Recalcification stabilizes cadmium but magnifies phosphorus limitation in wastewater-irrigated calcareous soil.

Long-term wastewater irrigation leads to the loss of calcium carbonate (CaCO3) in the tillage layer of calcareous land, which irreversibly damages the soil's ability to retain cadmium (Cd). In this study, we selected calcareous agricultural soil irrigated with wastewater for over 50 years to examine the recalcification effects of sugar beet factory lime (SBFL) at doses of 0%, 2.5%, 5%, and 10%. We found that SBFL promoted Cd transformation in the soil from active exchangeable species to more stable carbonate-bonded and residual species, which the X-ray diffraction patterns also confirmed results that CdSO4 reduced while CdS and CaCdCO3 increased. Correspondingly, the soil bioavailable Cd concentration was significantly reduced by 65.6-84.7%. The Cd concentrations in maize roots and shoots were significantly reduced by 11.7-50.6% and 13.0-70.0%, respectively, thereby promoting maize growth. Nevertheless, SBFL also increased the proportion of plant-unavailable phosphorus (P) in Ca8-P and Ca10-P by 4.3-13.0% and 10.7-25.9%, respectively, reducing the plant-available P (Olsen P) content by 5.2-22.1%. Consequently, soil P-acquiring associated enzyme (alkaline phosphatase) activity and microbial (Proteobacteria, Bacteroidota, and Actinobacteria) community abundance significantly increased. Our findings showed that adding SBFL to wastewater-irrigated calcareous soil stabilized Cd, but exacerbated P limitation. Therefore, it is necessary to alleviate P limitations in the practice of recalcifying degraded calcareous land.

Comparison of organic and synthetic amendments for poplar phytomanagement in copper and lead-contaminated calcareous soil.

Soil remediation can be achieved through organic and synthetic amendments, but the differences in the phytomanagement of trace metal-contaminated land are unclear. We conducted an outdoor microcosm experiment to simulate the effects of organic amendment citric acid and synthetic amendments EDTA and EGTA on poplar phytomanagement of copper (Cu)- and lead (Pb)-contaminated calcareous land at doses of 0, 1, 3, and 9 mmol kg-1. We found that soil-bioavailable Cu and Pb contents increased by 2.11-27.27 and 1.48-269 times compared to the control, respectively. Additionally, synthetic amendments had a long-lasting (within 25 days) effect on metal bioavailability relative to organic amendments. Consequently, organic amendments increased the root Cu and Pb contents by 2.68-48.61% and 6.60-49.51%, respectively, whereas synthetic amendments increased them by 65.94-260% and 12.50-103%. The Cu and Pb contents in the leaves were lower than those in the roots, and increased significantly by 47.04-179% and 237-601%, respectively, only under synthetic amendments. Interestingly, none of the amendments increased the Cu and Pb content in poplar stems (<5 mg kg-1), which remained within the normal range for terrestrial plants. Regardless of the type and addition level, the amendments did not affect poplar growth. Nevertheless, synthetic amendments caused a significant redistribution of metals (Cu: 22-32%; Pb: 23-53%) from the topsoil into the subsoil within the root zone at medium and high levels relative to organic amendments. Therefore, organic and synthetic amendments can assist poplar phytomanagement with a phytostabilization strategy for Cu- and Pb-contaminated calcareous land and obtain marketable wood biomass. Moreover, collecting leaf litter is crucial when using synthetic amendments at optimum concentration levels.

Tradeoffs between pH, dissolved organic carbon, and mineral ions regulate cadmium uptake by Solanum hyperaccumulators in calcareous soil.

Soil solution pH and dissolved organic carbon (DOC) influence cadmium (Cd) uptake by hyperaccumulators but their tradeoff in calcareous soils is unclear. This study investigated the mechanisms of Solanum nigrum L. and Solanum alatum Moench in calcareous soil using a combination of concentration gradient experiments (0.6-100 mg Cd kg-1) and soil solution composition analysis. The results showed that the soil solution pH of S. nigrum remained stable despite Cd stress. On average, the soil solution pH of S. alatum was 0.23 units higher than that of S. nigrum, although pH decreased significantly under high Cd stress. In addition, the concentrations of potassium (K) and calcium (Ca) in the soil solution of S. nigrum increased and decreased under low and high levels of Cd stress, respectively. In S. alatum, the K and Ca concentrations in the soil solution generally increased with increasing Cd stress levels. Moreover, the level of DOC in the soil solution of both plants was higher under Cd stress compared to the control, and a gradually increasing trend with Cd stress level was observed in S. alatum. Consequently, the bioconcentration factors of the roots (2.62-19.35) and shoots (1.20-9.59) of both plants were >1, while the translocation factors were <1, showing an obstacle of Solanum hyperaccumulators in transferring Cd into their aboveground parts. Redundancy analysis revealed that the Cd concentration in S. nigrum roots was significantly negatively correlated with the soil solutions of K and Ca. In contrast, Cd concentrations in S. alatum roots and shoots were significantly positively correlated with soil solution DOC, K, and Ca but negatively correlated with pH. Our results suggest that calcareous soil neutralizes the acidity of released protons but does not affect cation exchange, inhibiting DOC in assisting the translocation of Cd within plants.

Exploring the safe utilization strategy of calcareous agricultural land irrigated with wastewater for over 50 years.

The trace element (TE) contamination of farmland caused by wastewater irrigation threatens food security and food safety. We selected a typical calcareous soil area in western China that has been irrigated with wastewater for >50 years to explore safe use strategies for flax farmland contaminated by cadmium (Cd) and arsenic (As). We found that Cd and As were mainly accumulated in flax roots rather than seeds. However, regardless of the type of TE and acceptor, direct ingestion of the flaxseed would seriously endanger human health (hazard quotient >1). According to the results of redundancy analysis and Pearson correlation analysis, the concentration of Cd and As in flaxseed depended on the concentration of soil total TE, Olsen phosphorus, dissolved organic carbon, soil organic matter, and active calcium carbonate (CaCO3). This was largely because the pH and total CaCO3 content in topsoil of flax farmland decreased by 1.05 units and 37 %, respectively, compared with their background levels before wastewater irrigation. Interestingly, after pressing, Cd and As in flaxseed transferred to flaxseed oil were 3.87-10.55 % and 17.21-30.48 %, respectively, which led to an acceptable risk of adults and children (hazard quotient <1) consuming flaxseed oil. Our results suggest that with the production of flaxseed oil as the goal, the long-term wastewater-irrigated calcareous land can be safely utilized while obtaining income.

Phosphorus supply level is more important than wheat variety in safe utilization of cadmium-contaminated calcareous soil.

Screening and cultivating crop varieties with low Cd accumulation is an effective way to safely utilize the Cd slightly contaminated soil. The characteristics and mechanism of Cd uptake by 13 wheat varieties in two calcareous soils with similar Cd contamination level but different P supply level were studied. The grain Cd concentration of almost all varieties in low-P soil was significantly higher than that in high-P soil and exceeded the maximum level of 0.2 mg kg-1 recommended by the Codex Alimentarius Commission. The pH value of low-P soil was significantly lower than that of high-P soil by 0.27 units, while leaf [Mn] (proxy for rhizosphere carboxylates) and the activities of soil alkaline phosphatase and phytase were significantly higher than those of high-P soil by 35%, 55%, and 286%, respectively. The exchangeable Cd concentration in low-P soil was 2.93 times higher than that in high-P soil, while the Cd concentration of oxides and organic species was significantly lower than that in high-P soil by 21% and 64%, respectively, collectively increasing soluble Cd concentration in low-P soil by 38%. In low-P calcareous soil, P mobilization induced the change of root-zone microenvironment, resulting in the mobilization of Cd.

Temporal changes of calcareous soil properties and their effects on cadmium uptake by wheat under wastewater irrigation for over 50 years.

Calcareous soil has a strong buffering capacity for neutralizing acid and stabilizing cadmium (Cd) because of the high calcium carbonate (CaCO3) content. However, it is not clear whether the buffering capacity of calcareous soil can be maintained after long-term wastewater irrigation. We selected a typical area in western China that has been irrigated with wastewater for over 50 years to study the temporal changes of soil properties and their effects on Cd uptake by wheat. The results showed that compared with the background level before the 1960s, the soil pH and CaCO3 content in 2018 were lower by 0.80 units and 35%, respectively, while the soil organic matter (SOM) content, Olsen phosphorus (P) content, and soil total Cd content in 2018 increased by 1.54, 13.05, and 164 times, respectively. Due to the significant decrease in the soil pH and CaCO3, the high load of soil total Cd and electrical conductivity, the low soil clay content, and the coupling of SOM with soil nitrogen and P, the input Cd was activated. Furthermore, the activated Cd was effectively taken up by wheat roots and transported to grains with the assistance of dissolved organic carbon. Our results highlight that long-term wastewater irrigation caused irreversible damage to soil buffering capacity, resulting in the Cd activation and the enhancement of Cd uptake by wheat.

Redistribution of calcium and sodium in calcareous soil profile and their effects on copper and lead uptake: A poplar-based phytomanagement.

Poplar serves as a phytostabilizator in phytomanagement of the trace metals (TMs) copper (Cu) and lead (Pb) contaminated land. In the process of long-term phytomanagement, it is not clear how the cycling of the mineral nutrients calcium (Ca) and sodium (Na) in calcareous soil will affect poplar remediation mechanisms. We selected a site contaminated by Cu and Pb and phytomanaged by Populus simonii Carr. stands of different ages (7, 14, and 28 years) to study the influencing mechanisms. The results showed that after afforestation, the Ca in the subsoil returned to the topsoil through fallen leaves, whereas the Na in the topsoil migrated downward to the subsoil by leaching, resulting in the redistribution of mineral nutrients in the soil profile. In addition, the Ca content in soil solution of the root-zone was significantly lower relative to that of the bulk soil, whereas the Na content in soil solution was significantly higher in all stands. As a result, because of the competitive adsorption of mineral nutrient and TM cations on the soil surface, the pool of bioavailable TM in root-zone soils did not significantly decrease with stand age. On the contrary, the TM content in poplar leaves (Cu: 31-37 mg kg-1; Pb: 62-84 mg kg-1) and litter (Cu: 230-790 mg kg-1; Pb: 394-1366 mg kg-1) increased significantly with stand age. Nevertheless, the TM content in poplar wood (Cu < 3 mg kg-1; Pb < 12 mg kg-1) remained at an extremely low level in all stands. Our results highlighted that strengthening leaf collection is necessary to eliminate ecological risks and ensure the safe production of poplar wood in the long-term phytomanagement of TM-contaminated land.

Impact of poplar-based phytomanagement on metal bioavailability in low-phosphorus calcareous soil with multi-metal contamination.

Bioavailability of trace metals (TMs) is the key component in the management of TM-contaminated soils; however, its impact mechanism is unclear in low-phosphorus (P) calcareous soils afforested by fast-growing tree species for a long duration (>10 years). We selected a site contaminated with multiple TMs and phytomanaged by poplar (Populus hopeiensis Hu & Chow) to study the impact mechanism of plant-soil interactions on TM bioavailability along a long-term chronosequence (i.e., 10, 15, 20, and 25 years). We found that phytomanagement significantly decreased soil organic carbon (SOC) content, soil total nitrogen (N) content, and soil C/P and N/P ratios with stand age, but did not significantly change soil total P content. In contrast, soil available P content significantly changed in rhizospheric soils compared with the bulk soil, suggesting the tight coupling between the amplification of P turnover and N availability. Soil pH in rhizospheric soils significantly decreased by 0.22 to 0.32 units, while calcium carbonate (CaCO3) content decreased by 14% to 39%, as compared with the bulk soil. Bioavailable concentrations of cadmium, lead, and zinc were positively correlated with soil available P, whereas bioavailable nickel concentration was negatively correlated with soil pH. Furthermore, TM bioavailability in rhizospheric soils significantly increased with stand age, regardless of the metal type. Our results suggest that P mobilization associated with SOC depletion induced soil acidification followed by CaCO3 dissolution, collectively leading to metal mobilization with stand age.

Ornamental hyperaccumulator Mirabilis jalapa L. phytoremediating combine contaminated soil enhanced by some chelators and surfactants.

Mirabilis jalapa L. is an ornamental plant of the composite family, which was found hyperaccumulating Cd. Due to its larger biomass, developed root system, root exudation, and microbial interactions, certain organic pollutants in its rhizosphere can be effectively degraded. Thus, M. jalapacan be used to co-remediate heavy metal and organic pollutant co-contaminated soil. The aim of this paper is to explore the remediation capacity of M. jalapa for Cd-PAHs co-contaminated soil in the presence of five chelators or surfactants. The concentrations of Cd and PAHs in collected soil samples were 0.85 mg kg-1 Cd and 1.138 mg kg-1 PAHs (16 kinds of priority control polycyclic aromatic hydrocarbons by USEPA). The chelators or surfactants of EDTA, EGTA, CA, TW80, and SA were respectively spiked to the pots according to the experiment design at 1 month before the plant harvested. The results showed that the capacity of Cd in shoot of M. jalapa was 7.99 µg pot-1 without any addition (CK4, M. jalapa in original soil without amendment). However, Cd capacity in shoot of M. jalapa was increased (p < 0.05) by 31.7%, 181.7%, and 107.4% in treatment of REGTA, RCA and REGTA + SA, respectively. As for the degradation of PAHs in soil, there was no significant decrease (p < 0.05) in the treatment of CK2 (original soil spiked with 0.9 SA without M. jalapa), CK3 (original soil spiked with 0.3 TW80 without M. jalapa), and CK4 compared to the control CK1 (original soil without M. jalapa and amendment). When amendments were added to soils with M. jalapa,the PAHs concentrations in soils significantly decreased (p < 0.05) by 21.7%, 23.8%, 27.0%, 19.8%, 21.8%, 31.2%, and 25.5% for the treatment of REDTA + SA, REDTA + T80, REGTA + SA, REGTA + T80, RCA + T80, RSA + T80 + EDTA, and RSA + T80 + CA, respectively. Basically, Cd capacity in shoot of M. jalapa was improved by chelators. PAHs degradation was caused by the existence of surfactants in rhizosphere of M. jalapa. But the roles of different chelators or surfactants were quite distinct. In short, the Cd capacity in the shoot and PAHs degradation in the rhizosphere of M. jalapa in the treatment of REGTA + SA were all significantly increased (p < 0.05), which was more practical for M. jalapa phytoremediating Cd-PAHs co-contaminated soil.

Temporal changes of metal bioavailability and extracellular enzyme activities in relation to afforestation of highly contaminated calcareous soil.

Metal bioavailability and extracellular enzyme activity are two important indicators of soil quality in metal-contaminated soil. However, it is unclear how the chronosequence effect modifies these two factors in highly contaminated calcareous soils undergoing afforestation. We used Populus simonii Carr. and the calciphilous Ulmus macrocarpa Hance as contrasting tree species to study the chronosequence effect. We found that afforestation significantly increased soil total nitrogen (N) content as well as soil carbon (C)/phosphorus (P) and N/P ratios, but decreased soil total P content and soil C/N ratio, regardless of the tree species and stand age, suggesting strong P limitation. However, available P did not change significantly with stand age. In both tree species, P mobilization depleted soil organic matter through the priming effect of dissolved organic carbon, whereas the decrease in soil pH in the U. macrocarpa stands enhanced CaCO3 dissolution, collectively reducing the capacity of the soil to immobilize metals, resulting in increased metal bioavailability with stand age. The activity of oxidase (dehydrogenase) was positively correlated with bioavailable zinc concentration, soil electrical conductivity, and soil total N content. Hydrolase activities (alkaline phosphatase, ß-glucosidase, and urease) were significantly positively correlated with the ratios of soil C/N and C/P, soil pH, and CaCO3, but negatively correlated with soil N/P ratio and bioavailable cadmium concentration. Increasing stand age was associated with the gradual recovery of oxidase activity and remarkable inhibition of hydrolase activity. Our results suggest that the combination of soil hydrolase activity and metal bioavailability can predict soil quality in the afforestation of highly contaminated soils.

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.

Heavy metal accumulation by poplar in calcareous soil with various degrees of multi-metal contamination: implications for phytoextraction and phytostabilization.

The object of this study was to assess the capacity of Populus alba L. var. pyramidalis Bunge for phytoremediation of heavy metals on calcareous soils contaminated with multiple metals. In a pot culture experiment, a multi-metal-contaminated calcareous soil was mixed at different ratios with an uncontaminated, but otherwise similar soil, to establish a gradient of soil metal contamination levels. In a field experiment, poplars with different stand ages (3, 5, and 7 years) were sampled randomly in a wastewater-irrigated field. The concentrations of cadmium (Cd), Cu, lead (Pb), and zinc (Zn) in the poplar tissues and soil were determined. The accumulation of Cd and Zn was greatest in the leaves of P. pyramidalis, while Cu and Pb mainly accumulated in the roots. In the pot experiment, the highest tissue concentrations of Cd (40.76 mg kg(-1)), Cu (8.21 mg kg(-1)), Pb (41.62 mg kg(-1)), and Zn (696 mg kg(-1)) were all noted in the multi-metal-contaminated soil. Although extremely high levels of Cd and Zn accumulated in the leaves, phytoextraction using P. pyramidalis may take at least 24 and 16 years for Cd and Zn, respectively. The foliar concentrations of Cu and Pb were always within the normal ranges and were never higher than 8 and 5 mg kg(-1), respectively. The field experiment also revealed that the concentrations of all four metals in the bark were significantly higher than that in the wood. In addition, the tissue metal concentrations, together with the NH4NO3-extractable concentrations of metals in the root zone, decreased as the stand age increased. P. pyramidalis is suitable for phytostabilization of calcareous soils contaminated with multiple metals, but collection of the litter fall would be necessary due to the relatively high foliar concentrations of Cd and Zn.

Chemical-assisted phytoremediation of CD-PAHs contaminated soils using Solanum nigrum L.

A well-characterized cadmium (Cd) hyperaccumulating plant Solanum nigrum was grown in Cd and polycyclic aromatic hydrocarbons (PAHs) co-contaminated soil that was repeatedly amended with chemicals, including EDTA, cysteine (CY), salicylic acid (Sa), and Tween 80 (TW80), to test individual and combined treatment effects on phytoremediation of Cd-PAHs contaminated soils. Plant growth was negatively affected by exogenous chemicals except for EDTA. S. nigrum could accumulate Cd in tissues without assistant chemicals, while there was no visible effect on the degradation of PAHs. Cysteine had significant effects on phytoextraction of Cd and the highest metal extraction ratio (1.27%) was observed in 0.9 mmol/kg CY treatment. Both salicylic acid and Tween 80 had stimulative effects on the degradation of PAHs and there was the maximal degradation rate (52.6%) of total PAHs while 0.9 mmol/kg Sa was applied. Furthermore, the combined treatment T(0.1EDTA+0.9CY+0.5TW80) and T(0.5EDTA+0.9CY+03Sa) could not only increase the accumulation of Cd in plant tissues, but also promote the degradation of PAHs. These results indicated that S. nigrum might be effective in phytoextracting Cd and enhancing the biodegradation of PAHs in the co-contaminated soils with assistant chemicals.

[Effects of illumination and seed-soaking reagent on seed germination of Solanum nigrum].

To explore a rapid seed germination method for hyperaccumulator Solanum nigrum, a germination experiment with different illumination and seed-soaking treatments was conducted in constant temperature box and greenhouse, with filter as burgeon base. Under illumination, the germination rate was about 5 times high of that without illumination (P < 0.05), indicating that illumination was one of the prerequisites for the seed germination of S. nigrum. All test seed-soaking reagents could significantly improve the germination rate of S. nigrum (P < 0.05), with the best effect of H2O2. The seeds treated with H2O2 had the shortest germination time. The germination rate of seeds soaked but without cleaning was 2-3 times as high as that of seeds soaked and cleaned with water.

Seed germination of a newly discovered hyperaccumulator Solanum nigrum L. affected by illumination and seed-soaking reagent.

Solanum nigrum is a newly found Cd-hyperaccumulator which showed very high remediation efficiency in polluted soil. Seed germination experiments with different illumination and seed-soaking reagents were conducted in constant temperature box and greenhouse with soil as burgeon base. The results showed that the germination rate with alternating light/dark photoperiod was about twice of that without lighting (p < 0.05), suggesting that illumination is one of the important conditions for seed germination of S. nigrum. All treatments with seed-soaking reagents significantly increased the seed germination rate of S. nigrum (p < 0.05). Treatment with H2O2 (0.1%) had the shortest germination time. The germination rate of seeds that were not washed in water following soaking was 2-3 times higher than that of seeds that were washed after soaking.

Bidens tripartite L.: a Cd-accumulator confirmed by pot culture and site sampling experiment.

Characteristics of accumulation and tolerance of cadmium (Cd) in Bidens tripartite L. were investigated to identify Cd-accumulating properties. In this study, pot culture experiment and site sampling experiments were conducted to assess whether this plant is a heavy metal hyperaccumulator or accumulator. The results indicated that the Cd enrichment factor (concentration in plant/soil) and Cd translocation factor (concentration in shoot/root) of B. tripartite was principally >1 in pot culture and concentration gradient experiments. Shoot biomass was not reduced significantly (p<0.05) compared to the controls. However, the Cd concentration in B. tripartite shoots was not higher than 100 mg kg(-1), the threshold concentration for a Cd-hyperaccumulator. In the site sampling experiment, B. tripartite also showed Cd-accumulator properties. Based on these results, B. tripartite could be identified as a Cd-accumulator. Thus, B. tripartite should only be considered as a Cd-accumulator.

Hyperaccumulative property comparison of 24 weed species to heavy metals using a pot culture experiment.

The screening of hyperaccumulators is still very much needed for phytoremediation. With properties such as strong tolerance to adverse environment, fast growing and highly reproductive rate, weed species may be an ideal plant for phytoremediation. The objectives of this study were to examine the tolerance and hyperaccumulative characteristics of 24 species in 9 families to Cd, Pb, Cu and Zn by using the outdoor pot-culture experiment. In the screening experiment, only Conyza canadensis and Rorippa globosa displayed Cd-hyperaccumulative characteristics. In a further concentration gradient experiment, C. canadensis was affirmed that it is not a Cd hyperaccumulator. Only R. globosa, indicated all Cd hyperaccumulative characteristics, especially Cd concentration in its stems and leaves were higher than 100 mg/kg, the minimum Cd concentration what a Cd-hyperaccumulator should accumulate. Thus, R. globosa was further validated as a Cd-hyperaccumulator.

Identification of a Cd accumulator Conyza canadensis.

One of key steps of phytoremediating heavy metal contaminated soils is still the identification of hyperaccumulator and accumulator. In a former published article, Conyza canadensis L. Cronq. expressed some basic properties of Cd-hyperaccumulators. In this study, concentration gradient experiment and two sample-analyzing experiments were used to identify whether this plant is a Cd-hyperaccumulator. When grown on soil spiked with Cd at the rate of 10 and 25 mg kg(-1) in concentration gradient experiment, C. canadensis had both Cd enrichment factor (EF) and Cd translocation factor (TF) greater than 1, while the shoot biomass did not differ significantly as compared to the control. On the other hand, with Cd-spiking rates of 10 and 25 mg kg(-1), the Cd concentration in the shoot did not exceed 100 mg kg(-1), which is considered as the minimum shoot Cd concentration to qualify as a hyperaccumulator. In the sample-analysis experiments from a Pb-Zn mine area and wastewater irrigation region, C. canadensis also showed Cd-accumulator characteristics. Based on the results accomplished, we propose C. canadensis as a Cd-accumulator.