Phytoextraction of Cd and Zn from agricultural soils by Salix ssp. and intercropping of Salix caprea and Arabidopsis halleri

Contamination of agricultural topsoils with Cd above guideline values is of concern in many countries throughout the world. Extraction of metals from contaminated soils using high-biomass, metal-accumulating Salix sp. has been proposed as a low-cost, gentle remediation strategy, but reasonable phytoextraction rates remain to be demonstrated. In an outdoor pot experiment we assessed the phytoextraction potential for Cd and Zn of four willow species (Salix caprea, S. fragilis, S. × smithiana, S. × dasyclados) and intercropping of S. caprea with the hyperaccumulator Arabidopsis halleri on three moderately contaminated, agricultural soils. Large concentrations of Cd (250 mg kg⁻¹) and Zn (3,300 mg kg⁻¹) were determined in leaves of Salix × smithiana grown on a soil containing 13.4 mg kg⁻¹ Cd and 955 mg kg⁻¹ Zn, resulting in bioaccumulation factors of 27 (Cd) and 3 (Zn). Total removal of up to 20% Cd and 5% Zn after three vegetation periods were shown for Salix × smithiana closely followed by S. caprea, S. fragilis and S. × dasyclados. While total Cd concentrations in soils were reduced by up to 20%, 1 M NH₄NO₃-extractable metal concentrations did not significantly decrease within 3 years. Intercropping of S. caprea and A. halleri partly increased total removal of Zn, but did not enhance total Cd extraction compared to single plantings of S. caprea after two vegetation periods.     

Soil pH Effects on Uptake of Cd and Zn by Thlaspi caerulescens

For phytoextraction to be successful and viable in environmental remediation, strategies that can optimize plant uptake must be identified. Thlaspi caerulescens is an important hyperaccumulator of Cd and Zn, whether adjusting soil pH is an efficient way to enhance metal uptake by T. caerulescens must by clarified. This study used two soils differing in levels of Cd and Zn, which were adjusted to six different pH levels. Thlaspi caerulescens tissue metal concentrations and 0.1 M Sr(NO₃)₂ extractable soil metal concentrations were measured. The soluble metal form of both Cd and Zn was greatly increased with decreasing pH. Lowering pH significantly influenced plant metal uptake. For the high metal soil, highest plant biomass was at the lowest soil pH (4.74). The highest shoot metal concentration was at the second lowest pH (5.27). For low metal soil, due to low pH induced Al and Mn toxicity, both plant growth and metal uptake was greatest at intermediate pH levels. The extraordinary Cd phytoextraction ability of T. caerulescens was further demonstrated in this experiment. In the optimum pH treatments, Thlaspi caerulescens extracted 40% and 36% of total Cd in the low and high metal soils, respectively, with just one planting. Overall, decreasing pH is an effective strategy to enhance phytoextraction. But different soils had various responses to acidification treatment and a different optimum pH may exist. This pH should be identified to avoid unnecessarily extreme acidification of soils.     

Variation in the uptake of Arsenic, Cadmium, Lead, and Zinc by different species of willows Salix spp. grown in contaminated soils

The experiment assessed the variability of in seven clones of willow plants of high biomass production (Salix smithiana S-218, Salix smithiana S-150, Salix viminalis S-519, Salix alba S-464, Salix ’Pyramidalis’ S-141, Salix dasyclados S-406, Salix rubens S-391). They were planted in a pots for three vegetation periods in three soils differing in the total content of risk elements. Comparing the calculated relative decrease of total metal contents in soils, the phytoextraction potential of willows was obtained for cadmium (Cd) and zinc (Zn), moderately contaminated Cambisol and uncontaminated Chernozem, where aboveground biomass removed about 30% Cd and 5% Zn of the total element content, respectively. The clones showed variability in removing Cd and Zn, depending on soil type and contamination level: S. smithiana (S-150) and S. rubens (S-391) demonstrated the highest phytoextraction effect for Cd and Zn. For lead (Pb) and arsenic (As), the ability to accumulate the aboveground biomass of willows was found to be negligible in both soils. The results confirmed that willow plants show promising results for several elements, mainly for mobile ones like cadmium and zinc in moderate levels of contamination. The differences in accumulation among the clones seemed to be affected more by the properties of clones, not by the soil element concentrations or soil properties. However, confirmation and verification of the results in field conditions as well as more detailed investigation of the mechanisms of cadmium uptake in rhizosphere of willow plants will be determined by further research.     

Field Evaluation of Willow Under Short Rotation Coppice for Phytomanagement of Metal-Polluted Agricultural Soils

Short rotation coppice (SRC) of willow and poplar might be a promising phytoremediation option since it uses fast growing, high biomass producing tree species with often a sufficient metal uptake. This study evaluates growth, metal uptake and extraction potentials of eight willow clones (Belders, Belgisch Rood, Christina, Inger, Jorr, Loden, Tora and Zwarte Driebast) on a metal-contaminated agricultural soil, with total cadmium (Cd) and zinc (Zn) concentrations of 6.5 ± 0.8 and 377 ± 69 mg kg^?1 soil, respectively. Although, during the first cycle, on average generally low productivity levels (3.7 ton DM (dry matter) ha^?1 y^?1) were obtained on this sandy soil, certain clones exhibited quite acceptable productivity levels (e.g. Zwarte Driebast 12.5 ton DM ha^?1 y^?1). Even at low biomass productivity levels, SRC of willow showed promising removal potentials of 72 g Cd and 2.0 kg Zn ha^?1 y^?1, which is much higher than e.g. energy maize or rapeseed grown on the same soil. Cd and Zn removal can be increased by 40% if leaves are harvested as well. Nevertheless, nowadays the wood price remains the most critical factor in order to implement SRC as an acceptable, economically feasible alternative crop on metal-contaminated agricultural soils.     

Potential of Salix as phytoextractor for Cd on moderately contaminated soils

Biomass production and Cd uptake in Salix (willow) is generally high. The aim of this study was to quantify the amounts of Cd in different plant and soil compartments in commercial Salix stands grown on different soil types and to use these data for estimation of the long-term effects of Cd removal by Salix at stem harvest on the soil Cd content by calculating Cd balances. Ten commercial short rotation willow coppice stands, planted with clone 78183 of Salix viminalis (L.) were investigated in order to include three different soil types with respect to pH. Stands with high stem biomass production generally had lower Cd concentrations in the plant and grew on more alkaline soils with low Cd availability when compared to stands with low stem biomass production. The highest output of Cd by Salix is therefore not necessarily achieved by stands with the highest yields or the highest Cd concentrations, as both these parameters have to be considered. The net removal of Cd from the plough layer by the Salix crop under different conditions varied between 2.6 and 16.5 g Cd ha^–1 year^–1. The long-term effects of Salix cultivation on soil Cd content, i.e. after 6–7 cutting cycles (25 years), were calculated and we reached the conclusion that there is a high potential in Salix to remove significant amounts of Cd at stem harvest. Salix can therefore presumably be used to counteract Cd pollution of agricultural soils in Sweden provided that the Cd in furnace ashes is taken care of.     

Predictability of the Zn and Cd phytoextraction efficiency of a Salix smithiana clone by DGT and conventional bioavailability assays

Aims

Phytomanagement of metal-polluted soils requires information on plant responses to metal availability in soil, but the predictability of metal accumulation in plant shoots and/or roots may be limited by metal toxicity and inherent shortfalls of the bioavailability assays.

Methods

We measured the uptake of Cd and Zn in a Salix smithiana clone grown in a pot experiment on soils with different characteristics and metal availabilities, determined by conventional soil single extractions (0.05 M Na₂-EDTA and 1 M NH₄NO₃), soil solution obtained by centrifugation, and diffusive gradients in thin films (DGT). The Cd and Zn phytoavailability after a 2-year phytoextraction by willow was assessed by metal accumulation in the straw of the following barley culture.

Results

The phytoextraction efficiency was largest on a moderately polluted acid soil. Biomass and shoot Zn concentrations of S. smithiana were better predicted by DGT-measured Zn concentrations in soil solution (C _DGT) than by Zn concentrations in the soil solution and extractable soil fractions. The weaker correlation for Cd in shoots may be related to relative Cd enrichment in the plant tissues. The metal accumulation in barley straw was unaffected or increased after a 2-year phytoextraction.

Conclusions

The shoot Zn and Cd removal of the tested Salix clone can be predicted by C _DGT concentrations and is highest on either calcareous or moderately polluted acid soils. Single extraction with NH₄NO₃ and the C _DGT value of Cd were not able to predict shoot Cd removal on the tested soils. Only shoot removal of Zn was predicted fairly well by the C _DGT value.     

Biomass growth variation and phytoextraction potential of four Salix varieties grown in contaminated soil amended with lime and wood ash

Abstract

In a greenhouse experiment, plant growth and copper (Cu) and zinc (Zn) uptake by four Salix cultivars grown in Cu and Zn contaminated soils collected from a mining area in Finland were tested to assess their suitability for phytoextraction. The cultivars displayed tolerance to heavily contaminated soils throughout the experiment. After uptake, total mean Cu concentrations in the leaves, shoots and roots in all cultivars and treatments ranged from 163 to 474?mg kg^?1 and mean Zn concentrations ranged from 776 to 1823?mg kg^?1. Lime and wood ash addition increased dry biomass growth (25–43%), chlorophyll fluorescence (Fv/Fm) values (3–6%), the translocation factor (TF) (15–60% for Cu; 10–25% for Zn), the bio-concentration factor (BCF) (40–85% for Cu; 70–120% for Zn), and metal uptake (55–70% for Cu; 50–65% for Zn) compared to unamended treatment across all cultivars. The results revealed that Salix cultivars have the potential to take up and accumulate significant amounts of Cu and Zn. Cultivar Klara (Salix viminalis × S. schwerinii × S. dasyclados) was found to be the most effective cultivar for phytoextraction since it displayed greater dry biomass production, Fv/Fm, TF, BCF values and uptake percentage rates of Cu and Zn compared to the other three cultivars. This study indicates that further research is needed to clarify the wider phytoextraction capabilities of different Salix cultivars.     

METAL UPTAKE AND ALLOCATION IN TREES GROWN ON CONTAMINATED LAND: IMPLICATIONS FOR BIOMASS PRODUCTION

Phytostabilization aims to reduce environmental and health risks arising from contaminated soil. To be economically attractive, plants used for phytostabilization should produce valuable biomass. This study investigated the biomass production and metal allocation to foliage and wood of willow (Salix viminalis L.), poplar (Populus monviso), birch (Betula pendula), and oak (Quercus robur) on five different soils contaminated with trace elements (TE), with varying high concentrations of Cu, Zn, Cd, and Pb as well as an uncontaminated control soil. In the treatment soils, the biomass was reduced in all species except oak. There was a significant negative correlation between biomass and foliar Cd and Zn concentrations, reaching up to 15 mg Cd kg^?1 and 2000 mg Zn kg ^?1 in willow leaves. Lead was the only TE with higher wood than foliage concentrations. The highest Pb accumulation occurred in birch with up to 135 mg kg ^?1 in wood and 78 mg kg ^?1 in foliage. Birch could be suitable for phytostabilization of soils with high Cd and Zn but low Pb concentrations, while poplars and willows could be used to stabilise soils with high Cu and Pb and low Zn and Cd concentrations.     

Phytoextraction of Risk Elements by Willow and Poplar Trees

To characterize the phytoextraction efficiency of two clones of willow trees (Salix x smithiana Willd., Salix rubens) and two clones of poplar trees (Populus nigra x maximowiczii, Populus nigra Wolterson) were planted in contaminated soil (0.4–2.0 mg Cd.kg^?1, 78–313 mg Zn.kg^?1, 21.3–118 mg Cu.kg^?1). Field experiment was carried out in Czech Republic. The study investigated their ability to accumulate heavy metals (Cd, Zn, and Cu) in harvestable plant parts. The poplars produced higher amount of biomass than willows. Both Salix clones accumulated higher amount of Cd, Zn and Cu in their biomass (maximum 6.8 mg Cd.kg^?1, 909 mg Zn.kg^?1, and 17.7 mg Cu.kg^?1) compared to Populus clones (maximum 2.06 mg Cd.kg^?1, 463 mg Zn.kg^?1, and 11.8 mg Cu.kg^?1). There were no significant differences between clones of individual species. BCs for Cd and Zn were greater than 1 (the highest in willow leaves). BCs values of Cu were very low. These results indicate that Salix is more suitable plant for phytoextraction of Cd and Zn than Populus. The Cu phytoextraction potential of Salix and Populus trees was not confirmed in this experiment due to low soil availability of this element.     

Novel Field Data on Phytoextraction: Pre-Cultivation With Salix Reduces Cadmium in Wheat Grains

Cadmium (Cd) is a health hazard, and up to 43% of human Cd intake comes from wheat products, since Cd accumulates in wheat grains. Salix spp. are high-accumulators of Cd and is suggested for Cd phytoextraction from agricultural soils. We demonstrate, in field, that Salix viminalis can remove Cd from agricultural soils and thereby reduce Cd accumulation in grains of wheat subsequently grown in a Salix-treated field. Four years of Salix cultivation reduce Cd concentration in the soil by up to 27% and in grains of the post-cultivated wheat by up to 33%. The higher the plant density of the Salix, the greater the Cd removal from the soil and the lower the Cd concentration in the grains of post-cultivated wheat, the Cd reduction remaining stable several years after Salix cultivation. The effect occurred in both sandy and clayey soil and in winter and spring bread wheat cultivars. Already one year of Salix cultivation significantly decrease Cd in post grown wheat grains. With this field experiment we have demonstrated that phytoextraction can reduce accumulation of a pollutant in post-cultivated wheat and that phytoextraction has no other observed effect on post-cultivated crops than reduced uptake of the removed pollutant.     

Heavy Metal Uptake by Willow Clones from Sewage Sludge-Treated Soil: The Potential for Phytoremediation

Willow (Salix spp.) has shown potential for use in the phytoremediation of soil contaminated with heavy metals. In particular, it can be grown in short rotation coppice systems to produce biomass that can be used for energy production. Twenty different species or varieties of willow, grown over 2 years (1995 to 1997) on a soil that was highly contaminated with heavy metals due to long-term sewage sludge disposal, showed considerable variation in survival, biomass production and metal uptake. The willows could be divided into two groups after the first harvest. One group had relatively low Ni and Cu in the bark and high Cd and Zn in the wood, with a good survival rate and biomass production. This group partitioned Cu, Cd, and Zn into the wood tissue from the bark, whereas Ni was excluded. The second group had relatively high Ni and Cu in the bark and low Cd and Zn in the wood and performed poorly in terms of survival and biomass production. Of the 20 types of willow used, 11 showed potential for use in phytoremediation, combining good survival and biomass production with high metal uptake. Of the others, 2 failed to survive until the second harvest and the other 7 had very poor survival rates.     

Contribution of Ectomycorrhizal Fungi to Cadmium Uptake of Poplars and Willows from a Heavily Polluted Soil

Phytoextraction has been proposed in recent years as an environmentally and cost-efficient treatment technique for the remediation of heavy-metal contaminated sites. In particular, plants that are fast growing, metal accumulating, and economically interesting, such as sunflowers or trees, recently became more important in research on phytoextraction. Heavy metal uptake of trees can be strongly influenced by ectomycorrhizal fungi. We investigated the possibility of enhancing phytoextraction of Cd by willows (Salix viminalis) and poplars (Populus canadensis) in association with three well known ectomycorrhizal fungi (Hebeloma crustuliniforme, Paxillus involutus and Pisolithus tinctorius). A pot experiment was conducted using Cd polluted soil from a contaminated site. Four replicates of each combination of fungus and tree species, and controls without fungal inoculum, were set up. After a growth period of 11 weeks, yields and Cd concentrations in roots, stems, and leaves were measured. In addition, the total Cd uptake, the transfer to roots, and the translocation to stems and leaves were calculated. The association of P. canadensis with P. involutus led to a highly significant increase of Cd concentrations, in particular in the leaves, which contained 2.74 ± 0.34 mg Cd per kg dry matter. Compared to the control this is an enhancement of nearly 100%. The fungi also significantly enhanced the translocation from the roots to the leaves, leading to a concentration ratio (leaves/roots) of 0.32 ± 0.06 compared to 0.20 ± 0.02 of the control plants. Additionally, P. involutus significantly enhanced the total Cd extraction by P. canadensis. Similar effects were not observed by other fungi or in association with S. viminalis.     

Reduction of Cadmium Availability to Tobacco (Nicotiana tabacum) Plants using Soil Amendments in Low Cadmium-contaminated Agricultural Soils: A Pot Experiment

Cadmium (Cd) concentration in field-grown tobacco leaves usually ranges from < 0.5 to 5 mg Cd kg^–1 dry matter (DM). Reducing bioavailability of soil Cd by adding amendments to the soil could be suitable to mitigate Cd uptake by tobacco plants. However, little is known on the effect of inorganic amendments on agricultural soils with low Cd concentrations. Therefore, we performed a pot experiment with tobacco plants that were grown during 56 days in two neutral to alkaline agricultural soils with low total Cd concentrations (soil 1 = 0.4, soil 2 = 0.7 mg kg^–1). Both soils were amended or not with 1 or 5% of sepiolite, zeolite, hydroxyapatite and apatite II™. Major and trace elements were measured in mid-stalk position leaves. Soil metals were measured in a DTPA soil extraction to assess the effect of the amendments on metal bioavailability. Some amendments significantly reduced Cd concentration in tobacco leaves, but the effect differed between the two soils tested. In soil 1, the use of zeolite at the 1% dose was the most efficient, reducing the average Cd concentration from 0.6 to 0.4 mg kg^–1. In soil 2, the 5% hydroxyapatite treatment led to the maximal reduction in Cd concentration (50%), with an average final Cd concentration in leaves of 0.7 mg kg^–1 (control: 1.5 mg kg^–1). There was a dose effect for some amendments in soil 2 (containing more Cd), suggesting a reduced efficiency of the amendment at the lowest addition rate. DTPA extractable Cd and Zn measured at the end of the pot experiment were correlated to the metal concentrations in tobacco leaves suggesting that (1) the reduction in leaf Cd concentration was due to a reduction in metal availability to tobacco and (2) DTPA may be a suitable extractant to estimate Cd availability to tobacco plants in these two soils. In addition, a batch experiment was performed with the same soils to test a larger number of amendments, including the four tested in the pot experiment. Results were compared to those of the pot experiment to assess whether a batch experiment may predict the efficiency of an amendment on a given soil. It gave results compatible with those from the pot experiment except for the sepiolite and highlighted the broad range of potential amendments available for heavy metal remediation in crop plants.     

Phytoextraction Potential of Poplar (Populus alba L. var. pyramidalis Bunge) from Calcareous Agricultural Soils Contaminated by Cadmium

To investigate the phytoextraction potential of Populus alba L. var. pyramidalis Bunge for cadmium (Cd) contaminated calcareous soils, a concentration gradient experiment and a field sampling experiment (involving poplars of different ages) were conducted. The translocation factors for all experiments and treatments were greater than 1. The bioconcentration factor decreased from 2.37 to 0.25 with increasing soil Cd concentration in the concentration gradient experiment and generally decreased with stand age under field conditions. The Cd concentrations in P. pyramidalis organs decreased in the order of leaves > stems > roots. The shoot biomass production in the concentration gradient experiment was not significantly reduced with soil Cd concentrations up to or slightly over 50 mg kg^–1. The results show that the phytoextraction efficiency of P. pyramidalis depends on both the soil Cd concentration and the tree age. Populus pyramidalis is most suitable for remediation of slightly Cd contaminated calcareous soils through the combined harvest of stems and leaves under actual field conditions.     

Phytoremediation of Metal Contaminated Soil Using Willow: Exploiting Plant-Associated Bacteria to Improve Biomass Production and Metal Uptake

Short rotation coppice (SRC) of willow and poplar is proposed for economic valorization and concurrently as remediation strategy for metal contaminated land in northeast-Belgium. However, metal phytoextraction appears insufficient to effectuate rapid reduction of soil metal contents. To increase both biomass production and metal accumulation of SRC, two strategies are proposed: (i) in situ selection of the best performing clones and (ii) bioaugmentation of these clones with beneficial plant-associated bacteria. Based on field data, two experimental willow clones, a Salix viminalis and a Salix alba x alba clone, were selected. Compared to the best performing commercial clones, considerable increases in stem metal extraction were achieved (up to 74% for Cd and 91% for Zn). From the selected clones, plant-associated bacteria were isolated and identified. All strains were subsequently screened for their plant growth-promoting and metal uptake enhancing traits. Five strains were selected for a greenhouse inoculation experiment with the selected clones planted in Cd-Zn-Pb contaminated soil. Extraction potential tended to increase after inoculation of S. viminalis plants with a Rahnella sp. strain due to a significantly increased twig biomass. However, although bacterial strains showing beneficial traits in vitro were used for inoculation, increments in extraction potential were not always observed.     

Phytoextraction of cadmium with Thlaspi caerulescens

The in situ phytoextraction of cadmium from soils can only be achieved using plants that are both tolerant to high Cd concentrations and able to extract sufficient amounts of the metal. However, very few plant species are capable of remediating Cd polluted soils in a reasonable time frame. This paper aims to show that the population of the hyperaccumulator Thlaspi caerulescens J. & C. Presl. from Viviez (south of France), which has a high Cd-accumulating capability, is an efficient tool to remove Cd from contaminated soils. Roots of T. caerulescensViviez proliferate in hot spots of metals in soils which is particularly advantageous because of heterogeneity of the distribution of metal in polluted soils. Isotopic techniques showed that plants from this population acquire Cd from the same pools as non-accumulating species, but that it was much more efficient than non-hyperaccumulators at removing the metal from the soil labile pool. This is due: to (i) a specific rooting strategy, and (ii) a high uptake rate resulting from the existence in this population of Cd-specific transport channels or carriers in the root membrane. Growth and overall extraction can be improved with appropriate N fertilisation, supplied either as mineral fertilisers or uncontaminated sewage sludge. Selecting bigger plants is possible from within a suitable Cd-accumulating population to improve the phytoextraction process. Growing the Cd-accumulating populations results in a reduction in the availability of Cd and Zn as shown with field and lysimeter experiments conducted for several years. As a result, on a practical aspect, Cd hyperaccumulating populations of T. caerulescens may be used as a tool to efficiently reduce the availability of Cd in soils, providing appropriate populations are used.     

Comparison of NTA and Elemental Sulfur as Potential Soil Amendments in Phytoremediation

The effect of NTA (nitrilotriacetic acid) and elemental sulfur (S), two soil amendments suggested for the enhancement of metal phytoavailability in phytoextraction, on heavy metal uptake by Nicotiana tabacum (tobacco) and Zea mays (maize) were studied and compared in two Zn-, Cu-, Cd -, and Pb-contaminated soils from northern Switzerland. Experiments were performed in the greenhouse with topsoil (0 to 20 cm) material from two locations, Dornach and Rafz. The Dornach soil was calcareous and had been contaminated by dust emissions from a nearby brass metal smelter. The Rafz soil, free of carbonates, had been polluted by former sewage sludge application. Soil amendments with S increased the solubility (NaNO₃ extraction) of Zn and Cd about 10-fold in Dornach soil and up to 30-fold in Rafz soil after 55 days. Zn and Cd removal by N. tabacum and Z. mays, however, increased only about 5.5- and 2.5-fold in these treatments in Rafz soil, respectively, while in the Dornach soil only a slight increase for Cd was found. Repeated NTA application increased soluble Zn, Cu, and Cd about 100-, 20-, and 19-fold in the Dornach soil and 13-, 4-, and 2-fold in the Rafz soil shortly after application. Soluble Pb was increased by NTA up to 50-fold in Rafz soil. After 90 days soluble heavy metal concentrations were only slightly elevated in both soils. Again, however, Zn, Cd, and Cu removal by N. tabacum and Z. mays increased only about 1.5- to 2.5-fold in the two soils, whereas Pb removal by N. tabacum increased about fivefold in the Rafz soil as a result of NTA application     

Heavy metal uptake by arbuscular mycorrhizas of Elsholtzia splendens and the potential for phytoremediation of contaminated soil

A pot culture experiment and a field experiment were carried out separately to study heavy metal (HM) uptake from soil contaminated with Cu, Zn, Pb and Cd by Elsholtzia splendens Nakai ex F. Maekawa inoculated with arbuscular mycorrhizal (AM) fungi and the potential for phytoremediation. The HM-contaminated soil in the pot experiment was collected from the field experiment site. Two AM fungal inocula, MI containing only one AM fungal strain, Glomus caledonium 90036, and M II consisting of Gigaspora margarita ZJ37, Gigaspora decipens ZJ38, Scutellospora gilmori ZJ39, Acaulospora spp. andGlomus spp., were applied to the soil under unsterilized conditions. In the pot experiment, the plants were harvested after 24 weeks of growth. Mycorrhizal colonization rate, plant dry weight (DW) and P, Cu, Zn, Pb, Cd concentrations were determined. MI-treated plants had higher mycorrhizal colonization rates than MII-treated plants. Both MI and MII increased shoot and root DW, and MII was more effective than MI. In shoots, the highest P, Cu, Zn and Pb concentrations were all observed in the plants treated with MII, while MI decreased Zn and Pb concentrations and increased P but did not alter Cu, and Cd concentrations were not affected by either of two inocula. In roots, MII increased P, Zn, Pb concentrations but did not alter Cu and Cd, and MI did not affect P, Cu, Zn, Pb, Cd concentrations. Cu, Zn, Pb, Cd uptake into shoots and roots all increased in MII-treated plants, while in MI-treated plants, Cu and Zn uptake into shoots and Cu, Zn, Pb, Cd into roots increased but Pb and Cd uptake into shoots decreased. In general, MII was more effective than MI in promoting plant growth and HM uptake. The field experiment following the pot experiment was carried out to investigate the effects of MII under field conditions. The 45-day-old nonmycorrhizal and MII-colonized seedlings of E. splendens were transplanted to HM-contaminated plots and harvested after 5 months. MII-inoculation increased shoot DW and shoot P, Cu, Zn, Pb concentrations significantly but did not alter shoot Cd concentrations, which led to higher uptake of Cu, Zn, Pb, Cd by E. splendens shoots. These results indicate that the AM fungal consortium represented by MII can benefit phytoextraction of HMs and therefore play a role in phytoremediation of HM-contaminated soils.     

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.     

Dynamics of plant metal uptake and metal changes in whole soil and soil particle fractions during repeated phytoextraction

Aims

Phytoextration of metal polluted soils using hyperaccumulators is a promising technology but requires long term successive cropping. This study investigated the dynamics of plant metal uptake and changes in soil metals over a long remediation time.

Methods

A soil slightly polluted with metals (S1) was mixed with highly polluted soil (S4) to give two intermediate pollution levels (S2, S3). The four resulting soils were repeatedly phyto-extracted using nine successive crops of Cd/Zn-hyperaccumulator Sedum plumbizincicola over a period of 4 years.

Results

Shoot Cd concentration decreased with harvest time in all soils but shoot Zn declined in S1 only. Similar shoot Zn concentrations were found in S2, S3 and S4 although these soils differed markedly in metal availability, and their available metals decreased during phytoextraction. A possible explanation is that plant active acquisition ability served to maintain plant metal uptake. Plant uptake resulted in the largest decrease in the acid-soluble metal fraction followed by reducible metals. Oxidisable and residual fractions were less available to plants. The coarse soil particle fractions made the major contribution to metal decline overall than the fine fractions.

Conclusion

Sedum plumbizincicola maintained long term metal uptake and the coarse soil particles played the most important role in phytoextraction.