Metallothioneins BcMT1 and BcMT2 from Brassica campestris enhance tolerance to cadmium and copper and decrease production of reactive oxygen species in Arabidopsis thaliana

Aims

Metallothioneins are cysteine-rich, metal-binding proteins, but their exact functions are not fully understood. In this study, we isolated two metallothionein genes, BcMT1 and BcMT2 from Brassica campestris to increase our understanding of metal tolerance mechanisms in Brassica plants.

Methods

Semi-quantitative RT-PCR was used to analyze expression of the two BcMTs genes. BcMT1 and BcMT2 were ectopically expressed in Arabidopsis thaliana. Quantitative real-time RT-PCR and GUS-staining method were used to select transgenic Arabidopsis plants. Cd and Cu concentrations were analyzed by flame atomic absorption spectrometry. Histochemical detection of H₂O₂ and O₂ ^?? were conducted by 3,3-diaminobenzidine and nitroblue tetrazoliu-staining methods.

Results

BcMT1 is expressed predominantly in roots, whereas BcMT2 is expressed mainly in leaves of B. campestris. Expression of BcMT1 was induced by both Cd and Cu, but expression of BcMT2 was enhanced only by Cd. Ectopic expression of BcMT1 and BcMT2 in Arabidopsis thaliana enhanced the tolerance to Cd and Cu and increased the Cu concentration in the shoots of the transgenic plants. Transgenic Arabidopsis accumulated less reactive oxygen species (ROS) than wild-type plants.

Conclusions

BcMT1 and BcMT2 increased Cd and Cu tolerance in transgenic Arabidopsis, and decreased production of Cd- and Cu-induced ROS, thereby protecting plants from oxidative damage.     

Molecular cloning and characterization of a Brassica juncea yellow stripe-like gene, BjYSL7, whose overexpression increases heavy metal tolerance of tobacco

Key message

BjYSL7 encodes a plasma-localized metal–NA transporter and has transport Fe(II)–NA complexes activity. BjYSL7 is involved in the transport of Cd and Ni from roots to shoots.

Abstract

Heavy metal transporters play a key role in regulating metal accumulation and transport in plants. In this study, we isolated a novel member of the yellow stripe-like (YSL) gene family BjYSL7 from the hyperaccumulator Brassica juncea. BjYSL7 is composed of 688 amino acids with 12 putative transmembrane domains and is over 90 % identical to TcYSL7 and AtYSL7. Real-time PCR analysis revealed that BjYSL7 mRNA was mainly expressed in the stem under normal condition. The expression of BjYSL7 was found to be up-regulated by 127.1-, 12.7-, and 3.4-fold in roots and 6.5-, 4.3-, and 2.8-fold in shoots under FeSO₄, NiCl₂, and CdCl₂ stresses, respectively. We have demonstrated that BjYSL7 is a Fe(II)–NA influx transporter by yeast functional complementation. Moreover, a BjYSL7::enhanced green fluorescent protein (EGFP) fusion localized to the plasma membrane of onion epidermal cells. The BjYSL7-overexpressing transgenic tobacco plants exhibited longer root lengths, lower relative inhibition rate of lengths and superior root hair development compared to that of wild-type (WT) plants in the presence of CdCl₂ and NiCl₂. Furthermore, the concentrations of Cd and Ni in shoots of BjYSL7-overexpressing plants are significantly higher than that of WT plants. Compared with WT plants, BjYSL7-overexpressing plants exhibited Fe concentrations that were higher in the shoots and seeds and lower in the roots. Taken together, these results suggest that BjYSL7 might be involved in the transport of Fe, Cd and Ni to the shoot and improving heavy metal resistance in plants.     

Cold treatment enhances lead resistance in Arabidopsis

As sessile organisms, plants usually experience several stresses simultaneously. It was shown that stress cross-tolerance may be induced by different stressors, including biotic factors as well as heavy metal, hypoxia, ultraviolet-B radiation, heat, high salt, drought, and cold stresses. However, it is unclear whether there is a cross-tolerance toward cold and lead (Pb) stresses in Arabidopsis. In this study, we showed that cold pretreatment enhanced Pb(II) resistance in Arabidopsis, as indicated by lower reduction of root length, fresh weight, and chlorophyll content in the cold-treated plants than the control ones. In the cold-treated seedlings, lower Pb contents were detected in roots and shoots in comparison to the control. This was associated, at least in part, with the activation of the expression of AtPDR12 gene, a pump excluding Pb(II) and/or Pb(II)-containing toxic compounds from the cytoplasm to the exterior of the cell. This finding was further supported by genetic evidence showing that cold treatment was unable to enhance resistance of atpdr12 mutant to Pb(II) stress but could enhance Pb(II) resistance of the wild type. In addition, we also found that cold-induced enhanced Pb(II) resistance was glutathione-independent. Taken together, all these results suggest that cold treatment enhanced Pb(II) resistance in Arabidopsis, at least in part, by activating the expression of AtPDR12 gene.     

The Arabidopsis Ethylene-Insensitive 2 gene is required for lead resistance

The Arabidopsis Ethylene-Insensitive 2 (EIN2) gene has been shown to be involved in the regulation of abiotic and biotic stresses, including ozone stress, high salt, oxidative stress and disease resistance. However, little is known about the role of EIN2 gene in lead (Pb) resistance in Arabidopsis. In this study, we showed that EIN2 gene is required for Pb(II) resistance in Arabidopsis. EIN2 gene was induced by Pb(II) treatment, and the ein2-1 mutant showed enhanced sensitivity to Pb(II). A higher Pb content was detected in ein2-1 plants than in wild-type plants when subjected to Pb(II) treatment, which was associated, at least in part, with reduction in expression of AtPDR12 gene, a pump excluding Pb(II) and/or Pb(II)-containing toxic compounds from the cytoplasm. Moreover, the ein2-1 mutation also impaired glutathione (GSH)-dependent Pb(II) resistance, which was related to constitutive reduction of express of GSH1 gene involved in GSH synthesis and consequently reduced GSH content. Taken together, all these results suggest that EIN2 gene mediates Pb(II) resistance, at least in part, through two distinct mechanisms, a GSH-dependent mechanism and a GSH-independent AtPDR12-mediated mechanism.     

Does radial oxygen loss and iron plaque formation on roots alter Cd and Pb uptake and distribution in rice plant tissues?

Background and Aims

Metal (e.g. Cd and Pb) pollution in agricultural soils and crops have aroused considerable attention in recent years. This study aimed to evaluate the effects of ROL and Fe plaque on Cd and Pb accumulation and distribution in the rice plant.

Methods

A rhizobag experiment was employed to investigate the correlations among radial oxygen loss (ROL), Fe plaque formation and uptake and distribution of Cd and Pb in 25 rice cultivars.

Results

Large differences between the cultivars were found in rates of ROL (1.55 to 6.88 mmol O₂ kg^?1 root d.w. h^?1), Fe plaque formation (Fe: 6,117–48,167 mg kg^?1; Mn: 127–1,089 mg kg^?1), heavy metals in shoot (Cd: 0.13–0.35 mg kg^?1; Pb: 4.8–8.1 mg kg^?1) and root tissues (Cd: 1.1–3.5 mg kg^?1; Pb: 45–199 mg kg^?1), and in Fe plaque (Cd: 0.54–2.6 mg kg^?1; Pb: 102–708 mg kg^?1). Rates of ROL were positively correlated with Fe plaque formation and metal deposition on root surfaces, but negatively correlated with metal transfer factors of root/plaque and distributions in shoot and root tissues.

Conclusions

ROL-induced Fe plaque promotes metal deposition on to root surfaces, leading to a limitation of Cd and Pb transfer and distribution in rice plant tissues.     

Integration of metal chemical forms and subcellular partitioning to understand metal toxicity in two lettuce (Lactuca sativa L.) cultivars

Aims

Metal chemical forms and subcellular partitioning model (SPM) in organisms can provide valuable insights into metal toxicity.

Methods

Two cultivars of lettuce (Lactuca sativa L.) were grown in Cd and Cu contaminated soils and chemical forms and subcellular distribution of Cd and Cu within the lettuce shoots were determined.

Results

Examination of the inhibition of superoxide dismutase (SOD) and catalase (CAT) activities, as well as the production of H₂O₂ showed that Lactuca sativa L. var. longifolia is more sensitive to metal-stress than is Lactuca sativa L. var. crispa. In L. crispa, the majority of accumulated Cd was in the pectate- and protein-integrated forms (53.7–62.9 %), while in L. longifolia, a higher proportion of the Cd was in the water soluble forms (33.0–39.2 %) and in the organelles fraction – these forms being potentially associated with toxicity. The chemically-based chemical form approach agreed closely with independent biologically-based SPM, as demonstrated by their significant linear relationships.

Conclusions

This study provides a first step towards the integration of chemical form approach and SPM into a common mechanistic framework, which is important for predicting the likelihood of toxic effects of metals in the environment of interest.     

Modelling of Cd, Cu, Pb and Zn transport in metal contaminated soil and their uptake by willow (Salix × smithiana) using HYDRUS-2D program

Aims

The main aim of this study was to validate the HYDRUS-2D model for the simulation of Cd, Cu, Pb and Zn transport within a soil column and their accumulation by willows.

Methods

A simulation of metal transport and uptake by willow was implemented using the HYDRUS-2D code. Two scenarios of the column experiment were compared: soil (C) and soil with planted willows (V). Seeping water, soil water content and actual transpiration were measured. Metal contents in soil water and willows were analysed.

Results

The single-porosity model (applied for isotropic soil media) was sufficient for scenario C. The single-porosity (applied for anisotropic soil media) and dual-porosity models (characterizing non-equilibrium water flow) were explored in scenario V. Measured cumulative Cd and Zn uptake showed a 20- and 10-times higher accumulation, respectively, in comparison with the modelled ones. On the other hand, modelled cumulative Pb uptake was reduced by reducing the maximum value of the root uptake concentration c _Root .

Conclusions

The HYDRUS-2D program was usable for modelling of Cd, Cu, Pb and Zn transport and their willow uptake. Additionally, consideration of dual-porosity soil media as well as anisotropy was suitable for the experiment with roots presence in the soil.     

NH4 + induces antioxidant cellular machinery and provides resistance to salt stress in citrus plants

Key message

NH ₄ ⁺ acts as a mild oxidative stressor, which triggers antioxidant cellular machinery and provide resistance to salinity.

Abstract

NH₄ ⁺ nutrition in Carrizo citrange (Citrus sinensis L. Osbeck × Poncirus trifoliata L) plants acts as an inducer of resistance against salinity conditions. NH₄ ⁺ treatment triggers mild chronic stress that primes plant defence responses by stress imprinting and confers protection against subsequent salt stress. In this work, we studied the influence of NH₄ ⁺ nutrition on antioxidant enzymatic activities and metabolites involved in detoxification of reactive oxygen species (ROS) to clarify their involvement in NH₄ ⁺-mediated salt resistance. Our results showed that NH₄ ⁺ nutrition induces in citrus plants high levels of H₂O₂, strongly inhibits superoxide dismutase (SOD) and glutathione reductase (GR) activities, and leads to higher content of oxidised glutathione (GSSG) than in control plants in the absence of salt, thus providing evidence to confirm mild stress induced by NH₄ ⁺ nutrition. However, upon salinity, plants grown with NH₄ ⁺ (N-NH₄ ⁺ plants) showed a reduction of H₂O₂ levels in parallel to an increase of catalase (CAT), SOD, and GR activities compared with the control plants. Moreover, N-NH₄ ⁺ plants were able to keep high levels of reduced glutathione (GSH) upon salinity and were able to induce glutathione-S-transferase (GST) and phospholipid hydroperoxide glutathione peroxidise (PHGPx) mRNA accumulation. Based on this evidence, we confirm that sublethal concentrations of NH₄ ⁺ might act as a mild oxidative stressor, which triggers antioxidant cellular machinery that can provide resistance to subsequent salt stress.     

Nitrate nutrition enhances nickel accumulation and toxicity in Arabidopsis plants

Background and aims

Nickel (Ni) has become a major heavy metal contaminant. The form of nitrogen nutrition remarkably affects IRT1 expression in roots. IRT1 has an activity of transporting Ni²⁺ into root cells. Therefore, nitrogen-form may affect Ni accumulation and toxicity in plants. The assumption was investigated in this study.

Methods

The Arabidopsis plants were treated in Ni-contained growth solutions with either nitrate (NO₃ ^?) or ammonium (NH₄ ⁺) as the sole N source. After 7-day treatments, Ni concentration, IRT1 expression, Ni-induced toxic symptoms and oxidative stress in plants were analyzed.

Results

The NO₃ ^?-fed plants contained a higher Ni concentration, had a greater IRT1 expression in roots, and developed more severe toxic symptoms in the youngest fully expanded leaves, compared with the NH₄ ⁺-fed plants. The Ni-induced growth inhibition was also more significant in NO₃ ^?-fed plants. Interestingly, Ni exposure resulted in greater hydrogen peroxide (H₂O₂) and superoxide radical (O₂ ^{. ?}) accumulations, more severe lipid peroxidation and more cell death in NO₃ ^?-fed plants, whereas the opposite was true for NH₄ ⁺-fed plants. Furthermore, the Ni-enhanced peroxidase (POD) and superoxide dismutase (SOD) activities were greater in NO₃ ^?-fed plants

Conclusion

NO₃ ^? nutrition promotes Ni uptake, and enhances Ni-induced growth inhibition and oxidative stress in plants compared with NH₄ ⁺ nutrition.     

Heme oxygenase is involved in H2O2-induced lateral root formation in apocynin-treated rice

Key message

Apocynin is a natural organic compound structurally related to vanillin. We demonstrated that hydrogen peroxide and heme oxygenase participated in apocynin-induced lateral root formation in rice.

Abstract

Apocynin, also known as acetovanillone, is a natural organic compound structurally related to vanillin. Information concerning the effect of apocynin on plants is limited. In this study, we examined the effect of apocynin on lateral root (LR) formation in rice. Treatment with apocynin induced LR formation and increased H₂O₂ production, but had no effect on nitric oxide production. Diphenyleneiodonium chloride, an inhibitor of H₂O₂ generating NADPH oxidase, was effective in reducing apocynin-induced H₂O₂ production and LR formation. Apocynin treatment also increased superoxide dismutase activity and decreased catalase activity. H₂O₂ application was able to increase the number of LRs. Moreover, H₂O₂ production caused by H₂O₂ and apocynin was localized in the root area corresponding to the LR emergence. Treatment with H₂O₂ and apocynin also increased heme oxygenase (HO) activity and induced OsHO1 mRNA expression. Lateral root formation and HO activity induced by H₂O₂ and apocynin were reduced by Zn protoporphyrin IX (the specific inhibitor of HO). Our data suggest that both H₂O₂ and HO are required for apocynin-induced LR formation in rice.     

Overexpression of tomato GDP-l-galactose phosphorylase gene in tobacco improves tolerance to chilling stress

Key message

The overexpression of tomato GDP- l -galactose phosphorylase gene enhanced tolerance to chilling stress and reduced photoinhibition of photosystems I and II in transgenic tobacco.

Abstract

Chilling stress is a crucial factor that limits the geographical distribution and yield of chilling-sensitive plants. Ascorbate (AsA) protects plants by scavenging reactive oxygen species and reduces photoinhibition by promoting the conversion of violaxanthin to zeaxanthin in the xanthophyll cycle to dissipate excess excitation energy. Possible mechanisms of AsA for plant photoprotection under chilling stress were investigated by isolating the tomato GDP-l-galactose phosphorylase gene (SlGGP) and producing transgenic tobacco plants with overexpression of SlGGP. The transgenic plants subjected to chilling stress accumulated less H₂O₂, demonstrated lower levels of ion leakage and malondialdehyde, and acquired higher net photosynthetic rate, higher maximum photochemical efficiency of PSII, and higher D1 protein content compared with the wild-type (WT) plants. The transgenic plants subjected to chilling stress also showed higher GDP-l-galactose phosphorylase activity, increased AsA content as well as ascorbate peroxidase and oxidizable P700 activities than WT plants. Thus, SlGGP overexpression is crucial in promoting AsA synthesis and alleviating photoinhibition of two photosystems.     

Exogenous spermidine enhances Hydrocharis dubia cadmium tolerance

The effects of exogenous spermidine (Spd) on arginine decarboxylase (ADC), ornithine decarboxylase (ODC), polyamine oxidase (PAO), and diamine oxidase (DAO) activities, the rate of superoxide radical (O ₂ ^·? ) generation and polyamine (PA), malondialdehyde (MDA), and H₂O₂ contents in Hydrocharis dubia (Bl.) Backer leaves under cadmium (Cd) toxicity were studied after 6-day treatment. Cd stress increased putrescine (Put) level and lowered spermidine (Spd) and spermine (Spm) levels. In addition, the activities of ADC, DAO, and PAO were increased, while that of ODC was decreased. Exogenous application of Spd markedly reversed these Cd-induced effects. It also significantly reduced the generation of O ₂ ^·? and H₂O₂ and prevented lipid peroxidation. These results suggest that exogenous Spd can enhance the tolerance of H. dubia to Cd. The maintenance of PA homeostasis was necessary for plant metal tolerance.     

The response of the Rhizobium leguminosarum bv. trifolii wild-type and exopolysaccharide-deficient mutants to oxidative stress

Aims

The aim of this study was investigation of the response of R. leguminosarum bv. trifolii wild-type and its two rosR and pssA mutant strains impaired in exopolysaccharide (EPS) synthesis to oxidative stress conditions caused by two prooxidants: a superoxide anion generator- menadione (MQ) and hydrogen peroxide (H₂O₂).

Methods

The levels of enzymatic (catalase, superoxide dismutase, pectinase and β-glucosidase) and non-enzymatic (superoxide anion generator, formaldehyde, phenolic compounds) biomarkers were monitored using biochemical methods in both the supernatants and rhizobial cells after treatment with 0.3?mM MQ and 1.5?mM H₂O₂. The viability of bacterial cells was estimated using fluorescent dyes and confocal laser scanning microscopy. In addition, the effect of prooxidants on symbiosis of the R. leguminosarum bv. trifolii strains with clover was established.

Results

The tested stress factors significantly changed enzymatic patterns of the rhizobial strains, and the wild-type strain proved to be more resistant to these prooxidants than both pssA and rosR mutants. Significantly higher activities of both catalase and superoxide dismutase have been detected in these mutants in comparison to the wild-type strain. H₂O₂ and MQ also increased the levels of pectinase and β-glucosidase activities in the tested strains. Moreover, pre-incubation of R. leguminosarum bv. trifolii strains with the prooxidants negatively affected the viability of bacterial cells and the number of nodules elicited on clover plants.

Conclusions

EPS produced in large amounts by R. leguminosarum bv. trifolii plays a significant protective role as a barrier against oxidative stress factors and during symbiotic interactions with clover plants.     

Variation in cadmium accumulation and translocation among peanut cultivars as affected by iron deficiency

Purpose

The current study aimed to test the hypothesis that the variations in shoot Cd accumulation among peanut cultivars was ascribed to the difference in capacity of competition with Fe transport, xylem loading and transpiration.

Methods

A hydroponics experiment was conducted to determine the plant biomass, gas exchange, and Cd accumulation in Fe-sufficient or -deficient plants of 12 peanut cultivars, at low Cd level (0.2 μM CdCl₂).

Results

Peanut varied among cultivars in morpho-physiological response to Cd stress as well as Cd accumulation, translocation and distribution. Qishan 208 and Xvhua 13 showed a higher capacity for accumulating Cd in their shoots. Fe deficiency increased the concentration and amount of Cd in plant organs, but decreased TF _{root to shoot} and TF _{root to stem}, while TF _{stem to leaf} remained unaffected. Fe deficiency-induced increase rates of Cd concentration and total Cd amount in roots and leaves were negatively correlated with the values in Fe-sufficient plants. Transpiration rate was positively correlated with leaf Cd concentration, TF _{root to shoot}, TF _{root to stem} and TF _{stem to leaf}.

Conclusions

The difference in shoot Cd concentration among peanut cultivars was mainly ascribed to the difference in Fe transport system, xylem loading capacity and transpiration.     

Predictability of the Zn and Cd phytoextraction efficiency of a Salix smithiana clone by DGT and conventional bioavailability assays

Aims

Phytomanagement of metal-polluted soils requires information on plant responses to metal availability in soil, but the predictability of metal accumulation in plant shoots and/or roots may be limited by metal toxicity and inherent shortfalls of the bioavailability assays.

Methods

We measured the uptake of Cd and Zn in a Salix smithiana clone grown in a pot experiment on soils with different characteristics and metal availabilities, determined by conventional soil single extractions (0.05 M Na₂-EDTA and 1 M NH₄NO₃), soil solution obtained by centrifugation, and diffusive gradients in thin films (DGT). The Cd and Zn phytoavailability after a 2-year phytoextraction by willow was assessed by metal accumulation in the straw of the following barley culture.

Results

The phytoextraction efficiency was largest on a moderately polluted acid soil. Biomass and shoot Zn concentrations of S. smithiana were better predicted by DGT-measured Zn concentrations in soil solution (C _DGT) than by Zn concentrations in the soil solution and extractable soil fractions. The weaker correlation for Cd in shoots may be related to relative Cd enrichment in the plant tissues. The metal accumulation in barley straw was unaffected or increased after a 2-year phytoextraction.

Conclusions

The shoot Zn and Cd removal of the tested Salix clone can be predicted by C _DGT concentrations and is highest on either calcareous or moderately polluted acid soils. Single extraction with NH₄NO₃ and the C _DGT value of Cd were not able to predict shoot Cd removal on the tested soils. Only shoot removal of Zn was predicted fairly well by the C _DGT value.     

Impact of active transport and transpiration on nickel and cadmium accumulation in the leaves of the Ni-hyperaccumulator Leptoplax emarginata: a biophysical approach

Aims

The primary aim of this study was to investigate the impact of active nickel and cadmium transport, transpiration and shoot biomass production on Ni and Cd accumulation in the leaves of the Ni-hyperaccumulator Leptoplax emarginata. A secondary objective was to observe the effects of various concentrations of nickel and cadmium in solutions on the plant growth and ecophysiological characteristics of these plants. Finally, the study sought to identify possible nickel and cadmium concentration gradients in solution as a function of the root distance.

Methods

The Intact Plant Transpiration Stream Concentration Factor (TSCF=xylem/solution solute concentration ratio) was determined for both Ni and Cd and for the selected intact transpiring Ni-hyperaccumulator Leptoplax emarginata, cultivated on two contrasting fertilized and Ni-Cd-contaminated sandy porous media (rhizotrons with central root compartments, linked to Mariotte tubes operated at ?1?kPa). IPTSCF_Ni and IPTSCF_Cd were calculated as the ratios between the hyperaccumulator plant’s nickel or cadmium mass in the leaves and the nickel or cadmium concentration in solution by the volume of water transpired during the period of culture. Plant growth characteristics and gas exchanges were also recorded.

Results

IPTSCF values were much greater than 1 (IPTSCF_Ni?=?5.2?±?0.9 and IPTSCF_Cd?=?4.4?±?0.6) whatever the amount of available Ni and Cd. This characterized a predominantly active plant metal uptake. Moreover, biological regulation was reported: plant growth and transpiration were significantly lower for hyperaccumulator plants cultivated in sand which was rich in available Ni and Cd, than for hyperaccumulator plants cultivated in topsoil, poor in available Ni and Cd. In the soil rhizosphere, capillary flow was related to transpiration and a depletion pattern was developed for Ni and sometimes for Cd.

Conclusions

Overall, the Intact Plant Transpiration Stream Concentration Factor appeared to be a relevant metal bioconcentration factor taking into account the predominant type of metal transport from roots to leaves, plant growth and transpiration coupling and metal availability. IPTSCF_Ni and IPTSCF_Cd values were much greater than 1 and similar whatever the amount of available Ni and Cd. This characterized a predominantly active plant combining Ni and Cd uptake and biological regulations dependent of the Ni and Cd concentrations in solution.     

Increased prevalence of tumor-infiltrating regulatory T cells is closely related to their lower sensitivity to H2O2-induced apoptosis in gastric and esophageal cancer

Purpose and experimental design

Although an increase in regulatory T cells (Tregs) is observed in tumor microenvironments, the underlying mechanism is not fully clarified. Since it was suggested that Tregs showed a lower sensitivity toward oxidative stress in comparison with conventional T cells, in the present study, we investigated the H₂O₂ production and apoptosis of Tregs in gastric and esophageal cancer tissues, employing flow cytometric analysis using fresh samples (n = 93) and immunohistochemical analysis (n = 203).

Results

The increased tumor-infiltrating Tregs coexisted with elevated H₂O₂ production according to disease progression. The grade of apoptosis in Tregs was less pronounced than that in conventional T cells, and there was a positive correlation between H₂O₂ production and the grade of apoptosis in conventional T cells, while there was no correlation between H₂O₂ production and the grade of apoptosis in Tregs. Moreover, Tregs were less sensitive to H₂O₂-induced apoptosis compared with conventional T cells in vitro.

Conclusions

We have demonstrated that the increased prevalence of tumor-infiltrating Tregs closely related to their lower sensitivity to H₂O₂-induced apoptosis.     

Influence of CdCl2 and CdSO4 supplementation on Cd distribution and ligand environment in leaves of the Cd hyperaccumulator Noccaea (Thlaspi) praecox

Background and aims

The aim was to investigate whether different Cd salts in the nutrient solution of the Cd/Zn hyperaccumulator Noccaea (Thlaspi) praecox alter leaf Cd distribution and Cd ligand environment, and plant fitness.

Methods

Plants were grown for 8 weeks with 100/300 μM CdCl₂ or CdSO₄. Leaf biomass, and total chlorophyll, anthocyanin, Cd, Cl, S and P concentrations were monitored. Cd localisation and ligand environment in leaves were analysed using quantitative synchrotron-based micro-X-ray fluorescence imaging, and Cd K-edge X-ray absorption fine structure and Cd L₃-edge micro-X-ray absorption near-edge structure measurements.

Results

Cd uptake and plant fitness were comparable for CdCl₂ and CdSO₄ treatments, and depended on applied Cd concentration. In all treatments, Cd preferentially accumulated with high concentrations of Cl in vacuoles of large vacuolarised epidermal cells, bound mainly to oxygen-based (O)-ligands. In the mesophyll of CdCl₂? treated plants, Cd was preferentially sequestered in vacuoles, while for CdSO₄, Cd accumulated preferentially in the apoplast. In the symplast, O-ligands increased with increasing Cd concentrations; in the apoplast, sulphur-based (S)-ligands prevailed.

Conclusions

Cd partitioning between leaf mesophyll apoplast and symplast and the Cd ligand environment in N. praecox depend on the Cd salt type and concentration added to the nutrient solution.     

Characterization of an Arabidopsis cadmium-resistant mutant <Emphasis Type="Italic">cdr3</Emphasis>-<Emphasis Type="Italic">1D</Emphasis> reveals a link between heavy metal resistance as well as seed development and flowering

A lot of studies have identified many key genes involved in heavy metal detoxification and tolerance in plants; however, our understanding of its molecular mechanisms is far from complete. To gain insight into the regulatory mechanisms for heavy metal detoxification and tolerance, we performed a mutant screen for identifying Arabidopsis (Arabidopsis thaliana) cadmium (Cd)-resistant mutants. A Cd-resistant mutant cdr3-1D (c a d mium-r esistant) was isolated because of its increased root growth and fresh weight in Cd stress, and genetic analysis showed that cdr3-1D is a single dominant nuclear mutation. Compared with the wild type, the cdr3-1D mutant was more resistant to heavy metals Cd, Pb, and copper as well as hydrogen peroxide. Moreover, we also observed that seeds of the cdr3-1D mutant were larger than those of wild type, and that cdr3-1D displayed early flowering compared with wild type. A lower Cd/Pb content was detected in cdr3-1D plants than in wild-type plants when subjected to Cd/Pb treatment, which was associated, at least in part, with increase of expression of AtPDR8/AtPDR12, a pump excluding Cd/Pb and/or Cd/Pb-containing toxic compounds from the cytoplasm, respectively. In addition, enhanced Cd/Pb resistance of the cdr3-1D mutant was partially glutathione (GSH) dependent, which was related to increase of expression of GSH1 gene involved in GSH synthesis and consequently increased GSH content. Taken together, our results provide genetic evidence indicating that CDR3 is involved in the regulation of heavy metal resistance as well as seed development and flowering.