Root uptake and shoot accumulation of cadmium by lettuce at various Cd:Zn ratios in nutrient solution.

Cadmium (Cd) is toxic to animals and humans after it accumulates over decades in the kidney cortex. Food crops grown in Cd-contaminated soils are the primary sources of excessive Cd entry into humans. Although plant available Zn concentration in soil is an important factor which can greatly reduce Cd uptake by plant roots and its translocation into the edible parts, Cd:Zn ratio is suggested to be a more important factor in comparison with Zn concentration alone in determining Cd uptake by plants. In the present study, the physiological mechanisms of Cd absorption by roots and its translocation to leaves of lettuce (Lactuca sativa L.) at various Cd:Zn ratios in the rooting media were investigated. For this purpose, seedlings of hydroponically-grown lettuce were exposed to combinations of four Zn (0, 12.5, 50 and 100µM) and four Cd (0, 0.5, 1 and 10µM) concentrations providing different ratios of Cd:Zn. At each level of Cd, decreasing the Cd:Zn ratio by increasing Zn concentration in the nutrient solution caused significant reduction of root symplastic Cd and also reduced Cd loading into the xylem and Cd transport to and accumulation in leaves. The highest root symplastic Cd (1087mg/kg-1 Dry Weight [DW]) and shoot Cd concentrations (64mg/kg-1 DW) were observed at the highest Cd:Zn ratio of = 0.8 (Zn = 12.5, Cd = 10). At the Cd:Zn ratios of ≤ 0.01, shoot Cd concentration was less than the Detection Limit (< 0.02mg/kg DW). Decreasing Cd:Zn ratio in nutrient solution was accompanied with significant increase in root apoplastic Cd and decrease in the root symplastic Cd. According to the obtained results, at the Cd:Zn ratio equal to 0.01 and less, Cd concentration in lettuce shoots decreased to < 0.02mg/kg.

Potential Adherence of Flue Gas Desulfurization Gypsum to Forage as a Consideration for Excessive Ingestion by Ruminants.

Gypsum (calcium sulfate dihydrate, CaSO⋅2HO) has long been used to improve soils and crop production, and its use has recently been encouraged by the USDA-NRCS for soil conservation through a new Conservation Practice Standard: Code 333. However, there is concern regarding the adverse effects of excessive direct ingestion of sulfate in gypsum by ruminants. The standard requires ruminants to be removed from grazing after application until after a rainfall, but there has been no research documenting gypsum adherence to forage or the potential for rainfall to reduce gypsum adherence. A study was established to examine the adherence and persistence of gypsum on different forage species. Two forages (bermudagrass [ L.] and tall fescue [ Schreb.]) were examined after gypsum applications at rates of 0, 1, and 5 Mg ha. The forage was sampled immediately after application, 1 wk after application, after a 1.5-cm rain, and after a 3.3-cm rain. Immediately after gypsum application, more gypsum adhered to the tall fescue (27.9 g gypsum kg) compared with bermudagrass (8.6 g gypsum kg), likely due to differences in the leaf structure. This represents S concentrations of 0.16 and 0.52% for any grazing ruminants feeding exclusively on the bermudagrass and tall fescue pastures. On succeeding sampling dates, substantial amounts of gypsum persisted only on the wider-leaved tall fescue. With tall fescue, a difference in gypsum adherence could be observed after a 1.5-cm rain, but no significant difference was observed between the gypsum application and the control after an additional 3.3-cm rain. Results indicate that care should be observed with grazing after gypsum application, especially on wide-leaved forages. However, using application rates within normal agronomic beneficial use guidelines (NRCS standard 333), negative results from direct ingestion of gypsum are not likely if grazing is discontinued several weeks and until a rainfall event occurs.

Towards practical cadmium phytoextraction with Noccaea caerulescens.

A series of field trials were conducted to investigate the potential of Noccaea caerulescens F.K. Mey [syn. Thlaspi caerulescens J &C Presl. (see Koch and Al-Shehbaz 2004)] populations (genotypes) derived from southern France to phytoextract localized Cd/Zn contamination in Thailand. Soil treatments included pH variation and fertilization level and application of fungicide. N. caerulescens populations were transplanted to the field plots three months after germination and harvested in May, prior to the onset of seasonal rains. During this period growth was rapid with shoot biomass ranging from 0.93-2.2 g plant(-1) (280-650 kg ha(-1)) DW. Shoot Cd and Zn concentrations for the four populations evaluated ranged from 460-600 and 2600-2900 mg kg(-1) DW respectively. Cadmium and Zn Translocation Factors (shoot/root) for the populations tested ranged from 0.91-1.0 and 1.7-2.1 and Bioaccumulation Factors ranged from 12-15 and 1.2-1.3. We conclude that optimizing the use of fungicidal sprays, acidic soil pH, planting density and increasing the effective cropping period will increase rates of Cd and Zn removal enough to facilitate practical Cd phytoextraction from rice paddy soils in Thailand.

Phytotoxicity of zinc and manganese to seedlings grown in soil contaminated by zinc smelting.

Historic emissions from two zinc smelters have injured the forest on Blue Mountain near Palmerton, Pennsylvania, USA. Seedlings of soybeans and five tree species were grown in a greenhouse in a series of mixtures of smelter-contaminated and reference soils and then phytotoxic thresholds were calculated. As little as 10% Palmerton soil mixed with reference soil killed or greatly stunted seedlings of most species. Zinc was the principal cause of the phytotoxicity to the tree seedlings, although Mn and Cd may also have been phytotoxic in the most contaminated soil mixtures. Calcium deficiency seemed to play a role in the observed phytotoxicity. Exposed soybeans showed symptoms of Mn toxicity. A test of the effect of liming on remediation of the Zn and Mn phytotoxicity caused a striking decrease in Sr-nitrate extractable metals in soils and demonstrated that liming was critical to remediation and restoration.

Evaluation of plant growth regulators to increase nickel phytoextraction by Alyssum species.

Recent studies have shown that application of phytohormones to shoots of Alyssum murale increased biomass production but did not increase Ni shoot concentration. Increased biomass and Ni phytoextraction efficiency is useful to achieve economically viable phytomining. The objective of this study was to evaluate the effect of two types of phytohormones on the Ni phytoextraction capacity of four Alyssum species. Two different commercially available phytohormones (Cytokin and Promalin) based on cytokinins and/or gibberellins were applied on shoot biomass of four Ni hyperaccumulating Alyssum species (A. corsicum, A. malacitanum, A. murale, and A. pintodasilvae). Cytokin was applied in two concentrations and promalin in one concentration. The application of phytohormones had no clear positive effect on biomass production, Ni accumulation and Ni phytoextraction efficiency in the studied Alyssum species. A. malacitanum was the only species in which a significantly negative effect of these treatments was observed (in Ni uptake). A slightly positive response to promalin treatment was observed in the biomass production and Ni phytoextraction efficiency of A. corsicum. Although this effect was not significant it does indicate a potential application of these approaches to improve phytoextraction ability. Further studies will be needed to identify the most adequate phytohormone treatment as well as the appropriate concentrations and application times.

Exogenous cytokinin treatments of an Ni hyper-accumulator, Alyssum murale, grown in a serpentine soil: implications for phytoextraction.

Application of exogenous plant growth regulators was examined as a viable technique to increase the efficiency of plant metal extraction from contaminated soils. The aim of this study was to investigate the alteration of Ni phytoextraction by Alyssum murale, a Ni hyperaccumulator, following the application of cytokinins. The following parameters were investigated: Ni accumulation, plant growth, gas exchange, stomata behavior and the concentration of nonprotein thiols (glutathione, y-Glu-Cys, and phytochelatins). In a pot experiment, A. murale plants grown in a serpentine soil were treated with a mix of naturally occurring cytokinins. Results showed that Ni accumulation in plants ranged from 4000 to 7000 mg kg(-1) confirming the hyper-accumulation ability from the soil used. Cytokinin treatments produced a significant increase in plant biomass and transpiration rate whereas no significant variation in Ni accumulation or the concentration of non-protein thiols was observed. The results suggest that A. murale is a plant species sensitive to cytokinin treatment and that cytokinin treatment is potentially useful in increasing the phytoextraction capability by increasing biomass. Moreover, for first time, evidence was obtained that the Ni hyperaccumulation mechanism is independent of water flux and transpiration rate.

Ovine ruminal microbes are capable of biotransforming hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX).

Bioremediation is of great interest in the detoxification of soil contaminated with residues from explosives such as hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). Although there are numerous forms of in situ and ex situ bioremediation, ruminants would provide the option of an in situ bioreactor that could be transported to the site of contamination. Bovine rumen fluid has been previously shown to transform 2,4,6-trinitrotoluene (TNT), a similar compound, in 4 h. In this study, RDX incubated in whole ovine rumen fluid was nearly eliminated within 4 h. Whole ovine rumen fluid was then inoculated into five different types of media to select for archaeal and bacterial organisms capable of RDX biotransformation. Cultures containing 30 µg mL(-1) RDX were transferred each time the RDX concentration decreased to 5 µg mL(-1) or less. Time point samples were analyzed for RDX biotransformation by HPLC. The two fastest transforming enrichments were in methanogenic and low nitrogen basal media. After 21 days, DNA was extracted from all enrichments able to partially or completely transform RDX in 7 days or less. To understand microbial diversity, 16S rRNA-gene-targeted denaturing gradient gel electrophoresis (DGGE) fingerprinting was conducted. Cloning and sequencing of partial 16S rRNA fragments were performed on both low nitrogen basal and methanogenic media enrichments. Phylogenetic analysis revealed similar homologies to eight different bacterial and one archaeal genera classified under the phyla Firmicutes, Actinobacteria, and Euryarchaeota. After continuing enrichment for RDX degraders for 1 year, two consortia remained: one that transformed RDX in 4 days and one which had slowed after 2 months of transfers without RDX. DGGE comparison of the slower transforming consortium to the faster one showed identical banding patterns except one band. Homology matches to clones from the two consortia identified the same uncultured Clostridia genus in both; Sporanaerobacter acetigenes was identified only in the consortia able to completely transform RDX. This is the first study to examine the rumen as a potential bioremediation tool for soils contaminated with RDX, as well as to discover S. acetigenes in the rumen and its potential ability to metabolize this energetic compound.

Absorption, tissue distribution, and elimination of residues after 2,4,6-trinitro[14C]toluene administration to sheep.

The compound 2,4,6-trinitrotoluene (TNT) is a persistent contaminant of some industrial and military sites. Biological bioremediation techniques typically rely on the immobilization of TNT reduction products rather than on TNT mineralization. We hypothesized that sheep ruminal microbes would be suitable for TNT destruction after phytoremediation of TNT-contaminated soils by cool-season grasses. Therefore we investigated the fate of [14C]TNT in ruminating sheep to determine the utility of ruminant animals as a portion of the bioremediation process. Three wether sheep were dosed with 35.5 mg each of dietary unlabeled TNT for 21 consecutive days. On day 22 sheep (41.9 +/- 3.0 kg) were orally dosed with 35.5 mg of [14C]TNT (129 microCi; 99.1% radiochemical purity). Blood, urine, and feces were collected at regular intervals for 72 h. At slaughter, tissues were quantitatively collected. Tissues and blood were analyzed for total radioactive residues (TRR); excreta were analyzed for TRR, bound residues, and TNT metabolites. Plasma radioactivity peaked within 1 h of dosing and was essentially depleted within 18 h. Approximately 76% of the radiocarbon was excreted in feces, 17% in urine, with 5% being retained in the gastrointestinal tract and 1% retained in tissues. Parent TNT, dinitroamino metabolites, and diaminonitro metabolites were not detected in excreta. Ruminal and fecal radioactivity was essentially nonextractable using ethyl acetate, acetone, and methanol; covalent binding of fecal radioactive residues was evenly distributed among extractable organic molecules (i.e., soluble organic matter, soluble carbohydrate, protein, lipid, and nucleic acid fractions) and undigested fibers (cellulose, hemicellulose, and lignin). This study demonstrated that TNT reduction within the ruminant gastrointestinal tract leads to substantial immobilization of residues to organic matter, a fate similar to TNT in other strongly reducing environments.

Selection and breeding of plant cultivars to minimize cadmium accumulation.

Natural variation occurs in the uptake and distribution of essential and nonessential trace elements among crop species and among cultivars within species. Such variation can be responsible for trace element deficiencies and toxicities, which in turn can affect the quality of food. Plant breeding can be an important tool to both increase the concentration of desirable trace elements and reduce that of potentially harmful trace elements such as cadmium (Cd). Selection programs for a low-Cd content of various crops, including durum wheat, sunflower, rice and soybean have been established and low-Cd durum wheat cultivars and sunflower hybrids have been developed. In durum wheat (Triticum turgidum L. var durum), low-Cd concentration is controlled by a single dominant gene. The trait is highly heritable, and incorporation of the low-Cd allele can help to reduce the average grain Cd to levels below proposed international limits. The allele for low-Cd concentration does not appear to affect major economic traits and should not cause problems when incorporated into durum cultivars. The cost of Cd selection in a breeding program is initially large both in terms of Cd determination and reduced progress towards development of other economic traits, but declines as more breeding lines in the program carry the low-Cd trait and are utilized in new crosses. Production of low-Cd crop cultivars can be used as a tool to reduce the risk of movement of Cd into the human diet.

Hyperaccumulator Alyssum murale relies on a different metal storage mechanism for cobalt than for nickel.

The nickel (Ni) hyperaccumulator Alyssum murale has been developed as a commercial crop for phytoremediation/phytomining Ni from metal-enriched soils. Here, metal co-tolerance, accumulation and localization were investigated for A. murale exposed to metal co-contaminants. A. murale was irrigated with Ni-enriched nutrient solutions containing basal or elevated concentrations of cobalt (Co) or zinc (Zn). Metal localization and elemental associations were investigated in situ with synchrotron X-ray microfluorescence (SXRF) and computed-microtomography (CMT). A. murale hyperaccumulated Ni and Co (> 1000 microg g(-1) dry weight) from mixed-metal systems. Zinc was not hyperaccumulated. Elevated Co or Zn concentrations did not alter Ni accumulation or localization. SXRF images showed uniform Ni distribution in leaves and preferential localization of Co near leaf tips/margins. CMT images revealed that leaf epidermal tissue was enriched with Ni but devoid of Co, that Co was localized in the apoplasm of leaf ground tissue and that Co was sequestered on leaf surfaces near the tips/margins. Cobalt-rich mineral precipitate(s) form on leaves of Co-treated A. murale. Specialized biochemical processes linked with Ni (hyper)tolerance in A. murale do not confer (hyper)tolerance to Co. A. murale relies on a different metal storage mechanism for Co (exocellular sequestration) than for Ni (vacuolar sequestration).

Trace element chemistry in residual-treated soil: key concepts and metal bioavailability.

Trace element solubility and availability in land-applied residuals is governed by fundamental chemical reactions between metal constituents, soil, and residual components. Iron, aluminum, and manganese oxides; organic matter; and phosphates, carbonates, and sulfides are important sinks for trace elements in soil-residual systems. The pH of the soil-residual system is often the most important chemical property governing trace element sorption, precipitation, solubility, and availability. Trace element phytoavailability in residual-treated soils is often estimated using soil extraction methods. However, spectroscopic studies show that sequential extraction methods may not be accurate in perturbed soil-residual systems. Plant bioassay is the best method to measure the effect of residuals on phytoavailability. Key concepts used to describe phytoavailability are (i) the salt effect, (ii) the plateau effect, and (iii) the soil-plant barrier. Metal availability in soil from metal-salt addition is greater than availability in soil from addition of metal-containing residuals. Plant metal content displays plateaus at high residual loadings corresponding to the residual's metal concentration and sorption capacity. The soil-plant barrier limits transmission of many trace elements through the food chain, although Cd (an important human health concern) can bypass the soil-plant barrier. Results from many studies that support these key concepts provide a basis of our understanding of the relationship between trace element chemistry and phytoavailability in residual-treated soils. Research is needed to (i) determine mechanisms for trace element retention of soil-residual systems, (ii) determine the effect of residuals on ecological receptors and the ability of residuals to reduce ecotoxicity in metal-contaminated soil, and (iii) predict the long-term bioavailability of trace elements in soil-residual systems.

Phenotypic characterization of microbes in the rhizosphere of Alyssum murale.

Metal hyperaccumulator plants like Alyssum murale are used for phytoremediation of Ni contaminated soils. Soil microorganisms are known to play an important role in nutrient acquisition for plants, however, little is known about the rhizosphere microorganisms of hyperaccumulators. Fresh and dry weight, and Ni and Fe concentrations in plant shoots were higher when A. murale was grown in non-sterilized compared to sterilized soils. The analysis of microbial populations in the rhizosphere of A. murale and in bulk soils demonstrated that microbial numbers were affected by the presence of the plant. Significantly higher numbers of culturable actinomycetes, bacteria and fungi were found in the rhizosphere compared to bulk soil. A higher percent of Ni-resistant bacteria were also found in the rhizosphere compared to bulk soil. Percentage of acid producing bacteria was higher among the rhizosphere isolates compared to isolates from bulk soil. However, proportions of siderophore producing and phosphate solubilizing bacteria were not affected by the presence of the plant. We hypothesize that microbes in the rhizosphere of A. murale were capable of reducing soil pH leading to an increase in metal uptake by this hyperaccumulator.

A modified risk assessment to establish molybdenum standards for land application of biosolids.

The USEPA standards (40 CFR Part 503) for the use or disposal of sewage sludge (biosolids) derived risk-based numerical values for Mo for the biosolids --> land --> plant --> animal pathway (Pathway 6). Following legal challenge, most Mo numerical standards were withdrawn, pending additional field-generated data using modern biosolids (Mo concentrations <75 mg kg(-1) and a reassessment of this pathway. This paper presents a reevaluation of biosolids Mo data, refinement of the risk assessment algorithms, and a reassessment of Mo-induced hypocuprosis from land application of biosolids. Forage Mo uptake coefficients (UC) are derived from field studies, many of which used modern biosolids applied to numerous soil types, with varying soil pH values, and supporting various crops. Typical cattle diet scenarios are used to calculate a diet-weighted UC value that realistically represents forage Mo exposure to cattle. Recent biosolids use data are employed to estimate the fraction of animal forage (FC) likely to be affected by biosolids applications nationally. Field data are used to estimate long-term Mo leaching and a leaching correction factor (LC) is used to adjust cumulative biosolids application limits. The modified UC and new FC and LC factors are used in a new algorithm to calculate biosolids Mo Pathway 6 risk. The resulting numerical standards for Mo are cumulative limit (RPc)=40 kg Mo ha(-1), and alternate pollutant limit (APL) = 40 mg Mo kg(-1) We regard the modifications to algorithms and parameters and calculations as conservative, and believe that the risk of Mo-induced hypocuprosis from biosolids Mo is small. Providing adequate Cu mineral supplements, standard procedure in proper herd management, would augment the conservatism of the new risk assessment.

Manure phosphorus extractability as affected by aluminum- and iron by-products and aerobic composting.

Shifts in manure phosphorus (P) chemical forms and pool sizes induced by water treatment residuals and industrial mineral by-products are largely undefined. We conducted a manure P fractionation study to determine mechanisms of reduction of dissolved reactive phosphorus (DRP) in poultry manure upon mineral by-product additions. The effects of composting on the P immobilization efficacy of the by-products were determined using laboratory self-heating composting simulators. The mineral by-products included an aluminum-water treatment residual (Al-WTR) and an iron-rich titanium-processing by-product. The noncomposted manure averaged 0.11 g g(-1) of total P as DRP forms. The by-products significantly reduced manure DRP, by an average of 39 and 48% in the Al- and the Fe-treated manure, respectively. The by-products also reduced the 0.5 M NH4F-extractable phosphorus (FEP) fraction. Shifts in P forms between FEP and 0.1 M NaOH-extractable phosphorus (SHEP) depended upon the Al and Fe contents of the by-products while the combined FEP + SHEP pool remained constant. Phosphate sorption measurements supported the observations that the Fe-rich by-product was more effective at reducing manure DRP and enhancing the formation of SHEP forms at the expense of FEP than the Al-WTR. Composting had no effect on the efficacy of either by-product to reduce DRP. Potential mechanisms of enhanced P stabilization in treated manure upon composting included chemical shifts from the DRP and FEP fractions to the citrate-bicarbonate-dithionite extractable P fraction. Thus, the choice of P immobilization agents affected the stability of immobilized P forms and should be taken into consideration in developing manure processing and nutrient stabilization methods.

Influence of the zinc hyperaccumulator Thlaspi caerulescens J. & C. Presl. and the nonmetal accumulator Trifolium pratense L. on soil microbial populations.

Metal hyperaccumulator plants like Thlaspi caerulescens J. & C. Presl. are used for phytoremediation of contaminated soils. Since little is known about the rhizosphere of hyperaccumulators, the influence of T. caerulescens was compared with the effects of Trifolium pratense L. on soil microbes. High- and low-metal soils were collected near a zinc smelter in Palmerton, Penn. Soil pH was adjusted to 5.8 and 6.8 by the addition of Ca(OH)2. Liming increased bacterial populations and decreased metal toxicity to levels allowing growth of both plants. The effects of the plants on total (culturable) bacteria, total fungi, as well as cadmium- and zinc-resistant populations were assessed in nonrhizosphere and rhizosphere soil. Both plants increased microbial populations in rhizosphere soil compared with nonrhizosphere soil. Microbial populations were higher in soils planted with T. pratense, but higher ratios of metal-resistant bacteria were found in the presence of T. caerulescens. We hypothesize that T. caerutescens acidifies its rhizosphere. Soil acidification in the rhizosphere of T. caerulescens would affect metal uptake by increasing available metals around the roots and consequently, increase the selection for metal-resistant bacteria. Soil acidification may be part of the hyperaccumulation process enhancing metal uptake from soil.

Mineral status of female rats affects the absorption and organ distribution of dietary cadmium derived from edible sunflower kernels (Helianthus annuus L.).

The intake of food cadmium (Cd) in microg/day over time can increase the body burden of this element. Some human populations that consume subsistence rice-based diets low in calcium (Ca), iron (Fe), and zinc (Zn) are more susceptible to Cd poisoning than populations that consume more nutritious diets. This study determined the effects of marginal deficiencies of these essential elements on the absorption and organ retention of Cd from a natural food that contains Cd, edible sunflower kernels (Helianthus annuus L.; SFK). Weanling female rats were fed diets containing 20% SFK in a 2x2x2 factorial design with marginal and adequate amounts of Ca, Zn, and Fe. Marginal Zn (11 mg/kg) and Fe (13 mg/kg), and Cd (0.18 mg/kg) were derived solely from 20% SFK. These amounts of Fe and Zn represented 39 and 90% of the NRC requirement for the rat, respectively. The marginal dietary Ca concentration (2.5 g/kg) was one-half the NRC requirement. After 5 weeks on the experiment, rats were fed 1 g of their respective diets containing SFK extrinsically labeled with 37 kBq 109Cd, and absorption was determined by whole-body counting techniques. Rats were then killed and organs collected for 109Cd assays. No effect of treatment on weight gain was observed; however, when dietary Zn was low, feeding marginal Ca elevated Cd absorption by 50% (P<0.05) over those fed adequate Ca and Zn. Feeding marginal Fe elevated Cd absorption >2.5-fold (P<0.001) over those fed adequate Fe. In contrast, the naturally occurring Zn in SFK that provided 90% of the rat's requirement was enough to deter excessive absorption of Cd and enough to alone prevent significant elevation of organ Cd. Organ content of 109Cd and Cd followed the same general pattern as whole-body absorption. These data show that marginal nutritional deficiencies of Ca and Fe can readily enhance the body burden of Cd that comes from the diet. Also, some natural competitors of Cd, such as Zn, contained in foods can independently minimize Cd absorption.

Amelioration of nickel phytotoxicity in muck and mineral soils.

In situ remediation (phytostabilization) is a cost-effective solution for restoring the productivity of metal-contaminated soils and protection of food chains. A pot experiment with wheat (Triticum aestivum L.), oat (Avena sativa L.), and redbeet (Beta vulgaris L.) was conducted to test the ability of limestone and hydrous ferric oxide (HFO) to ameliorate Ni phytotoxicity in two soils contaminated by particulate emissions from a nickel refinery. Quarry muck (Terric Haplohemist; 72% organic matter) contained 2210 mg kg(-1) of total Ni. The mineral soil, Welland silt loam (Typic Epiaquoll), was more contaminated (2930 mg Ni kg(-1)). Both soils were very strongly acidic, allowing the soil Ni to be soluble and phytotoxic. Nickel phytotoxicity of the untreated muck soil was not very pronounced and could be easily confused with symptoms of Mn deficiency that occurred in this soil even with Mn fertilization. Severe nickel phytotoxicity of the untreated mineral soil prevented any growth of redbeet, the most sensitive crop; even wheat, a relatively Ni-resistant species, was severely damaged. White banding indicative of Ni phytotoxicity was present on oat and wheat leaves grown on the acidic mineral soil. Soil Ni extracted with diethylenetriaminepentaacetic acid (DTPA) and 0.01 M Sr(NO3)2 was indicative of the ameliorative effect of amendments and correlated well with Ni concentrations in plant shoots. Making soils calcareous was an effective treatment to reduce plant-available Ni and remediate Ni phytotoxicity of these soils to all crops tested. The ameliorative effect of HFO was crop-specific and much less pronounced.

Effects of metal phytoextraction practices on the indigenous community of arbuscular mycorrhizal fungi at a metal-contaminated landfill.

Phytoextraction involves use of plants to remove toxic metals from soil. We examined the effects of phytoextraction practices with three plant species (Silene vulgaris, Thlaspi caerulescens, and Zea mays) and a factorial variation of soil amendments (either an ammonium or nitrate source of nitrogen and the presence or absence of an elemental sulfur supplement) on arbuscular mycorrhizal (AM) fungi (Glomales, Zygomycetes) at a moderately metal-contaminated landfill located in St. Paul, Minn. Specifically, we tested whether the applied treatments affected the density of glomalean spores and AM root colonization in maize. Glomalean fungi from the landfill were grouped into two morphotypes characterized by either light-colored spores (LCS) or dark-colored spores (DCS). Dominant species of the LCS morphotype were Glomus mosseae and an unidentified Glomus sp., whereas the DCS morphotype was dominated by Glomus constrictum. The density of spores of the LCS morphotype from the phytoremediated area was lower than the density of these spores in the untreated landfill soil. Within the experimental area, spore density of the LCS morphotype in the rhizosphere of mycorrhizal maize was significantly higher than in rhizospheres of nonmycorrhizal S. vulgaris or T. caerulescens. Sulfur supplement increased vesicular root colonization in maize and exerted a negative effect on spore density in maize rhizosphere. We conclude that phytoextraction practices, e.g., the choice of plant species and soil amendments, may have a great impact on the quantity and species composition of glomalean propagules as well as on mycorrhiza functioning during long-term metal-remediation treatments.

Phytoremediation of soil metals.

The phytoremediation of metal-contaminated soils offers a low-cost method for soil remediation and some extracted metals may be recycled for value. Both the phytoextraction of metals and the phytovolatilization of Se or Hg by plants offer great promise for commercial development. Natural metal hyperaccumulator phenotype is much more important than high-yield ability when using plants to remove metals from contaminated soils. The hypertolerance of metals is the key plant characteristic required for hyperaccumulation; vacuolar compartmentalization appears to be the source of hypertolerance of natural hyperaccumulator plants. Alternatively, soil Pb and Cr6+ may be inactivated in the soil by plants and soil amendments (phytostabilization). Little molecular understanding of plant activities critical to phytoremediation has been achieved, but recent progress in characterizing Fe, Cd and Zn uptake by Arabidopsis and yeast mutants indicates strategies for developing transgenic improved phytoremediation cultivars for commercial use.

Direct Measurement of 59Fe-Labeled Fe2+ Influx in Roots of Pea Using a Chelator Buffer System to Control Free Fe2+ in Solution.

Fe2+ transport in plants has been difficult to quantify because of the inability to control Fe2+ activity in aerated solutions and non-specific binding of Fe to cell walls. In this study, a Fe(II)-3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine-4[prime]4"-disulfonic acid buffer system was used to control free Fe2+ in uptake solutions. Additionally, desorption methodologies were developed to adequately remove nonspecifically bound Fe from the root apoplasm. This enabled us to quantify unidirectional Fe2+ influx via radiotracer (59Fe) uptake in roots of pea (Pisum sativum cv Sparkle) and its single gene mutant brz, an Fe hyperaccumulator. Fe influx into roots was dramatically inhibited by low temperature, indicating that the measured Fe accumulation in these roots was due to true influx across the plasma membrane rather than nonspecific binding to the root apoplasm. Both Fe2+ influx and Fe translocation to the shoots were stimulated by Fe deficiency in Sparkle. Additionally, brz, a mutant that constitutively exhibits high ferric reductase activity, exhibited higher Fe2+ influx rates than +Fe-grown Sparkle. These results suggest that either Fe deficiency triggers the induction of the Fe2+ transporter or that the enhanced ferric reductase activity somehow stimulates the activity of the existing Fe2+ transport protein.