soils from a serpentinite quarry were determined. The Cd, Pb and Zn pseudo-total and extractable contents were determined in soils developed in the Pb/Zn abandoned mine. Using a biotest, the chronic toxicity of the soil samples on higher plants was determined. Festuca ovina L., Cytisus scoparius (L.) Link., Sinapis alba L. and Brassica juncea L. were selected, the first two because they are spontaneous plants in the study areas and the last two because they have heavy metal phytoremediation potential.

Results and discussion

Pseudo-total contents of Co, Cr and Ni in the serpentinite quarry soils and of Zn, Pb and Cd in the Zn/Pb mine soils exceed generic reference levels. CaCl₂ is the reactant that extracts the highest proportion of Co, Cr and Ni in the quarry soils and EDTA the largest proportion of Pb Zn and Cd content in the mine soils. The germination index values based on seed germination and root elongation bioassays revealed increasing plant sensitivity to the mine soils in the following order: B. juncea?<?S. alba?<?F. ovina?<?C. scoparius. The wide range of GI values indicates that the response of test plants to soil heavy metals depended on their concentrations and soil characteristics, especially pH and organic matter content.

Conclusions

The pollution index indicates severe Cd, Pb and Zn contamination in the mine soils, as well as high Cr and Ni and moderate Co contamination in the serpentinite quarry soils. The performed biotests were suitable for identifying toxic soils and showed that the studied soils are toxic to the spontaneous plants, more to C. scoparius than to F. ovina. They also indicate that the mine soils are more toxic than the quarry soils for both species.

     

Regulation and function of AtNRAMP4 metal transporter protein

The NRAMP gene family encodes integral membrane proteins mediating the transport of a broad range of transition metals in bacteria, fungi, plants, and animals. We studied the regulation of AtNRAMP4 in Arabidopsis. In a previous study, we showed that AtNRAMP3 and AtNRAMP4 transport manganese (Mn), iron (Fe), and cadmium (Cd). In this study, we show that, in contrast to AtNRAMP3, AtNRAMP4 complements the growth phenotype of the zrt1zrt2 Zn uptake deficient yeast mutant. In a previous study, we have shown that, under Fe starvation, AtNRAMP4 mRNA levels are up-regulated in Arabidopsis. To analyze the regulation of AtNRAMP4 at the protein level, we generated specific antibodies against AtNRAMP4 protein. The antiserum was able to recognize a tagged version of AtNRAMP4 expressed in yeast. The antibody did not reveal any change in AtNRAMP4 protein level upon Fe starvation in Arabidopsis thaliana ecotype Columbia plants. In AtNRAMP4 overexpressing plants, high levels of AtNRAMP4 protein could be detected. AtNRAMP4 overexpressing plants display cadmium hypersensitivity in a medium containing 50 μm FeEDTA as Fe source. However, despite the constitutive accumulation of AtNRAMP4 protein, AtNRAMP4 over-expressing plants did not display Cd hypersensitivity under high Fe supply (100 μm FeHBED). AtNRAMP4 over-expressing lines displayed the same sensitivity to Zn as controls under all conditions tested. Our results suggest a translational level for the regulation of AtNRAMP4. Over-expression of AtNRAMP4 in Arabidopsis thaliana confers a slight hypersensitivity to Cd but not to Zn.     

Heavy metal species,mobility and availability in soils

The ecological effects of heavy metals in soils are closely related to the content and speciation of the elements in the solid and liquid phases of soils. Methods for the determination of metal species in both phases are described and critically evaluated. In connection with the possible binding mechanisms of heavy metals in soils the concept of specific and non-specific adsorption is discussed. Recent results indicate that the adsorption of heavy metals on soil particles is not only restricted to the formation of surface complexes but can also take place in the interior of minerals. Diffusion processes of heavy metals into soil minerals are described for goethite. In order to assess metal mobility and availability in soils, the concentration and speciation of metals in the soil solution or in comparable aeqeous equilibrium solutions of soil samples and the transfer of metals from solid pools to the liquid phase and vice versa have to be assessed. Methods for the determination of the amount of plant available heavy metals are described and the associated problems discussed.     

Particle size fractionation and metal distribution in sewage sludges

A sequential step filtration technique has been used to investigate the different particle size fractions of suspended solids in several raw, activated and digested sewage sludge samples. Liquid sludge was passed through filters with progressively smaller pore sizes, from 100 µm down to 0.2 µm. The concentrations of Cd, Cu, Ni, Pb, and Zn associated with each particle size fraction were also determined. The particle size distributions indicated that there was a greater proportion of larger particles in the raw and activated sludges than in the digested sludges. Generally in activated sludge the metals tended to be associated with the large particles of > 100 µm, whereas in raw and particularly digested sludges the metals were found in the smaller particle size fractions from 20 µm, down to 2.5 µm. Calculating specific associations in terms of mg kg^?1 the 8 to 20 µm range appears important in complexing metals. This was especially evident when comparing the sludges from the same treatment works. Generally distributions were more widespread when results were expressed as mg kg^?1 as opposed to mg L^?1.     

EPR spectra of humic acids and their metal complexes

On the basis of ESR spectra, humic substances are believed to be hydroquinone type polymer radicals with considerable cation exchange and redox capacity. All 3d-transition metal humates appear to be ionic high spin complexes. Manganese (II) ions doped in raw peats and peat humic acids are coordinated octahedrally with six oxygen-containing functional groups, e.g., carboxylate, phenolic, hydroxyl, carbonyl ligands, whereas copper (II) ions are in square planar arrangement with two carboxylate and two aliphatic nitrogen ligands. Doping with vanadyl(II) ions results in a square pyramidal arrangement with four oxygen-containing ligands. Diamagnetic manganese(VII), chromium(VI), molybdenum (VI) and vanadium(V) oxoanions in acidic solution are reduced by humic acids into paramagnetic manganese(II), chromium(III), molybdenum(V) and vanadium(IV) ions. Diamagnetic copper(I) ions, on the other hand, are oxidized by humic acids into paramagnetic copper(II) ions.In contrast to polyronic acid gels with outer-sphere structure, 3d-transition metal ions are generally bound to humic acids as inner-sphere chelate complexes.     

The influence of compost addition on heavy metal distribution between operationally defined geochemical fractions and on metal accumulation in plant

Purpose

Metal distribution patterns among geochemical fractions are informative for metal phytoavailability. Compost added to polluted soils may adsorb metals on the less phytoavailable fractions. A bioassay experiment was conducted to establish possible correlations between metal concentrations in different soil fractions and metal contents in edible plant parts and to investigate the influence of different compost loads on heavy metal availability to plants.

Materials and methods

Chinese cabbage plants were grown in pots with sandy and clayey soils and soils mixed with different doses of biosolid compost spiked with soluble heavy metal salts (Cd, Cu, and Pb). The metals’ distribution pattern in the soil and mixed samples was determined by sequential extraction procedure (modified BCR protocol). The studied fractions, from most to least bioavailable, were water-extractable (WE), exchangeable-adsorbed (EXC), associated with carbonates and acetic acid-soluble forms (CARB), occluded by reducible (hydro)oxides of Fe and Mn (RO), and associated with organic matter (OM) and a residual fraction (RES). Metal concentrations in soil extracts and in the digested plant tissue were measured by ICP-AES.

Results and discussion

The highest compost doses (72 and 115 Mg ha^?1) enhanced cabbage yield significantly. No excessive phytoaccumulation of metals was observed in plants grown in the clayey soil or its mixtures with compost. The compost dose of 72 Mg ha^?1 was optimal in decreasing Cu accumulation by plants grown in sandy soil, and 28.8 Mg ha^?1 was found to be effective in reducing Cd and Pb uptake. Metals were accumulated in plants primarily from the WE, EXC, and CARB fractions, whereas other fractions decreased phytoaccumulation. Compost addition suppressed heavy metal mobility, but different fractions were active in pollutant sorption, depending on soil type and metal.

Conclusions

Compost addition increased metal proportions in the RO and OM fractions, reducing metal phytoavailability. This is especially important for sandy soils with low adsorption ability and higher vulnerability to metal pollution than clayey soils. A compost dose of 20% v/v (or 28.8 Mg ha^?1) effectively reduced plant accumulation of Cd and Pb. We propose using the first three steps of the modified BCR protocol as a three-step sequential-extraction procedure for the most phytoavailable fractions of heavy metal: WE, EXC, and CARB.     

Spatial dependence and bioavailability of metal fractions in paddy fields on metal concentrations in rice grain at a regional scale

Purpose  

Although the bioavailability of heavy metals has been widely investigated, little information is available on the spatial correlations of heavy metals in soil–rice systems at a regional scale. A study of heavy metals in soil–rice systems at a present rice production area could provide valuable information on the safety of rice production and provide guidelines beneficial to agriculture management and strategic sustainable agriculture in China and other rapidly developing regions in the world. The overall goals of this study were to identify the characteristics of metal fractions and their bioavailability to rice plants in the paddy fields of a present rice production region.     

Evaluation of metal and microbial contamination in botanical supplements

The sale of botanical dietary supplements in the United States is on the rise. However, limited studies have been conducted on the safety of these supplements. There are reports on the presence of undesired metals in some of the botanical dietary supplements. In this study, echinacea, garlic, ginkgo, ginseng, grape seed extract, kava kava, saw palmetto, and St. John's wort supplements manufactured by Nature's Way, Meijer, GNC, Nutrilite, Solaray, Sundown and Natrol, have been analyzed for lead, mercury, cadmium, arsenic, uranium, chromium, vanadium, copper, zinc, molybdenum, palladium, tin, antimony, thallium, and tungsten using inductively coupled plasma mass spectrometry. All samples were devoid of mercury contamination. Results indicated that the botanical supplements analyzed did not contain unacceptable concentrations of these metals. These supplements were also evaluated for microbial contamination, and most samples analyzed showed the presence of bacteria or fungi or both. Microbes were not counted nor were microbial counts determined in these samples.