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Contamination of ground water and soil with toxic metalloids like arsenic (As) poses a serious hazard to the global agricultural food production. One of the best ways to restrict entry of As into the food chain is selection of germplasms which accrue extremely low level of As in grains. Here, we screened diverse maize genotypes under high arsenite (100 μM AsIII) stress and identified PMI-PV-9 and PMI-PV-3 as AsIII-tolerant and -sensitive maize genotype respectively. Expression of genes associated with As uptake, vacuolar sequestration, biosynthesis of phytochelatins, root-to-shoot translocation, in vivo ROS generation, fine tuning of antioxidant defense system, DNA and membrane damage, H 2 O 2 and superoxide anion (O 2 •- ) levels were compared among the selected genotypes. PMI-PV-9 plants performed much better than PMI-PV-3 in terms of plant growth with no visible symptom of As toxicity. Susceptibility of PMI-PV-3 to AsIII stress may be attributed to comparatively low expression of genes involved in phytochelatins (PCs) biosynthesis. Concomitant decrease in ABCC1 expression might be another key factor for futile sequestration of AsIII into root vacuoles. Moreover, up-regulation of ZmNIP3;1 might contribute in high root-to-leaf As translocation. Substantial spike in H 2 O 2 , O 2 •- and MDA levels indicates that PMI-PV-3 plants have experienced more oxidative stress than PMI-PV-9 plants. Appearance of prominent deep brown and dark blue spots/stripes on leaves as revealed after DAB and NBT staining respectively suggest severe oxidative burst in PMI-PV-3 plants. Marked reduction in DHAR and MDAR activity rendered PMI-PV-3 cells to recycle ascorbate pool ineffectively, which might have exacerbated their susceptibility to AsIII stress. In a nutshell, incompetent PCs mediated detoxification system and disruption of cellular redox homeostasis owing to feeble antioxidant defence system resulting oxidative burst might be the prime reasons behind reduced performance of PMI-PV-3 plants under AsIII stress., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Access to clean water, hygiene, and sanitation is becoming an increasingly pressing global demand, particularly owing to rapid population growth and urbanization. Phytoremediation utilizes a highly conserved phytochelatin in plants, which captures hazardous heavy metal ions from aquatic environments and sequesters them in vacuoles. Herein, we report the design of phytochelatin-inspired copolymers containing carboxylate and thiolate moieties. Titration calorimetry results indicate that the coexistence of both moieties is essential for the excellent Cd 2+ ion-capturing capacity of the copolymers. The obtained dissociation constant, K D  ~ 1 nM for Cd 2+ ion, is four-to-five orders of magnitude higher than that for peptides mimicking the sequence of endogenous phytochelatin. Furthermore, infrared and nuclear magnetic resonance spectroscopy results unravel the mechanism underlying complex formation at the molecular level. The grafting of 0.1 g bio-inspired copolymers onto silica microparticles and cellulose membranes helps concentrate the copolymer-coated microparticles in ≈3 mL volume to remove Cd 2+ ions from 0.3 L of water within 1 h to the drinking water level (<0.03 µM). The obtained results suggest that hyperconfinement of bio-inspired polymers in flow-through systems can be applied for the highly selective removal of harmful contaminants from the environmental water., (© 2024. The Author(s).)

To analyze the mechanism of copper accumulation in the marine alga Ulva compressa , it was cultivated with 10 μM of copper, with 10 μM of copper and increasing concentrations of a sulfide donor (NaHS) for 0 to 7 days, and with 10 μM of copper and a concentration of the sulfide acceptor (hypotaurine) for 5 days. The level of intracellular copper was determined as well as the level of glutathione (GSH) and phytochelatins (PCs) and the expression of metallothioneins (UcMTs). The level of intracellular copper in the algae treated with copper increased at day 1, slightly increased until day 5 and remained unchanged until day 7. The level of copper in the algae cultivated with copper and 100 or 200 μM of NaHS continuously increased until day 7 and the copper level was higher in the algae cultivated with 200 μM of NaHS compared to 100 μM of NaHS. In contrast, the level of intracellular copper decreased in the algae treated with copper and hypotaurine. The level of intracellular copper did not correlate with the level of GSH or with the expression of UcMTs, and PCs were not detected in response to copper, or copper and NaHS. Algae treated with copper and with copper and 200 μM of NaHS for 5 days were visualized by TEM and the elemental composition of electrondense particles was analyzed by EDXS. The algae treated with copper showed electrondense particles containing copper and sulfur, but not nitrogen, and they were mainly located in the chloroplast, but also in the cytoplasm. The algae treated with copper and NaHS showed a higher level of electrondense particles containing copper and sulfur, but not nitrogen, and they were located in the chloroplast, and in the cytoplasm. Thus, copper is accumulated as copper sulfide insoluble particles, and not bound to GSH, PCs or UcMTs, in the marine alga U. compressa .

The pollution caused by heavy metals (HMs) represents a global concern due to their serious environmental threat. Photosynthetic cyanobacteria have a natural niche and the ability to remediate HMs such as cadmium. However, their practical application is hindered by a low tolerance to HMs and issues related to recycling. In response to these challenges, this study focuses on the development and evaluation of engineered cyanobacteria-based living materials for HMs bioremediation. Genes encoding phytochelatins ( P CSs) and metallothioneins ( M Ts) were introduced into the model cyanobacterium Synechocystis sp. PCC 6803 , creating PM/6803. The strain exhibited improved tolerance to multiple HMs and effectively removed a combination of Cd 2+ , Zn 2+ , and Cu 2+ . Using Cd 2+ as a representative, PM/6803 achieved a bioremediation rate of approximately 21 μg of Cd 2+ /OD 750 under the given test conditions. To facilitate its controllable application, PM/6803 was encapsulated using sodium alginate-based hydrogels (PM/6803@SA) to create "living materials" with different shapes. This system was feasible, biocompatible, and effective for removing Cd 2+ under simulated conditions of zebrafish and mice models. Briefly, in vitro application of PM/6803@SA efficiently rescued zebrafish from polluted water containing Cd 2+ , while in vivo use of PM/6803@SA significantly decreased the Cd 2+ content in mice bodies and restored their active behavior. The study offers feasible strategies for HMs bioremediation using the interesting biomaterials of engineered cyanobacteria both in vitro and in vivo.

An unprecedented and direct PS-MS (paper spray ionization mass spectrometry) method was proposed for the detection of native peptides, that is, glutathiones (GSHs), homoglutathiones (hGSHs), and phytochelatins (PCs), in basil (Ocimum basilicum L.) roots before and after cadmium exposure. The roots were submitted to cold maceration followed by sonication with formic acid as the extractor solvent for sample preparation. PS-MS was used to analyze such extracts in the positive mode, and the results allowed for the detection of several GSHs, hGSHs, and PCs. Some of these PCs were not distinguished in the control samples, that is, basil roots not exposed to cadmium. Other PCs were noticed in both types of roots, uncontaminated and cadmium-contaminated, but the intensities were higher in the former samples. Moreover, long-time exposure to cadmium stimulated the formation of some of these PCs and their cadmium complexes. The results, therefore, provided some crucial insights into the defense mechanism of plants against an external stress condition due to exposure to a toxic heavy metal. The present study represents a promising alternative to investigate other crucial physiological processes in plants submitted to assorted stress conditions., (© 2024 John Wiley & Sons Ltd.)

Cadmium (Cd) and lead (Pb) are among the most toxic heavy metals affecting human health and crop yield. Suaeda maritima (L.) Dumort is an obligate halophyte that is well adapted to saline soil. The inbuilt salinity tolerance mechanisms of halophytes help them to survive in heavy metal-contaminated rhizospheric soil. In the present study, growth and ionomic responses, reactive oxygen species (ROS) accumulation, modulations of phytochelatins, antioxidative defense, and metabolomic responses were studied in S. maritima imposed to Cd and Pb stresses with an aim to elucidate Cd and Pb tolerance mechanisms and phytoremediation potential of this halophyte. Our results showed a reduction of biomass in S. maritima, which may serve as an energy conservation strategy for survival under heavy metal stress. The increased accumulation of ROS with concomitant higher expression of various antioxidative enzymes suggests the efficient scavenging of ROS. The metabolite profiling revealed significant up-regulation of sugars, sugar alcohols, amino acids, polyphenols, and organic acids under Cd and Pb stresses suggesting their possible role in osmotic balance, ionic homeostasis, ROS scavenging, and signal transduction for stress tolerance. In S. maritima, the translocation factors (Tf) are <1 in both Cd and Pb treatments, which indicates that this halophyte has high phytostabilization potential for Cd and Pb in roots and through restricted translocation of heavy metal ions to the aboveground part. The findings of this study offer comprehensive information on Cd and Pb tolerance mechanisms in S. maritima and suggest that this halophyte can detoxify the HMs through physiological, ionic, antioxidative, and metabolic regulations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

The Arabidopsis oligopeptide transporter AtOPT6 is reportedly involved in the long-distance transport of thiol compounds into sink organs. In the present study, transgenic Arabidopsis lines overexpressing AtOPT6 under the control of a phloem-specific promoter, sucrose-proton symporter 2 (pSUC2), were analyzed for thiol and cadmium (Cd) distribution during the reproductive stage, both with and without Cd exposure. Phloem specific AtOPT6-overexpressing lines did not exhibit an evident impact on bolting time. In the absence of Cd exposure, these transgenic lines showed significantly enhanced transport of endogenous glutathione into siliques, accompanied by a reduction in the glutathione content of flowers and roots during the reproductive stage. Additionally, exposure of the roots of the phloem specific AtOPT6-overexpressing lines to Cd altered the distribution of thiol compounds, resulting in an increase in the content of phytochelatins in sink organs, contributing to a significant elevation of Cd contents in reproductive sink. Our findings confirm the crucial role of AtOPT6 in unloading phytochelatin-Cd conjugates from the phloem into sink organ., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Arsenic (As) speciation analysis is scientifically relevant due to the pivotal role the As chemical form plays in toxicity, which, in turn, directly influences the effect it has on the environment. The objective of this study was to develop and optimize a method tailored for studying As compounds in plant samples. Different extraction procedures and HPLC methods were explored to assess their efficiency, determine mass balance, and improve the resolution of compounds in the chromatograms. Conventionally applied anion-exchange chromatography facilitated the separation of well-documented As compounds in the extracts corresponding to 19 to 82% of As present in extracts. To gain insight into compounds which remain undetectable by anion chromatography (18 to 81% of As in the extracts), but still possibly metabolically relevant, we explored an alternative chromatographic approach. The procedure of sample purification and preconcentration through solid-phase extraction, facilitating the detection of those minor As compounds, was developed. The system was further refined to achieve an online 2D-RP-HPLC system, which was employed to analyze the extracts more comprehensively with ICP and ESI MS. Using this newly developed method, As(III)-phytochelatins, along with other arseno-thio-compounds, were detected and identified in extracts derived from the tree roots of seedlings grown in the presence of As(III) and As(V), and a group of arseno lipids was detected in the roots of plants exposed to As(V).

Phytochelatins (PCs) are poly-Cys peptides containing a repeating γ-Glu-Cys motif synthesized in plants, algae, certain fungi, and worms by PC synthase from reduced glutathione. It has been shown that an excess of toxic metal ions induces their biosynthesis and that they are responsible for the detoxification process. Little is known about their participation in essential metal binding under nontoxic, basal conditions under which PC synthase is active. This study presents spectroscopic and thermodynamic interactions with the PC2-PC5 series, mainly focusing on the relations between Zn(II) complex stability and cellular Zn(II) availability. The investigations employed mass spectrometry, UV-vis spectroscopy, potentiometry, competition assays with zinc probes, and isothermal titration calorimetry (ITC). All peptides form ZnL complexes, while ZnL 2 was found only for PC2, containing two to four sulfur donors in the coordination sphere. Binuclear species typical of Cd(II)-PC complexes are not formed in the case of Zn(II). Results demonstrate that the affinity for Zn(II) increases linearly from PC2 to PC4, ranging from micro- to low-picomolar. Further elongation does not significantly increase the stability. Stability elevation is driven mainly by entropic factors related to the chelate effect and conformational restriction rather than enthalpic factors related to the increasing number of sulfur donors. The affinity of the investigated PCs falls within the range of exchangeable Zn(II) concentrations (hundreds of pM) observed in plants, supporting for the first time a role of PCs both in buffering and in muffling cytosolic Zn(II) concentrations under normal conditions, not exposed to zinc excess, where short PCs have been identified in numerous studies. Furthermore, we found that Cd(II)-PC complexes demonstrate significantly higher metal capacities due to the formation of polynuclear species, which are lacking for Zn(II), supporting the role of PCs in Cd(II) storage (detoxification) and Zn(II) buffering and muffling. Our results on phytochelatins' coordination chemistry and thermodynamics are important for zinc biology and understanding the molecular basis of cadmium toxicity, leaving room for future studies.

ATP-binding cassette transporter B6 (ABCB6), a protein essential for heme biosynthesis in mitochondria, also functions as a heavy metal efflux pump. Here, we present cryo-electron microscopy structures of human ABCB6 bound to a cadmium Cd(II) ion in the presence of antioxidant thiol peptides glutathione (GSH) and phytochelatin 2 (PC2) at resolutions of 3.2 and 3.1 Å, respectively. The overall folding of the two structures resembles the inward-facing apo state but with less separation between the two halves of the transporter. Two GSH molecules are symmetrically bound to the Cd(II) ion in a bent conformation, with the central cysteine protruding towards the metal. The N-terminal glutamate and C-terminal glycine of GSH do not directly interact with Cd(II) but contribute to neutralizing positive charges of the binding cavity by forming hydrogen bonds and van der Waals interactions with nearby residues. In the presence of PC2, Cd(II) binding to ABCB6 is similar to that observed with GSH, except that two cysteine residues of each PC2 molecule participate in Cd(II) coordination to form a tetrathiolate. Structural comparison of human ABCB6 and its homologous Atm-type transporters indicate that their distinct substrate specificity might be attributed to variations in the capping residues situated at the top of the substrate-binding cavity., (© 2024. The Author(s).)

Phytohormones, particularly cytokinin trans -zeatin ( t Z), were studied for their impact on the green alga Desmodesmus armatus under cadmium (Cd) stress, focusing on growth, metal accumulation, and stress response mechanisms. Using atomic absorption spectroscopy for the Cd level and high-performance liquid chromatography for photosynthetic pigments and phytochelatins, along with spectrophotometry for antioxidants and liquid chromatography-mass spectrometry for phytohormones, we found that t Z enhances Cd uptake in D. armatus , potentially improving phycoremediation of aquatic environments. Cytokinin mitigates Cd toxicity by regulating internal phytohormone levels and activating metal tolerance pathways, increasing phytochelatin synthase activity and phytochelatin accumulation essential for Cd sequestration. Treatment with t Z and Cd also resulted in increased cell proliferation, photosynthetic pigment and antioxidant levels, and antioxidant enzyme activities, reducing oxidative stress. This suggests that cytokinin-mediated mechanisms in D. armatus enhance its capacity for Cd uptake and tolerance, offering promising avenues for more effective aquatic phycoremediation techniques.

Arsenic (As) is a heavy metal that is toxic to both plants and animals. Silicon nanoparticles (SiNPs) can alleviate the detrimental effects of heavy metals on plants, but the underlying mechanisms remain unclear. The study aims to synthesize SiNPs and reveal how they promote plant health in Arsenic-polluted soil. 0 and 100% v/v SiNPs were applied to soil, and Arsenic 0 and 3.2 g/ml were applied twice. Maize growth was monitored until maturity. Small, irregular, spherical, smooth, and non-agglomerated SiNPs with a peak absorbance of 400 nm were synthesized from Pycreus polystachyos. The SiNPs (100%) assisted in the development of a deep, prolific root structure that aided hydraulic conductance and gave mechanical support to the maize plant under As stress. Thus, there was a 40-50% increase in growth, tripled yield weights, and accelerated flowering, fruiting, and senescence. SiNPs caused immobilization (As(III)=SiNPs) of As in the soil and induced root exudates Phytochelatins (PCs) (desGly-PC 2 and Oxidized Glutathione) which may lead to formation of SiNPs=As(III)-PCs complexes and sequestration of As in the plant biomass. Moreover, SiNPs may alleviate Arsenic stress by serving as co-enzymes that activate the antioxidant-defensive mechanisms of the shoot and root. Thus, above 70%, most reactive ROS (OH) were scavenged, which was evident in the reduced MDA content that strengthened the plasma membrane to support selective ion absorption of SiNPs in place of Arsenic. We conclude that SiNPs can alleviate As stress through sequestration with PCs, improve root hydraulic conductance, antioxidant activity, and membrane stability in maize plants, and could be a potential tool to promote heavy metal stress resilience in the field., (© 2024. The Author(s).)

Triclosan has been extensively used as a preservative in cosmetics and personal care products. However, its accumulation represents a real environmental threat. Thus, its phytotoxic impact needs more consideration. Our study was conducted to highlight the phytotoxic effect of triclosan on the growth, ROS homeostasis, and detoxification metabolism of two different plant species i.e., legumes (Glycine max) and grass (Avena sativa). Moreover, we investigated the potentiality of plant growth-promoting bacteria (ST-PGPB) in mitigating the phytotoxic effect of triclosan. Triclosan induced biomass (fresh and dry weights) reduction in both plants, but to a higher extent in oats. This decline was associated with a noticeable increment in the oxidative damage (e.g., MDA and H 2 O 2 ) and detoxification metabolites such as metallothionein (MTC), phytochelatins (PCs), and glutathione-S-transferase (GST). This elevation was associated with a remarkable reduction in both enzymatic and non-enzymatic antioxidants. On the other hand, the bioactive strain of ST-PGPB, Salinicoccus sp. JzA1 significantly alleviated the harmful effect of triclosan on both soybean and oat plants by enhancing their biomass, photosynthesis, as well as levels of minerals (K, Ca, P, Mn, and Zn). In parallel, a striking quenching in oxidative damage and an obvious improvement in non-enzymatic (polyphenols, tocopherols, flavonoids) and enzymatic antioxidants were observed. Furthermore, Salinicoccus sp. JzA1 augmented the detoxification metabolism by enhancing the levels of phytochelatins, metallothionein, and glutathione-S-transferase (GST) activity in a species-specific manner which is more apparent in soybean rather than in oat plants. To this end, stress mitigating impact of Salinicoccus sp. JzA1 provides a basis to improve the resilience of crop species under cosmetics and personal care products toxicity., Competing Interests: Declaration of competing interest All the authors disclosed that there is no any financial or personal relationship that may be perceived as influencing their work., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Cd pollution-safe cultivar (Cd-PSC) is a feasible strategy to minimize Cd contamination in leafy vegetables. The shoot Cd concentrations of 23 Lactuca sativa cultivars under Cd stress ranged from 0.124 to 2.155 mg·kg -1 with a maximum cultivar difference of 8 folds. Typical Cd-PSC C16 (L) and high-Cd-accumulating cultivar C13 (H) were screened to investigate the mechanisms of Cd accumulations in L. sativa through determining Cd concentrations, Cd subcellular distributions, phytochelatin profiles, and phytochelatin biosynthesis-related genes' expressions. Higher Cd distribution in a heat stable fraction in C13 (H) indicated that the high Cd accumulation trait of C13 (H) mainly depended on the Cd-phytochelatin complexes. Root phytochelatin concentrations were significantly elevated in C13 (H) (5.83 folds) than in C16 (L) (2.69 folds) ( p < 0.05) under Cd stress. Significantly downregulated expressions of glutathione S-transferase rather than the regulation of phytochelatin synthesis genes in the root of C13 (H) might be responsible for sufficient glutathione supply for phytochelatins synthesis. These findings suggested that phytochelatin elevation in C13 (H) would favor the Cd root to shoot transportation, which provides new insights into the phytochelatin-related cultivar-dependent Cd accumulating characteristic in L. sativa .

The potential of arsenic (As) tolerant and sensitive varieties of wheat (Triticum aestivum L.) has yet to be explored despite of alarming situation of arsenic toxicity. To fill this gap, the study aimed to explore the role of antioxidants, phytochelatins, and ascorbate-glutathione for As tolerance in wheat. A total of eight varieties were exposed to different arsenate treatments (0, 1, 5, 10, 50, 100, 200, 500, 1000, 2000, and 10,000 μM) initially to screen effective treatment as well as contrasting varieties via Weibull distribution frequency for further analysis. The Weibull analysis found 200 μM as the most effective treatment in the present study. Selected varieties were analyzed for accumulation of total As and As speciation, oxidative stress (malondialdehyde, hydrogen peroxide), antioxidants (superoxide dismutase, catalase, peroxidase), phytochelatins, and ascorbate-glutathione cycle (glutathione-S-transferase, glutathione reductase, glutathione peroxidase, ascorbate peroxidase). Tolerant varieties showed less accumulation and translocation of total As, arsenate, and arsenite to the shoots compared with sensitive varieties under 200 μM treatment. Low concentration in tolerant varieties correlated with better growth and development response. Tolerant varieties showed higher induction of metabolites (glutathione, phytochelatins) compared to sensitive ones. Furthermore, tolerant varieties showed better performance of antioxidant and ascorbate-glutathione cycle enzymes in response to As exposure. The findings of the present study provided great insight into the wheat tolerance mechanism upon As exposure between contrasting varieties., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Selenium (Se) elevates the antioxidant ability of rice against cadmium (Cd) stress, but previous studies only focused on the variation in antioxidant enzymes or nonenzymatic substances induced by Se under Cd stress and ignored the relationships between different antioxidant parameters during the interaction. Here, hydroponic experiments with rice were performed by adding both Cd and Se at doses in the range of 0-50 μM to explore the physiological responses of rice and their relationships in the presence of different levels of Se and Cd. Exogenous Cd markedly promoted the activity of antioxidant enzymes with the exception of catalase (CAT) and the concentration of nonenzymatic substances in aerial parts. Se enhanced the antioxidant capacity by improving the activities of all the enzymes tested in this study and increasing the concentrations of nonenzymatic compounds. The couplings among different antioxidant substances within paddy rice were then determined based on cluster and linear fitting results and their metabolic process and physiological functions. The findings specifically highlight that couplings among the ascorbic acid (AsA)-glutathione (GSH) cycle, glutathione synthase (GS)-phytochelatin synthetase (PCS) coupling system and glutathione peroxidase (GPX)-superoxide dismutase (SOD) coupling system in aerial parts helps protect plants from Cd stress. These coupling systems form likely due to the fact that one enzyme generated a product that could be the substrate for another enzyme. Noticeably, such coupling systems do not emerge in roots because the stronger damage to roots than other organs activates the ascorbate peroxidase (APX)-GPX-CAT and PCS-GS-SOD systems with distinct functions and structures. This study provides new insights into the detoxification mechanisms of rice caused by the combined effect of Se and Cd., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Two non-metallicolous and metallicolous populations of harmel plants were compared regarding the role of proline, cysteine, reducing sugars, hydrogen peroxide (H 2 O 2 ), glutathione, thiol compounds, organic acids, total free amino acids, and lipid peroxidation in detoxification and tolerance of silver stress (0, 1, 2.5, 5, 10 ppm Ag). The results of the present research state that the effects of Ag were increased total free amino acids, glutathione, organic acids, proline, reducing sugars, thiol compounds, and cysteine, so the accumulation of these compounds was higher in metallicolous populations than non-metallicolous. On the other hand, non-metallicolous populations showed higher content of lipid peroxidation and H 2 O 2 than metallicolous populations under Ag stresses. Also, the accumulation of phytochelatins (PC) was observed with increasing Ag concentration, which shows that compared to glutathione, non-protein thiols have a higher concentration. The number of organic acids (malic acids, fumaric, oxalic, and citric) except acetic acid increased in the leaves of harmel in both populations. According to the results of this research, the harmel metallophilic population has a crucial role in the tolerance and detoxification of Ag stress, so the antioxidant responses of the plant against Ag stress in the non-metallicolous population were lower than the metallicolous population. Based on the above results, it can be concluded that the harmel plant has a detoxification mechanism to deal with high concentrations of Ag., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Phytoremediation using plants is an environmentally friendly and cost-effective strategy for removing cadmium (Cd) from soil. Plants used for phytoremediation must have a high Cd accumulation capacity and strong Cd tolerance. Therefore, understanding the molecular mechanism of Cd tolerance and accumulation in plants is of great interest. In response to Cd exposure, plants produce various thio-rich compounds, such as glutathione, phytochelatins, and metallothioneins, which play important roles in Cd immobilization, sequestration, and detoxification. Therefore, sulfur (S) metabolism is crucial for Cd tolerance and accumulation. In this study, we report that the overexpression of low-S responsive genes, LSU1 and LSU2, confers Cd tolerance in Arabidopsis. First, LSU1 and LSU2 promoted S assimilation under Cd stress. Second, LSU1 and LSU2 inhibited the biosynthesis and promoted the degradation of aliphatic glucosinolates, which could limit the consumption and enhance the release of S, thus, facilitating the production of the S-rich metabolites, glutathione, phytochelatins, and metallothioneins. We further demonstrated that the Cd tolerance mediated by LSU1 and LSU2 was dependent on the myrosinases BGLU28 and BGLU30, which catalyze the degradation of aliphatic glucosinolates. In addition, the overexpression of LSU1 and LSU2 improved Cd accumulation, which has great potential for the phytoremediation of Cd-contaminated soil., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Cadmium (Cd) uptake from polluted soils inhibits plant growth and disturbs physiological processes, at least partly due to disturbances in the cellular redox environment. Although the sulfur-containing antioxidant glutathione is important in maintaining redox homeostasis, its role as an antioxidant can be overruled by its involvement in Cd chelation as a phytochelatin precursor. Following Cd exposure, plants rapidly invest in phytochelatin production, thereby disturbing the redox environment by transiently depleting glutathione concentrations. Consequently, a network of signalling responses is initiated, in which the phytohormone ethylene is an important player involved in the recovery of glutathione levels. Furthermore, these responses are intricately connected to organellar stress signalling and autophagy, and contribute to cell fate determination. In general, this may pave the way for acclimation (e.g. restoration of glutathione levels and organellar homeostasis) and plant tolerance in the case of mild stress conditions. This review addresses connections between these players and discusses the possible involvement of the gasotransmitter hydrogen sulfide in plant acclimation to Cd exposure., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Soil contamination with trace metals and metalloids can cause toxicity to plants and threaten food safety and human health. Plants have evolved sophisticated mechanisms to cope with excess trace metals and metalloids in soils, including chelation and vacuolar sequestration. Sulfur-containing compounds, such as glutathione and phytochelatins, play a crucial role in their detoxification, and sulfur uptake and assimilation are regulated in response to the stress of toxic trace metals and metalloids. This review focuses on the multi-level connections between sulfur homeostasis in plants and responses to such stresses, especially those imposed by arsenic and cadmium. We consider recent progress in understanding the regulation of biosynthesis of glutathione and phytochelatins and of the sensing mechanism of sulfur homeostasis for tolerance of trace metals and metalloids in plants. We also discuss the roles of glutathione and phytochelatins in controlling the accumulation and distribution of arsenic and cadmium in plants, and possible strategies for manipulating sulfur metabolism to limit their accumulation in food crops., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Rice is known to accumulate cadmium (Cd) in its grains, causing a severe threat to billions of people worldwide. The possible phytotoxicity and mechanism of 50-200 mg/L hydroxyapatite NPs (nHA), iron oxide NPs (nFe2O3) or nano zero valent iron (nZVI) co-exposed with Cd (100 μM) in rice seedlings were investigated. Three types of nanoparticles significantly reduced the bioaccumulation of Cd in rice shoots by 16-63%, with nZVI showing the greatest effect, followed by nHA and nFe2O3. A decrease in Cd content in the roots was observed only in the nZVI treatment, with values ranging from 8 to 19%. Correspondingly, nZVI showed the best results in promoting plant growth, increasing rice plant height, shoot and root biomass by 13%, 29% and 42%. In vitro studies showed that nZVI reduced the content of Cd in the solution by 20-52% through adsorption, which might have contributed to the immobilization of Cd in root. Importantly, the nZVI treatment resulted in 267% more iron plaques on the root surface, which acted as a barrier to hinder the entry of Cd. Moreover, all three nanoparticles significantly reduced the oxidative stress induced by Cd by regulating phytohormones, phytochelatin, inorganic homeostasis and the expression of genes associated with Cd uptake and transport. Overall, this study elucidates for the first time the multiple complementing mechanisms for some nanoparticles to reduce Cd uptake and transport in rice and provides theoretical basis for applying nanoparticles for reducing Cd accumulation in edible plants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Sweet sorghum deploys tremendous potential for phytoremediation of cadmium (Cd)-polluted soils. Nitrate increases Cd accumulation in sweet sorghum, but the mechanism underlying this is still elusive. Sulfur-containing metabolites have been corroborated to play important roles in Cd tolerance in plants. Thus, whether sulfur metabolism contributed to nitrate-increased Cd accumulation in sweet sorghum was investigated in the present study. Two-way ANOVA analysis showed that most sulfur-containing metabolites concentrations and relevant enzymes activities were regulated by nitrate, Cd and interplay of nitrate and Cd. By using grey correlation analysis and Pearson correlation coefficient, Cd accumulation in shoots as affected by nitrate was also mainly ascribed to sulfur metabolism. ATP sulfurylase (ATPS) activities and non-protein thiol (NPT) concentrations in leaves were the two prominent factors that positively correlated with Cd accumulation in shoots. Excess nitrate elevated ATPS activities in leaves which contributed to increased NPT and phytochelatins (PCs) concentrations in leaves. Nitrate enhanced Cd accumulation in shoots of sweet sorghum under a low level of Cd treatment. Intriguingly, Cd accumulation in shoots of sweet sorghum was similar between a low level and a high level of Cd treatment. Principal Components Analysis (PCA) based on 34 parameters failed to separate the low Cd treatment from the high Cd treatment either, suggesting sweet sorghum is exclusively suitable for phytoremediation of slight Cd-polluted arable lands. Taken together, enhanced Cd accumulation in shoots of sweet sorghum by excess nitrate application is closely correlated with sulfur metabolism containing elevated ATPS activities, NPT and PCs concentrations in leaves., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Phytochelatins (PCs) are small cysteine-rich peptides capable of binding metal(loid)s via SH-groups. Although the biosynthesis of PCs can be induced in vivo by various metal(loid)s, PCs are mainly involved in the detoxification of cadmium and arsenic (III), as well as mercury, zinc, lead, and copper ions, which have high affinities for S-containing ligands. The present review provides a comprehensive account of the recent data on PC biosynthesis, structure, and role in metal(loid) transport and sequestration in the vacuoles of plant cells. A comparative analysis of PC accumulation in hyperaccumulator plants, which accumulate metal(loid)s in their shoots, and in the excluders, which accumulate metal(loid)s in their roots, investigates the question of whether the endogenous PC concentration determines a plant's tolerance to metal(loid)s. Summarizing the available data, it can be concluded that PCs are not involved in metal(loid) hyperaccumulation machinery, though they play a key role in metal(loid) homeostasis. Unraveling the physiological role of metal(loid)-binding ligands is a fundamental problem of modern molecular biology, plant physiology, ionomics, and toxicology, and is important for the development of technologies used in phytoremediation, biofortification, and phytomining.

Phytochelatins (PCs) are class III metallothioneins in plants. They are low molecular-weight polypeptides rich in cysteine residues which can bind to metal ions and affect the physiological metabolism in plants. Unlike other types of metallothioneins, PCs are not the product of gene coding but are synthesized by phytochelatin synthase ( PCS ) based on glutathione (GSH). The chemical formula of phytochelatin is a mixture of (γ-Glu-Cys) n -Gly (n = 2-11) and is influenced by many factors during synthesis. Phytochelatin-like ( PCL ) is a gene-encoded peptide (Met-(α-Glu-Cys) 11 -Gly) designed by our laboratory whose amino acid sequence mimics that of a natural phytochelatin. This study investigated how PCL expression in transgenic plants affects resistance to Cd and Cd accumulation. Under Cd 2+ stress, transgenic plants were proven to perform significantly better than the wild-type (WT), regarding morphological traits and antioxidant abilities, but accumulated Cd to higher levels, notably in the roots. Fluorescence microscopy showed that PCL localized in the cytoplasm and nucleus.

Heavy metals present in the environment can cause a variety of injury symptoms in various organisms including lichens. Most studies examined metal-induced stress under controlled laboratory conditions, and little is known about actual response of lichens in their natural habitat. This study aims to recognize the effect of heavy metal accumulation (total and intracellular) on lichen physiological and biochemical parameters specifically related to the functioning of fungal component. Cladonia rei was used as a model species due to its common occurrence both in unpolluted and extremely polluted sites. We observed a decline in the fungal metabolism which was expressed by a decrease in ergosterol content and an increase in cell membrane damage as a result of increased Zn, Cd, Cu and Ni accumulation. Additionally, the results indicated that increased accumulation of xenobiotics (Pb and As) caused reduction of glutathione (GSH) concentrations and increased membrane lipid peroxidation. Therefore, we conclude that GSH does not provide high oxidative stress protection in C. rei which is somewhat against its insensitivity to pollution. The reduced pool of GSH could be explained by its oxidation to glutathione disulphide induced by heavy metal stress or its use for phytochelatin (PC) synthesis. The content of secondary metabolites was not related to heavy metal accumulation and remained at a relatively stable level. This indicates that the decline in the physiological condition did not weaken the mycobiont of C. rei enough to inhibit the synthesis of secondary metabolites and their precursors were supplied at a sufficient level. Thus, the potential function of main secondary metabolites as extracellular metal immobilizers and antioxidants is still possible even in individuals growing at extremely polluted sites. Despite the evident heavy metal stress, C. rei copes well and spreads easily through extremely polluted environments, which underlines its unique pioneering abilities in highly disturbed sites., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Chromium (Cr) contamination reduces crop productivity worldwide. On the other hand, the expected increase in the future CO 2 levels (eCO 2 ) would improve plant growth under diverse growth conditions. However, the synergetic effect of eCO 2 has not been investigated at both physiological and biochemical levels in Cr-contaminated soil. This study aims to analyze the mitigating effect of eCO 2 on Cr VI phytotoxicity in two rice cultivars (Giza 181 and Sakha 106). Plants are exposed to different Cr concentrations (0, 200 and 400 mg Cr/kg Soil) at ambient (aCO 2 ) and eCO 2 (410 and 620 ppm, respectively). Unlike the stress parameters (MDA, H2O2 and protein oxidation), growth and photosynthetic reactions significantly dropped with increasing Cr concentration. However, in eCO 2 conditions, plants were able to mitigate the Cr stress by inducing antioxidants as well as higher concentrations of phytochelatins to detoxify Cr. Notably, the expression levels of the genes involved in mineral nutrition i.e., OsNRAMP1, OsRT1, OsHMA3, OsLCT1 and iron chelate reductase were upregulated in Cr-stressed Giza 181 plants grown under eCO 2 . Mainly in Sakha 106, eCO 2 induced ascorbate-glutathione (ASC/GSH)-mediated antioxidative defense system. The present study brings the first ever comprehensive assessment of how future eCO 2 differentially mitigated Cr toxicity in rice., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Lead (Pb), like other heavy metals, is not essentially required for optimal plant growth; however, plants uptake it from the soil, which poses an adverse effect on growth and yield. Asparagine (Asp) and thiourea (Thi) are known to assuage the negative impacts of heavy metal pollution on plant growth; however, combined application of Asp and Thi has rarely been tested to discern if it could improve wheat yield under Pb stress. Thus, this experimentation tested the role of individual and combined applications of Asp (40 mM) and Thi (400 mg/L) in improving wheat growth under lead (Pb as PbCl 2 , 0.1 mM) stress. Lead stress significantly reduced plant growth, chlorophyll contents and photosystem system II (PSII) efficiency, whereas it increased Pb accumulation in the leaves and roots, leaf proline contents, phytochelatins, and oxidative stress related attributes. The sole or combined application of Asp and Thi increased the vital antioxidant biomolecules/enzymes, including reduced glutathione (GSH), ascorbic acid (AsA), ascorbate peroxsidase (APX), catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST), dehydroascorbate reductase (DHAR), and glutathione reductase (GR). Furthermore, the sole or the combined application of Asp and Thi modulated nitrogen metabolism by stimulating the activities of nitrate and nitrite reductase, glutamate synthase (GOGAT) and glutamine synthetase (GS). Asp and Thi together led to improve plant growth and vital physiological processes, but lowered down Pb accumulation compared to those by their sole application. The results suggest that Asp and Thi synergistically can improve wheat growth under Pb-toxicity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Masson SAS.)

Industrial wastes and fertilizers can introduce excessive levels of nickel (Ni) into the environment, potentially causing threats to plants, animals, as well as human beings. However, the number of studies on the effects of Ni toxicity on nodules is fairly limited. To address this issue, the effects of increasing Ni concentration on alfalfa nodules were assessed at chemical, biochemical, and transcriptomic levels. For this purpose, plants were grown in soils supplied with Ni (control, 0 mg/kg; C1, 50 mg/kg; C2, 150 mg/kg; C3, 250 mg/kg; and C4, 500 mg/kg) for 90 days. Ni loads in leaves, roots, and nodules were monitored after the exposure period. A set of biochemical biomarkers of oxidative stress was determined in nodules including antioxidants and metal homeostasis as well as lipid peroxidation. Gene expression levels of the main targets involved in oxidative stress and metal homeostasis were assessed. Our data indicated a high concentration of Ni in leaves, roots, and nodules where values reached 25.64 ± 3.04 mg/kg, 83.23 ± 5.16 mg/kg, and 125.71 ± 4.53 mg/kg in dry weight, respectively. Moreover, a significant increase in nodule biomass was observed in plants exposed to C4 in comparison to control treatment and percentage increased by 63%. Then, lipid peroxidation increased with a rate of 95% in nodules exposed to C4. Enzymatic activities were enhanced remarkably, suggesting the occurrence of oxidative stress, with increased superoxide dismutase (SOD), glutathione reductase (GR), and ascorbate peroxidase (APX). Our results showed also a significant upregulation of SOD, GR and APX genes in nodules. Nodule homoglutathione (HGSH) levels increased with the different Ni concentrations, with a remarkable decrease of glutathione S-transferase (GST) activity and glutathione (GSH) content for the highest Ni concentration with 43% and 52% reduction, respectively. The phytochelatin (PC) and metallothionein (MT) concentrations increased in nodules, which implied the triggering of a cellular protection mechanism for coping with Ni toxicity. The results suggested that Ni promotes a drastic oxidative stress in alfalfa nodules, yet the expression of MT and PC to reduce Ni toxicity could be used as Ni stress bioindicators. Our findings provide new insights into the central role of alfalfa nodules in limiting the harmful effects of soil pollution. Therefore, nodules co-expressing antioxidant enzymes may have high phytoremediation potential., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Sumoylation is a posttranslational modification (PTM) in which SUMO (small ubiquitin-like modifier) is covalently conjugated to protein substrates via a range of enzymes. SUMO E3 ligase SIZ1 is involved in mediating several essential or nonessential element-responsive SUMO conjugations in Arabidopsis. However, whether SIZ1 is involved in the cadmium (Cd) response remains to be identified. In this study, we found that SIZ1 positively regulates plant Cd tolerance. The loss-of-function siz1-2 mutant exhibited impaired resistance to Cd exposure and accumulated more reactive oxygen species (ROS). Moreover, the transcription of GSH1, GSH2, PCS1, and PCS2 was suppressed while the accumulation of Cd was enhanced in the siz1-2 mutant under Cd exposure. Further analysis revealed that the higher Cd sensitivity of the siz1-2 mutant was partially rescued by the overexpression of GSH1. Consistently, Cd stress stimulated the accumulation of SUMO1 conjugates in wild-type plants but not in the siz1-2 mutant. Together, our results demonstrate that Cd-induced SIZ1 activates GSH- and PC synthesis-related gene expression to increase the synthesis of GSH- and PCs, thereby leading to higher Cd tolerance in plants., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Lead (Pb) is one of the most harmful, toxic pollutants to the ecological environment and humans. Centipedegrass, a fast-growing warm-season turfgrass, is excellent for Pb pollution remediation. Exogenous low-molecular-weight organic acid (LMWOA) treatment is a promising approach for assisted phytoremediation. However, the effects of this treatment on the tolerance and Pb accumulation of centipedegrass are unclear. This study investigated these effects on the physiological growth response and Pb accumulation distribution characteristics of centipedegrass. Applications of 400 μM citric acid (CA), malic acid (MA) and tartaric acid (TA) significantly reduced membrane lipid peroxidation levels of leaves and improved biomass production of Pb-stressed plants. These treatments mainly increased peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) activities and enhanced free protein (Pro), ascorbic acid (AsA) and phytochelatins (PCs) contents, ultimately improving the Pb tolerance of centipedegrass. Their promoting effects decreased as follows: TA>CA>MA. All the treatments decreased root Pb concentrations and increased stem and leaf Pb concentrations, thus increasing total Pb accumulation and TF values. MA had the best and worst effects on Pb accumulation and Pb transportation, respectively. CA had the best and worst effects on Pb transportation and Pb accumulation, respectively. TA exhibited strong effects on both Pb accumulation and transport. Furthermore, all treatments changed the subcellular Pb distribution patterns and distribution models of the chemical forms of Pb in each tissue. The root Pb concentration was more highly correlated with the Pb subcellular fraction distribution pattern, while the stem and leaf Pb concentrations were more highly correlated with the distribution models of the chemical forms of Pb. Overall, TA improved plant Pb tolerance best and promoted both Pb absorption and transportation well and is considered the best candidate for Pb-contaminated soil remediation with centipedegrass. This study provides a new idea for Pb-contaminated soil remediation with centipedegrass combined with LMWOAs., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Key Message: An organomercurial phenylmercury activates AtPCS1, an enzyme known for detoxification of inorganic metal(loid) ions in Arabidopsis and the induced metal-chelating peptides phytochelatins are essential for detoxification of phenylmercury. Small thiol-rich peptides phytochelatins (PCs) and their synthases (PCSs) are crucial for plants to mitigate the stress derived from various metal(loid) ions in their inorganic form including inorganic mercury [Hg(II)]. However, the possible roles of the PC/PCS system in organic mercury detoxification in plants remain elusive. We found that an organomercury phenylmercury (PheHg) induced PC synthesis in Arabidopsis thaliana plants as Hg(II), whereas methylmercury did not. The analyses of AtPCS1 mutant plants and in vitro assays using the AtPCS1-recombinant protein demonstrated that AtPCS1, the major PCS in A. thaliana, was responsible for the PheHg-responsive PC synthesis. AtPCS1 mutants cad1-3 and cad1-6, and the double mutant of PC-metal(loid) complex transporters AtABCC1 and AtABCC2 showed enhanced sensitivity to PheHg as well as to Hg(II). The hypersensitivity of cad1-3 to PheHg stress was complemented by the own-promoter-driven expression of AtPCS1-GFP. The confocal microscopy of the complementation lines showed that the AtPCS1-GFP was preferentially expressed in epidermal cells of the mature and elongation zones, and the outer-most layer of the lateral root cap cells in the meristematic zone. Moreover, in vitro PC-metal binding assay demonstrated that binding affinity between PC and PheHg was comparable to Hg(II). However, plant ionomic profiles, as well as root morphology under PheHg and Hg(II) stress, were divergent. These results suggest that PheHg phytotoxicity is different from Hg(II), but AtPCS1-mediated PC synthesis, complex formation, and vacuolar sequestration by AtABCC1 and AtABCC2 are similarly functional for both PheHg and Hg(II) detoxification in root surficial cell types., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Phytochelatins (PCs) are cysteine-rich small peptides, enzymatically synthesized from reduced glutathione (GSH) by cytosolic enzyme phytochelatin synthase (PCS). The open reading frame (ORF) of the phytochelatin synthase CaPCS2 gene from the microalgae Chlamydomonas acidophila was heterologously expressed in Escherichia coli strain DH5α, to analyze its role in protection against various abiotic agents that cause cellular stress. The transformed E. coli strain showed increased tolerance to exposure to different heavy metals (HMs) and arsenic (As), as well as to acidic pH and exposure to UVB, salt, or perchlorate. In addition to metal detoxification activity, new functions have also been reported for PCS and PCs. According to the results obtained in this work, the heterologous expression of CaPCS2 in E. coli provides protection against oxidative stress produced by metals and exposure to different ROS-inducing agents. However, the function of this PCS is not related to HM bioaccumulation.

Globally, many people have been suffering from arsenic poisoning. Arsenate (AsV) exposure to twelve rice cultivars caused growth retardation, triggered production of As-chelatin biopeptides and altered activities of antioxidants along with increase in ascorbate (AsA)-glutathione (GSH) contents as a protective measure. The effects were more conspicuous in cvs. Swarnadhan, Tulaipanji, Pusa basmati, Badshabhog, Tulsibhog and IR-20 to attenuate oxidative-overload mediated adversities. Contrastingly, in cvs. Bhutmuri, Kumargore, Binni, Vijaya, TN-1 and IR-64, effects were less conspicuous in terms of alterations in the said variables due to reduced generation of oxidative stress. Under As(V) imposition, the protective role of phytochelatins (PCs) were recorded where peaks height and levels of PCs (PC2, PC3 and PC4) were elevated significantly in the test seedlings with an endeavour to detoxify cells by sequestering arsenic-phytochelatin (As-PC) complex into vacuole that resulted in reprogramming of antioxidants network. Additionally, scatter plot correlation matrices, color-coded heat map analysis and regression slopes demonstrated varied adaptive responses of test cultivars, where cvs. Bhutmuri, Kumargore, Binni, Vijaya, TN-1 and IR-64 found tolerant against As(V) toxicity. Results were further justified by hierarchical clustering. These findings could help to grow identified tolerant rice cultivars in As-prone soil with sustainable growth and productivity after proper agricultural execution., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Arsenic (As) is known to be one of the most toxic metalloids for humans and plants; however, little is known about the use of silicon (Si) and titanium dioxide (TiO 2 ) nanoparticles (NPs) in reducing As toxicity in rice (Oryza sativa L.). The experiment was conducted to examine the effects of Si-NPs (50 and 100 mg/L), TiO 2 -NPs (25 and 50 mg/L) and As (50 µM) on growth, photosynthetic pigments, antioxidant defense system, glyoxalase system, expression of Si/As transporters, and genes involved in As sequestration in rice under hydroponic conditions. The results revealed that Si- and TiO 2 -NPs by upregulating the activity of antioxidant enzymes and glyoxalase cycle reduced hydrogen peroxide, methylglyoxal, malondialdehyde, and electrolyte leakage, and thus protected the photosynthetic apparatus and improved plant growth under As stress. By increasing the expression of GSH1, PCS, and ABC1 genes, Si- and TiO 2 -NPs increased leaf and root accumulation of glutathione and phytochelatins and sequestered As in vacuoles, which protected plant cells from As toxicity. Si-NPs diminished As uptake and increased Si uptake in As-exposed rice plants by modulating the expression of Si/As transporters (Lsi1, Lsi2, and Lsi6). The results depicted that 100 mg/L Si-NPs treatment had the highest positive effect on plant growth and tolerance under As stress compared to other treatments. In general, Si- and TiO 2 -NPs augmented the growth of rice under As stress through different strategies, which can be used to design effective fertilizers to enhance the crop growth and yield in areas contaminated with toxic metals., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Phytochelatins (PCs) are nonribosomal thiol-rich oligopeptides synthetized from glutathione (GSH) in a γ-glutamylcysteinyl transpeptidation reaction catalyzed by PC synthases (PCSs). Ubiquitous in plant and present in some invertebrates, PCSs are involved in metal detoxification and homeostasis. The PCS-like enzyme from the cyanobacterium Nostoc sp. (NsPCS) is considered to be an evolutionary precursor enzyme of genuine PCSs because it shows sufficient sequence similarity for homology to the catalytic domain of the eukaryotic PCSs and shares the peptidase activity consisting in the deglycination of GSH. In this work, we investigate the catalytic mechanism of NsPCS by combining structural, spectroscopic, thermodynamic, and theoretical techniques. We report several crystal structures of NsPCS capturing different states of the catalyzed chemical reaction: (i) the structure of the wild-type enzyme (wt-NsPCS); (ii) the high-resolution structure of the γ-glutamyl-cysteine acyl-enzyme intermediate (acyl-NsPCS); and (iii) the structure of an inactive variant of NsPCS, with the catalytic cysteine mutated into serine (C70S-NsPCS). We characterize NsPCS as a relatively slow enzyme whose activity is sensitive to the redox state of the substrate. Namely, NsPCS is active with reduced glutathione (GSH), but is inhibited by oxidized glutathione (GSSG) because the cleavage product is not released from the enzyme. Our biophysical analysis led us to suggest that the biological function of NsPCS is being a part of a redox sensing system. In addition, we propose a mechanism how PCS-like enzymes may have evolved toward genuine PCS enzymes.

Main Conclusion: A critical investigation into arsenic uptake and transportation, its phytotoxic effects, and defense strategies including complex signaling cascades and regulatory networks in plants. The metalloid arsenic (As) is a leading pollutant of soil and water. It easily finds its way into the food chain through plants, more precisely crops, a common diet source for humans resulting in serious health risks. Prolonged As exposure causes detrimental effects in plants and is diaphanously observed through numerous physiological, biochemical, and molecular attributes. Different inorganic and organic As species enter into the plant system via a variety of transporters e.g., phosphate transporters, aquaporins, etc. Therefore, plants tend to accumulate elevated levels of As which leads to severe phytotoxic damages including anomalies in biomolecules like protein, lipid, and DNA. To combat this, plants employ quite a few mitigation strategies such as efficient As efflux from the cell, iron plaque formation, regulation of As transporters, and intracellular chelation with an array of thiol-rich molecules such as phytochelatin, glutathione, and metallothionein followed by vacuolar compartmentalization of As through various vacuolar transporters. Moreover, the antioxidant machinery is also implicated to nullify the perilous outcomes of the metalloid. The stress ascribed by the metalloid also marks the commencement of multiple signaling cascades. This whole complicated system is indeed controlled by several transcription factors and microRNAs. This review aims to understand, in general, the plant-soil-arsenic interaction, effects of As in plants, As uptake mechanisms and its dynamics, and multifarious As detoxification mechanisms in plants. A major portion of this article is also devoted to understanding and deciphering the nexus between As stress-responsive mechanisms and its underlying complex interconnected regulatory networks., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Soil contamination with toxic metalloids, such as arsenic, can represent a substantial human health and environmental risk. Some plants are thought to tolerate soil toxicity using root exudation, however, the nature of this response to arsenic remains largely unknown. Here, white lupin plants were exposed to arsenic in a semi-hydroponic system and their exudates were profiled using untargeted liquid chromatography-tandem mass spectrometry. Arsenic concentrations up to 1 ppm were tolerated and led to the accumulation of 12.9 μg As g -1 dry weight (DW) and 411 μg As g -1 DW in above-ground and belowground tissues, respectively. From 193 exuded metabolites, 34 were significantly differentially abundant due to 1 ppm arsenic, including depletion of glutathione disulphide and enrichment of phytochelatins and coumarins. Significant enrichment of phytochelatins in exudates of arsenic-treated plants was further confirmed using exudate sampling with strict root exclusion. The chemical tolerance toolkit in white lupin included nutrient acquisition metabolites as well as phytochelatins, the major intracellular metal-binding detoxification oligopeptides which have not been previously reported as having an extracellular role. These findings highlight the value of untargeted metabolite profiling approaches to reveal the unexpected and inform strategies to mitigate anthropogenic pollution in soils around the world., (© 2021 John Wiley & Sons Ltd.)

Rutin is a flavonoid with strong antioxidative effects on plant metabolism that facilitates resistance to environmental stress. The effect of foliar rutin on cadmium (Cd) uptake in Amaranthus hypochondriacus (K472) was studied. The results showed that a foliar spray of rutin alleviated Cd toxicity, promoted plant growth, improved Cd transfer to and storage in aerial plant parts and Cd accumulation with positive effects over time. A rutin concentration of 1.5 mg/mL showed the strongest promotion effect: the biomass and Cd content were increased at 13 days by 68.62% and 405.54% compared to 3 days, respectively, whereas a high concentration of rutin (5 mg/mL) inhibited plant growth and hindered Cd absorption. Two stages of Cd detoxification were identified in K472 after appropriate rutin application. First, an antioxidant system including an enzymatic antioxidant (superoxide dismutase [SOD]) and nonenzymatic antioxidants (glutathione [GSH] and flavonoids) was activated to enhance plant stress resistance. Quercetin and phytochelatin (PC) synthesis were then enhanced to perform detoxification synergistically with the antioxidant system to improve stress tolerance and achieve stable Cd detoxification. The results demonstrated that appropriately prolonging the application time of exogenous rutin to K472 is an effective way to improve the Cd remediation efficiency.

Plants detoxify toxic metal(loid)s by accumulating diverse metabolites. Beside scavenging excess reactive oxygen species (ROS) induced by metal(loid)s, some metabolites chelate metal(loid) ions. Classically, thiol-containing compounds, especially glutathione (GSH) and phytochelatins (PCs) are thought to be the major chelators that conjugate with metal(loid)s in the cytoplasm followed by transport and sequestration in the vacuole. In addition to this classical detoxification pathway, a role for secondary metabolites in metal(loid) detoxification has recently emerged. In particular, anthocyanins, a kind of flavonoids with ROS scavenging potential, contribute to enhanced arsenic tolerance in several plant species. Evidence is accumulating that, in analogy to GSH and PCs, anthocyanins may conjugate with arsenic followed by vacuolar sequestration in the detoxification event. Exogenous application or endogenous accumulation of anthocyanins enhances arsenic tolerance, leading to improved plant growth and productivity. The application of some plant hormones and signaling molecules stimulates endogenous anthocyanin synthesis which confers tolerance to arsenic stress. Anthocyanin biosynthesis is transcriptionally regulated by several transcription factors, including myeloblastosis (MYBs). The light-regulated transcription factor elongated hypocotyl 5 (HY5) also affects anthocyanin biosynthesis, but its role in arsenic tolerance remains elusive. Here, we review the mechanism of arsenic detoxification in plants and the potential role of anthocyanins in arsenic tolerance beyond the classical points of view. Our analysis proposes that anthocyanin manipulation in crop plants may ensure sustainable crop yield and food safety in the marginal lands prone to arsenic pollution., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

For a better understanding of metal-ligand interaction and its function in cells, we developed an easy, sensitive, and high-throughput method to quantify ligand-metal(loid) binding affinity under physiological conditions by combining ligand-attached affinity beads and inductively coupled plasma-optical emission spectrometry (ICP-OES). Glutathione (GSH) and two phytochelatins (PC2 and PC3, small peptides with different numbers of free thiols) were employed as model ligands and attached to hydrophilic beads. The principle of the assay resembles that of affinity purification of proteins in biochemistry: metals binding to the ligand on the beads and the rest in the buffer are separated by a spin column and quantified by ICP-OES. The binding assay using the GSH-attached beads and various metal(loid)s suggested the different affinity of the metal-GSH interactions, in accordance with the order of the Irving-Williams series and the reported stability constants. The binding assay using PC2 or PC3-attached beads suggested positive binding between PCs and Ni(II), Cu(II), Zn(II), Cd(II), and As(III) in accordance with the number of thiols in PC2 and PC3. We then conducted the competition assay using Cd(II), Mn(II), Fe(II), Cu(II), and Zn(II), and the results suggested a better binding affinity of PC2 with Cd(II) than with the essential metals. Another competition assay using PC2 and GSH suggested a robust binding affinity between PCs and Cd(II) compared to GSH and Cd(II). These results suggested the dominance of PC-Cd complex formation in vitro, supporting the physiological importance of PCs for the detoxification of cadmium in vivo. We also discuss the potential application of the assay., (© The Author(s) 2021. Published by Oxford University Press.)

Plants do not always have the genetic capacity to tolerate high levels of arsenic (As), which may not only arrest their growth but pose potential health risks through dietary bioaccumulation. Meanwhile, the interplay between the tomato plants and As-NO-driven molecular cell dynamics is obscure. Accordingly, seedlings were treated with As (10 mg/L) alone or in combination with 100 μM sodium nitroprusside (SNP, NO donor) and 200 μM 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO, NO scavenger). Sodium nitroprusside immobilized As in the roots and reduced the shoot translocation by up-regulating the transcriptional expression of the PCS, GSH1, MT2, and ABC1. SNP further restored the growth retardation through modulating the chlorophyll and proline metabolism, increasing NO accumulation and stomatal conductance along with clear crosstalk between the antioxidant activity as well as glyoxalase I and II leading to endogenous H 2 O 2 and MG reduction. Higher PCs and glutathione accumulation helped protect photosynthetic apparatus; however, cPTIO reversed the protective effects of SNP, confirming the role of NO in the As toxicity alleviation., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

Biscutella auriculata L. is a plant that belongs to the Brassicaceae family and it has been found growing in a metal-contaminated area of the San Quíntín mine (Ciudad Real, Spain). The purpose of this work was to evaluate the mechanisms that allow this plant to tolerate high concentrations of copper. Seedlings were grown in a semi-hydroponic system for 15 days under 125 μM of Cu (NO 3 ) 2 . Exposure to copper resulted in growth inhibition and reduction in the photosynthetic parameters. Copper was mainly accumulated in vascular tissue and vacuoles of the roots and only a minor proportion was transferred to the shoot. Biothiol analysis showed a greater enhancement of reduced glutathione in leaves and increases of phytochelatins (PC2 and PC3) in both leaves and roots. Copper treatment induced oxidative stress, which triggered a response of the enzymatic and non-enzymatic antioxidant mechanisms. The results show that B. auriculata is able to tolerate high metal levels through the activation of specific mechanisms to neutralize the oxidative stress produced and also by metal sequestration through phytochelatins. The preferential accumulation of copper in roots provides clues for further studies on the use of this plant for phytostabilization and environmental recovery purposes in Cu-contaminated areas., (© 2020 Scandinavian Plant Physiology Society.)

Brassica rapa is an important leafy vegetable that can potentially accumulate high concentrations of cadmium (Cd), posing a risk to human health. The aim of the present study was to identify cadmium detoxifying molecular mechanisms in B. rapa using a functional cloning strategy. A cDNA library constructed from roots of B. rapa plants treated with Cd was transformed into the Cd sensitive yeast mutant strain DTY167 that lacks the yeast cadmium factor (YCF1), and resistant yeast clones were selected on Cd containing media. Two hundred genes potentially conferring cadmium resistance were rescued from the surviving yeast clones and sequenced. Sequencing analysis revealed that genes encoding for metallothionein (MT)1, MT2a, MT2b and MT3, and phytochelatin synthase (PCS)1 and PCS2 accounted for 35.5%, 28.5%, 4%, 11.3%, 18.7% and 2%, respectively of the genes identified. MTs and PCSs expressing DTY167 cells showed resistance to Cd as well as to Zn. PCS1 expressing yeast cells were also more resistant to Pb compared to those expressing MTs or PCS2. RT-PCR results showed that Cd treatment strongly induced the expression levels of MTs in the root and shoot. Furthermore, the different MTs and PCSs exhibited tissue specific expression. The results indicate that MTs and PCS genes potentially play a central role in detoxifying Cd and other toxic metals in B. rapa., Competing Interests: The authors declare that no conflicting interests exist.

Cadmium ion (Cd 2+ ) is a common environmental pollutant with high biotoxicity. Interestingly, the Cd 2+ biotoxicity can be alleviated by the coexisting selenite (SeO 3 2- ), which induces the formation of cadmium selenide-rich nanoparticles (CdSe NPs) under the function of thiol-capping peptides. However, the detailed biochemical mechanisms by which Cd and Se are synergistically transformed into CdSe NPs in living organisms remain unclear so far. Here, we shed light on the molecular basis of such biotransformation processes in Caenorhabditis elegans by focusing on the roles of several key thiol-capping peptides. By monitoring the compositional and structural changes of the Cd and Se species and the genetic-level responses of nematodes, we revealed the specific roles of glutathione (GSH) and phytochelatins (PCs) in mediating the CdSe NP formation. With the aid of in vitro bioassembly assay and density functional theory calculations, the detailed Cd-Se interaction pathways were further deciphered: the ingested Cd binds predominantly to GSH and PCs in sequence, then further interacts with selenocysteine to form tetrahedral-structured PC2-Cd 2 -Sec 2 complex, and ultimately grows into CdSe NPs. This work provides molecular-level insights into the Cd-Se interaction in C. elegans and lays a basis for controlling the ecological and health risks of heavy metals in polluted environment.

Rice grains typically contain high levels of toxic arsenic but low levels of the essential micronutrient selenium. Anthropogenic arsenic contamination of paddy soils exacerbates arsenic toxicity in rice crops resulting in substantial yield losses. Here, we report the identification of the gain-of-function arsenite tolerant 1 (astol1) mutant of rice that benefits from enhanced sulfur and selenium assimilation, arsenic tolerance, and decreased arsenic accumulation in grains. The astol1 mutation promotes the physical interaction of the chloroplast-localized O-acetylserine (thiol) lyase protein with its interaction partner serine-acetyltransferase in the cysteine synthase complex. Activation of the serine-acetyltransferase in this complex promotes the uptake of sulfate and selenium and enhances the production of cysteine, glutathione, and phytochelatins, resulting in increased tolerance and decreased translocation of arsenic to grains. Our findings uncover the pivotal sensing-function of the cysteine synthase complex in plastids for optimizing stress resilience and grain quality by regulating a fundamental macronutrient assimilation pathway.

Metal tolerance in marine diatoms vary between morphotypes, strains, and species due to their long-term adaptations to stochastic environments. The mechanisms underlying this highly variable trait remain a matter of interest in ecotoxicology. In this study, we used several cutting-edge techniques, including a non-invasive micro-test technique, atomic force microscopy, and X-ray photoelectron spectroscopy to examine cadmium (Cd) accumulation and tolerance in the three morphotypes of Phaeodactylum tricornutum. Subcellular Cd distribution, metal transporter expression, and glutathione and phytochelatin activity were also analyzed to characterize the morphology-dependent Cd homeostasis and detoxification. We found that the oval morphotype accumulated more Cd, but was also more Cd tolerant than the other morphotypes. The greater surface binding of Cd to the oval morphotype is attributable to its smaller spherical form, rougher cell surface, and lower surface potential. Moreover, the oval morphotype was less permeable to Cd ions and contained higher phytochelatin and glutathione levels, which explained its higher metal tolerance. Our study offers new explanations for diatom's adaptations to changing environments that may contribute to its evolutionary success., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Although Cd is threatening to the environment, animal, and human, the eco-friendly approach to mitigate the Cd-toxicity in alfalfa was barely studied. Therefore, this study aims at elucidating the role of S, a crucial macroelement, in alleviating Cd toxicity in alfalfa plants. The supplementation of S in Cd-stressed alfalfa reversed the detrimental effect on plant biomass, chlorophyll synthesis, and protein concentration. Interestingly, S surplus restored the photosynthetic kinetics, such as Fv/Fm, Pi&#95;ABS, and Mo values in leaves of Cd-stressed alfalfa. Further, Cd-induced adverse effect on membrane stability, cell viability, and redox status was restored due to S under Cd stress. The exogenous S not only increased S status and the expression of sulfate transporters (MsSULRT1;2 and MsSULTR1;3), but also decreased the Cd concentration in the shoot by retaining elevated Cd in root tissue. Further analysis revealed the upregulation of MsGS (glutathione synthetase) and MsPCS1 (phytochelatin synthase) genes along with the increased concentration of glutathione and phytochelatin, predominantly in roots subjected to S surplus under Cd stress. The subcellular Cd analysis showed elevated Cd in the cell wall but not in the vacuole. It suggests that S-induced elevated glutathione enables the phytochelatin to bind with excess Cd leading to subcellular sequestration in the cell wall of roots. Also, S stimulates the S-metabolites and GR enzyme that coordinately counteracts Cd-induced oxidative damage. These findings can be utilized to popularize the application of S and to perform breeding/transgenic experiments to develop Cd-free forage crops., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Using an ideal parental line to breed hybrid rice with low cadmium (Cd) accumulation in grain is an environmental-friendly approach to reduce the risk of Cd contamination in field. A grain low-Cd rice line YaHui2816 has stably low Cd in grain but strong Cd accumulation in straw, revealing specific pattern of its straw-grain Cd remobilization is beneficial to effectively breed hybrid rice for safe production as well as phytoremediation. In this study, a pot experiment was conducted to investigate Cd partitioning of YaHui2816 at different stages by comparison with a common rice C268A. The Cd from lower nodes and leaves was restricted in nodeⅡ, this Cd retention led to weak Cd transport from straw to ear in YaHui2816. Cd concentration in nodeⅡ of YaHui2816 was 1.56-fold and 7.36-fold higher than C268A at filling and mature stages. Thus, glutathione (GSH) and phytochelatin (PC) concentration, PC synthase (OsPCS1), GSH S-Transferase (OsGST) and Cd-remobilization associated genes were measured at filling stage. OsHMA2 and OsCCX2 were up-regulated in nodeⅡ of YaHui2816, relative expression of them were significantly higher than C268A. PCs participated in Cd remobilization process, remarkable PC increase in nodeⅡ of YaHui2816 was found in response to Cd treatment under regulation of OsPCS1 and OsGST of which PC 2 was dominant form. Taken as a whole, the Cd retention in nodeⅡ of YaHui2816 acts as a 'firewall' to restrain Cd transport to grain. This work provides more insight to understand possible function of alleles for reducing Cd concentration in grain as well as strengthening Cd accumulation in straw of YaHui2816., (Copyright © 2020 Elsevier Inc. All rights reserved.)

In recent decades, nanoscale zero valent iron (nZVI) has been found to be a promising approach for heavy metal remediation. This study is the first report highlighting the role of nZVI to ameliorate Cadmium (Cd) stress in rice along with its effects in expressions of transporter genes, agronomic parameters and grain nutrient status. Initially, 3 concentration of Cd (10, 50, 250 μM) and nZVI (50, 100, 200 mg L -1 ) were selected. PCA analysis based on growth parameters, photosynthetic pigment contents and lipid peroxidation rate confirmed that 100 mg L -1 nZVI was most suitable for remediation of 10 μM Cd. It was evident that, nZVI can alleviate Cd-induced toxic effects by enhancing antioxidant defense mechanisms and other physiological processes in plants. nZVI treated rice seedlings also showed upregulation of phytochelatins which aided in Cd chelation within vacuoles. Study of root morphology with scanning electron microscopy and ROS imaging with confocal microscopy confirmed that nZVI could alleviate oxidative stress due to Cd uptake. In nZVI treated rice seedlings, gene expressions of iron (Fe) transporters (like, IRT1,IRT2,YSL2,YSL15) which are responsible for both Fe and Cd uptake were significantly down-regulated whereas, OsVIT1 and OsCAX4 genes were over expressed which lead to sequestration of Cd in vacuoles. Cd localization assay with leadmium proved that Cd translocation was reduced with nZVI treatment. To further validate our findings a pot experiment was carried out where it was found that nZVI could immobilize Cd in soil prevented accumulation of Cd in rice grains in addition to improving yield., Competing Interests: Declaration of competing interest The authors declare no competing financial interests., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Nickel is the most prevailing metal allergen with the highest sensitization rate among the "TOP 25" contact allergens and can affect about 15% of the human population. It is an essential trace metal in plants, animals, and humans. However, the environmental levels of nickel are considerably higher than what is needed for human life. Exposure to high levels of nickel can lead to skin allergies, lung fibrosis, and carcinogenesis. Few existing studies have closely examined the toxicity of nickel, let alone investigated the effective detoxification pathways. Here, we developed a high-throughput screening platform to comprehensively evaluate the nickel toxicity in wild-type C. elegans and explore the underlying detoxification mechanisms in transgenic nematodes. We demonstrated that nickel exerted multiple toxic effects on growth, brood size, feeding, and locomotion in C. elegans. Of which, brood size is the most sensitive endpoint. Nickel was found to first bind to phytochelatin (PC) after entering the worms' body and this PC-Ni complex was further transported by the ABC transporter, CeHMT-1, into the coelomocytes for further detoxification. Our study also demonstrated that the high-throughput screening platform is a promising system for evaluation and investigation of the ecological risks of heavy metals., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)