Enhanced Cd extraction of oilseed rape (Brassica napus) by plant growth-promoting bacteria isolated from Cd hyperaccumulator Sedum alfredii Hance

Four plant growth-promoting bacteria (PGPB) were used as study materials, among them two heavy metal-tolerant rhizosphere strains SrN1 (Arthrobacter sp.) and SrN9 (Bacillus altitudinis) were isolated from rhizosphere soil, while two endophytic strains SaN1 (Bacillus megaterium) and SaMR12 (Sphingomonas) were identified from roots of the cadmium (Cd)/zinc (Zn) hyperaccumulator Sedum alfredii Hance. A pot experiment was carried out to investigate the effects of these PGPB on plant growth and Cd accumulation of oilseed rape (Brassica napus) plants grown on aged Cd-spiked soil. The results showed that the four PGPB significantly boosted oilseed rape shoot biomass production, improved soil and plant analyzer development (SPAD) value, enhanced Cd uptake of plant and Cd translocation to the leaves. By fluorescent in situ hybridization (FISH) and green fluorescent protein (GFP), we demonstrated the studied S. alfredii endophytic bacterium SaMR12 were able to colonize successfully in the B. napus roots. However, all four PGPB could increase seed Cd accumulation. Due to its potential to enhance Cd uptake by the plant and to restrict Cd accumulation in the seeds, SaMR12 was selected as the most promising microbial partner of B. napus when setting up a plant–microbe fortified remediation system.     

Effects of inoculation of a plant growth promoting rhizobacterium <Emphasis Type="Italic">Burkholderia</Emphasis> sp. D54 on plant growth and metal uptake by a hyperaccumulator <Emphasis Type="Italic">Sedum alfredii</Emphasis> Hance grown on multiple metal contaminated soil

Batch experiments were designed to characterize a multiple metal resistant bacterium Burkholderia sp. D54 isolated from metal contaminated soils in the Dabaoshan Mine in South China, and a follow-up experiment was conducted to investigate the effects of inoculating the isolate on plant growth and metal uptake by Sedum alfredii Hance grown on soils collected from a heavily contaminated paddy field in Daxing County, Guangxi Zhuang Automounous Region, Southwest China. Our experiments showed that strain D54 produced indole acetic acid (IAA), siderophores, 1-aminocyclopropane-1-carboxylate (ACC) deaminase, and solubilizing inorganic phosphate and solubilized insoluble metal bearing minerals. Bacterial inoculation significantly enhanced S. alfredii biomass production, and increased both shoot and root Cd concentration, but induced little variation in root/shoot Pb concentration and shoot Zn concentration. Despite this, the total shoot and root uptake of Cd, Pb and Zn in S. alfredii inoculated with D54 increased greatly compared to the non-inoculated controls. It was concluded that inoculation with strain D54 could help S. alfredii grow better on metal contaminated soils, produce more biomass, and remove more metals from soil, which implies improved efficiency of phytoextraction from metal contaminated soil. The knowledge gained from the present experiments constitutes an important advancement in understanding of the interaction between plant growth-promoting bacteria and hyperaccumulators with regard to plant ability to grow and remove the multiple heavy metals from soils.     

Bacteria associated with yellow lupine grown on a metal‐contaminated soil: in vitro screening and in vivo evaluation for their potential to enhance Cd phytoextraction

In order to stimulate selection for plant‐associated bacteria with the potential to improve Cd phytoextraction, yellow lupine plants were grown on a metal‐contaminated field soil. It was hypothesised that growing these plants on this contaminated soil, which is a source of bacteria possessing different traits to cope with Cd, could enhance colonisation of lupine with potential plant‐associated bacteria that could then be inoculated in Cd‐exposed plants to reduce Cd phytotoxicity and enhance Cd uptake. All cultivable bacteria from rhizosphere, root and stem were isolated and genotypically and phenotypically characterised. Many of the rhizobacteria and root endophytes produce siderophores, organic acids, indole‐3‐acetic acid (IAA) and aminocyclopropane‐1‐carboxylate (ACC) deaminase, as well as being resistant to Cd and Zn. Most of the stem endophytes could produce organic acids (73.8%) and IAA (74.3%), however, only a minor fraction (up to 0.7%) were Cd or Zn resistant or could produce siderophores or ACC deaminase. A siderophore‐ and ACC deaminase‐producing, highly Cd‐resistant Rhizobium sp. from the rhizosphere, a siderophore‐, organic acid‐, IAA‐ and ACC deaminase‐producing highly Cd‐resistant Pseudomonas sp. colonising the roots, a highly Cd‐ and Zn‐resistant organic acid and IAA‐producing Clavibacter sp. present in the stem, and a consortium composed of these three strains were inoculated into non‐exposed and Cd‐exposed yellow lupine plants. Although all selected strains possessed promising in vitro characteristics to improve Cd phytoextraction, inoculation of none of the strains (i) reduced Cd phytotoxicity nor (ii) strongly affected plant Cd uptake. This work highlights that in vitro characterisation of bacteria is not sufficient to predict the in vivo behaviour of bacteria in interaction with their host plants.     

Phytoextraction of Metals and Rhizoremediation of PAHs in Co-Contaminated Soil by Co-Planting of Sedum Alfredii with Ryegrass (Lolium Perenne) or Castor (Ricinus Communis)

A pot experiment was conducted to investigate the potential for phytoextraction of heavy metals and rhizoremediation of polycyclic aromatic hydrocarbons (PAHs) in co-contaminated soil by co-planting a cadmium/zinc (Cd/Zn) hyperaccumulator and lead (Pb) accumulator Sedum alfredii with ryegrass (Lolium perenne) or castor (Ricinus communis). Co-planting with castor decreased the shoot biomass of S. alfredii as compared to that in monoculture. Cadmium concentration in S. alfredii shoot significantly decreased when grown with ryegrass or castor as compared to that in monoculture. However, no reduction of Zn or Pb concentration in S. alfredii shoot was detected in co-planting treatments. Total removal of either Cd, Zn, or Pb by plants was similar across S. alfredii monoculture or co-planting with ryegrass or castor, except enhanced Pb removal in S. alfredii and ryegrass co-planting treatment. Co-planting of S. alfredii with ryegrass or castor significantly enhanced the pyrene and anthracene dissipation as compared to that in the bare soil or S. alfredii monoculture. This appears to be due to the increased soil microbial population and activities in both co-planting treatments. Co-planting of S. alfredii with ryegrass or castor provides a promising strategy to mitigate both metal and PAH contaminants from co-contaminated soils.     

Isolation and characterization endophytic bacteria from hyperaccumulator Sedum alfredii Hance and their potential to promote phytoextraction of zinc polluted soil

The aim of this study was to isolate and characterize endophytic bacteria from roots, stems and leaves of Zn/Cd hyperaccumulator Sedum alfredii. Endophytic bacteria were observed in roots, stems and leave of S. alfredii, with a significantly higher density in roots, followed by leave and stems. A total of fourteen bacterial endophytes were isolated and are closely related phylogenetically to Pseudomonas, Bacillus, Stenotrophomonas, Acinetobacte by 16S rRNA sequence analysis. Most of the endophytic bacteria were found to exhibit high Zn and Cd resistance characteristics, but difference existed among this isolates. The fourteen endophytic bacteria all had the capacity to produce IAA. Moreover, strains VI₈L₁, VI₈L₂, VI₈L₄, VI₈R₂, VI₈R₃ and II₂R₃ could solubilize Ca₃(PO₄)₂, strains VI₈L₂, II₈L₄ and VI₈R₂ could produce siderophore, and strains VI₈L₂ and VI₈R₃ had the capacity of nitrogen fixation. Both plate and broth assay proved that strain VI₈L₁, VI₈L₂, II₈L₄ and VI₈R₂ were able to effectively solubilize ZnCO₃ and Zn₃(PO₄)₂. The filtrate liquid media after growth of strains VI₈L₁, VI₈L₂, II₈L₄ and VI₈R₂ extracted much higher Zn from artificially ZnCO₃ and Zn₃(PO₄)₂ contaminated soils than those extracted by axenic SMS broth, and the filtrates of the culture media supporting growth of strains VI₈L₂, II₈L₄ and VI₈R₂ also extracted significantly greater quantities of Zn from the Dabaoshan contaminated soils. This Zn mobilizing, plant growth promoting and metal resistant endophytic bacteria may offer promise as inoculants to increase soil Zn bioavailability and improve growth and Zn accumulation by S. alfredii.     

Integrated Fertilization Regimes Boost Heavy Metals Accumulation and Biomass of <i>Sedum alfredii</i> Hance

The hyperaccumulator Sedum alfredii Hance (S. alfredii) may be employed for zinc (Zn) and cadmium (Cd)-polluted soil remediation. However, the low phytoremediation efficiency, related to the low biomass production, limits its use with that purpose. In this experiment, nitrogen (N), phosphorus (P), and potassium (K) fertilizers, and organic manure were applied to investigate the phytoremediation ability of S. alfredii. Hydroponic and pot experiments were conducted using Zn-Cd polluted soil. The hydroponic experiment indicated that appropriate fertilizer application could increase (p < 0.05) the amount of accumulated Zn and Cd in S. alfredii. When N supply ranged from 0.5 to 2.5 mmol L⁻¹, it could improve growth and accumulation of Zn and Cd in whole plants of S. alfredii. The 1 mmol L^-1 N was an optimal N dosage for shoot biomass production and Cd accumulation in shoots, while the 2.5 mmol L^-1 was an optimal N dosage for Zn accumulation in shoots. Both low (<0.05 mmol L^-1) and high (>0.8 mmol L^-1) P supply decreased growth, and Zn/Cd accumulation in whole plants of the studied species. The 0.1 mmol L^-1 P was an optimal dosage for S. alfredii biomass production and Zn/Cd accumulation in shoots. The supply levels within the range from 0.3 to 1 mmol L^-1 K could significantly improve the biomass production of S. alfredii and its capability to accumulate Zn and Cd in the biomass. The 0.5 mmol L^-1 K was an optimal dosage for the whole biomass production and Zn accumulation in shoots, while the 1 mmol L^-1 was an optimal K dosage for Zn accumulation in shoots, which was 17.2% higher than the control. Moreover, the soil pot experiment showed that the combination of organic (fermented manure) and inorganic fertilizers made significant effects on the Zn and Cd-polluted soil remediation by S. alfredii. These effects varied, however, with the application of different proportions of N, P, K and organic matter. The Zn accumulation by S. alfredii reached the highest efficiency ability under the highest fertilizer mixing rate (N: 50 mg kg^-1, P: 40 mg kg^-1, K: 100 mg kg^-1, organic matter: 1%). Even more, S. alfredii showed the strongest ability to accumulate Cd with a lower fertilizer mixing rate (N: 25mg kg^-1, P: 20mg kg^-1, K: 50 mg kg^-1, organic matter: 0.5%).

     

Elevated CO2 improves root growth and cadmium accumulation in the hyperaccumulator Sedum alfredii

Aims

This study examined the effect of elevated CO₂ on plant growth, root morphology and Cd accumulation in S. alfredii, and assessed the possibility of using elevated CO₂ as fertilizer to enhance phytoremediation efficiency of Cd-contaminated soil by S. alfredii.

Methods

Both soil pot culture and hydroponic experiments were carried out to characterize plant biomass, root morphological parameters, and cadmium uptake in S. alfredii grown under ambient (350 μL L^?1) or elevated (800 μL L^?1) CO₂.

Results

Elevated CO₂ prompted the growth of S. alfredii, shoot and root biomass were increased by 24.6–36.7% and 35.0–52.1%, respectively, as compared with plants grown in ambient CO₂. After 10 days growth in medium containing 50 μM Cd under elevated CO₂, the development of lateral roots and root hairs were stimulated, additionally, root length, surface area, root volume and tip number were increased significantly, especially for the finest diameter roots. The total Cd uptake per pot was significantly greater under elevated CO₂ than under ambient CO₂. After 60 d growth, Cd phytoextraction efficiency was increased significantly in the elevated CO₂ treatment.

Conclusions

Results suggested that the use of elevated CO₂ may be a useful way to improve phytoremediation efficiency of Cd-contaminated soil by S. alfredii.     

Root foraging for zinc and cadmium requirement in the Zn/Cd hyperaccumulator plant Sedum alfredii

Positive root response to metals may enhance metal accumulation for greater requirement in hyperaccumulators. The effects of spatially heterogeneous Zn/Cd addition on root allocation, metal accumulation, and growth of the Zn/Cd hyperaccumulator Sedum alfredii were assessed in a pot experiment. Young shoots of S. alfredii were grown with or without supplied Zn/Cd. Two concentrations were used of each metal, and each metal concentration had one homogeneous and two heterogeneous treatments. Growth increased by 1.6–3.2 times with the increasing overall dose of Zn/Cd addition, and shoot biomass was positively correlated with shoot Zn/Cd concentration (P?<?0.001). In all heterogeneous treatments, the plants consistently allocated approximately 90% of root biomass to the metal-enriched patches, and shoot Zn/Cd contents were greater than or similar to those in the homogeneous treatment at each metal concentration. Plants in the control treatment showed symptoms of Zn deficiency, although their shoots had Zn concentrations 100-fold higher than the critical deficiency value for most plants. We conclude that S. alfredii has evolved root foraging mechanisms associated with its greater requirements for Zn/Cd. These results could have important implications both for phytoremediation and for investigation of positive role of Cd in higher plants.     

Colonization behavior of bacterium Burkholderia cepacia inside the Oryza sativa roots visualized using green fluorescent protein reporter

Colonization behavior of endophytic bacteria Burkholderia cepacia strains RRE-3 and RRE-5 was studied in the seedlings of rice variety NDR97 using confocal laser scanning microscopy under controlled laboratory and greenhouse conditions. For studying colonization pattern, bacterial strains were tagged with pHRGFPGUS plasmid. The role of bacterial strains (both gfp/gus-tagged and untagged) in growth promotion was also studied. After coming into contact with the host root system the bacteria showed an irregular spreading. Dense colonization was observed on the primary and secondary roots and also on the junction of emergence of the lateral roots. Results showed that the colonization pattern of Burkholderia cepacia strains was similar to that of other endophytic bacteria isolated from non-legumes. Burkholderia cepacia got entry inside the root at the sites of emergence of lateral roots, without formation of infection threads as in the case of symbiotic rhizobacteria. Observations suggested that the endophytic bacterial strains RRE-3 and RRE-5 entered inside the rice roots in a progressive manner. Bacteria were found to line up along the intercellular spaces of adjoining epidermal cells adjacent to the lateral root junction, indicating endophytic colonization pattern of Burkholderia cepacia strains. Experiments with the rice seedlings inoculated with RRE-3 and RRE-5 strains revealed that both strains enhanced plant growth considerably when observed under laboratory and greenhouse conditions and produced significantly higher plant biomass. No considerable difference was observed between the gfp/gus-tagged and non-gfp/gus-tagged strains in the plant growth experiments both in the laboratory and greenhouse conditions.     

The Endophytic Bacterium,Sphingomonas SaMR12, Improves the Potential for Zinc Phytoremediation by Its Host,Sedum alfredii

The endophytic bacterium, Sphingomonas SaMR12, isolated from Sedum alfredii Hance, appears to increase plant biomass and zinc-extraction from contaminated soil; however, the mechanism by which this occurs is not clear. Here, the ability of SaMR12 to promote zinc extraction and its effects on root morphology and exudation were examined in hydroponics. Zinc treatment increased shoot biomass by 30 to 45%, and by a further 10 to 19% when combined with SaMR12 inoculation. Zinc treatment also increased zinc accumulation modestly and this too was enhanced with SaMR12. Both biomass and zinc levels increased in a dose-dependent manner with significant effects seen at 50 µM zinc and apparent saturation at 500 µM. Zinc and the endophyte also increased levels of auxin but not at 50 µM and zinc increased levels of superoxide and hydrogen peroxide but mainly at 500 µM. As for root morphology, SaMR12 increased root branching, the number of root tips, and surface area. Zinc and SaMR12 also increased the exudation of oxalic acid. For most assays the effects of the endophyte and zinc were additive, with the notable exception of SaMR12 strongly reducing the production of reactive oxygen species at 500 µM zinc. Taken together, these results suggest that the promotion of growth and zinc uptake by exposure to zinc and to SaMR12 are independent of reactive oxygen and do not involve increases in auxin.     

Soil nitrogen and microbial biomass carbon dynamics in native forests and derived agricultural land uses in a humid tropical climate of India

Sedum alfredii, a cadmium (Cd) and zinc (Zn) hyperaccumulator at a mine located in Qu Zhou City, Zhejiang Province, China, can accumulate Cd and Zn exceeding 1,000 and 10,000 mg kg^?1, respectively in its shoot (dry weight) when growing under metal-contaminated habitats. Several strains of bacteria were isolated from the rhizosphere of S. alfredii thriving in different Pb/Zn mines in Hunan Province and Zhejiang Province, China, which can resist high levels of heavy metals. Among the different strains isolated, Burkholderia cepacia showed the highest ability in mobilizing Cd and Zn as well as resisting high concentrations of soluble Zn (500 mg L^?1). The soluble Zn concentration in the medium increased from 13 to 72 and 99% (p?<?0.001) after bacterial inoculation in the medium supplemented with insoluble zinc oxide and zinc carbonate, respectively, while pH dropped from 7 to 2.93. The soluble Cd concentration was also increased from 8 to 96% (p?<?0.001), and pH decreased from 7 to 2.65. Short-chain organic acids were also analyzed and the results indicated that oxalic acid, tartaric acid, formic acid and acetic acid had a significant correlation (p?<?0.001) with the concentrations of Cd and Zn being mobilized during the assay. The present results implicated that certain bacteria associated with metal hyperaccumulators could contribute significantly in mobilizing heavy metals, which would enhance the phytoextraction process.     

Culturable bacteria from Zn‐ and Cd‐accumulating Salix caprea with differential effects on plant growth and heavy metal availability

Aims: To characterize bacteria associated with Zn/Cd‐accumulating Salix caprea regarding their potential to support heavy metal phytoextraction. Methods and Results: Three different media allowed the isolation of 44 rhizosphere strains and 44 endophytes, resistant to Zn/Cd and mostly affiliated with Proteobacteria, Actinobacteria and Bacteroidetes/Chlorobi. 1‐Aminocyclopropane‐1‐carboxylic acid deaminase (ACCD), indole acetic acid and siderophore production were detected in 41, 23 and 50% of the rhizosphere isolates and in 9, 55 and 2% of the endophytes, respectively. Fifteen rhizosphere bacteria and five endophytes were further tested for the production of metal‐mobilizing metabolites by extracting contaminated soil with filtrates from liquid cultures. Four Actinobacteria mobilized Zn and/or Cd. The other strains immobilized Cd or both metals. An ACCD‐ and siderophore‐producing, Zn/Cd‐immobilizing rhizosphere isolate (Burkholderia sp.) and a Zn/Cd‐mobilizing Actinobacterium endophyte were inoculated onto S. caprea. The rhizosphere isolate reduced metal uptake in roots, whereas the endophyte enhanced metal accumulation in leaves. Plant growth was not promoted. Conclusions: Metal mobilization experiments predicted bacterial effects on S. caprea more reliably than standard tests for plant growth‐promoting activities. Significance and Impact of the Study: Bacteria, particularly Actinobacteria, associated with heavy metal‐accumulating Salix have the potential to increase metal uptake, which can be predicted by mobilization experiments and may be applicable in phytoremediation.     

Increased plant growth and copper uptake of host and non-host plants by metal-resistant and plant growth-promoting endophytic bacteria

The effects of inoculation with two metal-resistant and plant growth-promoting endophytic bacteria (Burkholderia sp. GL12 and Bacillus megaterium JL35) were evaluated on the plant growth and Cu uptake in their host Elsholtzia splendens and non-host Brassica napus plants grown in natural Cu-contaminated soil. The two strains showed a high level of ACC deaminase activities. In pot experiments, inoculation with strain GL12 significantly increased root and above-ground tissue dry weights of both plants, consequently increasing the total Cu uptake of E. splendens and Brassica napus by 132% and 48.2% respectively. Inoculation with strain JL35 was found to significantly increase not only the biomass of B. napus, consequently increasing the total Cu uptake of B. napus by 31.3%, but Cu concentration of E. splendens for above-ground tissues by 318% and roots by 69.7%, consequently increasing the total Cu uptake of E. splendens by 223%. The two strains could colonize the rhizosphere soils and root interiors of both plants. Notably, strain JL35 could colonize the shoot tissues and significantly increase the translocation factors and bioaccumulation factors of E. splendens. These results suggested that Burkholderia sp. GL12 and B. megaterium JL35 were valuable bacterial resource which had the potential in improving the efficiency of Cu phytoextraction by E. splendens and B. napus in a natural Cu-contaminated soil.     

ENDOPHYTIC BACTERIA FROM SEEDS OF NICOTIANA TABACUM CAN REDUCE CADMIUM PHYTOTOXICITY

Although endophytic bacteria seem to have a close association with their host plant, little is known about the influence of seed endophytic bacteria on initial plant development and on their interactions with plants under conditions of metal toxicity. In order to further elucidate this close relationship, we isolated endophytic bacteria from surface sterilized Nicotiana tabacum seeds that were collected from plants cultivated on a cadmium-(Cd) and zinc-enriched soil. Many of the isolated strains showed Cd tolerance. Sterilely grown tobacco plants were inoculated with either the endogenous microbial consortium, composed of cultivable and noncultivable strains; single strains; or defined consortia of the most representative cultivable strains. Subsequently, the effects of inoculation of endophytic bacteria on plant development and on metal and nutrient uptake were explored under conditions with and without exposure to Cd. In general, seed endophytes were found to have a positive effect on plant growth, as was illustrated by an increase in biomass production under conditions without Cd. In several cases, inoculation with endophytes resulted in improved biomass production under conditions of Cd stress, as well as in a higher plant Cd concentration and total plant Cd content compared to noninoculated plants. These results demonstrate the beneficial effects of seed endophytes on metal toxicity and accumulation, and suggest practical applications using inoculated seeds as a vector for plant beneficial bacteria.     

Long-term field phytoextraction of zinc/cadmium contaminated soil by Sedum plumbizincicola under different agronomic strategies

In two long-term field experiments the zinc (Zn)/cadmium (Cd) hyperaccumulator Sedum plumbizincicola (S. plumbizincicola) was examined to optimize the phytoextraction of metal contaminated soil by two agronomic strategies of intercropping with maize (Zea mays) and plant densities. Soil total Zn and Cd concentrations decreased markedly after long-term phytoextraction. But shoot biomass and Cd and Zn concentrations showed no significant difference with increasing remediation time. In the intercropping experiment the phytoremediation efficiency in the treatment “S. plumbizincicola intercropped with maize” was higher than in S. plumbizincicola monocropping, and Cd concentrations of corn were below the maximum national limit. In the plant density experiment the phytoremediation efficiency increased with increasing plant density and 440,000 plants ha^?1 gave the maximum rate. These results indicated that S. plumbizincicola at an appropriate planting density and intercropped with maize can achieve high remediation efficiency to contaminated soil without affecting the cereal crop productivity. This cropping system combines adequate agricultural production with soil heavy metal phytoextraction.     

Molecular Characterization of Endophytic Bacteria from Metal Hyperaccumulator Aquatic Plant (Eichhornia crassipes) and Its Role in Heavy Metal Removal

In this study, among a collection of heavy metals resistant endophytic bacterial strains isolated from aquatic hyperaccumulator plant (Eichhornia crassipes), one plant growth promoting endophytic bacteria (PGPE), SVUB4 was selected for its ability to utilize 1-aminocyclopropane-1-carboxylic acid (ACC) as the sole N source and accumulate different heavy metals. The SVUB4 strain was characterized as Enterobacter sp. on the basis of its 16S rDNA sequences. Assessment of the parameters of plant growth promotion revealed the intrinsic ability of the strain for the production of IAA, siderophore and solubilization of insoluble phosphate. Furthermore, plasmid DNA analysis of Enterobacter sp. strain SVUB4 indicated the presence of a single large plasmid element. The results of plasmid curing experiments demonstrated that the ability of this strain to grow in presence of Cd and Zn was encoded by the 98 kb plasmid, whereas the ability to grow in the presence of Pb appeared to be encoded by the chromosome. The Cd and Zn removal capacity of the respective metal sensitive strain (plasmidless) were about 36 and 45 μg/g-1 DW, respectively, while the removal capacity of the both metal by metal resistant strain (p SVUB4) showed a significantly higher Cd and Zn removal capacity of 153 and 228 μg/g^?1 DW, respectively. However, both strains exhibited a similar pattern of Pb accumulation. The present observation also showed that for wild-type strain SVUB4 (pSVUB4), the overall level of IAA production in the absence and in the presence of Cd²⁺ or Zn²⁺was approximately the same. Nevertheless, strain SVUB4M in this respect appeared to be more sensitive to heavy metals: a noticeable decrease in IAA production was observed under the effect of both metals, especially with Cd²⁺.     

Cleaning up of heavy metals-polluted water by a terrestrial hyperaccumulator Sedum alfredii Hance

Sedum alfredii Hance is a terrestrial zinc/cadmium （Zn/Cd）-hyperaccumulating and lead （Pb）-accumulating plant. Previous studies on S. alfredii were mostly focused on its physiological mechanism of heavy metal uptake and the application in phytoextraction of metals from contaminated soils. In this study, we evaluated the application potential of S. alfredii in the cleanup of heavy metals from contaminated lake water. Our research revealed that changing pH in lake water would not make particular difference on the final accumulation amount of heavy metals, because the acidic water environment negatively affected plant growth compared with the neutral and alkaline environments, but was more conducive for heavy metal absorption and accumulation. In addition, S. alfredii showed an increase of approximately 2.2-fold in dry weight （DW） when cultured with lake water for 25 d. At the same time, it accumulated approximately 5.0 mg/kg DW of Cd and 41.4 mg/kg DW of Pb. The absorption of heavy metals was highly effective during the first 10 d of culture. Also, the quality of lake water was greatly improved after only 2-d cleanup by S. aifredii. In general, this hyperaccumulator exhibits great potential for application in the cleanup of heavy metals-polluted waters.     

Root endophytes enhance stress‐tolerance of Cicuta virosa L. growing in a mining pond of eastern Japan

Cicuta virosa L. plants can grow in a pond subjected to heavy‐metal inputs at the Hitachi mine, eastern Japan. They accumulate heavy‐metal elements, especially high concentrations of zinc (Zn), in their roots. We focused on the role that root bacterial endophytes play in the heavy‐metal uptake of plants and the provision of heavy‐metal tolerance within plants. Our purpose was to clarify the effects of endophytes on: (i) Zn accumulation in C. virosa roots; (ii) growth of C. virosa seedlings; and (iii) heavy‐metal tolerance of C. virosa plants. Root endophytic Pseudomonas putida and Rhodopseudomonas sp., which induced the high production of Zn‐chelating compounds, were selected for the seedling inoculation test. The results of the inoculation test demonstrated that both strains of endophytes increased Zn accumulation in C. virosa roots by solubilizing Zn in the sediment. Both strains also increased the growth of seedlings by possible production of indole‐3‐acetic acid in the plant. The heavy‐metal tolerance of C. virosa seedlings was likely promoted by producing metal‐chelating compounds that detoxify the metals in the plant tissues, and by decreasing the heavy‐metal contents in the tissues via rapid seedling growth. Thus, such mutualistic interactions between plants and bacteria contribute to the persistence of C. virosa in this severe environment.     

The long-term variation of Cd and Zn hyperaccumulation by Noccaea spp and Arabidopsis halleri plants in both pot and field conditions

Three Cd and Zn hyperaccumulating plant species Noccaea caerulescens Noccaea praecox and Arabidopsis halleri (Brassicacceae) were cultivated in seven subsequent vegetation seasons in both pot and field conditions in soil highly contaminated with Cd, Pb, and Zn. The results confirmed the hyperaccumulation ability of both plant species, although A. halleri showed lower Cd uptake compared to N. caerulescens. Conversely, Pb phytoextraction was negligible for both species in this case. Because of the high variability in plant yield and element contents in the aboveground biomass of plants, great variation in Cd and Zn accumulation was observed during the experiment. The extraction ability in field conditions varied in the case of Cd from 0.2 to 2.9 kg ha^?1 (N. caerulescens) and up to 0.15 kg ha^?1 (A. halleri), and in the case of Zn from 0.2 to 6.4 kg ha^?1 (N. caerulescens) and up to 13.8 kg.ha^?1 (A. halleri). Taking into account the 20 cm root zone of the soil, the plants were able to extract up to 4.1% Cd and 0.2% Zn in one season. However, cropping measures should be optimized to improve and stabilize the long-term phytoextraction potential of these plants.     

Metal accumulation and arbuscular mycorrhizal status in metallicolous and nonmetallicolous populations of <Emphasis Type="Italic">Pteris vittata</Emphasis> L. and <Emphasis Type="Italic">Sedum alfredii</Emphasis> Hance

Although Pteris vittata L. and Sedum alfredii Hance have been identified as an As hyperaccumulator and a Zn/Cd hyperaccumulator, respectively, for a few years, variations in metal accumulation among populations and their arbuscular mycorrhizal (AM) status have not been fully explored. Six populations of P. vittata and four populations of S. alfredii from southeast China were investigated. Up to 1,373 As, 680 Pb, 376 Zn, 4.8 Cd, 169 Cu mg kg⁻¹ in fronds of P. vittata and 358 As, 2,290 Pb, 23,403 Zn, 708 Cd, 342 Cu mg kg⁻¹ in shoots of S. alfredii were detected. Constitutive properties of As and Zn hyperaccumulation in metallicolous populations of P. vittata and S. alfredii, respectively, were confirmed. However, Cd hyperaccumulation in S. alfredii varied among populations. The two hyperaccumulators varied in efficiency in taking up other heavy metals. Different metal tolerance strategies adopted by the two hyperaccumulators varied among plant species and metal species. Low to moderate levels of AM colonization in P. vittata (4.2–12.8%) and S. alfredii (8.5–45.8%) were observed at uncontaminated and metal-contaminated sites. The relationship between metal concentrations and AM colonization in the two hyperacumulators was also examined. The abundance of AM fungal spores ranged from 16 to 190 spores per 25 g soil. Glomus microaggregatum, Glomus mosseae, Glomus brohultii and Glomus geosporum were the most common species associated with both P. vittata and S. alfredii. To our knowledge, this is the first report of AM fungal status in rhizosphere of P. vittata and S. alfredii.