Contrasting patterns of genetic divergence in two sympatric pseudo-metallophytes: Rumex acetosa L. and Commelina communis L.

BACKGROUND: Patterns of genetic divergence between populations of facultative metallophytes have been investigated extensively. However, most previous investigations have focused on a single plant species making it unclear if genetic divergence shows common patterns or, conversely, is species-specific. The herbs Rumex acetosa L. and Commelina communis L. are two pseudo-metallophytes thriving in both normal and cupriferous soils along the middle and lower reaches of the Yangtze River in China. Their non-metallicolous and metallicolous populations are often sympatric thus providing an ideal opportunity for comparative estimation of genetic structures and divergence under the selective pressure derived from copper toxicity. RESULTS: In the present study, patterns of genetic divergence of R. acetosa and C. communis , including metal tolerance, genetic structure and genetic relationships between populations, were investigated and compared using hydroponic experiments, AFLP, ISSR and chloroplast genetic markers. Our results show a significant reduction in genetic diversity in metallicolous populations of C. communis but not in R. acetosa . Moreover, genetic differentiation is less in R. acetosa than in C. communis , the latter species also shows a clustering of its metallicolous populations. CONCLUSIONS: We propose that the genetic divergences apparent in R. acetosa and C. communis , and the contrasting responses of the two species to copper contamination, might be attributed to the differences in their intrinsic physiological and ecological properties. No simple and generalised conclusions on genetic divergence in pseudo-metallophytes can thus be drawn.

Effectiveness of amendments on re-acidification and heavy metal immobilization in an extremely acidic mine soil.

Previous studies have shown that the application of soil amendments is efficient in reducing acidity and heavy metal bioavailability in mine soils. However, it remains a challenge for environmentalists to predict accurately and control economically the re-acidification in re-vegetated mine soils. In this study, net acid generation (NAG) test and bioassay technique were employed to assess the effectiveness of the amendments [including lime, N-P-K (nitrogen, phosphorous and potassium) fertilizer, phosphate and river sediment] on re-acidification and heavy metal immobilization in an extremely acid (pH < 3) mine soil. Our results suggested that NAG test was a rapid and accurate approach to assess the effectiveness of the amendments on re-acidification potential of the mine soil. Interestingly, it was found that phosphate and river sediment played quite specific roles in preventing the re-acidification in the mine soil. In addition, the results also indicated that the addition of 25 t ha(-1) lime combined with river sediment (30%) might be an economical method to successfully control the acidification and re-acidification in the extremely acid mine soil, allowing the re-establishment of the plants. Collectively, our results implied that the combined use of NAG test and bioassay assessment was effective in evaluating a reclamation strategy for extremely acidic mine soils.

Cadmium tolerance and accumulation in cultivars of a high-biomass tropical tree (Averrhoa carambola) and its potential for phytoextraction.

Averrhoa carambola is a high-biomass tropical tree that has been identified as a Cd accumulator. In the present study, field survey, pot, and hydroponic experiments were conducted to investigate the variation of Cd tolerance and accumulation in cultivars of A. carambola as well as its potential for phytoextraction. In the field survey, it was found that concentrations of Cd in aerial tissues of A. carambola varied greatly among sites and cultivars. The Cd bioconcentration factors (BCFs) and Cd removals by the field-grown A. carambola differed significantly among sites but not among cultivars. Nonetheless, all four carambola cultivars investigated were able to accumulate considerably high concentrations of Cd in their shoots, which indicated that the 4-yr-old carambola stands could remove 0.3 to 51.8% of the total Cd content in the top 20-cm soil layer. When cultured in Cd-spiked soils, the carambola cultivar Hua-Di always showed higher Cd tolerance than the other cultivars; however, this tendency was not confirmed by hydroponic experiment. The Cd BCFs of cultivar Thailand grown in soils with 6 and 12 mg Cd kg(-1) were highest among cultivars, whereas this trend was reversed at 120 mg Cd kg(-1) treatment. Nevertheless, the pot- and hydroponics-grown carambola cultivars generally showed higher capacities to tolerate and accumulate Cd, compared with the control species. The present results indicate that a strong ability to tolerate and accumulate Cd seems to be a trait at the species level in A. carambola, although some degree of variances in both Cd tolerance and accumulation exists among cultivars.

Zinc, nickel and cadmium in carambolas marketed in Guangzhou and Hong Kong, China: implication for human health.

Carambola (Averrhoa carambola L.) is a popular juicy fruit throughout the tropical and subtropical world. This study was designed to quantify the levels of zinc (Zn), nickel (Ni) and cadmium (Cd) in carambolas marketed in southern China, and further to evaluate the potential health risk of human consumption of carambola. Zinc concentrations, ranging from 1.471 to 2.875 mg/kg (on fresh weight basis), were below the maximum permissible concentration for Zn in fruit of China (5 mg/kg). However, Ni concentrations (0.134-0.676 mg/kg) were considerably higher than the related recommendation values. Furthermore, Cd concentrations in 51% of the carambolas purchased from Guangzhou exceeded the maximum permissible concentration for Cd in fruit of China (0.03 mg/kg). Our results implicated that the consumption of 0.385 kg carambola contaminated by Cd per day would cause the tolerable daily intake (TDI) of Cd by the consumer to be exceeded. In addition, the remarkably high Ni concentrations in carambolas should also be of concern. The status of heavy metal contamination of carambola products marketed in the other regions and their implications for human health should be identified urgently by in-depth studies.

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.

Uptake and accumulation of arsenic by 11 Pteris taxa from southern China.

A field survey was conducted at a deserted arsenic (As) mine in Guangxi Province, China to explore new potential As hyperaccumulators. In addition, young plants of 11 Pteris taxa were grown in glasshouse conditions for 12 weeks on As-amended soils with 0, 50 and 200 mg As kg(-1). Results of the field survey showed that the fern Pteris fauriei accumulated over 1000 mg As kg(-1) in its fronds. Of the 11 Pteris taxa, Pteris aspericaulis, Pteris cretica var. nervosa, P. fauriei, Pteris multifida, P. multifida f. serrulata, and Pteris oshimensis were all found to hyperaccumulate As in addition to P. cretica 'Albo-Lineata' and Pteris vittata (already reported as As hyperaccumulators). However, Pteris ensiformis, Pteris semipinnata and Pteris setuloso-costulata showed no evidence of As hyperaccumulation. Results also revealed a constitutive property of As hyperaccumulation in different populations of P. cretica var. nervosa, P. multifida, P. oshimensis and P. vittata.

Cut-off net acid generation pH in predicting acid-forming potential in mine spoils.

Acidification of mine wastes can lead to a series of environmental problems, such as acid drainage, heavy metal mobilization, and ecosystem degradation. Prediction of acid-forming potential is one of the key steps in management of sulfide-bearing mine wastes. In this paper, the acid-forming potential of 180 mine waste samples collected from 17 mine sites in China were studied using a net acid generation (NAG) method. The samples contained different contents of total sulfur (ranging from 0.6 to 200 g kg(-1)), pyritic sulfur (ranging from 0 to 100 g kg(-1)), and acid neutralization capacity (ANC, ranging from -41 to 274 kg H2SO4 t(-1)). Samples with high acid-forming potential are generally due to their high sulfur content or low acid neutralization capacity. After the samples were oxidized by H2O2, the amounts of acid generation and the final NAG pH were measured. Results indicated that the final NAG pH gave a well-defined demarcation between acid-forming and non-acid-forming materials. Samples with final NAG pH >or= 5 could be classified as non-acid-forming materials, while those with NAG pH 2.5, but < 5, had low risk of being acid-forming. The confirmation of cut-off NAG pH will be used as a rapid and cost-effective operational monitoring tool for the in-pit prediction of acid-forming potential of mine wastes and classification of waste types.

Cadmium contamination in orchard soils and fruit trees and its potential health risk in Guangzhou, China.

This study examines cadmium (Cd) contamination in orchard soils and fruit trees in Guangzhou, China, and assesses its potential health risk. Soils and tissues samples of three species of fruit trees were collected from three orchards. The average soil Cd concentration was 1.27, 1.84 and 0.68 mg/kg in orchards I, II, and III, respectively. The carambola (Averrhoa carambola) accumulated exceptionally high concentrations of Cd (7.57, 10.84, 9.01 and 2.15 mg/kg dw in root, twig, leaf and fruit, respectively), being 6.0-24 times and 4.0-10 times the corresponding tissue Cd in the longan (Dimocarpus longan) and wampee (Clausena lansium), respectively. Furthermore, all Cd concentrations (0.04-0.25 mg Cd/kg fw) of the fruits exceeded the tolerance limit of cadmium in foods of PR China (0.03 mg/kg fw). Our results indicate that the carambola tree has high Cd accumulation capacity and might be a Cd accumulator; and its fruit, among the three species of fruits studied, also poses the highest potential health risk to local residents.

Long-term efficiency and stability of wetlands for treating wastewater of a lead/zinc mine and the concurrent ecosystem development.

A constructed wetland system in Guangdong Province, South of China has been used for treating Pb/Zn mine discharge since 1985. The performance in the purification of the mine discharge and the concurrent ecosystem development within the system during the period of 1985-2000 has been studied. The untreated wastewater contained rather high concentrations of cadmium (Cd) (0.05 mg L(-1)), lead (Pb) (11.5 mg L(-1)), and zinc (Zn) (14.5 mg L(-1)), which greatly exceed the upper limits for industrial wastewater discharge in China. The constructed wetland system effectively removed Cd by 94.00%, Pb by 99.04%, Zn by 97.30%, and total suspended solids (TSS) by 98.95% from the mine discharge over a long period (over 16 years) leading to significant improvement in water quality; it was also found that there were no significantly annual or monthly variations in pH values, As, Cd, Hg, Pb, and Zn concentrations in water collected from the outlet of the wetland. Moreover, diversity and abundance of living organisms, including protozoan, higher plants, terrestrial animals, and birds, increased gradually. The 16-year monitoring results showed a reciprocal relationship, at a certain extent, between restoration of the wetland ecosystem, in other words, the maturity of the wetland, and the long-term efficiency and stability on purifying heavy metal-contaminated wastewater.

Lead in paddy soils and rice plants and its potential health risk around Lechang lead/zinc mine, Guangdong, China.

As part of a project on phytoextraction of lead (Pb) in paddy soils around a lead/zinc (Pb/Zn) Mine in Lechang of Guangdong Province, South China, the concentration distribution of Pb in paddy soil-rice system was investigated, and its potential health risk to animal/human was evaluated. Total and diethylenetetraminepentaacetic acid (DTPA)-extractable Pb in soils averaged 1486 and 268 microg/g, respectively. According to sequential extraction procedure, soil Pb occurred primarily in the residual, carbonate and exchangeable fractions (30.2%, 26.7% and 19.1%, respectively). Lead extracted by the gastric juice simulation test (GJST) was 1068 microg/g and accounted for 75.4% of the total concentration. Mean Pb concentrations of 419 microg/g in rice root, 69.1 microg/g in whole straw, 51.0 microg/g in part straw (without two leaves near above the root), 44.9 microg/g in stalk, 21.9 microg/g in hull, 13.2 microg/g in grain with hull and 4.67microg/g in grain without hull (namely, unpolished rice) were found. Lead concentrations in both soil and rice plant were far above the corresponding tolerable levels. Lead daily intakes by local residents were 2.6 mg for adults and 1.2 mg for children, which were much higher than the allowable level. Thus, Pb in this area might pose a potential health risk to the local population.

Acidification of lead/zinc mine tailings and its effect on heavy metal mobility.

The acid-forming potential of lead/zinc (Pb/Zn) mine tailings at Lechang City of Guangdong Province was studied using both net acid generation (NAG) and acid-base accounting (ABA) methods. The pyritic and total sulfur contents of the tailings were 12.6% and 18.7%, respectively. The mean acid neutralization capacity (ANC) was 63.5 kg H2SO4/t while three oxidized tailings samples had an ANC less than zero. The NAG and net acid production potential (NAPP) values were 220 and 326 kg H2SO4/t, and both the NAG and NAPP results indicated that the tailings had a high acid-forming potential. NAG was more accurate than NAPP in predicting acid-forming potential of the tailings due to uncompleted oxidization of pyritic sulfur. Analysis of samples from two profile tests indicated that acidification mainly occurred at the surface (0-20 cm) and had little effects at deep layer of the tailings. Total concentrations of Pb, Zn, Cu, and Cd were increased greatly with depth at the acidified tailings profile, while heavy metal concentrations at different depths of nonacidified tailings profile were similar. The results indicated that depletion of heavy metals at the acidified surface was due to acidification. The diethylenetetramine pentaacetic acid (DTPA)-extractable Pb, Zn, Cu, and Cd concentrations of acidified tailings surface (0-20 cm) were significantly higher than those of nonacidified tailings, which further revealed that acidification enhanced the mobility of heavy metals in the tailings.

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.

Fluoride and aluminium concentrations of tea plants and tea products from Sichuan Province, PR China.

Some Tibetans in Sichuan Province in southwestern China have been suffering from fluorosis, due to drinking and eating tea with high fluoride (F) and aluminium (Al) contents. Tea plants, soils of tea plantations and tea products from Yaan, Gaoxian and Yibin Cities in Sichuan Province were investigated to evaluate the factors affecting F and Al contents in tea products. The F and Al concentrations of four commercial brands of brick teas were significantly higher than those of 11 brands of green teas. Chemical analysis indicated that total and available F and Al concentrations in tea plantation soils in Yaan and Gaoxian were within the normal range compared with acid soils in South China and tea soils in Fujian Province. Edaphic conditions did not contribute to the high F and Al concentrations in brick tea. Analysis of raw materials of brick tea indicated that old leaves were the major contributors to the high F and Al contents contained in brick tea. There were also great variations among different tea varieties in accumulating F and Al, and concentrations of F and Al in tea variety of Qianmei 303 were about 2-3-fold higher than the other three varieties. Selection of appropriate varieties would be important to lower F and Al contents in tea products.

Growth and metal accumulation in vetiver and two Sesbania species on lead/zinc mine tailings.

The lead (Pb)/zinc (Zn) tailings contained high concentrations of heavy metals (total Pb, Zn, Cu and Cd concentrations 4164, 4377, 35 and 32 mg kg(-1), respectively), and low contents of major nutrient elements (N, P, and K) and organic matter. A field trial was conducted to compare growth performance, metal accumulation of Vetiver (Vetiveria zizanioides) and two legume species (Sesbania rostrata and Sesbania sesban) grown on the tailings amended with domestic refuse and/or fertilizer. It was revealed that domestic refuse alone and the combination of domestic refuse and artificial fertilizer significantly improved the survival rates and growth of V. zizanioides and two Sesbania species, especially the combination. However, artificial fertilizer alone did not improve both the survival rate and growth performance of the plants grown on tailings. Roots of these species accumulated similar levels of heavy metals, but the shoots of two Sesbania species accumulated higher (3-4 folds) concentrations of Pb, Zn, Cu and Cd than shoots of V. zizanioides. Most of the heavy metals in V. zizanioides were accumulated in roots, and the translocation of metals from roots to shoots was restricted. Intercropping of V. zizanioides and S. rostrata did not show any beneficial effect on individual plant species, in terms of height, biomass, survival rate, and metal accumulation, possibly due to the rather short experimental period of 5 months.

Evaluation of major constraints to revegetation of lead/zinc mine tailings using bioassay techniques.

The residues from the extraction of lead/zinc (Pb/Zn) ores of most Pb/Zn mines are permanently stored in tailings ponds, which require revegetation to reduce their environmental impact. This can only be done if the main constraints on plant establishment are evaluated. This can readily be done by field and greenhouse studies. To test this, the properties of different tailings from Lechang Pb/Zn mine located at the north of Guangdong Province in southern China have been studied. Physical and chemical properties including concentrations of metals (Pb, Zn, Cd and Cu) in the tailings and soils collected from different sites have been measured. The results showed that tailings contain low nitrogen (0.016-0.075%), low-organic matter (0.58-1.78%), high salt (3.55-13.85 dS/m), and high total and diethylene-tetramine-pentaacetic acid (DTPA)-extractable metal concentrations (total: 1,019-1,642 microg g(-1) Pb, 3,078-6,773 microg g(-1) Zn, 8-23 microg g(-1) Cd, and 85-192 microg g(-1) Cu; DTPA-extractable: 59-178 microig g(-1) Pb, 21-200 microg g(-1) Zn, 0.30-1.5 mcirog g(-1) Cd, and 4.3-12 microg g(-1) Cu). Aqueous extracts of tailings/soils (10%, 20% and 30%, w/v) from different sites were prepared for testing their effects on seed germination and root elongation of a vegetable crop Brassica chinensis and a grass species Cynodon dactylon. It was found that root elongation provided a better evaluation of toxicity than seed germination. The ranking of toxicity using root elongation was: high-sulfur tailings > tailing dam > sparsely vegetated tailings > densely vegetated tailings > mountain soil for both plants. This order was consistent with DTPA-extractable Pb contents in the tailings and soils. B. chinensis seedlings were then grown in the mixtures of different proportions of tailings and farm soil for 4 weeks, and the results (dry weights of seedlings) were in line with the root elongation test. All these demonstrated that heavy metal toxicity, especially available Pb, low content of nutrient, and poor physical structure were major constraints on plant establishment and colonization on the Pb/Zn mine tailings.

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.

Phytoextraction of Cd-contaminated soil by carambola (Averrhoa carambola) in field trials.

Use of metal-accumulating woody species to extract metals from heavy metal contaminated soil has received more attention. While considerable studies have focused on the phytoextraction potential of willow (Salix spp.) and poplar (Populus spp.), similar information is rare for other woody species. Carambola (Averrhoa carambola) is a high-biomass tree and has been identified as a new Cd-accumulating species. The present study aimed to evaluate the Cd phytoextraction potential of carambola under field condition. After growing in a slightly Cd-contaminated site for about 170 d, the carambola stand initiated by seed-seedling with high planting density (encoded with "HD-1yr") attained a high shoot biomass yield of 18.6 t ha(-1) and extracted 213 g Cdha(-1), resulting in a 1.6-fold higher Cd removal efficiency than that of a contrasting stand established by grafted-seedling with low planting density (5.3% vs. 2%). That is, "HD-1yr" would remove 50% of the total soil Cd with 13yr, assuming that the Cd removal efficiency would not change over time. Further, one crop of "HD-1yr" significantly decreased (63-69%) the Cd uptake by subsequent vegetables. Among the four carambola stands established using grafted-seedling, the 2-yr-old stand exhibited the highest annual Cd removal efficiency (3.7%), which was yet lower than that of "HD-1yr". These results suggested that phytoextraction of Cd by carambola (especially for "HD-1yr" stand) presented a feasible option to clean up agricultural soils slightly contaminated by Cd.

Metal contamination in the sediment, pondweed, and snails of a stream receiving effluent from a lead/zinc mine in southern China.

This study assessed the potential of Potamogeton crispus and Pomacea canaliculata as biomonitors of sedimentary metal contamination. The results indicate P. crispus possesses several attributes of a biomonitor and its tissue concentrations of Cd, Pb and Zn may reflect the levels of sedimentary contamination by these metals. Although P. canaliculata can accumulate metals to high levels and serve as an indicator of metal contamination, its tissue metal concentrations did not correlate with those of the sediments or macrophytes.

Arsenic uptake and accumulation in fern species growing at arsenic-contaminated sites of southern China: field surveys.

Aiming at searching for new arsenic (As) hyperaccumulators, field surveys were conducted at 12 As-contaminated sites located in Guangxi and Guangdong Provinces of southern China. Samples of 24 fern species belonging to 16 genera and 11 families as well as their associated soils were collected and As concentrations in plant and soil samples were determined by inductively coupled plasma-atomic emission spectrometry (ICP-AES). The results show that among 24 fern species, Pteris multifida and P. oshimensis can (hyper)accumulate As in their fronds with high concentrations in addition to P. vittata and P. cretica var. nervosa, which have been previously identified as As hyperaccumulators. Total As concentrations in soils associated with P. multifida and P. oshimensis varied from 1262 to 47,235 mg kg(-1), but the DTPA-extractable As concentrations were relatively low, with a maximum of 65 mg kg(-1). Forty-four of 49 samples of P. multifida collected from five sites and 3 of 13 samples of P. oshimensis collected from one site accumulated over 1000 mg As kg(-1) in their fronds and As concentrations in the fronds were higher than those in the petioles and rhizoids. Although As concentrations in the fronds of P. oshimensis (789 mg kg(-1) averaged, range 301-2142 mg kg(-1)) were comparatively lower than those of P. multifida (1977 mg kg9-1), 624-4056 mg kg(-1)), its high aboveground biomass makes it more suitable for phytoremediating As-contaminated soils. Among all the species in Pteris genus studied, Pteris semipinnata accumulated only very low As concentration in its fronds (8 mg kg(-1), 1-18 mg kg(-1)). Further research is needed to study the differences in As uptake and accumulation among fern species in the same or other genera.