μM Cd Research Articles

Calcium and ethylene glycol tetraacetic acid mitigate toxicity and alteration of gene expression associated with cadmium stress in chickpea (Cicer arietinum L.) shoots.

In the aim to estimate the protective role of calcium (Ca) and ethylene glycol tetraacetic acid (EGTA) against cadmium (Cd)-induced damage, chickpea (Cicer arietinum L.) seeds were exposed to 200 μM Cd stress for 6 days or 3 days then subjected to co-treatment of the metal with either 100 mM CaCl2 or 100 μM EGTA for 3 additional days. The addition of Ca and EGTA improved seedling growth. This protecting effect was correlated to the alleviation of the metal-induced oxidative stress, exemplified by the reduction of hydrogen peroxide (H2O2) contents. Besides, Ca and EGTA stimulated thioredoxin (Trx) and thioredoxin reductase (NTR) activities (2.75- and 1.75-fold increase when compared to Cd-stressed, respectively) protecting, thereby, protein -SH groups from the Cd-mediated oxidation, and modulated ferredoxin (Fdx) activity to a control level. Moreover, Ca and EGTA reinstated the glutathione redox steady state, mainly via preserving a high level of glutathione reduced form (GSH). This effect coincided with the maintaining of the Cd-stimulated glutathione reductase (GR) activity and the decline of glutathione peroxidase (GPX, 43% lower than Cd-stressed shoots) activity. Ca and EGTA counteracted the inhibitory effect of Cd on the activity and gene expression of Cu/Zn-superoxide dismutase (Cu/Zn-SOD) isoenzyme and modulated the activities of catalase (CAT) and ascorbate peroxidase (APX). Overall, our results provided evidence that Ca and EGTA supplement could be a promising approach in the remediation of Cd-contaminated environment.

Combined transcriptomic, proteomic and biochemical approaches to identify the cadmium hyper-tolerance mechanism of turnip seedling leaves.

Cadmium (Cd) pollution is a prominent environment problem, and great interests have been developed towards the molecular mechanism of Cd accumulation in plants. In this study, we conducted combined transcriptomic, proteomic and biochemical approaches to explore the detoxification of a Cd-hyperaccumulating turnip landrace exposed to 5 μM (T5) and 25 μM (T25) Cd treatments. A total of 1090 and 2111 differentially expressed genes (DEGs) and 161 and 303 differentially expressed proteins (DEPs) were identified in turnips under T5 and T25, respectively. However, poor correlations were observed in expression changes between mRNA and protein levels. The enriched KEGG pathways of DEGs with a high proportion (> 80%) of upregulated genes were focused on the flavonoid biosynthesis, sulphur metabolism and glucosinolate biosynthesis pathways, whereas those of DEPs were enriched on the glutathione metabolism pathway. This result suggests that these pathways contribute to Cd detoxification in turnips. Furthermore, induced antioxidant enzymes, heat stock proteins and stimulated protein acetylation modification seemed to play important roles in Cd tolerance in turnips. In addition, several metal transporters were found responsible for the Cd accumulation capacity of turnips. This study may serve as a basis for breeding low-Cd-accumulating vegetables for foodstuff or high-Cd-abstracting plants for phytoremediation.

Comparative transcriptome analysis uncovers roles of hydrogen sulfide for alleviating cadmium toxicity in Tetrahymena thermophila

BackgroundCadmium (Cd) is a nonessential heavy metal with potentially deleterious effects on different organisms. The organisms have evolved sophisticated defense system to alleviate heavy metal toxicity. Hydrogen sulfide (H2S) effectively alleviates heavy metal toxicity in plants and reduces oxidative stress in mammals. However, the function of H2S for alleviating heavy metal toxicity in aquatic organisms remains less clear. Tetrahymena thermophila is an important model organism to evaluate toxic contaminants in an aquatic environment. In this study, the molecular roles of exogenously H2S application were explored by RNA sequencing under Cd stress in T. thermophila.ResultsThe exposure of 30 μM Cd resulted in T. thermophila growth inhibition, cell nigrescence, and malondialdehyde (MDA) content considerably increase. However, exogenous NaHS (donor of H2S, 70 μM) significantly alleviated the Cd-induced toxicity by inhibiting Cd absorbtion, promoting CdS nanoparticles formation and improving antioxidant system. Comparative transcriptome analysis showed that the expression levels of 9152 genes changed under Cd stress (4658 upregulated and 4494 downregulated). However, only 1359 genes were differentially expressed with NaHS treatment under Cd stress (1087 upregulated and 272 downregulated). The functional categories of the differentially expressed genes (DEGs) by gene ontology (GO) revealed that the transcripts involved in the oxidation–reduction process, oxidoreductase activity, glutathione peroxidase activity, and cell redox homeostasis were the considerable enrichments between Cd stress and NaHS treatment under Cd stress. Kyoto Encyclopedia of Genes and Genomes (KEGG) indicated that the carbon metabolism, glutathione metabolism, metabolism of xenobiotics by cytochrome P450, and ABC transporters were significantly differentially expressed components between Cd stress and NaHS treatment under Cd stress in T. thermophila. The relative expression levels of six DEGs were further confirmed through quantitative real-time polymerase chain reaction (qRT-PCR).ConclusionNaHS alleviated Cd stress mainly through inhibiting Cd absorbtion, promoting CdS nanoparticles formation, increasing oxidation resistance, and regulation of transport in free-living unicellular T. thermophila. These findings will expand our understanding for H2S functions in the freshwater protozoa.

Modulation of Key Physio-Biochemical and Ultrastructural Attributes after Synergistic Application of Zinc and Silicon on Rice under Cadmium Stress.

Excessive industrialization and the usage of pesticides plague the farming soils with heavy metals, reducing the quality of arable land. Assessing phytoavailability of cadmium (Cd) from growth medium to plant system is crucial and necessitates precise and timely monitoring of Cd to ensure food safety. Zinc (Zn) and silicon (Si) have singularly demonstrated the potential to ameliorate Cd toxicity and are important for agricultural production, human health, and environment in general. However, Zn-Si interaction on Cd toxicity alleviation, their effects and underlying mechanisms are still fragmentarily understood. Seven treatments were devised besides control to evaluate the single and combined effects of Zn and Si on the physio-biochemical attributes and ultrastructural fingerprints of Cd-treated rice genotypes, i.e., Cd tolerant “Xiushui-110” and Cd sensitive “HIPJ-1”. Supplementation of both Zn and Si promoted plant biomass, photosynthetic parameters, ionic balance, and improved chloroplast ultrastructure with minimized Cd uptake and malondialdehyde (MDA) content due to the activation of antioxidant enzymes in Cd stressed plants. The combined effects of 10 μM Zn and 15 μM Si on 15 μM Cd displayed a greater reduction in Cd uptake and root-leaf MDA content, while enhancing photosynthetic activity, superoxide dismutase (SOD) activity and root-leaf ultrastructure particularly in HIPJ-1, whilst Xiushui-110 had an overall higher leaf catalase (CAT) activity and a higher root length and shoot height was observed in both genotypes compared to the Cd 15 µM treatment. Alone and combined Zn and Si alleviation treatments reduced Cd translocation from the root to the stem for HIPJ-1 but not for Xiushui-110. Our results confer that Zn and Si singularly and in combination are highly effective in reducing tissue Cd content in both genotypes, the mechanism behind which could be the dilution effect of Cd due to improved biomass and competitive nature of Zn and Si, culminating in Cd toxicity alleviation. This study could open new avenues for characterizing interactive effects of simultaneously augmented nutrients in crops and provide a bench mark for crop scientists and farmers to improve Cd tolerance in rice.

Acute cadmium toxicity and post-stress recovery: Insights into coordinated and integrated response/recovery strategies of Anabaena sp. PCC 7120

Cyanobacteria, the first photoautotrophs have remarkable adaptive capabilities against most abiotic stresses, including Cd. A model cyanobacterium, Anabaena sp. PCC 7120 has been commonly used to understand cyanobacterial plasticity under different environmental stresses. However, very few studies have focused on the acute Cd toxicity. In this context, Anabaena was subjected to 100 μM Cd for 48 h (acute Cd stress, ACdS) and then transferred into the fresh medium for post-stress recovery (PSR). We further investigated the dynamics of morpho-ultrastructure, physiology, cytosolic proteome, thylakoidal complexes, chelators, and transporters after ACdS, as well as during early (ER), mid (MR), and late (LR) phases of PSR. The findings revealed that ACdS induced intracellular Cd accumulation and ROS production, altered morpho-ultrastructure, reduced photosynthetic pigments, and affected the structural organization of PSII, which subsequently hindered photosynthetic efficiency. Anabaena responded to ACdS and recovered during PSR by reprogramming the expression pattern of proteins/genes involved in cellular defense and repair; CO2 access, Calvin-Benson cycle, glycolysis, and pentose phosphate pathway; protein biosynthesis, folding, and degradation; regulatory functions; PSI-based cyclic electron flow; Cd chelation; and efflux. These modulations occurred in an integrated and coordinated manner that facilitated Anabaena to detoxify Cd and repair ACdS-induced cellular damage.

Identification of a new function of metallothionein-like gene OsMT1e for cadmium detoxification and potential phytoremediation

Cadmium (Cd) is a biologically non-essential and toxic heavy metal leaking to the environment via natural emission or anthropogenic activities, thereby contaminating crops and threatening human health. Metallothioneins (MTs) are a group of metal-binding proteins playing critical roles in metal allocation and homeostasis. In this study, we identified a novel function of OsMT1e from rice plants. OsMT1e was dominantly expressed in roots at all developmental stages and, to less extent, expressed in leaves at vegetative and seed filling stages. OsMT1e was mainly targeted to the nucleus and substantially induced by Cd exposure. Expression of OsMT1e in a yeast Cd-sensitive strain ycf1 conferred cellular tolerance to Cd, even though the ycf1 + OsMT1e cells accumulated more Cd than the control cells (ycf1 + pYES2). Both transgenic rice overexpressing (OX) and repressing OsMT1e by RNA interference (RNAi) were developed. Phenotypic analysis revealed that OsMT1e overexpression enhanced the rice growth concerning the increased shoot or root elongation, dry weight and chlorophyll contents, whereas the RNAi lines displayed a sensitive growth phenotype compared to wild-type. Assessment with 0.5, 2 and 10 μM Cd for two weeks revealed that the RNAi lines accumulated less Cd, while the OX lines had an increased Cd accumulation in root and shoot tissues. The contrasting Cd accumulation phenotypes between the OX and RNAi lines were further confirmed by a long-term study with 0.5 μM Cd for one month. Overall, the study unveiled a new function of OsMT1e in rice, which can be potentially used for engineering genotypes for phytoremediation or minimizing Cd in rice crops.

Phosphate and methionine affect cadmium uptake in valerian (Valeriana officinalis L.)

This study investigated the effects of exogenous methionine (Met) and different phosphate (PO4) concentrations on Cd uptake and translocation in Valeriana officinalis L. Seedlings were grown in nutrient solutions with three different concentrations of phosphate (900, 1200, and 1500 μM) for two weeks, then exposed for 4 days to 10 μM Cd, either in presence or absence of 400 μM methionine. The Met treatment decreased root Cd accumulation by up to 40%, while it enhanced Cd uptake into the shoots by 50%. In absence of Met, shoot Cd uptake was not affected by the level of phosphate application, although root Cd contents increased. The latter effect was entirely due to increased apoplastic Cd binding. In presence of Met, the Cd accumulation of both plant parts showed trends to increase with increasing phosphate level. In contrast to the treatments without Met, however, the phosphate effect on root Cd was due to increased symplastic root Cd allocation. The results suggest that the effects of Met on Cd uptake were due to the formation of mobile Cd-Met complexes, reducing phosphate-promoted Cd-retention in the apoplast and enhancing Cd transfer into the root symplast. Irrespective of the treatment, shoot Cd accumulation showed a close linear relationship to shoot mass, suggesting that convective transport with the transpirational water stream was the rate-governing uptake process. The results indicate that methionine supplementation could reduce Cd accumulation in valerian roots, which are the parts of this plant harvested for medicinal purposes, in Cd-contaminated soil, while phosphate would enhance it.

Effect of cell cycle synchronization on cadmium-induced apoptosis and necrosis in NRK-52E cells

ABSTRACT Heavy metal pollution is a problem that cannot be ignored. Due to the prevalence of cadmium in the environment and its harmful effects on humans, cadmium pollution has become a research hotspot recently. The mechanism of cadmium-induced toxicity has also drawn much attention and most studies have been conducted using whole cells, but the toxicological mechanism of cadmium remains unclear. In this study, we aimed to obtain NRK-52E cells at different growth stages by various methods and analyze the differences in cadmium toxicity. The results show that the cadmium sensitivity of cells in each phase was different and the late apoptotic rate was increased significantly after 5 µM Cd treatment. In addition, cadmium easily induces apoptosis of G0- and S-phase cells, as well as necrosis of S- and M-phase cells, but has no significant effect on G1-phase cells. Overall, we first explored the differences in the effects of cadmium on NRK-52E cells at various growth phases. Besides, the findings of this study might provide a theoretical basis for further exploration of the toxicological mechanism of cadmium.Abbreviations Cd: cadmium; CDK: cyclin-dependent kinases; DAPI 2-(4-amidinophenyl)-1H-indole-6-carboxamidine; TBST: Tris-buffered saline with Tween-20; PI: propidium iodide; DMEM: Dulbecco’s Modified Eagle Medium; BCA: bicinchoninic acid

Mechanisms of Cd and Cu induced toxicity in human gastric epithelial cells: Oxidative stress, cell cycle arrest and apoptosis

Cadmium (Cd) and copper (Cu) are widely present in foods. However, their adverse effects on human gastric epithelium are not fully understood. Here, human gastric epithelial cells (SGC-7901) were employed to study the toxicity and associated mechanisms of Cd + Cu co-exposure. Their effects on cell viability, morphology, oxidative damage, cell cycle, apoptosis, and the mRNA levels of antioxidases and cell cycle regulatory genes were investigated. Co-exposure to Cd (5 μM)/Cu (10 μM) induced >40% cell viability loss, whereas little effect on cell viability at <10 μM Cd or 40 μM Cu. Compared to individual exposure, co-exposure induced greater oxidative damage by elevating ROS (3.5 folds), malondialdehyde (2.3 folds) and expression of SOD1 and HO-1 besides inhibiting CAT, GPX1 and Nrf2. A marked S cell-cycle arrest was observed in co-exposure, evidenced by more cells staying in the S phase (36%), up-regulation of cyclins-dependent kinase (CDK4) and CDKs inhibitor (p21) and down-regulation of CDK2, CDK6 and p27. Furthermore, higher apoptosis (22%) with floated and round cells occurred in co-exposure group. Our data implicate the cytotoxicity of Cd + Cu co-exposure was higher than individual exposure, and individual assessment would underestimate their potential health risk. Oxidative stress and cell cycle arrest possibly played a role in Cd + Cu induced toxicity and apoptosis in SGC-7901 cells. Our data suggest the importance to reduce Cd in foods to decrease its adverse impacts on human digestive system.

Influence of Cd toxicity on subcellular distribution, chemical forms, and physiological responses of cell wall components towards short-term Cd stress in Solanum nigrum.

Solanum nigrum is a well-documented cadmium (Cd) hyperaccumulator; however, its Cd-induced tolerance capability and detoxification mechanism remain elusive. Hence, a short-term hydroponic experiment was performed in a multiplane glasshouse to determine the influence of Cd toxicity on subcellular distribution, chemical forms, and the physiological responses of cell wall towards Cd stress in a 4-week-old plant. The experiment was conducted following completely randomized design (CRD) with five treatments (n = 4 replicates). The results showed that Cd stress showed dose-dependent response towards growth inhibition. The subcellular distribution of Cd in S. nigrum was in the order of cell wall > soluble fractions > organelles, and Cd was predominantly extracted by 1 M NaCl (29.87~43.66%). The Cd contents in different plant tissues and cell wall components including pectin, hemicellulose 1 (HC1), hemicellulose 2 (HC2), and cellulose were increased with the increase in Cd concentrations; however, the percentage of Cd concentration decreased in pectin and cellulose. Results of the polysaccharide components such as uronic acid, total sugar contents, and pectin methylesterase (PME) activity showed Cd-induced dose-dependent increase relative to exposure Cd stress. The pectin methylesterase (PME) activity was significantly (p < 0.05) enhanced by 125.78% at 75 μM Cd in root, 105.78% and 73.63% at 100 μM Cd in stem and leaf, respectively. In addition, the esterification, amidation, and pectinase treatment of cell wall and Fourier transform infrared spectroscopy (FTIR) assay exhibited many functional groups that were involved in cell wall retention Cd, especially on carboxyl and hydroxyl groups of cell wall components that indicated that the -OH and -COOH groups of S. nigrum cell wall play a crucial role in Cd fixation. In summary, results of the current study will add a novel insight to understand mobilization/immobilization as well as detoxification mechanism of cadmium in S. nigrum.

Using multiple endpoints to assess the toxicity of cadmium and cobalt for chlorophycean Raphidocelis subcapitata

Metals may cause damage to the biota of contaminated environments. Moreover, using multiple endpoints in ecotoxicological studies is useful to better elucidate the mechanisms of toxicity of these compounds. Therefore, this study aimed to evaluate the effects of cadmium (Cd) and cobalt (Co) on growth, biochemical and photosynthetic parameters of the microalgae Raphidocelis subcapitata, through quantification of lipid classes composition, chlorophyll a (Chl a) content, maximum (ΦM) and effective (Φ’M) quantum yields and efficiency of the oxygen-evolving complex (OEC). Both metals affected the algal population growth, with an IC50–96h of 0.67 and 1.53 μM of Cd and Co, respectively. Moreover, the metals led to an increase in the total lipid content and reduced efficiency of OEC and ΦM. Cell density was the most sensitive endpoint to detect Cd toxicity after 96 h of treatment. Regarding Co, the photosynthetic parameters were the most affected and the total lipid content was the most sensitive endpoint as it was altered by the exposure to this metal in all concentrations. Cd led to increased contents of the lipid class wax esters (0.89 μM) and phospholipids (PL - at 0.89 and 1.11 μM) and decreased values of triglycerides (at 0.22 μM) and acetone-mobile polar lipids (AMPL - at 0.44 and 1.11 μM). The percentage of free fatty acids (FFA) and PL of microalgae exposed to Co increased, whereas AMPL decreased in all concentrations tested. We were able to detect differences between the toxicity mechanisms of each metal, especially how Co interferes in the microalgae at a biochemical level. Furthermore, to the best of our knowledge, this is the first study reporting Co effects in lipid classes of a freshwater Chlorophyceae. The damage caused by Cd and Co may reach higher trophic levels, causing potential damage to the aquatic communities as microalgae are primary producers and the base of the food chain.

Cadmium and molybdenum co-exposure triggers autophagy via CYP450s/ROS pathway in duck renal tubular epithelial cells

Cadmium (Cd) and excessive molybdenum (Mo) are detrimental to animals, but the combined nephrotoxic impacts of Cd and Mo on duck are still unclear. To evaluate the combined impacts of Cd and Mo on autophagy via Cytochrome P450s (CYP450s)/reactive oxygen species (ROS) pathway, duck renal tubular epithelial cells were treated with 3CdSO4·8H2O (4.0 μM Cd), (NH4)6Mo7O24·4H2O (500.0 μM Mo), butylated hydroxy anisole (BHA) (100.0 μM) and combination of Cd and Mo or Cd, Mo and BHA for 12 h, and combined cytotoxicity was investigated. The results indicated that Mo or/and Cd induced CYP1A1, CYP1B1, CYP2C9, CYP3A8 and CYP4B1 mRNA levels, decreased superoxide dismutase (SOD), catalase (CAT) activities and glutathione peroxidase (GSH-Px) content, and increased malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents. Besides, Mo or/and Cd elevated the number of autophagosome and microtubule-associated protein light chain 3 (LC3) puncta, upregulated mRNA levels of Beclin-1, LC3A, LC3B, Atg5 and adenosine 5′-monophosphate (AMP)-activated protein kinase α1 (AMPKα-1), inhibited Dynein, p62 and mammalian target of rapamycin (mTOR) mRNA levels, increased Beclin-1 and LC3II/LC3I protein levels. Moreover, the changes of these factors in Mo and Cd co-treated groups were more apparent. Additionally, BHA could efficiently alleviate the changes of above these indicators co-induced by Mo and Cd. Overall, these results manifest Cd and Mo co-exposure may synergistically trigger autophagy via CYP450s/ROS pathway in duck renal tubular epithelial cells.

Cadmium-stimulated invasion of rat liver cells during malignant transformation: Evidence of the involvement of oxidative stress/TET1-sensitive machinery

Cadmium (Cd) is recognized as a highly toxic heavy metal for humans in part because it is a multi-organ carcinogen. To clarify the mechanism of Cd carcinogenicity, we have established an experimental system using rat liver TRL1215 cells exposed to 2.5 μM Cd for 10 weeks and then cultured in Cd-free medium for an additional 4 weeks (total 14 weeks). Recently, we demonstrated, by using this experimental system, that 1) Cd stimulates cell invasion by suppression of apolipoprotein E (ApoE) expression, and 2) Cd induces DNA hypermethylation of the regulatory region of the ApoE gene. However, the underlying mechanism(s) as well as other potential genetic participants in the Cd-stimulated invasion are undefined. In the present work, we found that concurrent with enhanced invasion, Cd induced oxidative stress, coupled with the production of oxidative stress-sensitive metallothionein 2A (MT2A), which lead to down-modulation of ten-eleven translocation methylcytosine dioxygenase 1 (TET1: DNA demethylation) in addition to ApoE, without impacting DNA methyltransferases (DNMTs: DNA methylation) levels. Furthermore, the expression of tissue inhibitor of metalloproteinase 2 and 3 (TIMP2 and TIMP3) that are positively regulated by TET1, were decreased by Cd. The genes (ApoE/TET1/TIMP2/TIMP3) suppressed by Cd were further suppressed by hydroquinone (HQ; a reactive oxygen species [ROS] producer), whereas N-acetyl-l-cysteine (NAC; a ROS scavenger) prevented the suppression of their expression by HQ. In addition, NAC reversed their expression suppressed by Cd. Cd-stimulated cell invasion was clearly dampened by NAC in a concentration-dependent manner. Overall these findings suggest that 1) altered TET1 expression and activity together with ApoE are likely involved in the enhanced invasiveness due to Cd exposure, and 2) Cd down-regulation of TET1 likely evokes a reduction in ApoE expression (possible by DNA hypermethylation), and 3) anti-oxidants are effective in abrogation of the enhanced invasiveness that occurs concurrently with Cd-induced malignant transformation.

Boron supply alleviates cadmium toxicity in rice (Oryza sativa L.) by enhancing cadmium adsorption on cell wall and triggering antioxidant defense system in roots

Cadmium (Cd) pollution is a key concern globally that affects plant growth and productivity. Boron (B) is a micronutrient that helps in the formation of the primary cell wall (CW) and alleviates negative effects of toxic elements on plant growth. Nonetheless, knowledge about how B can reduce Cd toxicity in rice seedlings is not enough, particularly regarding CW-Cd adsorption. Therefore, the current experiment investigated the alleviative role of B on Cd toxicity in rice seedling. The experiment was carried out with 0 μM and 30 μM H3BO3 under 50 μM Cd toxicity in hydroponics. The results showed that Cd exposure alone inhibited plant growth parameters and caused lipid peroxidation. Moreover, Cd toxicity led to obvious visible toxicity symptoms on the leaves. However, increasing the availability of B alleviated Cd toxicity by reducing Cd concentration in plant tissues and improving antioxidative system. Moreover, cell wall pectin and hemicellulose adsorbed a significant amount of Cd. Fourier-Transform Infrared spectroscopy (FTIR) spectra exhibited that cell wall functional groups were increased by B application. Scanning electron microscopy (SEM) equipped with energy-dispersive X-ray (EDX) microanalysis confirmed the higher Cd binding onto CW. The findings of this investigation showed that B could mitigate Cd stress by decreasing Cd uptake and encouraging Cd adsorption on CW, and activation of the protective mechanisms. The present results might help to increase rice productivity on Cd polluted soils.

Serratia sp. CP-13 alleviates Cd toxicity by morpho-physio-biochemical improvements, antioxidative potential and diminished Cd uptake in Zea mays L. cultivars differing in Cd tolerance

Cadmium (Cd) is highly toxic for plant metabolic processes even in low concentration due to higher retention rates, longer half-life and non-biodegradable nature. The current study was designed to assess the bioremediation potential of Cd tolerant PGPR, Serratia sp. CP-13 together with two differentially Cd tolerant maize cultivars (MMRI-Yellow, Sahiwal-2002) selected amongst ten cultivars after screening. The maize cultivars were grown under different Cd treatments (0, 6, 12, 18, 24, 30 µM) in Petri plates both with and without Serratia sp. CP-13 inoculation. Treated plants were analyzed for their biomass accumulation, chlorophylls, carotenoids, proline, anthocyanin, protein, malondialdehyde (MDA), H2O2 as well as for antioxidants (POD, SOD, CAT) and mineral elements (Ca, Mg, Zn, K, Fe, Na, Cd). The maize cultivar MMRI-Yellow (tolerant) and Sahiwal-2002 (sensitive) exhibited significant reduction in leaf area, nutrient contents, plant biomass, activity of antioxidants, total proteins, photosynthetic pigments as well as flavonoids with increased production of H2O2, proline, MDA and relative membrane permeability (RMP) under Cd stress. However, this reduction was cultivar specific and recorded higher in cv. Sahiwal-2002 as compared to MMRI-Yellow. Application of Serratia sp. CP-13 significantly augmented plant biomass, photosynthetic pigments, antioxidative machinery, as well as flavonoids and proline while diminishing H2O2, RMP MDA production even under Cd stress in studied cultivars. Furthermore, CP-13 inoculation assisted the Cd stressed plants to sustain an optimal level of essential nutrients (Ca, Mg, Zn, K, Fe) except for Na and Cd which responded antagonistically. It was inferred that both inoculated maize cultivars exhibited better health and metabolism but substantial Cd tolerance was acquired by the sensitive cv. Sahiwal-2002 than the tolerant cv. MMRI-Yellow under applied Cd regimes. Furthermore, studied maize cultivars depicted maximum Cd tolerance in order of 30 < 24 < 18 < 12 < 6 < 0 µM Cd treatments under Serratia sp. CP-13 inoculation. Findings of current work highlighted the importance of Serratia sp. CP-13 and its inoculation impact on morpho-physio-biochemical attributes of maize growth under Cd dominant environment, which is likely an addition towards efficient approaches for bacterially-assisted Cd bioremediation and minimal Cd retention in edible plant parts.

Physiological and Transcriptomic Response of Grey Poplar (Populus ×canescens Aiton Sm.) to Cadmium Stress.

(1) Background: Populus ×canescens (Aiton) Sm. is a fast-growing woody plant belonging to the family Salicaceae. Two poplar genotypes characterized by unique phenotypic traits (TP11 and TP20) were chosen to be characterized and tested for a physiological and transcriptomic response to Cd stress. (2) Methods: A comparative analysis of the effects of exposure to high cadmium (Cd) concentrations (10 µM and 100 µM) of TP11 and TP20 was performed. (3) Results: Neither of the tested Cd concentration negatively affected plant growth; however, the chlorophyll content significantly decreased. The potassium (K) content was higher in the shoots than in the roots. The magnesium concentrations were only slightly affected by Cd treatment. The zinc content in the shoots of TP20 was lower than that in the shoots of TP11. Cd accumulation was higher in the roots than in the shoots. After 10 days of exposure, 10 µM Cd resulted in comparable amounts of Cd in the roots and shoots of TP20. The most significant change in transcript amount was observed in endochitinase 2, 12-oxophytodienoate reductase 1 and phi classglutathione S-transferase. (4) Conclusions: Our study provided new insights for effective assessing the ability of different poplar genotypes to tolerate Cd stress and underlying Cd tolerance.

Exogenous Glutathione Alleviation of Cd Toxicity in Italian Ryegrass (Lolium multiflorum) by Modulation of the Cd Absorption, Subcellular Distribution, and Chemical Form.

Subcellular fractions and the chemical forms of cadmium (Cd) reflect its level of toxicity to plants; however, these effects of exogenous glutathione (GSH) are poorly understood. We exposed two Italian ryegrass (Lolium multiflorum) cultivars (IdyII and Harukaze) to 50 µM Cd or 200 µM GSH to investigate the effect of GSH on the Cd uptake, subcellular compartments, and chemical forms. Cd significantly inhibited the plant growth, while GSH supplementation decreased this inhibition. The application of GSH significantly improved the Cd concentration in the roots but reduced that in the shoots and decreased the Cd translocation from root to shoot. The Cd concentration of the root in the cell wall was increased while the concentration in the soluble fraction was decreased when supplied with GSH. The inorganic form (80% ethanol for Cd extraction) in the roots was significantly reduced when treated with GSH. The Cd form extracted by 2% acetic acid (HAC) with low toxicity and immobility were greatly increased. In leaves, the application GSH decreased in any form of Cd form extracted. In conclusion, exogenous GSH decreased the translocation of Cd and alleviated Italian ryegrass Cd toxicity by accumulating more Cd in the root cell wall and immobilizing more Cd in lower toxicity fractions.

Human placental cell line HTR-8/SVneo accumulates cadmium by divalent metal transporters DMT1 and ZIP14

Cadmium (Cd) is a global pollutant that accumulates in the placenta and can cause placental dysfunction. Although iron transporters have been suggested to participate in placental Cd uptake, it is still unknown which transporters are actually involved in this process. We specifically aimed to study the role of three iron transporters in the uptake of Cd into the placental cell line HTR-8/SVneo. For this purpose, Divalent Metal Transporter (DMT)1 and ZRT/IRT like protein (ZIP)8 and ZIP14 were downregulated and changes in cellular Cd levels analysed in relation to controls. As clearly shown by the reduction of the Cd content by ∼60% in DMT1- and ZIP14-downregulated cells, the two proteins are essential for Cd accumulation in HTR-8/SVneo cells. Using a validated antibody, we show DMT1 to be localised in situ in trophoblast and stromal cells. We further wanted to investigate how placental cells cope with Cd loading and which metallothionein (MT) isoforms they express. Cd-exposed cells accumulate Cd in a dose-dependent manner and upregulate MT2A accordingly (up to 15-fold induction upon 5 μM CdCl2 treatment for 72 h). 5 μM Cd exposure for 72 h decreased cell number to 60%, an effect that was aggravated by MT2A depletion (cell number reduced to 30%) indicating additive effects. In conclusion, our data suggest that DMT1 and ZIP14 are required for Cd uptake into human placental cells that upregulate MT2A to store and detoxify the metal. Cd storage in the placenta reduces Cd transport to the fetus, which, however, could impair placental functions and fetal development.

Improved physiological defense responses by application of sodium nitroprusside in Isatis cappadocica Desv. under cadmium stress.

Isatis cappadocica is a well-known arsenic-hyperaccumulator, but there are no reports of its responses to cadmium (Cd). Nitric oxide (NO) is a signaling molecule, which induces cross-stress tolerance and mediates several physio-biochemical processes related to heavy metal toxicity. In this study, the effects of Cd and sodium nitroprusside (SNP as NO donor) on the growth, defense responses and Cd accumulation in I. cappadocica were investigated. When I. cappadocica was treated with 100 and 200 μM Cd, there was an insignificant inhibition of shoot growth. However, Cd stress at Cd400 treatment decreased significantly the dry weight of root and shoot by 73 and 38%, respectively, as compared to control. The application of SNP significantly improved the growth parameters and mitigated Cd toxicity. In addition, SNP decreased reactive oxygen species (ROS) production induced by Cd. The increased total thiol and glutathione (GSH) concentrations after SNP application may play a decisive role in maintaining cellular redox homeostasis, thereby protecting plants against oxidative damage under Cd stress. Bovine hemoglobin (Hb as NO scavenger) reduced the protective role of SNP, suggesting a major role of NO in the defensive effect of SNP. Furthermore, the reduction in shoot growth and the increase of oxidative damage were more severe after the addition of Hb, which confirms the protective role of NO against Cd-induced oxidative stress. The protective role of SNP in decreasing Cd-induced oxidative stress may be related to NO production, which can lead to stimulation of the thiols synthesis and improve defense system.

Influence of cadmium contamination on brain glial cells: consequences and bioindication possibilities

Cadmium (Cd) is a heavy metal that currently presents in almost all components of the environment. Cd is a ubiquitous pollutant that is constantly entering the environment from industry and agriculture, mining, forest fires and many more sources. Some occupational diseases have aftereffects associated with Cd cytotoxicity. Despite long-term studies of the toxic effects of Cd, its cytotoxicity of low doses and the chronic effects on the nerve tissue cells remain undiscovered. The results of determining the Cd neurotoxicity indicate a disturbance of the permeability of the blood-brain barrier, the accumulation of Cd in the brain and the deterioration of the functional activity of the central nervous system. One of the main cellular targets for Cd in the brain are astrocytes. Astrocytes provide nutrition and functional activity of neurons, as well as recovery of physical and metabolic damage. The cytoskeleton of astrocytes is built of glial fibrillary acidic protein (GFAP). GFAP participates in important functions of astrocytes and its condition reflects the astrocytes reactivity. The molecular mechanisms of the neurotoxic effects of Cd on the glial cytoskeleton remain unknown. Glioblastomas are widely used to study the cytotoxic mechanisms of various compounds, including heavy metals, as cellular models of astrocytes. Taking into account the role of oxidative stress in a cell damage, as well as the reactive response of glial cells, we study the influence of low doses of Cd on oxidative stress and expression of GFAP and glucose-6-phosphate dehydrogenase (G6PD) in U373GM cells. Doses of 2-10 μM Cd induced a dose-dependent increase in reactive oxygen species and lipid peroxidation products. The same doses inhibited the expression of the cytoskeletal marker of astrocytes (GFAP) and metabolic marker of glucose utilization (G6PD). The obtained results indicate a pronounced cytotoxic effect of low doses of Cd in the astrocytic cell model U373GM. In addition, the astroglial cytotoxicity of Cd may be mediated by oxidative damage, inhibition of glial intermediate filament expression, and glucose utilization disorders. These parameters can be promising biomarkers of toxic effects both for the assessment of human and animal health and for determining the state of the environment as a whole.