In current study, the enhanced efficiency of copper (Cu) phytoremediation potential of Calendula officinalis L. was investigated in a Cu-spiked calcareous soil, using foliar and soil application of humic acid. For this purpose, in a greenhouse experiment, seedlings of C. officinalis were transferred to Cu-spiked soils (0, 250 and 500 mg/kg) and treated separately with soil (soil drench) and foliar (spraying plant leaves) humic acid applications at different levels (0, 10, 20 μM). The humic acid treatments were applied 2 weeks after transferring plant, and eventually the various biochemical-physiological traits and phytoremediation indices of Cu in C. officinalis were measured at (specific) time points. According to the results, C. officinalis grew normally without any toxicity signs in Cu-spiked soils, however with increasing the Cu levels, the dry weight biomass decreased and antioxidant enzymes activities increased. Both foliar and soil humic acid application in Cu-spiked soils increased dry weight biomass, photosynthetic pigment contents, Cu concentration, and bioconcentration factor (BCF). Furthermore, the application of this organic substance, obviously moderated the Cu stress since the antioxidant enzymes activities reduced compared to the control. Based on the results, the obtained translocation factor (TF) and BCF values of Cu, which were >1, indicated that this plant is a Cu-hyperaccumulator, which could extract Cu via phytoextraction mechanism. Generally, the results of this study showed that, among the humic acid treatments, application of 20 μM (especially soil drench application) had the best effect on increasing Cu phytoremediation efficiency in the studied soil and it recommended to enhance the efficiency of Cu phytoremediation in calcareous soils.

International audience | Gentle remediation options (GRO) are based on the combined use of plants, associated microorganisms and soil amendments, which can potentially restore soil functions and quality. We studied the effects of three GRO (aided-phytostabilisation, in situ stabilisation and phytoexclusion, and aided-phytoextraction) on the soil microbial biomass and respiration, the activities of hydrolase enzymes involved in the biogeochemical cycles of C, N, P, and S, and bacterial community structure of trace element contaminated soils (TECS) from six field trials across Europe. Community structure was studied using denaturing gradient gel electrophoresis (DGGE) fingerprinting of Bacteria, α- and β-Proteobacteria, Actinobacteria and Streptomycetaceae, and sequencing of DGGE bands characteristic of specific treatments. The number of copies of genes involved in ammonia oxidation and denitrification were determined by qPCR. Phytomanagement increased soil microbial biomass at three sites and respiration at the Biogeco site (France). Enzyme activities were consistently higher in treated soils compared to untreated soils at the Biogeco site. At this site, microbial biomass increased from 696 to 2352 mg ATP kg-1 soil, respiration increased from 7.4 to 40.1 mg C-CO2 kg-1 soil d-1, and enzyme activities were 2-11-fold higher in treated soils compared to untreated soil. Phytomanagement induced shifts in the bacterial community structure at both, the total community and functional group levels, and generally increased the number of copies of genes involved in the N cycle (nirK, nirS, nosZ, and amoA). The influence of the main soil physico-chemical properties and trace element availability were assessed and eventual site-specific effects elucidated. Overall, our results demonstrate that phytomanagement of TECS influences soil biological activity in the long term.

International audience | Gentle remediation options (GRO) are based on the combined use of plants, associated microorganisms and soil amendments, which can potentially restore soil functions and quality. We studied the effects of three GRO (aided-phytostabilisation, in situ stabilisation and phytoexclusion, and aided-phytoextraction) on the soil microbial biomass and respiration, the activities of hydrolase enzymes involved in the biogeochemical cycles of C, N, P, and S, and bacterial community structure of trace element contaminated soils (TECS) from six field trials across Europe. Community structure was studied using denaturing gradient gel electrophoresis (DGGE) fingerprinting of Bacteria, α- and β-Proteobacteria, Actinobacteria and Streptomycetaceae, and sequencing of DGGE bands characteristic of specific treatments. The number of copies of genes involved in ammonia oxidation and denitrification were determined by qPCR. Phytomanagement increased soil microbial biomass at three sites and respiration at the Biogeco site (France). Enzyme activities were consistently higher in treated soils compared to untreated soils at the Biogeco site. At this site, microbial biomass increased from 696 to 2352 mg ATP kg-1 soil, respiration increased from 7.4 to 40.1 mg C-CO2 kg-1 soil d-1, and enzyme activities were 2-11-fold higher in treated soils compared to untreated soil. Phytomanagement induced shifts in the bacterial community structure at both, the total community and functional group levels, and generally increased the number of copies of genes involved in the N cycle (nirK, nirS, nosZ, and amoA). The influence of the main soil physico-chemical properties and trace element availability were assessed and eventual site-specific effects elucidated. Overall, our results demonstrate that phytomanagement of TECS influences soil biological activity in the long term.

International audience | Dark septate endophytes (DSEs) can improve plant stress tolerance by promoting growth and affecting element accumulation. Due to its ability to accumulate high Cd, Zn, and Ni concentrations in its shoots, Noccaea caerulescens is considered a promising candidate for phytoextraction in the field. However, the ability of DSEs to improve trace element (TE) phytoextraction with N. caerulescens has not yet been studied. The aim of this study was therefore to determine the ability of five DSE strains, previously isolated from poplar roots collected at different TE-contaminated sites, to improve plant development, mineral nutrient status, and metal accumulation by N. caerulescens during a pot experiment using two soils differing in their level of TE contamination. Microscopic observations revealed that the tested DSE strains effectively colonised the roots of N. caerulescens. In the highly contaminated (HC) soil, a threefold increase in root biomass was found in plants inoculated with the Leptodontidium sp. Pr30 strain compared to that in the non-inoculated condition; however, the plant nutrient status was not affected. In contrast, the two strains Phialophora mustea Pr27 and Leptodontidium sp. Me07 had positive effects on the mineral nutrient status of plants without significantly modifying their biomass. Compared to non-inoculated plants cultivated on HC soil, Pr27- and Pr30-inoculated plants extracted more Zn (+ 30%) and Cd (+ 90%), respectively. In conclusion, we demonstrated that the responses of N. caerulescens to DSE inoculation ranged from neutral to beneficial and we identified two strains (i.e. Leptodontidium sp. (Pr30) and Phialophora mustea (Pr27)) isolated from poplar that appeared promising as they increased the amounts of Zn and Cd extracted by improving plant growth and/or TE accumulation by N. caerulescens. These results generate interest in further characterising the DSEs that naturally colonise N. caerulescens and testing their ability to improve phytoextraction.

Soil Cd and Zn contamination has become a serious environmental problem. This work explored the performance of wood vinegar (WV) in enhancing the phytoextraction of Cd/Zn by hyperaccumulator Sedum alfredii Hance. Rhizosphere chemical properties, enzyme activities and bacterial community were analyzed to determine the mechanisms of metal accumulation in this process. Results demonstrated that, after 120 days growth, different times dilution of WV increased the shoot biomass of S. alfredii by 85.2%–148%. In addition, WV application significantly increased soil available Cd and Zn by lowing soil pH, which facilitated plant uptake. The optimal Cd and Zn phytoextraction occurred from the 100 times diluted WV (D100), which increased the Cd and Zn extraction by 188% and 164%, compared to CK. The 100 and 50 times diluted WV significantly increased soil total and available carbon, nitrogen and phosphorus, and enhancing enzyme activities of urease, acid phosphatase, invertase and protease by 10.1–21.4%, 29.1–42.7%,12.2–38.3% and 26.8–85.7%, respectively, compared to CK. High-throughput sequencing revealed that the D 100 significantly increased the bacterial diversity compared to CK. Soil bacterial compositions at phylum, family and genera level were changed by WV addition. Compared to CK, WV application increased the relative abundances of genus with plant growth promotion and metal mobilization function such as, Bacillus, Gemmatimonas, Streptomyces, Sphingomonas and Polycyclovorans, which was positively correlated to biomass, Cd/Zn concentrations and extractions by S. alfredii. Structural equation modeling analysis showed that, soil chemical properties, enzyme activities and bacterial abundance directly or indirectly contributed to the biomass promotion, Cd, and Zn extraction by S. alfredii. To sum up, WV improved phytoextraction efficiency by enhancing plant growth, Cd and Zn extraction and increasing soil nutrients, enzyme activities, and modifying bacterial community.

In this work, arsenic (As) accumulation and distribution over time in Pteris vittata young fronds from adult plants and in whole plantlets, grown on a highly contaminated As-soil, was determined by μ-XRF. A linear increase in As content up to 60 days was found in young fronds at different times, and a progressive distribution from the apex to the base of the fronds was observed. In whole plantlets, As signal was detectable from 9 to 20 days in the apex of a few fronds and fiddleheads. Later, up to 60 days, As was localized in all fronds, in the rhizome and in basal part of the roots. The dynamics of expression of As-related genes revealed a good correlation between As content and the level of the As (III)-antiporter PvACR3 transcript in plantlets roots and fronds and in young fronds. Moreover, the transcription of As (V)-related gametophytic genes PvGAPC1, PvOCT4 increases over time during As accumulation while PvGSTF1 is expressed only in roots. Here, we demonstrate the suitability of the μ-XRF technique to monitor As accumulation, which allowed us to propose that As is initially directly transported to fiddleheads and apex of fronds, is later distributed to the whole fronds and simultaneously accumulated in the rhizome and roots. We also provide indications on the expression of candidate genes possibly involved in As (hyper)accumulation.

Arsenic contamination of ground water is a worldwide issue, causing a number of ailments in humans. As an engineered and integrated solution, a hybrid vertical subsurface flow constructed wetland (VSSF-CW) amended with BCXZM composite (Bacillus XZM immobilized on rice husk biochar), was found effective for the bioremediation of arsenic contaminated water. Biological filter was prepared by amending top 3 cm of VSSF-CW bed with BCXZM. This filter scavenged ∼64% of total arsenic and removal efficiency of ∼95% was achieved by amended and planted (As + P + B) VSSF-CW, while non-amended (As + P) VSSF-CW showed a removal efficiency of ∼55%. The unplanted and amended (As + B) VSSF-CW showed a removal efficiency of ∼70%. The symbiotic association of Bacillus XZM, confirmed by SEM micrographs, significantly (p ≤ 0.05) reduced reactive oxygen species (ROS) and malondialdehyde (MDA) accumulation in Typha latifolia, hence, increasing the plant growth (2 folds). An increase in the indole acetic acid (IAA) and arsenic accumulation in plant was also observed in As + P + B system. The removal efficiency of the system was compromised after 4th consecutive cycle and 48 h was observed as optimum retention time. The FTIR-spectra showed the involvement of -N-H bond, carboxylic acids, –CH₂ stretching of –CH₂ and –CH₃, carbonyl groups, -C-H, C–O–P and C–O–C, sulphur/thiol and phosphate functional groups in the bio-sorption of arsenic by BCXZM filter. Our study is a first reported on the simultaneous phytoextraction and biosorption of arsenic in a hybrid VSSF-CW. It is proposed that BCXZM can be applied effectively in CWs for the bioremediation of arsenic contaminated water on large scale.

The work was conducted to establish contamination from improper disposal of hazardous wastes containing lead (Pb) and antimony (Sb) into nearby soils. Besides other elements in the affected area, the biological role of Sb, its behaviour in the pedosphere and uptake by plants and the food chain was considered. Wastes contained 139532 ± 9601 mg kg−1 (≈14%) Pb and 3645 ± 194 mg kg−1 (≈0.4%) Sb respectively and variability was extremely high at a decimetre scale. Dramatically high concentrations were also found for As, Cd, Cu, Mn, Ni, Sn and Zn. In adjacent natural soils metal(oid)s amounts decreased considerably (Pb 5034 ± 678 mg kg−1, Sb 112 mg kg−1) though largely exceeded the directives for a given soil use. Metal(oid)s potential mobility was assessed by using H2O→KNO3→EDTA sequential extractions, and EDTA extracts showed the highest concentration suggesting stable humus-metal complexes formation. Nevertheless, selected plants showed high absorption potential of the investigated elements. Pb and Sb values for Dittrichia viscosa grown in wastes was 899 ± 627 mg kg−1 and 37 ± 33 mg kg−1 respectively. The same plant showed 154 ± 99 mg kg−1 Pb and 8 ± 4 mg kg−1 Sb in natural soils. Helichrysum stoechas had 323 ± 305 mg kg−1 Pb, and 8 ± 3 mg kg−1 Sb. Vitis vinifera from alongside vineyards contained 129 ± 88 mg kg−1 Pb and 18 ± 9 mg kg−1 Sb, indicating ability for metal uptake and warning on metal diffusion through the food chain. The biological absorption coefficient (BAC) and the translocation factor (TF) assigned phytoextraction potential to Dittrichia viscosa and Foeniculum vulgare and phytostabilization potential to Helichrysum stoechas. Dissolved metal (oid)s in the analysed water strongly exceeded the current directive being a direct threat for livings. Data warned against the high contamination of the affected area in all its compartments. Even though native plants growing in metal-contaminated sites may have phytoremediation potential, high risk of metal diffusion may threat the whole ecosystem.

Phytoextraction using hyperaccumulator, Pteris vittata, to extract arsenic (As) from soil has been applied to large areas to achieve an As removal rate of 18% per year. However, remarkable difference among different studies and field practices has led to difficulties in the standardization of phytoextraction technology. In this study, data on As concentration in P. vittata and related environmental conditions were collected through literature search. A conceptual framework was proposed to guide the improvement of phytoextraction efficiency in the field. The following influencing factors of As concentration in this hyperaccumulator were identified: total As concentration in soil, soil available As, organic matter in soil, total potassium (K) concentration in soil, and annual rainfall. The geodetection results show that the main factors that affect As concentration in P. vittata include soil organic matter (q = 0.75), soil available As (q = 0.67), total K (q = 0.54), and rainfall (q = 0.42). The predictive models of As concentration in P. vittata were established separately for greenhouse and field conditions through multivariate linear stepwise regression method. Under greenhouse condition, soil available As was the most important influencing factor and could explain 41.4% of As concentration in P. vittata. Two dominant factors were detected in the field: soil available As concentration and average annual rainfall. The combination of these two factors gave better prediction results with R² = 0.762. The establishment of the model might help predict phytoextraction efficiency and contribute to technological standardization. The strategies that were used to promote As removal from soil by P. vittata were summarized and analyzed. Intercropping with suitable plants or a combination of different measures (e.g., phosphate fertilizer and water retention) was recommended in practice to increase As concentration in P. vittata.

Heavy metal pollution is becoming recurrent and threatens biota biosafety in many agricultural fields. Diverse solutions explore the application of amendments to enable remediation. Sulfur represents a nonmetallic chemical element that actively affects heavy metals phytoextraction, and promotes and alternatively mitigates soil functions. In this study, we conduct a meta-analysis to synthesize the current knowledge on the influence of sulfur amendments on plants heavy metals uptake from contaminated soil media. Random-effects model was used to summarize effect sizes from 524 data points extracted from 30 peer reviewed studies. The phytoextraction of cadmium, chromium and nickel were 1.6-, 3.3-, and 12.6-fold, respectively, higher when sulfur amendment was applied; while copper uptake was 0.3-fold lower. Irrespective of the sulfur type, heavy metal extraction increased with the raising sulfur stress. Individual organs showed significant differences of heavy metal uptake between sulfur applied and non-sulfur treatments, and combined organs did not. The heavy metals uptake in leaves and roots were higher in sulfur applied than non-sulfur applied treatments, while those in grain, husk, and stalks were lower. The heavy metals phytoextraction (response ratio) followed the order roots > leaves > stalk > grain > husk. Moreover, heavy metals uptake was 2-fold higher in the sulfur applied than the non-sulfur treatments under ideal (5.5–8) and alkaline conditions (8–14), and 0.2-fold lower under acidic pH (1–5.5). Cadmium, manganese and nickel, and chromium were the most extracted under sulfur application by Vicia sp., Sorghum sp. and Brassica sp., respectively; while chromium, manganese, and iron were the most uptake without sulfur amendments by Oryza sp., Zea sp. and Sorghum sp., respectively. Our study highlights that the influence of sulfur on heavy metal phytoextraction depends on the single or combined effects of sulfur stress intensity, sulfur compounds, plant organ, plant type, and soil pH condition.