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Background and aims Plant Ni uptake in aboveground biomass exceeding concentrations of 1000 µg g− 1 in dry weight is defined as Ni hyperaccumulation. Whether hyperaccumulators are capable of mobilizing larger Ni pools than non-accumulators is still debated and rhizosphere processes are still largely unknown. Aim of this study was to investigate rhizosphere processes and possible Ni mobilization by the Ni hyperaccumulator Odontarrhena chalcidica and to test Ni uptake in relation to a soil Ni gradient.Methods The Ni hyperaccumulator O. chalcidica was grown in a pot experiment on six soils showing a pseudo-total Ni and labile (DTPA-extractable) Ni gradient and on an additional soil showing high pseudo-total but low labile Ni. Soil pore water was sampled to monitor changes in soil solution ionome, pH, and dissolved organic carbon (DOC) along the experiment.Results Results showed that Ni and Fe concentrations, pH as well as DOC concentrations in pore water were significantly increased by O. chalcidica compared to unplanted soils. A positive correlation between Ni in shoots and pseudo-total concentrations and pH in soil was observed, although plant Ni concentrations did not clearly show the same linear pattern with soil available Ni.Conclusions This study shows a clear root-induced Ni and Fe mobilization in the rhizosphere of O. chalcidica and suggests a rhizosphere mechanism based on soil alkalinization and exudation of organic ligands. Furthermore, it was demonstrated that soil pH and pseudo-total Ni are better predictors of Ni plant uptake in O. chalcidica than labile soil Ni.

A method using diffusive gradients in thin films (DGT) for the accurate quantification of trace-level (μg L

Context Rock type may exert strong control on pedogenesis and resulting soil characteristics. Little is known on how different volcanic lithologies influence soil formation and nutrient supply and thereby affect ecosystem productivity in volcanic landscapes. Aims The aim of this study was to investigate lithologic control on the mentioned properties and processes. Methods We conducted a side-by-side comparison of two contrasting sites in close proximity on Alcedo Volcano, Galápagos, one on basaltic lava and the other on rhyolitic pumice. Key results The lava site was covered by lush, stratified woodland, whereas the pumice site was covered by grassland with only few plant species. The soil on lava was a Leptosol, whereas on pumice, an Andosol had formed. Both soils had similar pH (6.0–6.5 in H2O), but the soil organic carbon stocks at the lava site were almost twice as high as at the pumice site (13.1 ± 1.6 vs 7.6 ± 1.3 kg m−2 to 25 cm depth). The soil at the pumice site had considerably lower levels of Mehlich-3-extractable P, Ca, Mg, Cu, Zn and B. Soil microbes at the pumice site had lower biomass P (and a higher C:P ratio) and excreted more phosphatase. The grass biomass here also showed lower (and deficient) levels of P and other nutrients compared to the lava site. Conclusions and implications The contrasting lithologies and their bearings on soil formation and nutrient availability diversify the aboveground and belowground ecosystems and favour the development of distinct ecological niches in close proximity.

Soil P testing has been widely used to predict crop yields, P uptake, and fertilizer demands in agriculture. Diffusive gradients in thin films (DGT) provides a zero-sink soil P test which mimics diffusion-controlled plant uptake and has previously been found to predict P availability to crops better than conventional quantity-based P tests in highly weathered Australian, though not in European soils. Here we tested the performance of DGT and the Austrian and German standard P quantity test calcium acetate lactate (CAL) to explain the variation of crop yield and P uptake response of winter wheat (Triticum aestivum L.) and spring barley (Hordeum vulgare L.) in long-term P fertilization experiments at four different sites in eastern Austria. Phosphorus extracted with DGT (P-DGT) and CAL (P-CAL) correlated well in similar soils but not across sites with large variation in soil and site properties such as carbonate equivalent and water availability. The predictive power of DGT for barley (R2 = 0.42) and wheat grain yield (R2 = 0.32), and P uptake in wheat grains (R2 = 0.36) was clearly superior to that of the CAL, and less dependent on soil properties. The better performance of DGT compared to the quantity test is consistent with diffusion-limited P uptake in the water-limited cultivated soils of eastern Austria. The critical values of P deficiency derived from the Mitscherlich-type fits for barley and wheat at 80% relative yield are 64.9 and 26.2 µg L−1, respectively, consistent with differential P demands of the crops.

Toxic metal pollution requires significant adjustments in plant metabolism. Here, we show that the plant microbiota plays an important role in this process. The endophytic Sporobolomyces ruberrimus isolated from a serpentine population of Arabidopsis arenosa protected plants against excess metals. Coculture with its native host and Arabidopsis thaliana inhibited Fe and Ni uptake. It had no effect on host Zn and Cd uptake. Fe uptake inhibition was confirmed in wheat and rape. Our investigations show that, for the metal inhibitory effect, the interference of microorganisms in plant ethylene homeostasis is necessary. Application of an ethylene synthesis inhibitor, as well as loss-of-function mutations in canonical ethylene signalling genes, prevented metal uptake inhibition by the fungus. Coculture with S. ruberrimus significantly changed the expression of Fe homeostasis genes: IRT1, OPT3, OPT6, bHLH38 and bHLH39 in wild-type (WT) A. thaliana. The expression pattern of these genes in WT plants and in the ethylene signalling defective mutants significantly differed and coincided with the plant accumulation phenotype. Most notably, down-regulation of the expression of IRT1 solely in WT was necessary for the inhibition of metal uptake in plants. This study shows that microorganisms optimize plant Fe and Ni uptake by fine-tuning plant metal homeostasis.

Phytomining of nickel (Ni) refers to cropping of selected Ni hyperaccumulator plants on Ni-rich serpentine soils. In this study, the effect of different fertilization regimes on the Ni yield of Odo...

AimsMetal hyperaccumulation is a rare phenomenon described for an increasing number of plant taxa. In this study we investigated the root growth responses of the well-known nickel, zinc, cadmium hyperaccumulatorNoccaea caerulescensand of the metal tolerant (non-accumulator)Stellaria media, in order to observe root foraging vs avoidance responses to nickel. MethodsTo allow for observations of root growth and foraging preferences, two accessions ofNoccaea caerulescensand two accessions ofStellaria mediaorginating from high nickel and low nickel habitats were grown in rhizotrons with localized nickel enrichment.ResultsThe root density in the control and nickel-enriched soil areas in the rhizotrons with differentN. caerulescensaccessions had distinct responses: moderate nickel avoidance was recorded for the non-nickel accession, while a clear foraging response was observed inN. caerulescensfrom the nickel accession. In contrast, nickel rooting avoidance was observed for bothS. mediaaccessions and was more pronounced in the non-nickel accession.ConclusionsThis study shows thatN. caerulescensoriginating from different accessions responded differently to soil nickel enrichment, with the nickel accession ofN. caerulescensactively foraging for nickel, suggesting a physiological adaptation and demand for this metal. In contrast, a clear nickel avoidance response by a metal tolerant species, S. media, was observed in this study, a phenomenon which has not been previously described; this suggests that root avoidance responses might play a role in the adaptation of metal tolerant species to Ni-rich soils.

The remediation of Pb, Cd, and Zn contaminated soil by ex situ EDTA washing was investigated in two pot experiments. We tested the influence of (i) 0, 0.5, 1.0, and 1.5%wt zero-valent iron (ZVI) and (ii) a combination of 5%wt vermicompost, 2%wt biochar, and 1%wt ZVI on the metal availability in EDTA-washed soil using different soil extracts (Aqua regia, NH4NO3) and plant concentrations. We found that EDTA soil washing significantly reduced the total concentration of Pb, Cd, and Zn and significantly reduced the Cd and Zn plant uptake. Residual EDTA was detected in water extracts causing the formation of highly available Pb-EDTA complexes. While organic amendments had no significant effect on Pb behavior in washed soils, an amendment of ≥ 1%wt ZVI successfully reduced EDTA concentrations, Pb bioavailability, and plant uptake. Our results suggest that Pb-EDTA complexes adsorb to a Fe oxyhydroxide layer, quickly developing on the ZVI surface. The increase in ZVI application strongly decreases Zn concentrations in plant tissue, whereas the uptake of Cd was not reduced, but even slightly increased. Soil washing did not affect plant productivity and organic amendments improved biomass production. Supplementary Information The online version contains supplementary material available at 10.1007/s11270-021-05356-0.

Urban gardening has become increasingly popular, creating green oases in cities, however, many of these activities are undertaken in areas of high traffic density or on ex-brown field sites. As a consequence, there are still some barriers to the adoption of these urban gardening practices for food production. One of the public concerns is the transfer of urban pollutants such as heavy metals into the consumer’s food chain, however, city-wide data is often difficult and expensive to collect. In the citizen science project described herein, we conducted simple citizen-led common collaborative experiments in urban community gardens. These data provided information on the potential risk of heavy metal contaminants and ways in which to mitigate those risks in an urban gardening context. Generally, values were below guideline thresholds, however, at a few garden sites, soil trace metal concentrations (Pb, Cd, Zn) exceeded Austrian recommended limits. Moreover, only at two sites were plant trace metal concentrations shown to be above European food standards limits. Given the citizen’s positive response to the project, we suggest expanding this study to the whole of Vienna, giving newly established gardens a chance to predetermine the risks posed by their local soils.

Visualization and quantification of corrosion processes is essential in materials research. Here we present a new approach for 2D spatiotemporal imaging of metal corrosion dynamics in situ. The approach combines time-integrated Mg

Pteris vittata (PV) and Pteris quadriaurita (PQ) are reported to hyperaccumulate arsenic (As) when grown in As-rich soil. Yet, little is known about the impact of their unique As accumulation mechanisms on As transformations and cycling at the soil-root interface. Using a combined approach of two-dimensional (2D), sub-mm scale solute imaging of arsenite (AsIII), arsenate (AsV), phosphorus (P), manganese (Mn), iron (Fe) and oxygen (O2), we found localized patterns of AsIII/AsV redox transformations in the PV rhizosphere (AsIII/AsV ratio of 0.57) compared to bulk soil (AsIII/AsV ratio of ≤0.04). Our data indicate that the high As root uptake, translocation and accumulation from the As-rich experimental soil (2080 mg kg−1) to PV fronds (6986 mg kg−1) induced As detoxification via AsV reduction and AsIII root efflux, leading to AsIII accumulation and re-oxidation to AsV in the rhizosphere porewater. This As cycling mechanism is linked to the reduction of O2 and MnIII/IV (oxyhydr)oxides resulting in decreased O2 levels and increased Mn solubilization along roots. Compared to PV, we found 4-fold lower As translocation to PQ fronds (1611 mg kg−1), 2-fold lower AsV depletion in the PQ rhizosphere, and no AsIII efflux from PQ roots, suggesting that PQ efficiently controls As uptake to avoid toxic As levels in roots. Analysis of root exudates obtained from soil-grown PV showed that As acquisition by PV roots was not associated with phytic acid release. Our study demonstrates that two closely-related As-accumulating ferns have distinct mechanisms for As uptake modulating As cycling in As-rich environments., Environmental and Experimental Botany, 177, ISSN:0098-8472, ISSN:1873-7307

Phytomining of nickel (Ni) is based on the cropping of Ni hyperaccumulators on Ni-rich serpentine soils. The efficiency of this approach is dependent on shoot nickel concentration and harvestable biomass. In a field experiment conducted on an Austrian serpentine site, the phytomining efficiency of the two plant species Odontarrhena chalcidica (syn. Alyssum murale) and Noccaea goesingensis was evaluated. O. chalcidica was planted in three treatments: control, sulphur application (0.46 g S kg−1 soil) and intercropping with the legume Lotus corniculatus. For N. goesingensis the treatments control, high-density planting (110 plants m−2) and intercropping were implemented. Given the experimental set-up, shoot biomass, shoot Ni concentration and thus the total amount of harvested Ni were on average higher for O. chalcidica. The highest Ni yield was achieved with O. chalcidica, reaching 55 kg Ni ha−1 in the sulphur treatment. N. goesingensis showed the maximum yield in the high-density treatment with 36 kg Ni ha−1. However, high-density planting of N. goesingensis and sulphur application to O. chalcidica plots did not significantly increase the Ni yield compared to the control. Intercropping with L. corniculatus tended to decrease the shoot biomass of both species. Planting of the hyperaccumulators led to a decrease of DTPA-extractable Ni and to an increase of soil pH, with the exception of sulphur-amended plots. Likewise, rhizosphere soil pH was higher than bulk soil values. Our data suggest that in particular O. chalcidica is suitable for Ni phytomining on the tested site. Measures to further increase the Ni yield and to optimise crop management will be evaluated in follow-up experiments.

Petroleum contamination and its remediation via plant-based solutions have got increasing attention by environmental scientists and engineers. In the current study, the physiological and growth responses of two diesel-tolerant plant species (tolerance limit: 1500-2000 mg/kg), Italian ryegrass (Lolium multiflorum) and Birdsfoot trefoil (Lotus corniculatus), have been investigated in vegetable oil- and diesel oil-amended soils. A long-term (147-day) greenhouse pot experiment was conducted to differentiate the main focus of the study: physical and chemical effects of oil (vegetable and diesel) in freshly spiked soils via evaluating the plant performance and hydrocarbon degradation. Moreover, plant performance was evaluated in terms of seed germination, plant shoot biomass, physiological parameters, and root biomass. Addition of both diesel oil and vegetable oil in freshly spiked soils showed deleterious effects on seedling emergence, root/shoot biomass, and chlorophyll content of grass and legume plants. Italian ryegrass showed more sensitivity in terms of germination rate to both vegetable and diesel oil as compared to non-contaminated soils while Birdsfoot trefoil reduced the germination rate only in diesel oil-impacted soils. The results of the current study suggest that both physical and chemical effects of oil pose negative effects of plant growth and root development. This observation may explain the phenomenon of reduced plant growth in aged/weathered contaminated soils during rhizoremediation experiments.

Purpose: Rhizosphere processes are known to modify uptake of elements from soil, but limited information is available for hyperaccumulators. We investigated labile Ni fractions and their kinetics of replenishment in the rhizospheres of the Ni-hyperaccumulator Odontarrhena serpyliffolia, the Ni-excluder Holcus lanatus and in bulk soils collected at the same serpentine outcrop. Methods: Labile Ni fractions in rhizosphere and bulk soil were evaluated using conventional extractions and also by the Diffusive Gradients in Thin Films (DGT) technique. DGT data were used to predict the kinetics of Ni resupply using DIFS modelling. Chemical imaging of Ni distribution along roots using DGT coupled with laser-ablation was conducted. Results: Labile Ni forms were higher in both rhizosphere than in bulk soils, together with an increase in dissolved organic C, cation exchange capacity and the Ca/Mg ratio. Ni fractionation indicated a shift towards less stable Ni fractions in the rhizosphere, particularly in the hyperaccumulator. DIFS modelling showed that the rhizosphere of the excluder was able to sustain the initially lower soluble Ni concentration through replenishment from the solid phase, while Ni resupply in the rhizosphere of the hyperaccumulator was not sufficient to maintain the initially high concentrations of soluble Ni. However, the amount of DGT-labile Ni was higher in the rhizosphere of the hyperaccumulator compared to the excluder in all deployment times. Conclusion: Our data suggest that compounds derived from root activity, in particular DOC, are important controls of Ni availability to plants growing on serpentine soil.

While the Galapagos Islands have been renowned for their unique flora and fauna since the time of Charles Darwin, the soils of the isolated island chain have been mostly overlooked and little information on their heavy metal contents is available. The aim of this study was therefore to examine the total heavy metal (Cd, Co, Cr, Cu, Ni, Pb, U, Zn) contents of soils from the agricultural areas on islands Isabela, Santa Cruz and San Cristobal, and identify trends with duration of exposure to weathering processes. Additionally, the mobility of these elements was assessed using ammonium nitrate extraction. In general, levels of Cd, Co, Cr, Cu, Ni and Zn were high compared to other world locations, while Pb levels were low and U levels were similar. Ni, Co, Cr, and to a lesser extent Pb and U tended to accumulate with increasing weathering duration. Soil concentrations of Cd, Zn, Cu, and possibly Pb and U, may have been influenced by use of agrochemicals, particularly on Santa Cruz Island. Mobility of Cd displayed an increasing trend with soil age, while Ni mobility decreased. Many soils had total contents of Cd, Co, Cr, Cu, Ni and Zn above threshold values indicating possible ecological or health risks. Systematic examination of trace element contents in soils from pristine national park areas would further assist in the delineation of background levels and the development of soil quality standards to ensure crop quality, animal and human health on this unique island chain.

The reliable sampling of root exudates in soil grown plants is experimentally challenging. This study aimed at developing a citrate sampling and mapping technique with millimetre-resolution using DGT (diffusive gradients in thin films) ZrOH binding gels. Citrate adsorption kinetics, DGT capacity and stability of ZrOH gels were evaluated. ZrOH gels were applied to generate 2D maps of citrate exuded by white lupin roots grown in rhizotrosn in a phosphorus deficient soil. Citrate was adsorbed quantitatively and rapidly by the ZrOH gels, these gels can be stored after sampling for several weeks prior to analysis. The DGT capacity of the ZrOH gel for citrate depends on the ionic strength and the pH of the soil solution but was suitable for citrate sampling. 2D citrate maps of rhizotron grown plants have been generated for the first time at a millimetre resolution to measure an illustrated plant response to P fertilization. DGT-based citrate sampling is suitable for studying the root exudation in soil environments, at unprecedented spatial resolution. By changing binding material, the technique is also applicable to other exudate classes and might be used for the evaluation of whole root exudation crucial in specific cultivar breeding.HighlightWe present a novel, reliable, easy to use, non-destructive citrate sampling- and two-dimensional high-resolution imaging technique for soil grown plant roots.

Initial experiments using Mediterranean Ni-hyperaccumulator plants for the purpose of phytomining were carried out in the 1990s. In order to meet commercial phytoextraction requirements, a technology has been developed using hyperaccumulator species with adapted intensive agronomic practices on naturally Ni-rich soils. Ultramafic soils in the Balkans and other parts of Europe display a great variability in Ni concentrations and available Ni levels. In Albania, Vertisols are currently being used for low-productivity agriculture (pasture or arable land) on which phytomining could be included in cropping practices. In northwestern Greece (Pindus and Vourinos mountain regions), agricultural soils may occur on ultramafic Cambisols. In Spain and Austria, these soils are much more erratically distributed and are seldom used for crops and pastures. In the Balkans, Odontarrhena chalcidica (synonym Alyssum murale) occurs widely on these ultramafic soils and is a spontaneous weed that grows among other crops. Field studies across Europe have now been carried out outside Mediterranean areas and have evaluated the Ni-hyperaccumulator O. chalcidica, as well as two other species native to northwestern Greece (Bornmuellera emarginata and B. tymphaea). At each site, local hyperaccumulator plants were tested for comparison (Noccaea goesingense in Austria and Odontarrhena serpyllifolia s.l. in Spain), in the context of two recent EUfunded projects (by Agronickel and LIFEAgromine). Soil and crop management practices are being developed in order to optimize the Ni agromining process. Field studies have evaluated the potential benefits of fertilization regimes, crop selection, and cropping patterns (introducing agroecological practices), and bioaugmentation using plant-associated microorganisms.

The reliable sampling of root exudates in soil grown plants is experimentally challenging. This method shows how to sample citrate from rhizotron grown plants with ZrOH DGT gels to produce high resolution citrate imaging. The DGT technique is based on thin hydrogels with homogeneously distributed analyte-selective binding phases and is used for 2D visualization and quantification of the distribution of labile (i.e., reversibly adsorbed) analytes.

Microbacterium species have been isolated from a wide range of hosts and environments, including heavy metal-contaminated sites. Here, we present a comprehensive analysis on the phylogenetic distribution and the genetic potential of 70 Microbacterium belonging to 20 different species isolated from heavy metal-contaminated and non-contaminated sites with particular attention to secondary metabolites gene clusters. The analyzed Microbacterium species are divided in three main functional clades. They share a small core genome (331 gene families covering basic functions) pointing to high genetic diversity. The most common secondary metabolite gene clusters encode pathways for the production of terpenoids, type III polyketide synthases and non-ribosomal peptide synthetases, potentially responsible of the synthesis of siderophore-like compounds. In vitro tests showed that many Microbacterium strains produce siderophores, ACC deaminase, auxins (IAA) and are able to solubilize phosphate. Microbacterium isolates from heavy metal contaminated sites are on average more resistant to heavy metals and harbor more genes related to metal homeostasis (e.g., metalloregulators). On the other hand, the ability to increase the metal mobility in a contaminated soil through the secretion of specific molecules seems to be widespread among all. Despite the widespread capacity of strains to mobilize several metals, plants inoculated with selected Microbacterium isolates showed only slightly increased iron concentrations, whereas concentrations of zinc, cadmium and lead were decreased.

This work aimed to investigate the metal accumulation characteristics as well as biogeochemical changes in the rhizosphere and root foraging strategies of this plant species. Previous reports suggested that Noccaea rotundifolia ssp. cepaeifolia is a Zn, Cd and Pb hyperaccumulator. We used hydroponic, rhizobox and split-pot experiments for studying metal accumulation and related rhizosphere processes. Although this species accumulated up to 1250 mg Pb kg−1 and 27,000 mg Zn kg−1 in shoots, translocation factors

The determination of citrate exuded from soil-grown roots is very challenging due to its rapid microbial degradation and mineralization, sorption to the solid soil phase and ongoing release of organic molecules from organic matter breakdown. For this reason, our knowledge about citrate release is mainly based on experiments carried out in hydroponics. Results obtained in hydroponics cannot directly be transferred to soil-plant systems, as hydroponics represents an artificial environment. This study aimed to develop a localization and quantification technique for citrate exuded from soil-grown plant roots, based on diffusive gradients in thin film (DGT). Polyacrylamide gels containing precipitated zirconium hydroxide (ZrOH) were applied to the rhizosphere of soil grown plants, on which citrate is efficiently immobilized, thereby creating a zero sink to sample the citrate exuded from the roots. Citrate was eluted with 1 mL 0.5 mol L-1 NaOH from the ZrOH gel and quantified by ion chromatography. ZrOH gel discs were able to bind the citrate contained in 10 mL of 2.77 mg citrate L-1 solutions within a 4h uptake period. Elution efficiency was ~89%. ZrOH gel capacity at pH 8 was 200 µg per gel disc and 299 µg per gel disc at pH 4, which is sufficient to act as a zero sink for citrate released from plant roots. As a first exemplary method application, we grew white lupin plants in rhizotrons using a highly phosphorus deficient soil. ZrOH gel sheets were applied for 26 h onto cluster roots for citrate sampling following established DGT protocols. Gels were cut afterwards into 5×5 and 5×2 mm slices for obtaining a citrate exudation map. In both cases we were able to localize and quantify up to 7.89 µg citrate on individual gel slices, as well as to identify longitudinal and lateral citrate gradients around the cluster roots. Moreover, the characterization of ZrOH gels showed its suitability for citrate sampling in terms uptake kinetics and capacity. These results demonstrate that the developed method is suitable for citrate sampling and localization in a non-destructive way from soil-grown plant roots. As it is applicable to soil grown-roots and provides unprecedented spatial resolution, this sampling technique advances the experimental possibilities for researching root exudates considerably. Using suitable binding materials, this approach is also applicable to other carboxylates such as malate or oxalate and other compound classes such as phenolics, flavonolos etc. Furthermore, this technique can be combined with complementary imaging methods for mapping e.g. nutrients, contaminants, pH or enzyme activity distributions.

Nickel (Ni) is used in numerous industrial processes, with large amounts of Ni-rich industrial wastes produced, which are largely sent to landfill. Nickel recovery from waste materials that would otherwise be disposed is of particular interest. Nickel phytomining represents a new technology in which hyperaccumulator plants are cultivated on Ni-rich substrates for commercial metal recovery. The aim of this study was to investigate the possibility of Ni transfer from industrial waste into plant biomass, to support recovery processes from bio-ores. Different industrial galvanic sludges (containing 85–150 g kg−1 Ni) were converted into artificial substrates (i.e. technosols) and the Ni hyperaccumulator Odontarrhena chalcidica (formerly Alyssum murale) was cultivated on these Ni-rich matrices. A greenhouse pot experiment was conducted for three months including an ultramafic soil control and testing fertilized (NPK) and unfertilized replicates. The results showed that fertilization was effective in improving plant biomass for all the substrates and that O. chalcidica was capable of viably growing on technosols, producing a comparable biomass to O. chalcidica on the control (ultramafic soil). On all technosols, O. chalcidica achieved Ni shoot concentrations of more than >1000 mg Ni kg −1 and maximum Ni uptake was obtained from one of the technosols (26.8 g kg −1 Ni, unfertilized; 20.2 g kg −1 Ni, fertilized). Nickel accumulation from three of the technosols resulted to be comparable with the control ultramafic soil. This study demonstrated the feasibility of transferring Ni from toxic waste into the biomass of Odontarrhena chalcidica and that phytomining from galvanic sludge-derived technosols can provide similar Ni yields as from natural ultramafic soils.

The remediation of petroleum hydrocarbon-contaminated soils has remained a challenging issue for environmental scientists and engineers. Within the scope of bioremediation; rhizoremediation has emerged as the most suitable method for petroleum-impacted soils. It can be promoted by the proper selection of suitable plant-microbe combinations and its overall efficiency can be enhanced by adding suitable soil amendments. This review article summarizes the available knowledge on rhizoremediation with respect to plant selection, inoculation with specific microbial strains (hydrocarbon-degrading as well as plant growth-promoting microbes) and a variety of soil amendments for enhancing the remediation efficiency. A large set of organic and inorganic soil amendments has been presented for the discussion on potential increase of rhizospheric effects. Finally, we discussed a unique multi-process phytoremediation system (MPPS) for the remediation of petroleum-polluted soils along with its field applications.

Co-contaminations of soils with organic and inorganic pollutants are a frequent environmental problem. Due to their toxicity and recalcitrance, the heterogeneous pollutants may persist in soil. The hypothesis of this study was that degradation of polycyclic aromatic hydrocarbons (PAHs) is enhanced if heavy metals in soil are immobilized and their bioavailability reduced. For metal immobilization and enhanced biodegradation, distinct mineral and organic soil amendments (iron oxides, gravel sludge, biochar) were deployed in an incubation batch experiment. The second part of the experiment consisted of a greenhouse pot experiment applying fast-growing and pollution-tolerant woody plants (willow and black locust). Soil amendments initially immobilized NH4NO3-extractable zinc, cadmium, and lead; after 100 days of incubation, soil amendments showed reductions only for cadmium and a tendency to enhance arsenic mobility. In order to monitor the remediation success, a 13C-phenanthrene (PHE) label was applied. 13C-phospholipid fatty acid analysis (13C-PLFA) further enabled the identification of PHE-degrading soil microorganisms. Both experiments exhibited a similar PLFA profile. Gram-negative bacteria (esp. cy17:0, 16:1ω7 + 6, 18:1ω7c) were the most significant microbial group taking up 13C-PHE. Plants effectively increased the label uptake by gram-positive bacteria and increased the biomass of the fungal biomarker, although their contribution to the degradation process was minor. Plants tended to prolong PAH dissipation in soil; at the end of the experiment, however, all treatments showed equally low total PAH concentrations in soil. While black locust plants tended not to take up potentially toxic trace elements, willows accumulated them in their leaves. The results of this study show that the chosen treatments did not enhance the remediation of the experimental soil.

The availability of iron (Fe) is highly limited for plant uptake in calcareous soils. To overcome this challenge, plants have developed different strategies of Fe mobilization. Non-gramineous species employ a reduction-based strategy (strategy I), which is assisted by the release of Fe(III)-chelating and -reducing coumarins. In this study, the coumarin release by three strategy-I plant species of the Brassicaceae family, i.e. Brassica napus L., Raphanus sativus L. and Sinapis alba L., on calcareous soil was examined using the RHIZOtest approach. With this experimental setup, the plants were first grown hydroponically in a cylindrical growth unit for 12 days, which was then subsequently brought in contact with calcareous, Fe deficient soil for another 9 days. Root exudates were collected in both experimental phases. Total carbon (C) release, coumarin exudation rates, trace metal plant uptake as well as trace metal mobilization were determined in response to Fe deficiency after 3, 6 and 9 days of soil exposure. High total C exudation rates indicated a clear stress response upon Fe deficiency during the soil contact period. Six different coumarins were detected, with scopoletin being predominantly released by all three plant species during the hydroponic stage with zero Fe supply. The exudation rate of all detected coumarins was significantly lower during soil exposure and represented only a very small fraction of the total organic compounds released. Although the role of coumarins in Fe mobilization is very complex and still not completely understood, it seems that coumarins are less important in Fe mobilization by the three investigated plant species when grown on soil compared to phytosiderophore-releasing strategy-II plants. Our data further indicate that plants growing on a Fe-deficient soil release less coumarins than in an artificial nutrient solution setup with zero Fe supply, which highlights the importance of determining root exudation rates from soil-grown plants, since data from hydroponic studies are likely not representative for soil-grown plants.

The in situ stabilization of multielement-contaminated agricultural soils has limited effectiveness when using common single amendments. This study examined the use of drinking water treatment residues (WTR), based on (hydr)oxides of Fe, Al, or Mn, as a cost-effective solution to optimize the immobilization of metals (Cd, Pb, Zn) and As. Trace elements (TE) bioavailability was assessed under semi-controlled conditions in a pot study cultivating winter wheat (Triticum aestivum L. cv. Tiger) until maturity. An Fe-based WTR and a Mn-based WTR, applied at rates of 0.5 and 1% (m/m), were related to effects of lime marl (LM) application. Additionally, a bioassay with earthworms (Dendrobaena veneta) was conducted. Both bioassays were compared with measurements of NH4NO3-soluble, diffusive gradients in thin film (DGT)-available and soil solution TE concentrations, representing well-established surrogates for mimicking the bioavailable element fractions in soil. The application of the Fe-based WTR reduced As accumulation in vegetative wheat tissues (by up to 75%) and earthworms (by up to 41%), which corresponded with the findings from soil chemical analyses and improved plant growth and earthworm body weight. However, As concentrations in cereal grains were not affected, Cd or Pb accumulation by wheat was not mitigated, and Zn uptake was enhanced. By contrast, the Mn-based WTR effected the greatest reduction in Pb uptake, and lowered Cd transfer to wheat grain (by up to 25%). Neither the NH4NO3-soluble nor DGT-available concentrations matched with Cd and Zn accumulation in plants or earthworms, indicating interferences due to competition for binding sites according to the biotic ligand model. The results obtained in this study suggest that a bioassay with key species prior to field application should be mandatory when designing in situ stabilization options. The application of WTR to an agricultural soil strongly affected TE bioavailability to plants and earthworms. Low application rates tended to improve biomass production of biota. Higher application rates involved risks (e.g., P fixation, TE inputs), and none of the amendments tested could immobilize all targeted elements.

We investigated to which extent phytosiderophores (PS), released by grasses for the acquisition of iron, solubilize other metals in contaminated soils, and how this affects metal mobilization and uptake in wheat plants. A plant-based bioassay (‘RHIZOtest’) and batch extraction scheme were carried out for assessing metal mobilisation in soil, PS exudation and metal accumulation in wheat. Increased PS exudation was observed in Fe-deficient wheat, leading to enhanced Zn, Cu, Mn and Ni concentrations in wheat shoots on some soils. In contrast, plant Cd and Pb concentrations were not affected. Likewise, in the batch experiment, strongly increased extractable Cu, Ni and Zn concentrations were observed, in particular when 100 or 1000 μM PS were added. Our results suggest that Fe deficiency can enhance the accumulation of some metals in shoots of grass species. Although our results indicate that the risk of enhanced accumulation of Cd and Pb in Fe deficient wheat shoots is rather small, further experiments conducted on soil for the complete vegetation period would be needed to confirm this observation.

Phytomining could represent an innovative low-cost technology for the selective recovery of valuable trace elements from secondary resources. In this context the potential of phytomining from waste incineration bottom ash was tested in a pot experiment. Fresh bottom ash was acidified, leached to reduce salinity and amended with organic material to obtain a suitable substrate for plant growth. Two hyperaccumulator species, Alyssum serpyllifolium subsp. lusitanicum and Sedum plumbizincicola as well as three metal tolerant species, Brassica napus, B. juncea and Nicotiana tabacum were tested for their phytomining potential on the pre-treated and amended bottom ashes from municipal solid waste and hazardous waste incineration. The hyperaccumulators had severe difficulties to establish on the bottom ash and to produce sufficient biomass, likely due to salinity and Cu toxicity. Nevertheless, concentrations of Ni in A. serpyllifolium and Zn in S. plumbizincicola were high, but total metal removal was limited by the low biomass production and was clearly less than on metalliferous soils. The Brassica species proved to be more tolerant to salinity and high Cu concentrations and produced considerably higher biomass, but total metal removal was limited by rather low shoot concentrations. The observed limitations of the phytomining process along with currently low market prices of Ni and Zn suggest that further optimisation of the process is required in order to make phytomining economically feasible on the tested waste incineration bottom ashes.

International audience; Brownfield surface is expanding in Europe, but as often abandoned or underused, these areas become refuge for microbial, faunal and floral biodiversity. However, brownfield sites are generally contaminated, likely posing severe environmental risks. At a former wood preservation site contaminated with Cu, we evaluated the efficiency of compost and dolomitic limestone incorporation into the soil, followed by revegetation with Cu-tolerant grassy species, as a phytomanagement option to increase vegetation cover and plant diversity while reducing pollutant linkages. 7 years of phytomanagement enhanced natural revegetation through the improvement of soil physicochemical properties, particularly with compost-based amendments. The compost incorporation increased soil Cu solubility; however, no increment in Cu availability and a reduction in Cu-induced phytotoxicity were observed with the compost. The improved soil nutrient availability and the soil phytotoxicity mitigation in compost-amended soils facilitated over the 7 years the growth of beneficial plant colonists, including leguminous species, which can potentially promote essential soil functions. Soil treatments did not affect Cu uptake and translocation by plants and shoot Cu levels indicated no risk for the food chain. Overall, a long-term phytomanagement combining an initial amendment of compost and dolomitic limestone with the cultivation of Cu-tolerant grassy populations can ameliorate such Cu-contaminated soils, by mitigating risks induced by Cu excess, ultimately allowing the development of a meadow that can provide ecological and economic benefits in terms of ecosystem services.Agrostis capillaris; Agrostis gigantea; Metal pollution; Organic amendments; Phytotoxicity; Phytoremediation

Root transcriptomic profile was comparatively studied in a serpentine (TM) and a non-metallicolous (NTM) population of Noccaea goesingensis in order to investigate possible features of Ni hyperaccumulation. Both populations were characterised by contrasting Ni tolerance and accumulation capacity. The growth of the TM population was unaffected by metal excess, while the shoot biomass production in the NTM population was significantly lower in the presence of Ni in the culture medium. Nickel concentration was nearly six- and two-fold higher in the shoots than in the roots of the TM and NTM population, respectively. The comparison of root transcriptomes using the RNA-seq method indicated distinct responses to Ni treatment between tested ecotypes. Among differentially expressed genes, the expression of IRT1 and IRT2, encoding metal transporters, was upregulated in the TM population and downregulated/unchanged in the NTM ecotype. Furthermore, differences were observed among ethylene metabolism and response related genes. In the TM population, the expression of genes including ACS7, ACO5, ERF104 and ERF105 was upregulated, while in the NTM population, expression of these genes remained unchanged, thus suggesting a possible regulatory role of this hormone in Ni hyperaccumulation. The present results could serve as a starting point for further studies concerning the plant mechanisms responsible for Ni tolerance and accumulation.

Human activities lead to increasing concentration of the stable elements cesium (Cs) and strontium (Sr) and their radioactive isotopes in the food chain, where plants play an important part. Here we investigated Plantago major under the influence of long-term exposure to stable Cs and Sr. The plants were cultivated hydroponically in different concentrations of cesium sulfate (between 0.002 and 20 mM) and strontium nitrate (between 0.001 and 100 mM). Uptake of Cs and Sr into leaves was analyzed from extracts by inductively coupled plasma mass spectrometry (ICP-MS). It was increased with increasing external Cs and Sr concentrations. However, the efficiency of Cs and Sr transfer from solution to plants was higher for low external concentrations. Highest transfer factors were 6.78 for Cs and 71.13 for Sr. Accumulation of Sr was accompanied by a slight decrease of potassium (K) and calcium (Ca) in leaves, whereas the presence of Cs in the medium affected only uptake of K. The toxic effects of Cs and Sr were estimated from photosynthetic reactions and plant growth. In leaves, Cs and Sr affected the chlorophyll fluorescence even at their low concentrations. Low and high concentrations of both ions reduced dry weight and length of roots and leaves. The distribution of the elements between the different tissues of leaves and roots was investigated using Energy Dispersive X-Ray microanalysis (EDX) with scanning electron microscope (SEM). Overall, observations suggested differential patterns in accumulating Cs and Sr within the roots and leaves. When present in higher concentrations the amount of Cs and Sr transferred from environment to plants was sufficient to affect some physiological processes. The experimental model showed a potential for P. major to study the influence of radioactive contaminants and their removal from hotspots.

The pollutant input and the risk of contamination by long-term application of recycled P fertilizers (RPFs) in European agricultural soils were estimated by a mass balance approach. Calculations based on literature data were carried out for the potentially toxic elements (PTEs) Cd, Cr, Cu, Ni, Pb, Zn and for the persistent organic pollutants (POPs) PCBs, PAHs and polychlorinated dibenzo-dioxins and -furans. The PTE accumulation estimation during 200 years of fertilizer application, equivalent to 11 kg P ha−1 year−1, the mean P export via harvested products on European stockless organic farms, regarded soil background concentrations and proposed threshold concentrations. For PTEs, inputs were fertilizer application, atmospheric deposition and liming, output processes were leaching and crop harvest. The effect of varying site conditions was assessed by considering two precipitation excess (F) values and two soil pH values. For POPs, fertilizer application and the half-life time were considered. The PTE accumulation risk was low for most RPFs. For the analysed POPs no accumulation was found. The highest accumulation was found for all PTEs at pH 7 and F = 0.1 m year−1, the lowest at pH 5 and F = 0.3 m year−1. A high P concentration in fertilizers resulted in a low PTE flow per unit of P. Composts had the highest PTE accumulation risk due to lowest P contents. Struvite, meat and bone meal, digestate of catering waste, ash and biosolids would be better suited as P fertilizers. The use of fertilizers should be regulated based on their pollutant-to-nutrient ratio.

Gentle remediation options (GRO), i.e. in situ stabilisation, (aided) phytoextraction and (aided) phytostabilisation, were implemented at ten European sites contaminated with trace elements (TE) from various anthropogenic sources: mining, atmospheric fallout, landfill leachates, wood preservatives, dredged-sediments, and dumped wastes. To assess the performance of the GRO options, topsoil was collected from each field trial, potted, and cultivated with lettuce (Lactuca sativa L.) for 48 days. Shoot dry weight (DW) yield, photosynthesis efficiency and major element and TE concentrations in the soil pore water and lettuce shoots were measured.GRO implementation had a limited effect on TE concentrations in the soil pore water, although use of multivariate Co-inertia Analysis revealed a clear amelioration effect in phytomanaged soils. Phytomanagement increased shoot DW yield at all industrial and mine sites, whereas in agricultural soils improvements were produced in one out of five sites. Photosynthesis efficiency was less sensitive than changes in shoot biomass and did not discriminate changes in soil conditions.Based on lettuce shoot DW yield, compost amendment followed by phytoextraction yielded better results than phytostabilisation; moreover shoot ionome data proved that, depending on initial soil conditions, recurrent compost application may be required to maintain crop production with common shoot nutrient concentrations.

Characterisation of geochemical transformations and processes in soils with special focus on the rhizosphere is crucial for assessing metal(loid) bioavailability to plants during in situ immobilisation and phytostabilisation. In this study, the effects of nano zero-valent iron (nZVI) were investigated in terms of the immobilisation of As, Zn, Pb and Cd in two soil types and their potential uptake by plants using rhizobox experiments. Such system allowed monitoring the behaviour of trace elements in rooted and bulk soil compartments separately. Sunflower (Helianthus annuus L.) and ryegrass (Lolium perenne L.) were tested for As-rich (15.9 g As kg−1) and Zn-rich (4.1 g Zn kg−1) soil samples, respectively. The application of nZVI effectively lowered the uptake of all target risk elements into plant tissues. Efficient immobilisation of As was determined in the As-soil without a significant difference between plant and bulk soil compartments. Similarly, a significant decrease was determined for CaCl2-available fractions of Zn, Pb and Cd in nZVI-treated Zn-soil. The behaviour of As corresponded to changes in Eh, while Zn and Cd showed to be mainly pH-dependent. However, despite the observed stabilisation effect of nZVI, high amounts of As and Zn still remained available for plants. Furthermore, the accumulation of the target risk elements in roots and the overall effect of nZVI transformations in the rhizosphere were verified and visualised by SEM/EDS. The following immobilising mechanisms were suggested: (i) sorption onto both existing and newly formed Fe (hydr)oxides, (ii) formation of secondary Fe-As phases, and (iii) sorption onto Mn (hydr)oxides.

In soil, mixed contamination with potentially toxic trace elements and polycyclic aromatic hydrocarbons (PAHs) may persist for a long time due to strong adsorption to the soil matrix and to its toxicity to microorganism. We conducted an incubation batch experiment to test the effect of soil amendments (biochar, gravel sludge, iron oxides) on the immobilisation of trace elements. To monitor microbial degradation, a 13C-PHE (phenanthrene) label was introduced to soil for 13C-PLFA (phospholipid fatty acid) analysis. Soil amendments increased soil pH, reduced mobility of NH4NO3-extractable trace elements Cd and Zn, and increased mobile Cu. A small consortium of PHE degraders was identified mainly in the microbial groups of gram-negative bacteria and actinomycetes. The degradation process of PHE peaked 9days after incubation start. PAH concentrations remained constant in the soil within the 30-day incubation, except for the easily available 13C-PHE in the amended treatment. In order to test the effect of plants and soil amendments under more realistic conditions, we also conducted an outdoor pot experiment with black locust (Robinia pseudoacacia Nyirsegi). Furthermore, soil amendments increased the mobility of soil Cu and As and decreased the mobility of Cd, Pb and Sb. The uptake of trace elements to leaves was low. Σ 16 U.S. EPA PAHs were significantly reduced only in the combined treatment of black locust and soil amendments after 12months of plant growth. Soil amendment-assisted phytoremediation showed a high efficiency in PAH dissipation and may be a useful remediation technique for mixed contaminated soils.