Enhanced phytoremediation of 2,4-DNP-contaminated wastewater by Salix matsudana Koidz with MeJA pretreatment and associated mechanism.

2,4-Dinitrophenol (2,4-DNP) is recognized as an emerging contaminant due to its high toxicity and poor biodegradability, posing a threat to animals, plants, and human health. The efficient removal of 2,4-DNP remains a challenging issue in phytoremediation research, particularly because of its toxic effects on plants. To address this, a hydroponic simulation experiment was conducted to investigate the impact of adding exogenous methyl jasmonate (MeJA) on the tolerance and purification capabilities of Salix matsudana Koidz (S. matsudana) seedlings exposed to 2,4-DNP. The results indicated that the addition of exogenous MeJA mitigated the damage caused by 2,4-DNP to S. matsudana seedlings by enhancing the activity of antioxidant enzymes, reducing excess reactive oxygen species (ROS), lowering membrane lipid peroxidation, and minimizing membrane damage. Notably, the most effective alleviation was observed with the addition of 50 mg·L-1 MeJA. Furthermore, exogenous MeJA helped maintain the biomass indices of S. matsudana seedlings under 2,4-DNP stress and increased the removal efficiency of 2,4-DNP by these seedlings. Specifically, the addition of 50 mg·L-1 MeJA resulted in a removal percentage of 79.57%, which was 11.88% higher than that achieved with 2,4-DNP treatment. In conclusion, exogenous MeJA can improve the plant resistance and enhance 2,4-DNP phytoremediation.

Impacts of in-vitro zebularine treatment on genome-wide DNA methylation and transcriptomic profiles in Salix purpurea L.

Renewable energy resources such as biomass are crucial for a sustainable global society. Trees are a major source of lignocellulosic biomass, which can vary in response to different environmental factors owing to epigenetic regulation, such as DNA C-methylation. To investigate the effects of DNA methylation on plant development and wood formation, and its impacts on gene expression, with a focus on secondary cell wall (SCW)-associated genes, Salix purpurea plantlets were cloned from buds derived from a single hybrid tree for both treatment and control conditions. For the treatment condition, buds were exposed to 50 µM zebularine in vitro and a combined strategy of whole-genome bisulfite sequencing (WGBS) and RNA-seq was employed to examine the methylome and transcriptome profiles of different tissues collected at various time points under both conditions. Transcriptomic and methylome data revealed that most of the promoter and gene body demethylation had no marked effects on the expression profiles of genes. Nevertheless, gene expression tended to decrease with the increased methylation levels of genes with highly methylated promoters. Results indicated that demethylation is less evident in centromeric regions and sex chromosomes. Promoters of secondary cell wall-associated genes, such as 4-coumarate-CoA ligase-like and Rac-like GTP-binding protein RHO, were differentially methylated in the secondary xylem samples collected from two-month potted treated plants compared to control samples. Our results provide novel insights into DNA methylation and gene expression landscapes and a basis for investigating the epigenetic regulation of wood formation in S. purpurea as a model plant for bioenergy species.

Integrated physiological, transcriptomic and metabolomic analyses provide insights into phosphorus-mediated cadmium detoxification in Salix caprea roots.

Phosphorus (P) plays a crucial role in facilitating plant adaptation to cadmium (Cd) stress. However, the molecular mechanisms underlying P-mediated responses to Cd stress in roots remain elusive. This study investigates the effects of P on the growth, physiology, transcriptome, and metabolome of Salix caprea under Cd stress. The results indicate that Cd significantly inhibits plant growth, while sufficient P alleviates this inhibition. Under Cd exposure, P sufficiency resulted in increased Cd accumulation in roots, along with reduced oxidative stress levels (superoxide anion and hydrogen peroxide contents were reduced by 16.8% and 30.1%, respectively). This phenomenon can be attributed to the enhanced activities of antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT), as well as increased levels of antioxidants including ascorbic acid (AsA) and flavonoids under sufficient P conditions. A total of 4208 differentially expressed genes (DEGs) and 552 differentially accumulated metabolites (DAMs) were identified in the transcriptomic and metabolomic analyses, with 2596 DEGs and 113 DAMs identified among treatments with different P levels under Cd stress, respectively. Further combined analyses reveal the potential roles of several pathways in P-mediated Cd detoxification, including flavonoid biosynthesis, ascorbate biosynthesis, and plant hormone signal transduction pathways. Notably, sufficient P upregulates the expression of genes including HMA, ZIP, NRAMP and CAX, all predicted to localize to the cell membrane. This may elucidate the heightened Cd accumulation under sufficient P conditions. These findings provide insights into the roles of P in enhancing plant resistance to Cd stress and improving of phytoremediation.

Divergent responses of rhizosphere soil phosphorus fractions and biological features of Salix psammophila to fertilization strategies under cadmium contamination.

Soil oligotrophy in areas heavily contaminated with heavy metals poses a significant challenge to vegetation establishment and phytoremediation processes. Phosphorus (P) cycling plays a critical role in global biogeochemical cycles, but there is limited understanding of its response to varying fertilization strategies and its correlation with phytoremediation effectiveness. This study primarily investigated the effects of various fertilization strategies, including nitrogen (N, 300 mg·kg-1), P (100 mg·kg-1), NP (combined N and P at 300 mg·kg-1 and 100 mg·kg-1, respectively), and HP (high P, 300 mg·kg-1) application, on rhizosphere soil P fractions and P-solubilizing microbial community (harboring phoD and phoC genes, respectively) of Salix psammophila under cadmium contamination. Application of NP significantly enhanced plant growth and cadmium accumulation, whereas HP inhibited cadmium bioaccumulation but promoted its translocation. Compared to untreated soil, N application promoted P cycling, leading to increases of 141.9 %, 60.4 %, and 10.3 % in Resin-Pi, diluted HCl-Pi, and conc.HCl-Pi, respectively. P application decreased organic phosphorus (Po) fractions by 24.4 % - 225.8 %, but N incorporation mitigated the declining trend in Po and augmented alkaline phosphatase activity. Fertilization strategies significantly regulated phoC- or phoD-harboring bacterial community structure, but their differential nutrient demands resulted in distinct responses. The phoD-harboring bacteria exhibited higher diversity and network complexity, with numerous biomarkers and fertilizer-sensitive OTUs discovered across treatments. Structural equation modeling (SEM) analysis indicated that phytoremediation efficiency was directly affected by Pi fractions, and phoD-harboring bacteria exhibited stronger associations with Pi fractions than phoC-harboring bacteria. In conclusion, our results reveal potential pathways through which fertilization strategies influence phytoremediation by affecting the structure of P-solubilizing microbial community. Furthermore, our study emphasizes the importance of combined N and P application in promoting Cd accumulation in plants, with high P levels appearing as an ideal fertilization strategy for phytoremediation targeting the harvest of aboveground biomass.

Morphological and physiological responses of two willow species from different habitats to salt stress.

Plant salt tolerance is a complex mechanism, and different plant species have different strategies for surviving salt stress. In the present study, we analyzed and compared the morphological and physiological responses of two willow species (Salix linearistipularis and Salix matsudana) from different habitats to salt stress. S. linearistipularis exhibited higher seed germination rates and seedling root Na+ efflux than S. matsudana under salt stress. After salt treatment, S. linearistipularis leaves exhibited less Na+ accumulation, loss of water and chlorophyll, reduction in photosynthetic capacity, and damage to leaf cell structure than leaves of S. matsudana. Scanning electron microscopy combined with gas chromatography mass spectrometry showed that S. linearistipularis leaves had higher cuticular wax loads than S. matsudana leaves. Overall, our results showed that S. linearistipularis had higher salt tolerance than S. matsudana, which was associated with different morphological and physiological responses to salt stress. Furthermore, our study suggested that S. linearistipularis could be a promising tree species for saline-alkali land greening and improvement.

[Transcriptome analysis of Salix matsudana under cadmium stress].

With the expanded application of heavy metal cadmium, soil cadmium pollution is more and more serious. In this study, using Salix matsudana as a phytoremediation candidate, we observed changes of gene expression and metabolic pathway after 1, 7 and 30 days under 2.5 mg/L and 50 mg/L cadmium stress. The result of transcriptome sequencing showed that we obtained 102 595 Unigenes; 26 623 and 32 154 differentially expressed genes (DEG) in the same concentration and different stress time; 8 550, 3 444 and 11 428 DEG with different concentrations at the same time; 25 genes closely related to cadmium stress response were screened. The changes of genes expression (such as metallothionein, ABC transporter, zinc and manganese transporter) depended on both concentration of cadmium and exposure time. The expression of several genes was obviously up-regulated after cadmium stress, for example 3,6-deoxyinosinone ketolase (ROT3) in brassinolide synthesis pathway and flavonoid synthase (FLS), flavanone-3-hydroxylase (F3H) in the synthesis pathway of brassinolide. In addition, GO analysis shows that GO entries were mainly enriched in metabolic processes including cellular processes, membranes, membrane fractions, cells, cellular fractions, catalytic activation and binding proteins in response to cadmium stress, whose number would increase along with cadmium concentration and exposure time. The reliability of transcriptome information was verified by qPCR and physiological experimental data. Response mechanisms of S. matsudana after cadmium stress were analyzed by transcriptome sequencing, which provided theoretical guidance for remediation of cadmium pollution in soil by S. matsudana.

Characterisation of early responses in lead accumulation and localization of Salix babylonica L. roots.

BACKGROUND: Lead (Pb) is a harmful pollutant that disrupts normal functions from the cell to organ levels. Salix babylonica is characterized by high biomass productivity, high transpiration rates, and species specific Pb. Better understanding the accumulating and transporting Pb capability in shoots and roots of S. babylonica, the toxic effects of Pb and the subcellular distribution of Pb is very important. RESULTS: Pb exerted inhibitory effects on the roots and shoots growth at all Pb concentrations. According to the results utilizing inductively coupled plasma atomic emission spectrometry (ICP-AES), S. babylonica can be considered as a plant with great phytoextraction potentials as translocation factor (TF) value > 1 is observed in all treatment groups throughout the experiment. The Leadmium™ Green AM dye test results indicated that Pb ions initially entered elongation zone cells and accumulated in this area. Then, ions were gradually accumulated in the meristem zone. After 24 h of Pb exposure, Pb accumulated in the meristem zone. The scanning electron microscopy (SEM) and energy-dispersive X-ray analyses (EDXA) results confirmed the fluorescent probe observations and indicated that Pb was localized to the cell wall and cytoplasm. In transverse sections of the mature zone, Pb levels in the cell wall and cytoplasm of epidermal cells was the lowest compared to cortical and vessel cells, and an increasing trend in Pb content was detected in cortical cells from the epidermis to vascular cylinder. Similar results were shown in the Pb content in the cell wall and cytoplasm of the transverse sections of the meristem. Cell damage in the roots exposed to Pb was detected by propidium iodide (PI) staining, which was in agreement with the findings of Pb absorption in different zones of S. babylonica roots under Pb stress. CONCLUSION: S. babylonica L. is observed as a plant with great potential of Pb-accumulation and Pb-tolerance. The information obtained here of Pb accumulation and localization in S. babylonica roots can furthers our understanding of Pb-induced toxicity and its tolerance mechanisms, which will provide valuable and scientific information to phytoremediation investigations of other woody plants under Pb stress.

Effect of biochar and redmud amendment combinations on Salix triandra growth, metal(loid) accumulation and oxidative stress response.

Remediation of metal(loid) polluted soils is an important area of research nowadays. In particular, one remediation technique is much studied, phytomanagement. Phytomanagement combines amendment application and plant growth in order to reduce the risk posed by contaminants. Salicaceae plants showed tolerance towards metal(loid)s and the ability to accumulate high amounts of metal(loid)s in their tissue. Amendments are often applied to counterbalance the reduced soil fertility and high metal(loid) concentrations. Two amendments gathered attention over the last decades, biochar (product of biomass pyrolysis), which can be activated for better effects, and redmud (by-product of alumina production). Those two amendments showed ability to improve soil conditions and thus plant growth, although few studied their combined application. Moreover, since metal(loid)s are known to induce the overproduction of reactive oxygen species, it is important to measure the level of oxidative stress in the plant, to which plants respond using enzymatic and non-enzymatic systems. But no studies evaluate the response of Salicaceae plants to metal(loid) stress and amendment application at the biochemical level in a real soil condition. Therefore, a mesocosm study was set up to evaluate the effect of amending a mine soil with redmud combined to diverse biochars on the soil properties and Salix triandra growth, metal(loid) accumulation and stress marker levels. Results showed that all amendment combinations improved the soil fertility, reduced metal(loid) mobility and thus ameliorated Salix triandra growth, which accumulated metal(loid)s mainly in its roots. Moreover, among the different amendment combinations, Salix triandra plants still suffered from oxidative stress when grown on PG soil amended with redmud and chemical activated carbon, showing elevated levels of phenolic compounds and salicinoids and important antioxidant and enzymatic activities. Finally, one treatment showed levels of these stress markers similar or lower than the control, the combination of redmud with steam activated carbon. In conclusion, this treatment seemed a good solution in a phytomanagement strategy using Salix triandra, improving soil conditions and plant growth and reducing oxidative stress level in the plant roots.

Physiological and transcriptomic analysis provide novel insight into cobalt stress responses in willow.

Cobalt (Co) is an essential component of several enzymes and coenzymes in living organisms. Excess Co is highly toxic to plants. The knowledge of molecular response mechanisms to cobalt stress in plants is still limited, especially in woody plants. The responses of weeping willow (Salix babylonica) seedlings to Co stress were studied using morphological and physiochemical measurements and RNA-seq analysis. The physiological and biochemical indexes such as growth rate, the content of chlorophyll and soluble sugar, photosynthesis and peroxidase activity were all changed in willow seedlings under Co stress. The metal ion concentrations in willow including Cu, Zn and Mg were disturbed due to excess Co. Of 2002 differentially expressed genes (DEGs), 1165 were root-specific DEGs and 837 were stem and leaf-specific DEGs. Further analysis of DEGs showed there were multiple complex cascades in the response network at the transcriptome level under Co stress. Detailed elucidation of responses to oxidative stress, phytohormone signaling-related genes and transcription factors (TFs), and detoxification of excess cellular Co ion indicated the various defense mechanisms in plants respond to cobalt stress. Our findings provide new and comprehensive insights into the plant tolerance to excess Co stress.

Accumulation and effects of perfluoroalkyl substances in three hydroponically grown Salix L. species.

The potential of young rooted cuttings of three Salix L. species plants to accumulate a mixture of eleven perfluoroalkyl substances (PFASs), in particular, perfluoroalkyl acids (PFAAs), from the nutrient solution and their effects on plant growth and photosynthesis were assessed in an 8-day experiment. The growth rate of the willow plants exposed to the PFAA mixture was not much affected except for S. triandra. Regarding photosynthesis, the gas exchange parameters were affected more than those related to chlorophyll fluorescence, with significant increase of the net CO2 assimilation rate and parameters related to stomatal conductance. A decreasing trend in the PFAA concentration in leaves with increasing carbon chain length was observed, whereas long-chain PFAAs showed higher concentrations in roots. Accordingly, the foliage to root concentration factor highlighted that PFAAs with shorter carbon chain length (C ≤ 7) translocated and accumulated relatively more in leaves compared to roots. Removal efficiency of individual PFAAs for leaves and roots were comparatively higher with S. eleagnos and S. purpurea than S. triandra, with mean removal values at the whole plant level ranging around 10% of the amount initially spiked, suggesting their potential for phytoremediation of PFASs.

The effects of exogenous organic acids on the growth, photosynthesis and cellular ultrastructure of Salix variegata Franch. Under Cd stress.

We studied the effects of three organic acids (citric acid, tartaric acid and malic acid) on the biomass, photosynthetic pigment content and photosynthetic parameters of Salix variegata under Cd stress and observed the ultrastructure of mesophyll cells in each treatment. Cd stress significantly reduced photosynthesis by reducing the content of pigments and disrupting chloroplast structure, which consequently decreased the biomass. However, respective addition of three organic acids greatly increased the biomass of S. variegata under Cd stress. Among them, the effect of malic acid or tartaric acid on shoot and total biomass accumulation was greater than that of citric acid. The alleviation of biomass probably related with the photosynthetic process. Results revealed that treatment with each organic acid enhanced the net photosynthesis rate under Cd stress. Malic acid promoted plant growth and biomass by increasing the chlorophyll content and mitigating damage to the photosynthetic apparatus resulting from Cd stress. Tartaric acid had little impact on the photosynthetic pigment content, but it was important in mitigating the ultrastructural damage of plants caused by Cd. Addition of citric acid significantly increased the carotenoid as well as the number and volume of chloroplasts in mesophyll cells, while the mitigation of structural damage in the photosynthetic apparatus was weaker than that in tartaric acid or malic acid treatment. It is concluded that application of tartaric acid or malic acid is effective in increasing the growth potential of S. variegata under Cd stress and thus can be a promising approach for the phytoremediation of Cd-contaminated soil.

Lead transfer into the vegetation layer growing naturally in a Pb-contaminated site.

The lead was one of the main elements in the glazes used to colour ceramic tiles. Due to its presence, ceramic sludge has been a source of environmental pollution since this dangerous waste has been often spread into the soil without any measures of pollution control. These contaminated sites are often located close to industrial sites in the peri-urban areas, thus representing a considerable hazard to the human and ecosystem health. In this study, we investigated the lead transfer into the vegetation layer (Phragmites australis, Salix alba and Sambucus nigra) growing naturally along a Pb-contaminated ditch bank. The analysis showed a different lead accumulation among the species and their plant tissues. Salix trees were not affected by the Pb contamination, possibly because their roots mainly develop below the contaminated deposit. Differently, Sambucus accumulated high concentrations of lead in all plant tissues and fruits, representing a potential source of biomagnification. Phragmites accumulated large amounts of lead in the rhizomes and, considering its homogeneous distribution on the site, was used to map the contamination. Analysing the Pb concentration within plant tissues, we got at the same time information about the spread, the history of the contamination and the relative risks. Finally, we discussed the role of natural recolonizing plants for the soil pollution mitigation and their capacity on decreasing soil erosion and water run-off.

Modulation of the defence responses against Cd in willow species through a multifaceted analysis.

Willow, due to the extensive root system, high transpiration rates and ability to accumulate large amounts of cadmium, is considered particularly useful for green remediation practices. In this study two different willow species, Salix viminalis and Salix alba, were used to assess possible differences in their ability of cadmium accumulation and to analyse in detail the physiology of their response to treatments with this metal using a multidisciplinary approach. Plants were grown in hydroponics and treated with 0, 50 and 100 µM Cd2+ (CdCl2) for 7 and 14 days. Cadmium content, oxidative stress, both evaluated by biochemical and histochemical techniques, antioxidant response, leaf stomatal conductance and photosynthetic efficiency were measured in control and treated roots and/or leaves. The two willow species removed cadmium with a high efficiency from the growth solution; however, the highest contents of Cd recorded in plants grown in the presence of the lower Cd concentrations suggest a limited capacity of metal accumulation. No photochemical limitation characterised treated plants, probably due to the ability to store large amounts of Cd in the root compartment, with reduction of damage to the photosynthetic machinery. S. viminalis, able to uptake cadmium also in the root apical region, seemed to be a more efficient accumulator than S. alba and, thanks to a relatively higher antioxidant response, did not show a higher level of oxidative stress. On the basis of the above, the two plant species, in particular S. viminalis, are confirmed as useful for cadmium phytostabilisation/phytoextraction.

Growth, accumulation, and antioxidative responses of two Salix genotypes exposed to cadmium and lead in hydroponic culture.

Cd and Pb are a toxic environmental pollutant, and their elevated concentrations in the waters and soils could exert detriment effects on human health by food chain. In order to evaluate the capacity to heavy metal accumulation and the physiochemical responses of two Salix genotypes, a 35-day hydroponic seedling experiment was implemented with Salix matsudana Koidz. 'Shidi1' (A42) and Salix psammophila C. 'Huangpi1' (A94) under different concentrations of Cd (15 and 30 µM) or Pb (250 and 300 µM). The results showed that the biomass of A94 severely reduced more than that of A42. The accumulation ability of Cd in different plant organs followed the sequence of leaves > roots > stems. Pb primarily accumulated in the roots for both Salix genotypes (54.27 mg g-1 for A42 and 54.52 mg g-1 for A94). Translocation factors based on accumulation (TF') for Cd were more than 8.0, while TF's for Pb were less than 1.0 in both A42 and A94, implying they could be applied in the phytoremediation of Cd-contaminated sites due to their stronger ability to Cd phytoextraction. The stress of Cd or Pb significantly increased malondialdehyde (MDA) contents and increased photosynthetic rates in leaves of two Salix genotypes. Transpiration rates of willow were positively correlated with its Cd translocation. Both catalase (CAT) and peroxidase (POD) activities were suppressed, while the superoxide dismutase (SOD) was boosted with increasing Cd and Pb levels in the leaves and roots of the two willow genotypes, suggesting SOD plays an important role in the removal of ROS. The inconsistency of the changes in enzyme activity suggests that the integrated antioxidative mechanisms regulate the tolerance to Cd and Pb stress.

Biochar effect associated with compost and iron to promote Pb and As soil stabilization and Salix viminalis L. growth.

Soil pollution by metal(loid)s is one of the most significant problems in Europe. To remediate and potentially rehabilitate these contaminated sites, phytoremediation procedures are being put into place, often using amendments to help offset the extreme conditions of such soils. The aim of this study was to define the best amendment to use on the field. This was done by studying how the addition of three different amendments (biochar, compost and iron grit), alone or in combination, could affect: (i) soil physico-chemical properties, (ii) Salix viminalis growth, and (iii) metal(loid) stabilization. A 69 day-mesocosm study was thus set up using a former mine technosol, the three amendments applied alone or combined, and S. viminalis cuttings. The results showed that biochar and/or compost improved the soil fertility and the soil pore water characteristics, with reductions of acidity, metal(loid) mobility and toxicity, while iron grit amendment presented negative effects on such parameters. Such ameliorations allowed better plant growth and higher biomass production. In addition, stress indicators (leaf pigment content and root guaiacol peroxidase activity) showed a reduction in plant stress following biochar and/or compost application. Finally, among the different treatments, the use of compost or a biochar-compost combination showed better results in terms of improvement of soil conditions, increase in plant growth and reduced translocation of metal(loid)s towards upper parts, making these two treatments a valuable option for a field trial.

The impact of nanoparticles zero-valent iron (nZVI) and rhizosphere microorganisms on the phytoremediation ability of white willow and its response.

Soil contaminated with heavy metals (HMs) is a serious problem throughout the world that threatens all living organisms in the soil. Therefore, large-scale remediation is necessary. This study investigated a new combination of remediation techniques on heavy metal contaminated soil, phytoremediation, and soil amendment with nano-sized zero-valent iron (nZVI) and rhizosphere microorganisms. White willow (Salix alba L.) was grown for 160 days in pots containing Pb, Cu, and Cd and amended with 0, 150, and 300 (mg kg-1) of nZVI and rhizosphere microorganisms, including the arbuscular mycorrhizal fungus (AMF), Rhizophagus irregularis, and the plant growth promoting rhizobacteria (PGPR), Pseudomonas fluorescens. The results showed that inoculation with PGPR and AMF, particularly dual inoculation, improved plant growth as well as the physiological and biochemical parameters of white willow, and increased the bioconcentration factor (BCF) of Pb, Cu, and Cd. The low dose of nZVI significantly increased the root length and the leaf area of the seedlings and increased the BCF of Cd. In contrast, the high dose of nZVI had negative effects on the seedlings growth and the BCF of Pb and Cu, about - 32% and - 63%, respectively. Our results demonstrate that nZVI at low doses can improve plant performance in a phytoremediation context and that the use of beneficial rhizosphere microorganisms can minimize nZVI stress in plants and make them less susceptible to stress even under high dose conditions.

Effects of soil amendments on the growth response and phytoextraction capability of a willow variety (S. viminalis × S. schwerinii × S. dasyclados) grown in contaminated soils.

This study was conducted to evaluate the effects of lime and bisphosphonates (BPs) such as N10O chelate amendment on the growth, physiological and biochemical parameters, and phytoextraction potential of the willow variety Klara (Salix viminalis × S. schwerinii × S. dasyclados) grown in soils heavily contaminated with copper (Cu), nickel (Ni) and zinc (Zn). The plants were irrigated with tap or processed water (mine wastewater). The results suggest that the combined effects of the contaminated soil and processed water inhibited growth parameters, gas exchange parameters and chlorophyll fluorescence (Fv/Fm) values. In contrast, malondialdehyde (MDA) content, organic acids, total phenolic and total flavonoid contents, and the accumulation of metals/metalloids in the plant tissues were increased compared to the control. When the soil was supplemented with lime and N10O; growth, physiological, biochemical parameters, and resistance capacity were significantly higher compared to unamended soil treatments, especially in the contaminated soil treatments. The combined lime‒ and N10O‒amended soil treatment produced higher growth rates, resistance capacity, photosynthesis rates and phytoextraction efficiency levels relative to either the lime‒amended or the N10O‒amended soil treatments. This study provides practical evidence of the efficient chelate‒assisted phytoextraction capability of Klara and highlights its potential as a viable and inexpensive novel approach for in situ remediation of Cu‒, Ni‒ and Zn‒contaminated soils and mine wastewaters.

Effect of amendments on contaminated soil of multiple heavy metals and accumulation of heavy metals in plants.

The contamination of soil with heavy metals is a severe problem due to adverse impact of heavy metals on environmental safety and human health. It is essential to remediate soil contaminated with heavy metals. This study has evaluated the effects of pine biochar, kaolin, and triple super phosphate (TSP) on multiple heavy metals (Ni, Zn, Cu, and Cd) in contaminated soil and accumulation of heavy metals in plants. The amendments can reduce availability of heavy metals in soil by increasing pH, adsorption, complexation, or co-precipitation. Different amendments have variable effects on accumulation of heavy metals in plants and in soil due to its diverse mechanism of stability. The results showed that application of triple super phosphate (TSP) has significant reduced soil Cd exchangeable (EXC) fraction from 58.59 to 21.30%. Bound to carbonates (CAR) fraction decreased from 9.84 to 5.11%, and bound to Fe-Mn oxides (OX) fraction increased from 29.61 to 69.86%. The triple super phosphate (TSP) has the ability to stabilize Cu and especially Cd. However, triple super phosphate (TSP) has enhanced ecological risk of Zn and Ni. Application of pine biochar has significantly enhanced soil pH. The kaolin has significantly reduced EXC fraction of Cd and increased OX fraction of Cu. The amendments and heavy metals have not caused significant effect on SPAD value of Buxus microphylla Siebold & Zucc (B. microphylla). The triple super phosphate (TSP) has significant decreased biomass of B. microphylla and bamboo-williow (Salix sp.) by 24.91 and 57.43%, respectively. Pine biochar and kaolin have increased the accumulation of Zn and Cd in plants. It is concluded that triple super phosphate (TSP) was effective in remediation of Cd and kaolin was effective in remediation of Cd and Cu. Pine biochar was effective in remediation of Cd, Cu, and Zn.

Effects of ozone (O3) and ethylenediurea (EDU) on the ecological stoichiometry of a willow grown in a free-air exposure system.

Ground-level ozone (O3) concentrations have been elevating in the last century. While there has been a notable progress in understanding O3 effects on vegetation, O3 effects on ecological stoichiometry remain unclear, especially early in the oxidative stress. Ethyelenediurea (EDU) is a chemical compound widely applied in research projects as protectant of plants against O3 injury, however its mode of action remains unclear. To investigate O3 and EDU effects early in the stress, we sprayed willow (Salix sachalinensis) plants with 0, 200 or 400 mg EDU L-1, and exposed them to either low ambient O3 (AOZ) or elevated O3 (EOZ) levels during the daytime, for about one month, in a free air O3 controlled exposure (FACE); EDU treatment was repeated every nine days. We collected samples for analyses from basal, top, and shed leaves, before leaves develop visible O3 symptoms. We found that O3 altered the ecological stoichiometry, including impacts in nutrient resorption efficiency, early in the stress. The relation between P content and Fe content seemed to have a critical role in maintaining homeostasis in an effort to prevent O3-induced damage. Photosynthetic pigments and P content appeared to play an important role in EDU mode of action. This study provides novel insights on the stress biology which are of ecological and toxicological importance.

Effect of water cadmium concentration and water level on the growth performance of Salix triandroides cuttings.

The growth performance of Salix triandroides cuttings at three water cadmium (Cd) concentrations (0, 20, and 40 mg L-1) and three water levels (- 40 cm, water level 40 cm below the soil surface; 0 cm, water level even with the soil surface; and 40 cm, water level 40 cm above soil surface) was investigated to evaluate its potential in phytoextraction strategies. Compared to cuttings in the - 40 or 0 cm water levels, cuttings in the 40 cm water level showed significantly lower biomass, height, and adventitious root length and significantly fewer leaves and adventitious roots. However, these growth and morphological parameters were not different among the three water Cd concentrations. Water level decreased stomatal conduction and transpiration rate but showed no significant effects on chlorophyll concentration or photosynthetic rate. Chlorophyll concentration and stomatal conductance were higher at 40 mg L-1 Cd treatment than at 0 or 20 mg L-1 Cd treatment; yet, photosynthetic rate and transpiration rate were not different. Cd concentration in the leaves and stems increased as the water level increased, but the highest Cd concentration in the roots occurred in the 0 cm water level. As water Cd concentration increased, Cd concentration in the leaves, stems, and roots increased in all three water levels, except in stems in the - 40 cm water level. Under Cd stress, cuttings in the - 40 or 0 cm water levels were characterized by a higher bioaccumulation coefficient, but a lower translocation factor, than those in the 40 cm water level. However, the bioaccumulation coefficient increased with increasing water Cd concentration in all three water levels, as did the translocation factor in the 40 cm water level. The tolerance index for the cuttings was the same among all water levels and water Cd concentrations. The results clearly indicated that the low water level increased plant growth and Cd accumulation in underground parts, while the high water level decreased plant growth but increased Cd accumulation in aboveground parts.