Screening the wetland plant species Alisma plantago-aquatica, Carex rostrata and Phalaris arundinacea for innate tolerance to zinc and comparison with Eriophorum angustifolium and Festuca rubra Merlin

Several wetland plant species appear to have constitutive metal tolerance. In previous studies, populations from contaminated and non-contaminated sites of the wetland plants Typha latifolia, Phragmites australis, Glyceria fluitans and Eriophorum angustifolium were found to be tolerant to high concentrations of metals. This study screened three other species of wetland plants: Alisma plantago-aquatica, Carex rostrata and Phalaris arundinacea for innate tolerance to zinc. The degree of tolerance was compared to known zinc-tolerant E. angustifolium and Festuca rubra Merlin. It was found that A. plantago-aquatica and P. arundinacea did not posses innate tolerance to zinc, but that C. rostrata was able to tolerate elevated levels of zinc, at levels comparable to those tolerated by E. angustifolium and F. rubra Merlin. The findings support the theory that some wetland angiosperm species tend to be tolerant to exposure to high levels of metals, regardless of their origin.     

Accumulation of lead, zinc, copper and cadmium by 12 wetland plant species thriving in metal-contaminated sites in China

The concentrations of lead, zinc, copper and cadmium accumulated by 12 emergent-rooted wetland plant species including different populations of Leersia hexandra, Juncus effusus and Equisetum ramosisti were investigated in field conditions of China. The results showed that metal accumulation by wetland plants differed among species, populations and tissues. Populations grown in substrata with elevated metals contained significantly higher metals in plants. Metals accumulated by wetland plants were mostly distributed in root tissues, suggesting that an exclusion strategy for metal tolerance widely exists in them. That some species/populations could accumulate relatively high metal concentrations (far above the toxic concentration to plants) in their shoots indicates that internal detoxification metal tolerance mechanism(s) are also included. The factors affecting metal accumulation by wetland plants include metal concentrations, pH, and nutrient status in substrata. Mostly concentrations of Pb and Cu in both aboveground and underground tissues of the plants were significantly positively related to their total and/or DTPA-extractable fractions in substrata while negatively to soil N and P, respectively. The potential use of these wetland plants in phytoremediation is also discussed.     

利用芦竹修复重金属污染湿地的研究

研究了芦竹(Arundodonaxlinn)对Cu2+、Ni2+、Cr6+重金属污染湿地的响应和对该污染湿地的影响。实验结果表明,芦竹对这3种重金属离子有一定的耐受性,并有不同程度的吸收,Cu2+、Ni2+和Cr6+去除率分别为63.8%,42.3%和34.4%。芦竹在100mg/kg浓度的Cu2+、Ni2+污染湿地中生长正常,在低浓度(55mg/kg)Cr6+污染中能存活,但生长速度较慢。在100mg/kg浓度Cr6+污染湿地中,出现急性中毒现象,半月后致死。在重金属污染环境中,芦竹普遍出现失绿现象,但除高浓度Cr6+(100mg/kg)以外,都能正常存活,表现出较强的适应性。     

Effects of chromium and nickel on germination and growth in tolerant and non-tolerant populations of Echinochloa colona (L.) Link

The tolerance of populations of a grass, Echinochloa colona, growing abundantly on chromite minewaste dumps, was tested in two separate experiments. Seed-based experiments indicate that the populations growing naturally on uncontaminated sites, germinated better in nutrient solutions without metal than those collected from minewaste dumps. Metal tolerance indices were greater in the plant populations derived from metal contaminated sites and better growth of these plants was noted on mine spoil soil-mix in the ratio of 1:1; the percentage of seed germination and the rate of seedling growth, however, declined in a soil compost containing 25% mine spoil and 75% uncontaminated (control) soil. Populations of Echinochloa colona occurring naturally on chromite mine spoils, therefore, appear to have developed metal tolerance. It is maintained by a balanced and stable genetic system built up and adjusted by natural selection. Such material is very suitable to be used in restoration work designed to produce an effective vegetation cover to improve the derelict land and to reduce erosion. This finding might be useful in revegetation programmes on metalliferous minewastes.     

Specificity of metal tolerance and use of excluder metallophytes for the phytostabilization of metal polluted soils: the case of Silene paradoxa L.

This work was planned for providing useful information about the use of excluder metallophytes for phytostabilization of soils contaminated also with elements scarcely represented in the metalliferous environment of origin. To this aim, we investigated tolerance and accumulation of several different elements in a metallicolous and a nonmetallicolous population of Silene paradoxa through a hydroponic experiment. S. paradoxa metallicolous population showed increased tolerance not only to all the metals highly represented in the environment of origin but also to some of those scarcely present. Therefore, our results deposed in favor of the occurrence of the co-tolerance phenomenon in S. paradoxa for some elements. Metal accumulation was higher in the roots than in the shoots and lower in the metallicolous population than in the nonmetallicolous one, thus showing tolerance mechanisms to be based largely on metal exclusion. Anyway, the relative contribution of avoidance and of internal tolerance to metal tolerance was shown to be element-dependent. Present data revealed that metallicolous plants can effectively posses metal co-tolerances, which deserve to be investigated; as such, plants can actually represent a precious and exploitable tool also for the phytostabilization of soils contaminated with elements underrepresented in the environment of their origin.     

Colonisation of a Zn, Cd and Pb hyperaccumulator Thlaspi praecox Wulfen with indigenous arbuscular mycorrhizal fungal mixture induces changes in heavy metal and nutrient uptake

Plants of the Zn, Cd and Pb hyperaccumulator Thlaspi praecox Wulfen (Brassicaceae) inoculated or not with indigenous arbuscular mycorrhizal (AM) fungal mixture were grown in a highly Cd, Zn and Pb contaminated substrate in order to evaluate the functionality of symbiosis and assess the possible impact of AM colonisation on heavy metal uptake and tolerance. The results suggest AM development in the metal hyperaccumulating T. praecox is favoured at elevated nutrient demands, e.g. during the reproductive period. AM colonisation parameters positively correlated with total soil Cd and Pb. Colonised plants showed significantly improved nutrient and a decreased Cd and Zn uptake as revealed by TRXRF, thus confirming the functionality of the symbiosis. Reduced heavy metal uptake, especially at higher soil metal contents, indicates a changed metal tolerance strategy in colonised T. praecox plants. This is to our knowledge the first report on AM colonisation of the Zn, Cd and Pb hyperaccumulator T. praecox in a greenhouse experiment.     

Constitutional tolerance to heavy metals of a fiber crop, ramie (Boehmeria nivea), and its potential usage

It is observed that ramie (Boehmeria nivea), an economic fiber crop, can establish and colonize metal-contaminated sites in China. Metal tolerance and accumulation by ramie originating from 13 metal-contaminated and 4 “clean” sites in China were compared under field and hydroponic conditions. All selected populations and germplasms displayed good growth performance under diverse metal-contaminated habitats; while growth responses, metal accumulation and tolerance were similar among the 8 populations and 2 germplasms when exposed to solutions containing elevated As, Cd, Pb, or Zn in the laboratory. These revealed that ramie possesses a certain degree of constitutional metal tolerance. To our knowledge, this is the first report of constitutional metal tolerance possessed by a fiber crop. Ramie can be considered as a good candidate for both fiber production and phytoremediation of sites contaminated by multi-metals, as it accumulates relative low metal concentrations, but possesses both high biomass and high economic value.     

The arbuscular mycorrhizal fungus Glomus mosseae induces growth and metal accumulation changes in Cannabis sativa L

The effect of arbuscular mycorrhiza on heavy metal uptake and translocation was investigated in Cannabis sativa. Hemp was grown in the presence and absence of 100 microg g-1 Cd and Ni and 300 microg g-1 Cr(VI), and inoculated or not with the arbuscular mycorrhizal fungus Glomus mosseae. In our experimental condition, hemp growth was reduced in inoculated plants and the reduction was related to the degree of mycorrhization. The percentage of mycorrhizal colonisation was 42% and 9% in plants grown in non-contaminated and contaminated soil, suggesting a significant negative effect of high metal concentrations on plant infection by G. mosseae. Soil pH, metal bioavailability and plant metal uptake were not influenced by mycorrhization. The organ metal concentrations were not statistically different between inoculated and non-inoculated plants, apart from Ni which concentration was significantly higher in stem and leaf of inoculated plants grown in contaminated soil. The distribution of absorbed metals inside plant was related to the soil heavy metal concentrations: in plant grown in non-contaminated soil the greater part of absorbed Cr and Ni was found in shoots and no significant difference was determined between inoculated and non-inoculated plants. On the contrary, plants grown in artificially contaminated soil accumulated most metal in root organ. In this soil, mycorrhization significantly enhanced the translocation of all the three metals from root to shoot. The possibility to increase metal accumulation in shoot is very interesting for phytoextraction purpose, since most high producing biomass plants, such as non-mycorrhized hemp, retain most heavy metals in roots, limiting their application.     

The role of bacteria in the heavy metals removal and growth of Sedum alfredii Hance in an aqueous medium

This study was the first attempt to examine the possible role of the naturally occurring rhizospheric bacteria in heavy metal removal by Sedum alfredii Hance, a terrestrial Zn/Cd hyperaccumuluator, from Zn, Cd, Cu and Pb contaminated water using antibiotic ampicillin. Moreover, the toxicity symptom in plants under heavy metal stress expressed as total chlorophyll, chlorophyll a and b content, growth inhibition, root length, and N, P contents were studied, and the possible relationship among them were also discussed. These results indicate that rhizospheric bacteria may play an important role in the uptake of N and P by S. alfredii, and consequently result in the increase of Chlorophyll content in the leaves and plant biomass due to improved photosynthesis. At the same time, root length significantly decreased under the treatment with ampicillin, which suggested that rhizospheric bacteria appeared to protect the roots against heavy metal toxicity. The Pb, Zn, Cu and Cd concentrations in the roots, stems and leaves of S. alfredii were much higher than those exposed to ampicillin. Accordingly, metal concentrations in the contaminated water without ampicillin treatment were lower than those treated with ampicillin. These results suggest that the rhizospheric bacteria may be useful in plant tolerance to heavy metal toxicity, and also accelerate the metal removal from contaminated water.     

Chemical methods and phytoremediation of soil contaminated with heavy metals

The effects of chemical amendments (calcium carbonate (CC), steel sludge (SS) and furnace slag (FS)) on the growth and uptake of cadmium (Cd) by wetland rice, Chinese cabbage and wheat grown in a red soil contaminated with Cd were investigated using a pot experiment. The phytoremediation of heavy metal contaminated soil with vetiver grass was also studied in a field plot experiment. Results showed that treatments with CC, SS and FS decreased Cd uptake by wetland rice, Chinese cabbage and wheat by 23-95% compared with the unamended control. Among the three amendments, FS was the most efficient at suppressing Cd uptake by the plants, probably due to its higher content of available silicon (Si). The concentrations of zinc (Zn), lead (Pb) and Cd in the shoots of vetiver grass were 42-67%, 500-1200% and 120-260% higher in contaminated plots than in control, respectively. Cadmium accumulation by vetiver shoots was 218 g Cd/ha at a soil Cd concentration of 0.33 mg Cd/kg. It is suggested that heavy metal-contaminated soil could be remediated with a combination of chemical treatments and plants.     

Lead,zinc and copper accumulation and tolerance in populations of Paspalum distichum and Cynodon dactylon

Both Fankou and Lechang lead/zinc (Pb/Zn) mine tailings located at Guangdong Province contained high levels of total and DTPA-extractable Pb, Zn and Cu. Paspalum distichum and Cynodon dactylon were dominant species colonized naturally on the tailings. Lead, zinc and copper accumulation and tolerance of different populations of the two grasses growing on the tailings were investigated. Tillers of these populations including those from an uncontaminated area were subjected to the following concentrations: 5, 10, 20, 30 and 40 mg l(-1) Pb, 2.5, 5, 10, 20 and 30 mg (l-1) Zn, or 0.25, 0.50, 1 and 2 mg l(-1) Cu for 14 days, respectively, then tolerance index (TI) and EC50 (the concentrations of metals in solutions which reduce 50% of normal root growth) were calculated. The results indicated that both Lechang and Fankou populations of the two grasses showed a greater tolerance to the three metals than those growing on the uncontaminated area, which suggested that co-tolerant ecotypes have evolved in the two grasses. P. distichum collected from Fankou tailings had the highest tolerance to Cu while Lechang population the highest tolerance to Pb and Zn among the tested populations, and tolerance levels in P. distichum were related to metal concentrations in the plants. P. distichum had a better growth performance than C. dactylon when both of them were grown on the tailings sites. Tolerant populations of these species would serve as potential candidates for re-vegetation of wastelands contaminated with Pb, Zn and Cu.     

Zinc and cadmium accumulation and tolerance in populations of Sedum alfredii

To investigate the variation of Zn and Cd accumulation and tolerance of Sedum alfredii (a newly reported Zn/Cd hyperaccumulator), field surveys and hydroponic experiments were conducted among three populations of this species: two originating from old Pb/Zn mines in Zhejiang (ZJ) and Hunan (HN) Provinces and one from a "clean" site in Guangdong (GD) Province, China. Under field conditions, up to 12,524 and 12,253 mg kg(-1) Zn, and 1400 and 97 mg kg(-1) Cd in shoots of ZJ and HN plants were recorded respectively. Under hydroponic conditions, ZJ and HN plants accumulated significantly higher Zn and Cd in their leaves and stems, and possessed significantly higher Zn and Cd tolerance than GD plants. Among the two contaminated populations, ZJ plants showed higher Cd tolerance and accumulation (in leaves) than HN plants. The present results indicate that significant differences in Zn and Cd accumulation and tolerance exist in populations of S. alfredii.     

The effect of hydrological regime on the metal bioavailability for the wetland plant species Salix cinerea

The hydrological conditions on a site constitute one of the many factors that may affect the availability of potentially toxic trace metals for uptake by plants. Bioavailability of Cd, Mn and Zn in a contaminated dredged sediment-derived soil under different hydrological regimes was determined by measuring metal uptake by the wetland plant species Salix cinerea, both in field circumstances and in a greenhouse experiment. Longer submersion periods in the field caused lower Cd concentrations in leaves and bark. The wetland hydrological regime in the greenhouse experiment resulted in normal Cd and Zn concentrations in the leaves, while the upland hydrological regime resulted in elevated Cd and Zn concentrations in the leaves. Field observations and the greenhouse experiment suggest that a hydrological regime that creates or sustains a wetland is a potential management option that reduces metal bioavailability to willows. This would constitute a safe management option of metal-polluted, willow-dominated wetlands provided that wetland conditions can be maintained throughout the full growing season.     

Phytoextraction potential of two Rumex acetosa L. accessions collected from metalliferous and non-metalliferous sites: effect of fertilization

Metal tolerance and phytoextraction potential of two common sorrel (Rumex acetosa L.) accessions, collected from a Pb/Zn contaminated site (CS, Lanestosa) and an uncontaminated site (UCS, Larrauri), were studied in fertilized and non-fertilized pots prepared by combining soil samples from both sites in different proportions (i.e., 0%, 33%, 66% and 100% of Lanestosa contaminated soil). The original metalliferous mine soil contained 20480, 4950 and 14 mg kg(-1) of Zn, Pb and Cd, respectively. The microcosm experiment was carried out for two months under greenhouse controlled conditions. It was found that fertilization increased mean plant biomass of both accessions as well as their tolerance. However, only the CS accession survived all treatments even though its biomass decreased proportionally according to the percentage of contaminated mine soil present in the pots. This metallicolous accession would be useful for the revegetation and phytostabilization of mine soils. Due to its high concentration and bioavailability in the contaminated soil, the highest values of metal phytoextracted corresponded to Zn. The CS accession was capable of efficiently phytoextracting metal from the 100% mine soil, indeed reaching very promising phytoextraction rates in the fertilized pots (6.8 mg plant(-1) month(-1)), similar to the ones obtained with hyperaccumulator plants. It was concluded that fertilization is certainly worth being considered for phytoextraction and revegetation with native plants from metalliferous soils.     

Cd-tolerant Suillus luteus: A fungal insurance for pines exposed to Cd

Soil metal pollution can trigger evolutionary adaptation in soil-borne organisms. An in vitro screening test showed cadmium adaptation in populations of Suillus luteus (L.: Fr.) Roussel, an ectomycorrhizal fungus of pine trees. Cadmium stress was subsequently investigated in Scots pine (Pinus sylvestris L.) seedlings inoculated with a Cd-tolerant S. luteus, isolated from a heavy metal contaminated site, and compared to plants inoculated with a Cd-sensitive isolate from a non-polluted area. A dose-response experiment with mycorrhizal pines showed better plant protection by a Cd-adapted fungus: more fungal biomass and a higher nutrient uptake at high Cd exposure. In addition, less Cd was transferred to aboveground plant parts. Because of the key role of the ectomycorrhizal symbiosis for tree fitness, the evolution of Cd tolerance in an ectomycorrhizal partner such as S. luteus can be of major importance for the establishment of pine forests on Cd-contaminated soils.     

Differences in phosphorus translocation contributes to differential arsenic tolerance between plants of Borreria verticillata (Rubiaceae) from mine and non-mine sites

We have identified new arsenic-tolerant plant species Borreria verticillata (Rubiaceae) that has mine and non-mine populations at a highly contaminated site (CS) and an uncontaminated site (UCS), respectively, in Brazil. Plants of B. verticillata from both sites were cultivated at different As and P concentrations. At low P concentration, CS plants showed reduced As uptake, higher P translocation to shoots, higher constitutive levels of phenolic compounds in roots, and higher tolerance to this metalloid. At the lowest P and highest As concentration, CS plants showed higher biomass. In addition, CS plants showed higher P uptake in the absence of As, suggesting that more efficient P translocation could contribute more to tolerance than decreased As uptake. In contrast, at low P concentration, UCS plants showed higher As content in shoot and root, increase in phenol levels in roots, reduction in dry biomass, and decrease of the effective efficiency of photochemical reactions and the electron transport rate. Under higher P concentrations, the decrease in As uptake was similar in both populations. The differences between the two populations with respect to As and P uptake suggest that altered kinetic properties or expression of P transporters contribute to higher As tolerance in B. verticillata from CS. As a ruderal and As-tolerant plant, B. verticillata could be successfully used for the revegetation of contaminated soils.     

Transgenic poplar trees expressing yeast cadmium factor 1 exhibit the characteristics necessary for the phytoremediation of mine tailing soil

Genetic engineering of plants for phytoremediation is thought to be possible based on results using model plants expressing genes involved in heavy metal resistance, which improve the plant’s tolerance of heavy metals and accumulation capacity. The next step of progress in this technology requires the genetic engineering of plants that produce large amounts of biomass and the testing of these transgenic plants in contaminated soils. Thus, we transformed a sterile line of poplar Populus alba X P. tremula var. glandulosa with a heavy metal resistance gene, ScYCF1 (yeast cadmium factor 1), which encodes a transporter that sequesters toxic metal(loid)s into the vacuoles of budding yeast, and tested these transgenic plants in soil taken from a closed mine site contaminated with multiple toxic metal(loid)s under greenhouse and field conditions. The YCF1-expressing transgenic poplar plants exhibited enhanced growth, reduced toxicity symptoms, and increased Cd content in the aerial tissue compared to the non-transgenic plants. Furthermore, the plants accumulated increased amounts of Cd, Zn, and Pb in the root, because they could establish an extensive root system in mine tailing soil. These results suggest that the generation of YCF1-expressing transgenic poplar represents the first step towards producing plants for phytoremediation. The YCF1-expressing poplar may be useful for phytostabilization and phytoattenuation, especially in highly contaminated regions, where wild-type plants cannot survive.     

Prosopis juliflora--a green solution to decontaminate heavy metal (Cu and Cd) contaminated soils

Soil and plant samples (root and shoot) of Prosopis juliflora were collected in the vicinity of metal based foundry units in Coimbatore and assessed for their heavy metal content (Cu and Cd) to ascertain the use of P. juliflora as a green solution to decontaminate soils contaminated with Cu and Cd. The results showed that Cu and Cd content was much higher in plant components compared to their extractable level in the soil. Furthermore, there exist a strong correlation between the distance of the sources of industrial units and accumulation of heavy metals in plants. Accumulation of Cd in roots is comparatively higher than that of shoots. However, in case of Cu no such clear trend is seen. Considering the accumulation efficiency and tolerance of P. juliflora to Cd and Cu, this plant can be explored further for the decontamination of metal polluted soils. On the other hand, in view of heavy metal accumulate the practice of providing foliage and pods as fodder for live stock should be avoided.     

Effect of mixed cadmium,copper, nickel and zinc at different pHs upon alfalfa growth and heavy metal uptake

Alfalfa plants were grown in soil-pots contaminated with a mixture of Cd(II), Cu(II), Ni(II), and Zn(II), (at 50 mg/kg each) at pHs of 4.5, 5.8, and 7.1. The plants were fertilized using a nutrient solution, which was adjusted appropriately to the same pH. Plants in the control treatment were grown in the absence of the heavy metals mixture. The growth of the control plants was the same at the three pHs studied and the heavy metal stressed plants also showed similar behavior at each pHs. There were statistically significant differences (P<0.05) between the shoot length of the control treatment plants and the length of plants grown in the presence of the heavy metal mixture. Under the effects of the heavy metal mixture, nickel was the most accumulated element in the shoot tissue, with 437, 333, and 308 ppm at pH 7.1, 5.8, and 4.5, respectively. Cadmium was found to be second in accumulated concentrations with 202 ppm, 124 ppm, and 132 ppm at pH 7.1, 5.8, and 4.5, respectively, while zinc was third, followed by copper. The maximum relative uptakes (element in plant/element in soil-water-solution) were found to be 26 times for nickel, 23 times for cadmium, 12 times for zinc. and 6 times for copper. We considered these relations as indicative of the ability of alfalfa plants to take up elements from a soil matrix contaminated with a mixture of cadmium, copper, nickel, and zinc.