Phytochelatin Synthase Research Articles

Effects of SpGSH1 and SpPCS1 overexpression or co-overexpression on cadmium accumulation in yeast and Spirodela polyrhiza

Cadmium (Cd) is one of the most toxic elements to all organisms. Glutathione (GSH)-dependent phytochelatin (PC) synthesis pathway is considered an extremely important mechanism in Cd detoxification in plants. However, few studies have focused on the roles of glutamate-cysteine ligase (GSH1) and phytochelatin synthase (PCS1) in Cd accumulation and detoxification in plants. In this study, SpGSH1 and SpPCS1 were identified and cloned from Spirodela polyrhiza and analyzed their functions in yeast and S. polyrhiza via single- or dual-gene (SpGP1) overexpression. The findings of this study showed that SpGSH1, SpPCS1, and SpGP1 could dramatically rescue the growth of the yeast mutant Δycf1. In S. polyrhiza, SpGSH1 was located in the cytoplasm and could promote Mn and Ca accumulation. SpPCS1 was located in the cytoplasm and nucleus, mainly expressed in meristem regions, and promoted Cd, Fe, Mn, and Ca accumulation. SpGSH1 and SpPCS1 co-overexpression increased the Cd, Mn, and Ca contents. Based on the growth data of S. polyrhiza, it was recommended that biomass as the preferable indicator for assessing plant tolerance to Cd stress compared to frond number in duckweeds. Collectively, this study for the first time systematically elaborated the function of SpGSH1 and SpPCS1 for Cd detoxification in S. polyrhiza.

Phytochelatin Synthase: An In Silico Comparative Analysis in Cyanobacteria and Eukaryotic Microalgae.

Phytochelatins (PCs) are small cysteine-rich peptides involved in metal detoxification, not genetically encoded but enzymatically synthesized by phytochelatin synthases (PCSs) starting from glutathione. The constitutive PCS expression even in the absence of metal contamination, the wide phylogenetic distribution and the similarity between PCSs and the papain-type cysteine protease catalytic domain suggest a wide range of functions for PCSs. These proteins, widely studied in land plants, have not been fully analyzed in algae and cyanobacteria, although these organisms are the first to cope with heavy-metal stress in aquatic environments and can be exploited for phytoremediation. To fill this gap, we compared the features of the PCS proteins of different cyanobacterial and algal taxa by phylogenetic linkage. The analyzed sequences fall into two main, already known groups of PCS-like proteins. Contrary to previous assumptions, they are not classed as prokaryotic and eukaryotic sequences, but rather as sequences characterized by the alternative presence of asparagine and aspartic/glutamic acid residues in proximity of the catalytic cysteine. The presence of these enzymes with peculiar features suggests differences in their post-translational regulation related to cell/environmental requirements or different cell functions rather than to differences due to their belonging to different phylogenetic taxa.

Phytochelatins Bind Zn(II) with Micro- to Picomolar Affinities without the Formation of Binuclear Complexes, Exhibiting Zinc Buffering and Muffling Rather than Storing Functions.

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 ZnL2 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.

RBOH-dependent H2O2 burst induced by silicon dioxide nanoparticles establishes systemic acquired acclimation in quinoa under lead toxicity

The role of silicon dioxide nanoparticles (nSiO2) and hydrogen peroxide (H2O2) in heavy metal tolerance of plants has been well defined; however, their interaction together remains largely unknown. Therefore, two experiments were conducted to determine whether RBOH-dependent signaling is involved in nSiO2-induced systemic lead (Pb) acclimation in Chenopodium quinoa. In the first experiment, quinoa leaves were sprayed with nSiO2 (0, 2, 4 mM) under various Pb concentrations (0, 200 and 400 µM). After 12 h, an H2O2 burst, and higher transcription levels of RBOHD and PCS1 genes (encoding NADPH oxidase and phytochelatin synthase) were recorded in leaves of nSiO2-treated plants. After 15 days, Pb toxicity reduced growth parameters, and Chl a, Chl b contents and increased electrolyte leakage, and foliar spray with nSiO2 reversed this tendency. In the second experiment, foliar application of 4 mM nSiO2 minimized intracellular H2O2 and O2˙¯ and reduced malondialdehyde content and leaf Pb concentration under 400 µM Pb. While, this treatment increased the content of reduced ascorbate, and thiol compounds and enhanced the activities of superoxide dismutase and ascorbate peroxidase not only in leaves but also in untreated roots. However, H2O2 scavenging with DMTU and the inhibition of RBOH-dependent signaling by DPI aggravated Pb phytotoxicity and impaired the beneficial effects of nSiO2 on aforementioned attributes; thereby reduced tolerance index in both leaves and roots. Together, our pharmacological and molecular evidence for the first time unraveled that RBOH-dependent H2O2 burst plays vital role in triggering nSiO2-induced systemic acquired acclimation in Pb-stressed plants.

Intracellular and extracellular thiol-peptides modulate the response of Marchantia polymorpha to physiological needs, excess, and starvation of zinc, copper, and iron

Thiol-peptides, such as glutathione and phytochelatins, are believed to play a crucial role in regulating metal micronutrient needs (in particular, copper, iron, and zinc), and in detoxifying their excessive amounts. This study provides insight into the mechanisms involved in detoxification, starvation, and physiological control of the aforesaid metals in the liverwort Marchantia polymorpha. The results show that the phytochelatin synthase activity in gametophytes of M. polymorpha is induced mainly by zinc, followed by copper, and iron. Interestingly, the amount of phytochelatins produced seems to be controlled by the enzyme activity, rather than by the expression level of its gene. Moreover, when gametophytes are deprived of zinc, copper, and iron, phytochelatin levels decrease, compared with normal nutrient supply. Phytochelatins and glutathione, on the other hand, appear to play a key role in detoxification of excess zinc, copper, and iron, particularly when these thiol-peptides are released extracellularly. This suggests a potential novel function for phytochelatins and glutathione as detoxifying compounds also in the extracellular environment.

Mitigating Effect of Trans-Zeatin on Cadmium Toxicity in Desmodesmus armatus.

Phytohormones, particularly cytokinin trans-zeatin (tZ), 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 tZ 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 tZ 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.

Glycine-Rich RNA-Binding Protein AtGRP7 Functions in Nickel and Lead Tolerance in Arabidopsis.

Plant glycine-rich RNA-binding proteins (GRPs) play crucial roles in the response to environmental stresses. However, the functions of AtGRP7 in plants under heavy metal stress remain unclear. In the present study, in Arabidopsis, the transcript level of AtGRP7 was markedly increased by Ni but was decreased by Pb. AtGRP7-overexpressing plants improved Ni tolerance, whereas the knockout mutant (grp7) was more susceptible than the wild type to Ni. In addition, grp7 showed greatly enhanced Pb tolerance, whereas overexpression lines showed high Pb sensitivity. Ni accumulation was reduced in overexpression lines but increased in grp7, whereas Pb accumulation in grp7 was lower than that in overexpression lines. Ni induced glutathione synthase genes GS1 and GS2 in overexpression lines, whereas Pb increased metallothionein genes MT4a and MT4b and phytochelatin synthase genes PCS1 and PCS2 in grp7. Furthermore, Ni increased CuSOD1 and GR1 in grp7, whereas Pb significantly induced FeSOD1 and FeSOD2 in overexpression lines. The mRNA stability of GS2 and PCS1 was directly regulated by AtGRP7 under Ni and Pb, respectively. Collectively, these results indicate that AtGRP7 plays a crucial role in Ni and Pb tolerance by reducing Ni and Pb accumulation and the direct or indirect post-transcriptional regulation of genes related to heavy metal chelators and antioxidant enzymes.

Potential of Salvinia molesta as a Copper Phytoremediation Agent based on Gene Expression Analysis

This research explores the rapid environmental impact of the batik industry, particularly concerning batik waste pollution, especially from the heavy metal copper (Cu). Untreated batik waste can have adverse effects on the environment. The study highlights the potential of the water fern plant (Salvinia molesta) as a heavy metal phytoremediator, specifically for Cu, by activating phytochelatin synthase. The research aims to determine the most effective waste concentration for Cu absorption by water ferns, measure the reduction in Cu levels after phytoremediation treatment, evaluate post-phytoremediation water quality, and observe PCS gene expression in the roots and leaves of water ferns. A static method is employed with variations in waste concentrations. Data analysis utilizes one-way ANOVA for Cu level reduction and changes in water quality. The results indicate that the most effective wastewater concentration for absorbing heavy metal copper (Cu) using water fern (Salvinia molesta) is 2%. The highest reduction in heavy metal copper (Cu) concentration after phytoremediation treatment with water fern (Salvinia molesta) is 41.48%. Water quality post-phytoremediation treatment using water fern (Salvinia molesta) at all concentrations exhibits improvement with an increase in dissolved oxygen (DO) levels. The expression of the phytochelatin synthase (PCS) gene in the leaves and roots of water fern (Salvinia molesta) confirms the role of the PCS gene in binding heavy metal copper (Cu) to the plant's vacuole.

Characterization of a Nicotiana tabacum phytochelatin synthase 1 and its response to cadmium stress.

Phytochelatin synthase (PCS) is a critical enzyme involved in heavy metal detoxification in organisms. In this study, we aim to comprehensively investigate the molecular and functional characteristics of the PCS1 gene from Nicotiana tabacum by examining its enzymatic activity, tissue-specific expression pattern, Cd-induced expression, as well as the impact on Cd tolerance and accumulation. The results demonstrated that the amino acid sequence of NtPCS1 shared a highsimilarity in its N-terminal region with PCS from other species. The enzymaticactivity of NtPCS1 was found to be enhanced in the order Ag2+ > Cd2+ > Cu2+ > Pb2+ > Hg2+ > Fe2+ > Zn2+. In addition, RT-PCR data indicated that NtPCS1 gene is constitutively expressed, with the highest expression observed in flowers, and that its transcript levels are up-regulated by CdCl2. When tobacco overexpressing NtPCS1 (PCS1 lines) were grown under CdCl2 stress, they produced more phytochelatins (PCs) than WT plants, but this did not result in increased Cd accumulation. However, in a root growth assay, the PCS1 lines exhibited hypersensitivity to Cd. The overexpression of NtPCS1 itself does not appear to be the primary cause of this heightened sensitivity to Cd, as the Arabidopsis thaliana Atpcs1 mutant overexpressing NtPCS1 actually exhibited enhanced tolerance to Cd. Furthermore, the addition of exogenous glutathione (GSH) progressively reduced the Cd hypersensitivity of the PCS1 lines, with the hypersensitivity even being completely eliminated. Surprisingly, the application of exogenous GSH led to a remarkably enhanced Cd accumulation in the PCS1 lines. This study enriches our understanding of the molecular function of the NtPCS1 gene and suggests a promising avenue for Cd tolerance through the heterologous expression of PCS genes in different species.

Hormones and the antioxidant transduction pathway and gene expression, mediated by Serratia marcescens DB1, lessen the lethality of heavy metals (As, Ni, and Cr) in Oryza sativa L.

Microorganisms have recently gained recognition as efficient biological tool for reducing heavy metal toxicity in crops. In this experiment, we isolated a potent heavy metal (As, Ni, and Cr) resistant rhizobacterium Serratia marcescens DB1 and detected its plant growth promoting traits such as phosphate solubilization, gibberellin synthesis, organic acid production and amino acid regulation. Based on these findings, DB1 was further investigated for application in a rice var. Hwayeongbyeo subjected to 1 mM As, 4 mM Ni, and 4 mM Cr stress. The rice plants treated with Cr and Ni appeared healthy but were lethal, indicating unfitness for consumption due to toxic metal deposition, whereas the plants treated with > 1 mM As instantaneously died. Our results showed that DB1 inoculation significantly decreased metal accumulation in the rice shoots. Particularly, Cr uptake dropped by 16.55% and 22.12% in (Cr + DB1) and (Cr + As + Ni + DB1), respectively, As dropped by 48.90% and 35.82% in (As + DB1) and (Cr + As + Ni + DB1), respectively, and Ni dropped by 7.95% and 19.56% in (Ni + DB1) and (Cr + As + Ni + DB1), respectively. These findings were further validated by gene expression analysis results, which showed that DB1 inoculation significantly decreased the expression of OsPCS1 (a phytochelatin synthase gene), OsMTP1 (a metal transporting gene), and OsMTP5 (a gene for the expulsion of excess metal). Moreover, DB1 inoculation considerably enhanced the morphological growth of rice through modulation of endogenous phytohormones (abscisic acid, salicylic acid, and jasmonic acid) and uptake of essential elements such as K and P. These findings indicate that DB1 is an effective biofertilizer that can mitigate heavy metal toxicity in rice crops.

Integrated analysis of transcriptome, metabolome and physiology revealed the molecular mechanisms for elevated pH-mediated-mitigation of aluminum-toxicity in Citrus sinensis leaves

The contribution of aluminum (Al)-pH-interaction-responsive metabolites and genes to Al-tolerance in Citrus leaves is still poorly understood. Seedlings of ‘Xuegan’ (Citrus sinensis) were supplied with nutrient solution at a pH of 4.0 or 3.0 and an Al concentration of 1 or 0 mM for 18 weeks. Thereafter, we investigated the impacts of Al-pH interactions on plant growth; Al concentrations in leaves and roots; and metabolome, transcriptome, and related physiological indices in leaves. Elevated pH ameliorated Al-toxicity-induced reduction of seedling (leaf) growth. Here, we screened 867 upregulated and 1041 downregulated genes, and 43 [eight primary metabolites (PMs) + 35 secondary metabolites (SMs)] and 28 downregulated (23 PMs + five SMs) in pH 3.0 + 1 mM Al-treated leaves (P3AL) vs. pH 3.0 + 0 mM Al-treated leaves (P3L); and 410 upregulated and 149 downregulated genes, and 75 upregulated (20 PMs + 55 SMs) and 32 downregulated (25 PMs + seven SMs) metabolites in pH 4.0 + 1 mM Al-treated leaves (P4AL) vs. pH 4.0 + 0 mM Al-treated leaves (P4L), implying that elevated pH ameliorated Al-toxic impacts on gene expression, but it enhanced the adaptability of metabolites to Al-stress. Further analysis suggested that raised pH-mediated-amelioration of leaf Al-stress involved the several aspects, including: (a) decreased accumulation of callose and low methylesterified pectin, increased accumulation of high methylesterified pectin, and enhanced cell wall stability; (b) less alterations of lipid metabolism and downregulation of PMs; (c) increased biosynthesis and accumulation of SMs; and (d) enhanced ability to maintain energy, phosphate and cell redox homeostasis, and less oxidative injury due to higher ability to maintain the balance between reactive oxygen species and aldehyde generation and detoxification. Some metabolic pathways such as glycolysis / gluconeogenesis and phenylpropanoid pathway; metabolites such as callose and phenolic compounds; and genes such as xyloglucan endotransglucosylase/hydrolases, purple acid phosphatases, phytochelatin synthase 3, and cysteine synthase, might involve in raised pH-induced amelioration of leaf Al-stress. Also, some similarities and differences existed between leaves and roots in Al-pH-interaction-responsive metabolites and genes.

The Effect of Cadmium on Plants in Terms of the Response of Gene Expression Level and Activity.

Cadmium (Cd) is a heavy metal that can cause damage to living organisms at different levels. Even at low concentrations, Cd can be toxic to plants, causing harm at multiple levels. As they are unable to move away from areas contaminated by Cd, plants have developed various defence mechanisms to protect themselves. Hyperaccumulators, which can accumulate and detoxify heavy metals more efficiently, are highly valued by scientists studying plant accumulation and detoxification mechanisms, as they provide a promising source of genes for developing plants suitable for phytoremediation techniques. So far, several genes have been identified as being upregulated when plants are exposed to Cd. These genes include genes encoding transcription factors such as iron-regulated transporter-like protein (ZIP), natural resistance associated macrophage protein (NRAMP) gene family, genes encoding phytochelatin synthases (PCs), superoxide dismutase (SOD) genes, heavy metal ATPase (HMA), cation diffusion facilitator gene family (CDF), Cd resistance gene family (PCR), ATP-binding cassette transporter gene family (ABC), the precursor 1-aminocyclopropane-1-carboxylic acid synthase (ACS) and precursor 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) multigene family are also influenced. Thanks to advances in omics sciences and transcriptome analysis, we are gaining more insights into the genes involved in Cd stress response. Recent studies have also shown that Cd can affect the expression of genes related to antioxidant enzymes, hormonal pathways, and energy metabolism.

Effect of Methyl Jasmonate on the Gene Expression, Encoding Non-Protein Thiol Enzymes in Wheat under Cadmium.

The effect of the phytohormone methyl jasmonate (MJ) on the expression of the TaGS1 and TaPCS1 genes encoding glutathione synthetase and phytochelatin synthase, respectively, which are key enzymes in the synthesis of glutathione and phytochelatins in wheat (cv. Moskovskaya 39), was studied. It was shown for the first time that pretreatment of plants by exogenous MJ (1 μM) leads to an increase in the accumulation of transcripts of the TaGS1 and TaPCS1 genes in leaves without cadmium effect. When cadmium (CdSO4, 100 μM) was added into the nutrient solution, the transcript level of the TaGS1 gene in the plants pretreated by MJ increased (compared to the untreated plants), whereas the transcript level of TaPCS1 remained unaffected. The pretreatment of plants with MJ leads to a lower accumulation of cadmium in the roots and leaves of wheat. At the same time, MJ had no effect on the linear growth parameters of plants, but had a positive effect on the accumulation of biomass under cadmium. It was assumed that MJ is involved in plant tolerance to cadmium by increasing the expression of the TaGS1 and TaPCS1 genes and, as a result, enhancing the synthesis of chelating compounds, as well as by reducing the supply of metal ions to plants.

Glutathione and phytochelatins jointly allow intracellular and extracellular detoxification of cadmium in the liverwort Marchantia polymorpha

Plants have evolved a set of mechanisms that control and respond to the uptake and accumulation of both essential and non-essential metals, including chelation and sequestration of these elements by thiol ligands, such as glutathione and phytochelatins. Indeed, such thiol peptides can chelate some metals, quickly form thiol-metal complexes, and segregate them in the vacuolar compartment. Reasonably, conceptually similar mechanisms can be assumed to be responsible for the transport of metal complexes ̶ in particular thiol-cadmium complexes ̶ across the plasma membrane, with the consequent release of this toxic metal in the extracellular environment. Such hypothesis, focusing on prevention and detoxification mechanisms, was here verified in axenically-grown gametophytes of the model liverwort Marchantia polymorpha, exposed to three different cadmium concentrations over five exposure times. From the data obtained, it can be deduced that the cell wall of M. polymorpha moderately reduced the influx of cadmium into the cells, since it was rapidly saturated by this metal. At an intracellular level, cadmium induced the activity, but not the gene expression, of the phytochelatin synthase enzyme, leading to synthesis of phytochelatins. Moreover, both glutathione and phytochelatins chelated cadmium at the cytosolic level and allowed its detoxification, possibly involving tonoplast transporters belonging to type-C ABC subfamily (Mp7g13860 and Mp4g11930). Likewise, cadmium, glutathione and phytochelatins were released extracellularly, thus highlighting a possible novel role in cadmium detoxification, in a pH-dependent manner. The overall results suggest that, in M. polymorpha, glutathione and phytochelatins can accomplish intracellular and extracellular detoxification of cadmium.

EFFECT OF METHYL JASMONATE ON THE GENE EXPRESSION, ENCODING NON-PROTEIN THIOL ENZYMES IN WHEAT UNDER CADMIUM

The effect of the phytohormone – methyl jasmonate (MJ) on the expression of the TaGS1 and TaPCS1 genes encoding glutathione synthetase and phytochelatin synthase, respectively, which are key enzymes in the synthesis of glutathione and phytochelatins in wheat (var. Moskovskaya 39), was studied. For the first time, it has been shown, that pretreatment of plants by exogenous MJ (1 μM) leads to an increase in the accumulation of transcripts of the TaGS1 and TaPCS1 genes in leaves without cadmium effect. When cadmium (CdSO4, 100 μM) was added into the nutrient solution, the transcript level of TaGS1 gene increased in plants pretreated by MJ (compared to untreated plants), while the transcript level of TaPCS1 remained unaffected. The pretreatment of plants by MJ leads to a lower accumulation of cadmium in the roots and leaves of wheat. At the same time, MJ had no effect on the linear growth parameters of plants, but had a positive effect on the accumulation of biomass under cadmium. It has been suggested that MJ is involved in plant tolerance to cadmium by increasing the expression of the TaGS1 and TaPCS1 genes and, as a result, enhancing the synthesis of chelating compounds, as well as by reducing the supply of metal ions to plants.

The role of phosphatidylcholine 34:1 in the occurrence, development and treatment of ulcerative colitis.

Lipid homeostasis is considered to be related to intestinal metabolic balance, while its role in the pathogenesis and treatment of ulcerative colitis (UC) remains largely unexplored. The present study aimed to identify the target lipids related to the occurrence, development and treatment of UC by comparing the lipidomics of UC patients, mice and colonic organoids with the corresponding healthy controls. Here, multi-dimensional lipidomics based on LC-QTOF/MS, LC-MS/MS and iMScope systems were constructed and used to decipher the alteration of lipidomic profiles. The results indicated that UC patients and mice were often accompanied by dysregulation of lipid homeostasis, in which triglycerides and phosphatidylcholines were significantly reduced. Notably, phosphatidylcholine 34:1 (PC34:1) was characterized by high abundance and closely correlation with UC disease. Our results also revealed that down-regulation of PC synthase PCYT1α and Pemt caused by UC modeling was the main factor leading to the reduction of PC34:1, and exogenous PC34:1 could greatly enhance the fumarate level via inhibiting the transformation of glutamate to N-acetylglutamate, thus exerting an anti-UC effect. Collectively, our study not only supplies common technologies and strategies for exploring lipid metabolism in mammals, but also provides opportunities for the discovery of therapeutic agents and biomarkers of UC.

Physiological and transcriptome analysis of response of soybean (Glycine max) to cadmium stress under elevated CO2 concentration

The continuous accumulation of Cd has long-lasting detrimental effects on plant growth and food safety. Although elevated CO2 concentration (EC) has been reported to reduce Cd accumulation and toxicity in plants, evidence on the functions of elevated CO2 concentration and its mechanisms in the possible alleviation of Cd toxicity in soybean are limited. Here, we used physiological and biochemical methods together with transcriptomic comparison to explore the effects of EC on Cd-stressed soybean. Under Cd stress, EC significantly increased the weight of roots and leaves, promoted the accumulations of proline, soluble sugars, and flavonoid. In addition, the enhancement of GSH activity and GST gene expressions promoted Cd detoxification. These defensive mechanisms reduced the contents of Cd2+, MDA, and H2O2 in soybean leaves. The up-regulation of genes encoding phytochelatin synthase, MTPs, NRAMP, and vacuoles protein storage might play vital roles in the transportation and compartmentalization process of Cd. The MAPK and some transcription factors such as bHLH, AP2/ERF, and WRKY showed changed expressions and might be engaged in mediation of stress response. These findings provide a boarder view on the regulatory mechanism of EC on Cd stress and provide numerous potential target genes for future engineering of Cd-tolerant cultivars in soybean breeding programs under climate changes scenarios.

Characterization of γ-glutamyltransferase- and phytochelatin synthase-mediated catabolism of glutathione and glutathione S-conjugates in Arabidopsis thaliana.

Glutathione (GSH, γ-L-glutamyl-L-cysteinyl-glycine) has been implicated in a multitude of cellular functions, such as protection of cells against oxidative stress, detoxification of xenobiotics via degradation of GSH S-conjugates, and disease resistance. Glutathione also serves as a precursor of phytochelatins, and thereby plays an essential role in heavy metal detoxification. The Arabidopsis genome encodes three functional γ-glutamyltransferase genes (AtGGT1, AtGGT2, AtGGT4) and two phytochelatin synthase genes (AtPCS1, AtPCS2). The function of plant GGT has not yet been clearly defined, although it is thought to be involved in GSH and GSH S-conjugate catabolism. On the other hand, besides its role in heavy metal detoxification, PCS has also been involved in GSH S-conjugate catabolism. Herein we describe the HPLC characterization of GSH and GSH S-conjugate catabolism in Arabidopsis mutants deficient in GSH biosynthesis (pad2-1/gsh1), atggt and atpcs1 T-DNA insertion mutants, atggt pad2-1, atggt atpcs1 double mutants, and the atggt1 atggt4 atpcs1 triple mutant. The results of our HPLC analysis confirm that AtGGT and AtPCS play important roles in two different pathways related with GSH and GSH S-conjugate (GS-bimane) catabolism in Arabidopsis.

Effects of Phytochelatin-like Gene on the Resistance and Enrichment of Cd2+ in Tobacco.

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 Cd2+ 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.

Physiological and transcriptomic study reveal SeNPs-mediated AsIII stress detoxification mechanisms involved modulation of antioxidants, metal transporters, and transcription factors in Glycine max L. (Merr.) roots

Physiological changes and genome-wide alteration in gene expression were performed in soybean (Glycine max [L.] Merr.) roots exposed to AsⅢ (25 μmol/L) alone and supplemented with selenium nanoparticles (SeNPs) at the concentration of 10 and 25 μmol/L at the V2 growth stage. Excessive arsenic in the root zone poses a potential threat to soybean yield, particularly to roots, due to the limited translocation of AsIII from root to shoot in the case of soybean. We hypothesized that SeNPs can relieve AsⅢ toxicity to soybean root by reducing the AsⅢ uptake and regulating the internal tolerance mechanism of the plants. Results accomplished that SeNPs had positive impact on soybean dry weight and roots parameters under AsⅢ stress. Then, we further evaluated physiological indexes, whole genome transcriptomic analysis and quantitative real-time PCR to elucidate the underlying mechanism of AsⅢ tolerance under SeNPs supplementation. Under the condition of AsⅢ-stress, SeNPs exposure significantly reduced the electrolyte leakage, O2−•, H2O2 and MDA accumulation while increasing the antioxidants level. The RNA-seq dataset revealed total of 5819 up and 7231 down expressed DEGs across all libraries. The number of exclusively regulated genes were higher under As + SeNP10 (4909) treatment than in the AsⅢ-alone (4830) and As + SeNP25 (3311) treatments. The KEGG and GO analyses revealed that stress responsive DEGs such as glutathione S-transferase, glutathione peroxidase, ascorbate, glutaredoxin, thioredoxin, and phytochelatins synthase are responsible for AsⅢ tolerance under the SeNPs supplementation. Similarly, sulfate transporter, and ABC transporters (ATP-binding cassettes) expression were induced, and aquaporin channels related DEGs expression were reduced under SeNPs application in AsⅢ exposure condition. Furthermore, the expression of molecular chaperones (HSP) and transcription factors (MYB, bZIP, bHLH, and HSFs) were increased in SeNPs treatment groups. These results provide vital information of AsⅢ tolerance mechanism in response to SeNPs in soybean. We suggest that functional characterization of these genes will help us learn more about the SeNPs responsive arsenic tolerance mechanism in soybean.