HEAVY METAL ACCUMULATION AND THE NATURE OF EDAPHIC ENDEMISM IN THE GENUS CALOCHORTUS (LILIACEAE)

Bulb and leaf tissues from five species in the genus Calochortus and the soils from collection sites were analyzed for cation and trace element concentrations. All five species, only three of which are considered ultramafic endemics, possessed high concentrations of nickel and copper in both the bulbs and leaves, but not at levels high enough (> 1% dry wt) to be considered hyperaccumulators. Only moderate to trace amounts of cobalt and chromium were detected in both plant tissues. Calcium-magnesium ratios, typically low in ultramafic soils and plants, were low in bulb tissue, but two to 10 times higher in leaf tissue. The ability to tolerate excessively high levels of nickel and other heavy metals may be a physiological exaptation of the genus Calochortus and not necessarily an evolutionary response by several species to life on an ultramafic substrate. The nature of “serpentine” endemism is discussed in the context of heavy metal accumulation by various plant species.     

Nicotianamine Over-accumulation Confers Resistance to Nickel in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Nicotianamine is a methionine derivative involved in iron homeostasis, able to bind various other metals in vitro. To investigate its role in vivo, we expressed a nicotianamine synthase cDNA (TcNAS1) isolated from the polymetallic hyperaccumulator Thlaspi caerulescens in Arabidopsis thaliana. Transgenic plants expressing TcNAS1 over-accumulated NA, up to 100-fold more than wild type plants. Furthermore, increased NA levels in different transgenic lines were quantitatively correlated with increased nickel tolerance. The tolerance to nickel is expressed at the cellular level in protoplast experiments and is associated with an increased NA content. We have also shown that the most NA-over accumulating line showed a high tolerance to nickel and a significant Ni accumulation in the leaves when grown on nickel-contaminated soil. Our results highlight a new potential role for nicotianamine in heavy metal tolerance at the cellular but also at the whole plant level, easily transposable to a non-tolerant non-hyperaccumulator species. These results open new perspectives for the modulation of nicotianamine content in plants for phytoremediation.     

Nickel localization in tissues of different hyperaccumulator species of Euphorbiaceae from ultramafic areas of Cuba

Pantropical species of the genera Phyllanthus and Euphorbia and the Cuban endemic genus Leucocroton from the Euphorbiaceae family, were selected for nickel localization microanalysis. Scanning Electron Microscopy coupled with Energy Dispersive X-ray Microanalysis (SEM-EDX) was used for qualitative detection of nickel in the selected Ni-hyperaccumulator species: Euphorbia helenae, Leucocroton linearifolius, L. flavicans Phyllanthus orbicularis, P. discolor and P. xpallidus, all collected from Cuban ultramafic soils. The leaves and stems from the Euphorbiaceae species analyzed were the organs with higher nickel accumulation. Elemental mapping of leaves and stem tissues from these species have been compared. The highest Ni concentrations were found in the laticifer tubes of stems and the epidermis tissues of leaves in all the analyzed species, suggesting a general pattern of the Euphorbiaceae family for nickel accumulation. The high nickel concentrations and its rather homogeneous distribution found in leaves of these Ni-hyperaccumulating plants suggest a possible role in protection mechanisms against environmental stress, such as UV irradiation.     

Response to cytosolic nickel of Slow Vacuolar channels in the hyperaccumulator plant <Emphasis Type="Italic">Alyssum bertolonii</Emphasis>

We applied the patch-clamp technique to investigate the transport properties of the Slow Vacuolar (SV) channel identified in leaf vacuoles of Alyssum bertolonii Desv., a nickel hyperaccumulator plant growing in serpentine soil of the northern Apennines (Italy). SV currents recorded in vacuoles from adult plants collected in their natural habitat showed high sensitivity towards cytosolic nickel. Dose-response analyses indicated half-maximal current inhibition at submicromolar concentrations, i.e. up to three orders of magnitude lower than previously reported values from other plant species. The voltage-dependent increase of residual currents at saturating nickel concentrations could be interpreted as relief of channel block by nickel permeation at high positive membrane potentials. Including young plants of A. bertolonii into the study, we found that SV channels from these plants did not display elevated nickel sensitivity. This difference may be related to age-dependent changes in nickel hyperaccumulation of A. bertolonii leaf cells.     

The vegetation of ultrabasic soils on the Isle of Rhum II. The causes of the debris

Summary

The relationship between the Rhum ultrabasic skeletal soils and their debris vegetation was investigated by plant analyses and field and laboratory experiments. Samples of Agrostis vinealis, Arenaria norvegica ssp norvegica, Calluna vulgaris, Festuca vivipara, Plantago maritima and Racomitrium lanuginosum from these soils usually had low concentrations of potassium and calcium, and high concentrations of sodium, magnesium (and high Mg/Ca quotients), iron and nickel. There were instances of very high iron concentrations (up to 22.4mg g^?1 in Plantago maritima), very high Mg/Ca quotients (up to 27.8 in Arenaria novegica spp. norvegica) and high nickel concentrations (up to 0.48mg g^?1 in Plantago maritima). A nutrient addition experiment which was set up in 1965 on an exposed barren area had in 1982 over twice as many species as originally recorded and a nearly complete plant cover. Shorter-term work has confirmed that nutrient availability limits the ultrabasic vegetation. An experiment on Agrostis vinealis in simulated soil solutions showed that a gabbro (non-ultrabasic) clone had a higher R.G.R. (relative growth rate), even at the higher of two experimental Mg/Ca quotients, than two peridotite (ultrabasic) clones and was moreover not significantly affected by the higher nickel concentration used. It is concluded that the low vegetation cover on the skeletal soils is maintained by low soil nutrients which might interact with the coarsely sandy texture in exacerbating the effects of periodic drought and frost heaving. There is no unequivocal evidence for plant toxicities associated with high magnesium or nickel in the Rhum soils.     

Evaluation of the phytoremediation potential of Arundo donax L. for nickel-contaminated soil

This study investigates the accumulation and distribution of nickel in Arundo donax L. parts to assess the potential use of this plant in phytoremediation of Ni-contaminated soils. The effect of ethylene diamine tetra-acetic acid (EDTA) and nutrient solution containing NPK on the plant was proped. A 35-day pot experiment was performed in the laboratory and the pots were irrigated with Ni-contaminated solution combined or not with EDTA and NPK. The growth of plants was evaluated at the end of the experiment. The accumulation of Ni was analyzed by atomic absorption spectroscopy (AAS). The obtained results indicate that the plant was able to survive with high Ni content. The growth and the concentrations of Ni in the plant tissues were less affected. In the absence of the amendments, Ni was accumulated in the stems and leaves. However, the addition of NPK significantly reduced Ni concentration in the stems and leaves. The application of EDTA enhanced Ni uptake in roots. The translocation factor (TF) was greater than 1, which categorizes A. donax L. as a great candidate for Ni phytoextraction. A. donax L. is suitable for phytoremediation of Ni. This investigation contributes to the studies on the potential of phytoremediation technologies in Algeria.     

Serpentine and non-serpentine Silene dioica plants do not differ in nickel tolerance

Serpentine soils are hostile to plant life. They are dry, contain high concentrations of nickel and have an unfavorable calcium/magnesium ratio. The dioecious plant Silene dioica (L.) Clairv. (Caryophyllaceae) is the most common herb on serpentine soils in the Swedish mountains. It also commonly grows on non-serpentine soils in the subalpine and coastal area. I have compared the germination frequency, plant establishment and growth of serpentine and subalpine non-serpentine populations in serpentine soil under greenhouse conditions. Further more I have studied the specific effect of nickel on root and shoot growth of serpentine and non-serpentine plants from the subalpine and coastal area in solutions with different concentrations of nickel. Plants from serpentine and non-serpentine populations grew well and in a similar fashion in serpentine soil. Moreover, S. dioica plants, irrespective of original habitat, tolerated enhanced concentrations of nickel when grown in solutions. An analysis of metal content in serpentine plants from natural populations shows that S. dioica has a higher nickel concentration in the roots than in the shoots. The growth studies show that S. dioica is constitutively adapted to serpentine, and that all populations have the genetic and ecological tolerance to grow on serpentine.     

The use of transgenic canola (Brassica napus) and plant growth-promoting bacteria to enhance plant biomass at a nickel-contaminated field site

The applicability of transgenic plants and plant growth-promoting bacteria to improve plant biomass accumulation as a phytoremediation strategy at a nickel (Ni)-contaminated field site was examined. Two crops of 4-day old non-transformed and transgenic canola (Brassica napus) seedlings in the presence and absence of Pseudomonas putida strain UW4 (crop #1) or P. putida strain HS-2 (crop #1 and 2) were transplanted at a Ni-contaminated field site in 2005. Overall, transgenic canola had increased growth but decreased shoot Ni concentrations compared to non-transformed canola, resulting in similar total Ni per plant. Under optimal growth conditions (crop #2), the addition of P. putida HS-2 significantly enhanced growth for non-transformed canola. Canola with P. putida HS-2 had trends of higher total Ni per plant than canola without P. putida HS-2, indicating the potential usefulness of this bacterium in phytoremediation strategies. Modifications to the planting methods may be required to increase plant Ni uptake.     

Ulteriori indagini sul rapporto tra nichel e acidi malico e malonico in Alyssum

Abstract

Further contribution on the relationship between nickel and malic and malonic acids in Alyssum.—Further research has been carried out on extracts of leaves and seeds of A. bertolonii Desv., grown on serpentine and normal garden soil, in order to investigate the nature of the nickel compounds present in this plant. On the purified extracts, obtained with water and formic acid, malic and malonic acids have been determined through gas-chromatography of the trimethyl-sililderivatives; Ni, Ca, Mg and K have been analysed by atomic absorption spectrophotometry.

Both malic and malonic acids are present in the leaves of the plants collected on serpentine and therefore rich in nickel, whilst they are present only in small amounts in the plants grown in garden soil. Seeds contain mainly high levels of malic acid. Remarkable amounts of malic acid, related to a high nickel content, have been observed also in the leaves of another nickel accumulator in the genus Alyssum, i.e. A. serpyllifolium Desf. ssp. lusitanicum Dudley et P. Silva, endemic of Portugal serpentines.     

<Emphasis Type="Italic">Chenopodium</Emphasis>: a prospective plant for phytoextraction

Leaf samples were collected from 40 accessions of Chenopodium spp. and assessed for six heavy metals (Fe, Zn, Cu, Ni, Cr and Cd) accumulation to explore the use of Chenopodium for phytoextraction of heavy metals. The results suggest that Chenopodium spp. have the ability to accumulate large quantities of heavy metals in the leaf tissues even when they are present in low concentrations in the soil. C. quinoa is a better accumulator of Ni, Cr and Cd than the rest of the species, while C. album accessions are good copper accumulators. Bioconcentration factor for chromium ranged from 0.36 (C. album “Chandanbathua”) to 6.57 (C. quinoa Ames 13719) with 13 accessions of C. quinoa scoring above the mean value. High heritability coupled with high genetic advance was recorded for Ni, Cr and Cd, which indicated a major role of additive gene action in the inheritance of these characters. Zinc showed significant positive association with iron (0.351**), nickel (0.659**), chromium (0.743**) and cadmium (0.288**). Nickel was significantly and negatively associated with copper (−0.663**), while it was positively and significantly correlated with chromium (0.682**) and cadmium (0.461**). Considering the accumulation efficiency of Chenopodium spp. with respect to heavy metals, this genus should be further explored for decontamination of metal polluted soils, with plant breeding playing an important role in evolving new plant types with higher capacity of heavy metal accumulation.     

The Effect of Recombinant Heavy Metal-Resistant Endophytic Bacteria on Heavy Metal Uptake by Their Host Plant

The ncc-nre nickel resistance system of Ralstonia metallidurans 31A was efficiently expressed in Burkholderia cepacia L.S.2.4 and Herbaspirillum seropedicae LMG2284. The heterologous expression of ncc-nre encoded nickel resistance was accompanied by nickel removal from the culture medium. B. cepacia L.S.2.4:: ncc-nre and H. seropedicae LMG2284::ncc-nre were able to remove 35 and 15% nickel, respectively. The capacity to remove nickel through sequestration or bio-precipitation processes and consequently lowering the free nickel concentration could offer interesting benefits for these endophytic bacteria and their host plants. Once inoculated in their host plant, they could possibly alter the nickel speciation and therefore decrease the free ions and thus toxic concentration for the plant metabolism. Lupinus luteus L, when grown on a nickel enriched substrate and inoculated with B. cepacia L.S.2.4::ncc-nre, showed a significant increase (30%) of nickel concentration in the roots, whereas the nickel concentration in the shoots remained comparable with that of the control plants. The inoculation of Lolium perenne (cv Atlas) with the nickel resistance derivative of H. seropedicae LMG2284::ncc-nre resulted in a significant decrease of the nickel concentration in the roots (11%) as well as in the shoots (14%). As this phenomenon was also observed in the Lolium perenne plants inoculated with the wild-type strain LMG2284, the nickel resistance characteristics probably are not responsible for the altered nickel uptake observed.     

Plant-soil relationships in a New Caledonian serpentine flora

Summary Elemental concentrations were studied in two New Caledonian hyper-accumulators of nickel and in the soils supporting these plants. The nickel content of Hybanthus austrocaledonicus was directly related to the total nickel content of the soil, and this species may therefore be used for biogeochemical prospecting. Nickel was correlated well with other elements of the iron family in non-lateritic ultrabasic soils but not in laterites. Both H. austrocaledonicus and Homalium kanaliense contained adequate amounts of potassium despite very low concentrations in the supporting soils. These low nutrient levels were further aggravated by antagonism to potassium uptake caused by other soil elements. Because of the paucity of relationships between nickel in H. kanaliense and elements in the soil, it is suggested that the high nickel levels in this plant may be controlled by organic constituents in this species. re]19760504     

Effect of cations,including heavy metals,on cadmium uptake by Lemna polyrhiza L.

Cations, including calcium, magnesium, potassium, sodium, copper, iron, nickel and zinc, inhibited (up to 40%) extracellular binding and intracellular uptake of cadmium by Lemna polyrhiza in solution culture. Test plants showed a high capacity of extracellular cadmium binding which was competitively inhibited by copper, nickel and zinc; however, calcium, magnesium and potassium caused non-competitive inhibition. Iron and sodium increased K _m and decreased V _max, thereby causing mixed inhibition of extracellular binding. Intracellular cadmium uptake displayed Michaelis-Menten kinetics. It was competitively inhibited by calcium, magnesium, iron, nickel and zinc. Monovalent cations (sodium and potassium) caused non-competitive and copper caused mixed inhibition of intracellular cadmium uptake. Thus, high levels of cations and metals in the external environment should be expected to lower the cadmium accumulation efficiency of L. polyrhiza.     

NICKEL UPTAKE BY CALIFORNIAN STREPTANTHUS AND CAULANTHUS WITH PARTICULAR REFERENCE TO THE HYPERACCUMULATOR S. POLYGALOIDES GRAY (BRASSICACEAE)

The nickel content of herbarium specimens of Californian Streptanthus and Caulanthus has been determined. Serpentine-tolerant species had nickel values of 10–100 μg/g (dry mass) except in the case of S. polygaloides Gray which had nickel values in the range 3,300–14,800 μg/g (0.33%–1.48%). This is the first hyperaccumulator of nickel found in the Americas. It is suggested that the later classification of S. polygaloides as the monotypic Microsemia polygaloides (Gray) Greene may be justified if the nickel accumulation data are used to reinforce the existing marked morphological differences from other Streptanthus.     

Effect of pH on the biosorption of nickel and other heavy metals by Pseudomonas fluorescens 4F39

Accumulation of heavy metals by Pseudomonas fluorescens 4F39 was rapid and pH-dependent. The affinity series for bacterial accumulation of metal cations decreased in the order Ni>>Hg>U>>As>Cu>Cd>Co>Cr>Pb. Metal cations were grouped into those whose accumulation increased as the pH increased, with a maximum accumulation at the pH before precipitation (Ni, Cu, Pb, Cd, Co), and those whose maximum accumulation was not associated with precipitation (Cr, As, U, Hg). High Ni²⁺ accumulation was studied. Electron microscopy indicated that at pH 9, Ni²⁺ accumulated on the cell surface as needle and hexagon-like precipitates, whose crystalline structure was confirmed by electron diffraction analysis and corresponded to two different orientations of the nickel hydroxide crystals. Crystals on cells showed marked anisotropy by X-ray powder diffraction, which differentiated them from crystals observed in nickel solution at pH 10 and 11 and from commercial nickel hydroxide. Nickel biosorption by Pseudomonas fluorescens 4F39 was a microprecipitation consequence of an ion exchange. Journal of Industrial Microbiology & Biotechnology (2000) 24, 146–151. Received 22 June 1999/ Accepted in revised form 04 December 1999     

The effect of cooling seawater effluents of a power plant on growth rate of cultured Gracilaria conferta (Rhodophyta)

The effect of cooling seawater effluents of a power plant on the growth rate of Gracilaria conferta in tanks has been studied, as a possible solution for the decrease in the winter growth rate in ambient seawater tanks. The Gracilaria cultures did not survive more than 2–8 weeks in the power plant effluents during the one-year-long repeated experiments. The major reason was the high accumulation of copper, iron, lead and chromium from the power plant effluents as compared to concentrations in Gracilaria cultured in ambient seawater. The survival increased and the copper accumulation decreased significantly when the effluents were passed through an Ulva biofilter.     

Seminal plasma trace metal levels in industrial workers

This study compares the seminal plasma trace metal levels of hospital workers with groups of industrial workers in a petroleum refinery, smelter, and chemical plant. The metals measured were the essential metals (copper, zinc, nickel, cobalt, and manganese) and the toxic metals (lead, cadmium, and aluminum). The group mean±SE metal level for each group (50 subjects per group) was calculated, and the statistical significance of the group mean differences of the industrial groups with the hospital group (control) was determined by the Student’s t-test. The differences observed in the smelter group were increased copper and zinc (p≤0.001) and decreased nickel, cobalt, and manganese (p≤0.001,≤0.01). The refinery group differences were increased copper, zinc, and nickel (p≤0.001) but decreased cobalt and manganese (p≤0.001). The chemical group differences were increased zinc (p≤0.001) and decreased cobalt (p≤0.001). The seminal plasma levels of the toxic metals lead and aluminum were increased in each of the industrial groups (p≤0.001). Concurrent differences were (1) decreased accumulation of nickel, cobalt, and manganese in the smelter group, (2) decreased cobalt and managanese in the refinery group, and (3) only decreased cobalt in the chemical group.     

Heterogeneity of epidermal cells in relation to nickel accumulation in hyperaccumulator plants belonging to the genus <Emphasis Type="Italic">Alyssum</Emphasis> L.

Epidermal cells of some plants are able to accumulate high levels of heavy metals (Zn, Ni, Cd). We studied this ability in plants in the genus Alyssum L. distinguished by tolerance to nickel (Ni). It was established that the predominant Ni accumulation occurred in epidermis, whereas in other tissues lower concentrations of the metal were revealed. It was also found that epidermal cells were characterized by heterogeneity in relation to Ni accumulation. The highest metal amount was accumulated in ordinary epidermal cells and in trichomes. Species-specific features of Ni distribution in leaf tissues in Alyssum spp. were shown. The reasons for the heterogeneity of epidermal cells in relation to Ni accumulation were discussed. We have attempted to resolve the contradictions encountered in the literature concerning the distribution and accumulation of Ni in the leaf tissues of plants belonging to the genus Alyssum L.     

Effect of long-term cultivation on resveratrol accumulation in a high-producing cell culture of Vitis amurensis

Resveratrol is a polyphenol, present in grapes, peanuts, and other plant sources, with a wide range of valuable biological activities. We established a Vitis amurensis cell culture accumulating high levels of resveratrol by introducing the rolB gene of Agrobacterium rhizogenes in the V. amurensis genome, and studied the stability of resveratrol accumulation during 27 months of continuous subculturing. This study demonstrates a decline in the high level of resveratrol production by the rolB transgenic cell line during its long-term cultivation. Elicitation of the rolB transgenic calli with methyl jasmonate and salicylic acid, which are known to stimulate the production of plant secondary metabolites, resulted in a recovery of resveratrol accumulation in the rolB transgenic cell culture, while the empty vector-transformed culture with trace starting content of resveratrol exhibited low inducibility to the treatment.     

Plant Screening for Chromium Phytoremediation

Thirty-six plant species of different agronomic importance, size, dry matter production, and tolerance to heavy metals were evaluated for Cr(III) and Cr(VI) uptake and accumulation as influenced by rate, form, source, and chelate application to a Cr-contaminated soil. There was a significant difference in the degree of tolerance, uptake, and accumulation of Cr among plant species. Sunflower (Helianthus annuus) was the least tolerant to Cr, and Bermudagrass (Cynodon dactylon) and switchgrass (Panicum virgatum) were the most tolerant. Indian mustard (Brassica juncea, cv 426308) and sunflower accumulated more Cr than other agricultural plant species. There was no inhibition of growth and little Cr accumulation in the presence of Cr(III) in soil, but most of the plant species that were treated with Cr(VI) hyperaccumulated Cr and died. EDTA chelate added to soil enhanced Cr(III) accumulation in some plants. The phytoremediation potential of the plant species tested was limited because Cr was accumulated in the plant roots and a high concentration in the shoots was toxic to plants. The difference in behavior between Cr(III) and Cr(VI) and their importance in soil and environment contamination should be the basis for remediation strategies.