Determination of arsenicals in mouse tissues after simulated exposure to arsenic from rice for sixteen weeks and the effects on histopathological features.

The accumulation of arsenic in rice has become a worldwide concern. In this study, dose-dependency in tissues (intestine, liver and kidney) and blood distribution of inorganic arsenicals and their methylated metabolites were investigated in male C57BL/6 mice exposed to four arsenic species (arsenite [iAs]III, arsenate [iAs]V, monomethylarsonate [MMA]V, and dimethylarsinate [DMA]V) at four doses (control [C]: 0 µg/g, simulation [S]: 0.91 µg/g, medium [M]: 9.1 µg/g and high [H]: 30 µg/g) according to the arsenical composition in rice for 8 and 16 weeks. No adverse effects were observed, while body weight gain decreased in group H. Increases in total arsenic concentrations (CtAs) and histopathological changes in the tissues occurred in all of the test groups. CtAs presented a tendency of kidney > intestine > liver > blood and were time-/dose-dependent in the liver and kidney in groups M and H. In the intestine and blood, abundant iAs (23%-28% in blood and 36%-49% in intestine) was detected in groups M and H, and CtAs decreased in group H from the 8th week to the 16th week. PMI decreased in the liver and SMI decreased in the kidney. These results indicate that the three tissues are injured through food arsenic. The intestine can also accumulate food arsenic, and the high arsenic dose will cause a deficiency in the absorbing function of the intestine. Thus, long-term exposure to arsenic-contaminated rice should be taken seriously attention.

Indigenous soil bacteria and the hyperaccumulator Pteris vittata mediate phytoremediation of soil contaminated with arsenic species.

Arsenic (As) is a pollutant of major concern worldwide, posing as a threat to both human health and the environment. Phytoremediation has been proposed as a viable mechanism to remediate As-contaminated soil environments. Pot experiments were performed to evaluate the phytoextraction efficiency of As by Pteris vittata, a known As hyperaccumulating fern, from soil amended with different concentrations of arsenate [As(V)] and arsenite [As(III)], the more common, inorganic As forms in soil. The greatest accumulation of As (13.3 ± 0.36 g/kg Dwt) was found in fronds of plants grown in soil spiked with 1.0 g As(V)/kg. The maximum As-bioaccumulation factor (27.3 ± 1.9) was achieved by plants grown in soil amended with 0.05 g As(V)/kg. A total of 864 bacterial cultures were isolated and examined for their ability to enhance phytoremediation of As-contaminated soils. Traits examined included tolerance to As (III and V), production of siderophores, and/or ability to solubilize calcium phosphate and indole acetic acid (IAA) production. A culture-based survey shows greater numbers of viable and As-resistant bacteria were found in the rhizosphere of As-grown plants compared to bulk and unplanted soils. The percentage of bacteria resistant to As(V) was greater (P < 0.0001) than those resistant to As(III) in culture medium containing 0.5, 1, 1.5, and 2 g As/L. Higher (P < 0.0001) percentages of siderophore producing (77%) and phosphate solubilizing (61%) bacteria were observed among cultures isolated from unplanted soil. About 5% (44 of 864) of the isolates were highly resistant to both As (III) and As (V) (2 g/L), and were examined for their As-transformation ability and IAA production. A great proportion of the isolates produced IAA (82%) and promoted As (V)-reduction (95%) or As(III)-oxidation (73%), and 71% exhibited dual capacity for both As(V) reduction and As(III) oxidation. Phylogenetic analysis indicated that 67, 23, and 10% of these isolates belonged to Proteobacteria, Actinobacteria, and Firmicutes, respectively. Analysis of the 16S rRNA gene sequences confirmed that these isolates were closely related to 12 genera and 25 species of bacteria and were dominated by members of the genus Pseudomonas (39%). These results show that these isolates could potentially be developed as inocula for enhancing plant uptake during large scale phytoremediation of As-impacted soils.

Interspecific biotransformation and detoxification of arsenic compounds in marine rotifer and copepod.

The toxicity of arsenic (As) has been reported to be different depending on their chemical forms. However, its toxicity mechanisms largely remain unknown. In this study, to investigate toxicity mechanism of As in marine zooplanktons, namely, the rotifer Brachionus plicatilis and the copepod Paracyclopina nana, metabolites of As were analyzed by using a high-performance liquid chromatography coupled with inductively coupled plasma mass spectrometry with in vivo toxicity and antioxidant responses in response to inorganic As, including arsenate (AsV) and arsenite (AsIII). While AsIII was more toxic than AsV in both organisms, the rotifer B. plicatilis exhibited stronger tolerance, compared to the copepod P. nana. The As speciation analysis revealed differences in biotransformation processes in two species with B. plicatilis having a more simplified process than P. nana, contributing to a better tolerance against As in the rotifer B. plicatilis compared to P. nana. Moreover, the levels of GSH content and the regulation of omega class glutathione S-transferases were different in response to oxidative stress between B. plicatilis and P. nana. These results suggest that the rotifer B. plicatilis has a unique survival strategy with more efficient biotransformation and antioxidant responses, compared to P. nana, conferring higher tolerance to As.

Biotransformation of dietary inorganic arsenic in a freshwater fish Carassius auratus and the unique association between arsenic dimethylation and oxidative damage.

The metabolic process and toxicity mechanism of dietary inorganic arsenic (iAs) in freshwater fish remain unclear to date. The present study conducted two iAs [arsenate (As(V)) and arsenite (As(III))] dietary exposures in freshwater fish crucian carp (Carassius auratus). The fish were fed on As supplemented artificial diets at nominal concentrations of 50 and 100 µg As(III) or As(V) g-1 (dry weight) for 10 d and 20 d. We found that the liver, kidney, spleen, and intestine of fish accumulated more As in As(V) feeding group than that in As(III), while the total As levels in muscle were similar between As(V) and As(III) group at the end of exposure. Reduction of As(V) to As(III) and oxidation of As(III) to As(V) occurred in fish fed with As(V) and As(III), respectively, indicating that toxicity of iAs was likely elevated or reduced when iAs was absorbed by fish before entering into human body through diet. Biomethylation to monomethylarsonic acid and dimethylarsinic acid and transformation to arsenocholine and arsenobetaine were also found in the fish. The linear regression analysis showed a positive correlation between secondary methylation index and the malondialdehyde content in tissues, highlighting the vital role of arsenic dimethylation in the oxidative damages in fish.

Two-generational effects and recovery of arsenic and arsenate on Daphnia magna in the presence of nano-TiO2.

The toxicity of arsenic (As) can be influenced by many environmental factors. Among them, nanomaterials can adsorb arsenic and alter its bioavailability in organisms. However, the studies on long-term effects of arsenic in the presence of nanoparticles are limited. Thus, the 21-d effect of titanium dioxide nanoparticles (nano-TiO2) on chronic toxicity of arsenic (arsenate and arsenite) was investigated in two generations of Daphnia magna. The exposed concentration of nano-TiO2 was 1 mg/L and the concentration of As(Ⅲ) or As(Ⅴ) was 0.2 mg/L which was lower than the 48 h-NOEC (no observed effect concentration). The survival, body length, average number of offspring and time of first brood were determined. Our results indicated that the exposure to nano-TiO2 and As during the parental generation can affect the health of offspring. Nano-TiO2 was found to significantly alleviate the mortality and reproduction inhibition of As on D. magna, and the alleviation of As(Ⅴ) was more prominent than that of As(Ⅲ). It is likely that nano-TiO2 alters the metabolism and adsorption condition of arsenic in the gastrointestinal tract of D. magna. Overall, these results indicate that the increase of arsenic adsorption onto nano-TiO2 in the gut of D. magna could alleviate the toxicity of arsenic. Nonetheless, further research should be conducted to study the influence of arsenic on the multi-generations of aquatic organisms, especially when it is coexisted with other substances.

Developmental neurotoxicity of inorganic arsenic exposure in Sprague-Dawley rats.

High levels of inorganic arsenic (iAs) exposure are associated with severe health effects. Less clear are effects of lower exposure levels on neurodevelopment. Relative to maternal intake, there is limited lactational transfer of arsenic in humans or rodents, yet there are few rodent studies which directly exposed preweaning animals. To more clearly determine iAs developmental neurotoxicity, 28 pregnant Sprague-Dawley rats were exposed to arsenate (AsV) via drinking water (0, 23.6, 47.7, 71.0 ppm) (n = 5-7/group) from gestational day (GD) 6 through GD 22 with targeted doses of 0, 2.33, 4.67, 7.00 mg/kg/day, respectively. Offspring were dosed by gavage daily with the same mg/kg AsV dose as intended for their dam from postnatal day (PND) 1 to 21. Gestational water intake was reduced at all AsV doses, but returned to control levels on lactational day (LD) 1 when control water was returned. Gestational body weight was reduced only at the highest dose on GD 22 and lactational body weight was unaffected. Food intake was unaffected. iAs exposure did not alter offspring body weight (PNDs 1-21) or age at fur development and bilateral ear opening. Incisor eruption, however, was significantly delayed in offspring of the 4.67 and 7.00 mg/kg groups. Further, all iAs groups were significantly delayed in bilateral eye opening. Righting reflex (PNDs 3-6) was unaffected, while slant board performance (PNDs 8-11) was significantly poorer at the highest dose. Brains of culled pups (PND 1) showed dose-dependent increases of iAs. There were no significant AsV-related effects on PND 21 brain regional concentrations of dopamine, DOPAC, HVA, 5-HT or 5-HIAA. These hazard identification results will guide the study designs of developmental iAs exposure at human-relevant levels essential for risk-assessment.

State of the science review of the health effects of inorganic arsenic: Perspectives for future research.

Human exposure to inorganic arsenic (iAs) is a global health issue. Although there is strong evidence for iAs-induced toxicity at higher levels of exposure, many epidemiological studies evaluating its effects at low exposure levels have reported mixed results. We comprehensively reviewed the literature and evaluated the scientific knowledge on human exposure to arsenic, mechanisms of action, systemic and carcinogenic effects, risk characterization, and regulatory guidelines. We identified areas where additional research is needed. These priority areas include: (1) further development of animal models of iAs carcinogenicity to identify molecular events involved in iAs carcinogenicity; (2) characterization of underlying mechanisms of iAs toxicity; (3) assessment of gender-specific susceptibilities and other factors that modulate arsenic metabolism; (4) sufficiently powered epidemiological studies to ascertain relationship between iAs exposure and reproductive/developmental effects; (5) evaluation of genetic/epigenetic determinants of iAs effects in children; and (6) epidemiological studies of people chronically exposed to low iAs concentrations.

Low arsenate influx rate and high phosphorus concentration in wheat (Triticum aestivum L.): A mechanism for arsenate tolerance in wheat plants.

Two wheat (Triticum aestivum L.) cultivars differing in arsenic (As)-tolerance were used to investigate the effects of phosphorus (P) concentration and nutrient solution pH on As(V) toxicity and As(V) uptake kinetics, and to illustrate the mechanism of As(V) tolerance in wheat seedlings. Low pH and low phosphate concentration enhanced wheat uptake of As, resulting in high As toxicity. The As(V)-tolerant cultivar MM45 exhibited higher relative root elongation than non-tolerant cultivar HM29 in all treatments, except that no genotypic difference was recorded for the solution P at 100 µmol L-1 or greater. Wheat seedling As(V) tolerance was positively correlated with P concentration in roots and shoots. In short-term (30 min) As(V)-uptake kinetics experiments, the maximum influx rate (Vmax) of As(V) increased with decreasing solution pH (from 7.0 to 6.0). Compared with HM29, although MM45 had lower Vmax, its Michaelis-Menten constant (Km) did not exceed that of HM29 in all treatments. The Vmax values of both cultivars were not significantly affected by phosphate treatments, except for HM29 which had significantly higher Vmax value in the presence of phosphate at pH 7.0. The Km values of the two cultivars increased by 9- to 20-fold when phosphate was present as opposed to absent from the uptake solution. This study showed that the Vmax values are mainly increased by high pH and As(V) uptake Km is mainly increased by phosphate presence. Decreased As(V) influx rates during early stages and increased P concentration in plant tissues are associated with increased As tolerance in wheat seedlings.

Statistical optimization of arsenic biosorption by microbial enzyme via Ca-alginate beads.

Bioremediation of arsenic using green technology via microbial enzymes has attracted scientists due to its simplicity and cost effectiveness. Statistical optimization of arsenate bioremediation was conducted by the enzyme arsenate reductase extracted from arsenic tolerant bacterium Pseudomonas alcaligenes. Response surface methodology based on Box-Behnken design matrix was performed to determine the optimal operational conditions of a multivariable system and their interactive effects on the bioremediation process. The highest biosorptive activity of 96.2 µg gm-1 of beads was achieved under optimized conditions (pH = 7.0; As (V) concentration = 1000 ppb; time = 2 h). SEM analysis showed the morphological changes on the surface of enzyme immobilized gluteraldehyde crosslinked Ca-alginate beads. The immobilized enzyme retained its activity for 8 cycles. ANOVA with a high correlation coefficient (R2 > 0.99) and lower "Prob > F"value (<0.0001) corroborated the second-order polynomial model for the biosorption process. This study on the adsorptive removal of As (V) by enzyme-loaded biosorbent revealed a possible way of its application in large scale treatment of As (V)-contaminated water bodies.

Arsenic uptake by arugula (Eruca vesicaria, L.) cultivars as affected by phosphate availability.

To assess the importance of variation among arugula (Eruca vesicaria subsp. sativa) cultivars in the ability to accumulate arsenic (As) in above-ground tissues, uptake of As by 16 cultivars was measured in the field and in hydroponic culture. In the field trial on soil contaminated by past pesticide use, As soil-plant uptake coefficients varied by a factor of 2.7 among different cultivars, approaching a value of one for the strongest accumulators. Compared to the field assay, hydroponically grown arugula accumulated much lower concentrations of As when nutrient solutions contained standard (high) concentrations of phosphate along with 1.0 mg L-1 As in the form of soluble arsenate. However, As accumulation was much greater in hydroponic culture using low-P nutrient solutions, an indication that phosphate strongly competed with arsenate for root uptake. Analysis of arugula roots after exposure to arsenate at 1.0 mg As L-1 and low phosphate revealed from 24 to 400 times greater As concentration in roots than tops, with S concentrations significantly greater in As-exposed than control roots. This indicated greater sulfate uptake by roots exposed to arsenate, and suggested that thiol-mediated As immobilization occurred in the roots which strongly restricted translocation to the tops.

Knocking Out OsPT4 Gene Decreases Arsenate Uptake by Rice Plants and Inorganic Arsenic Accumulation in Rice Grains.

Arsenic (As) accumulation in rice grains poses health risk to humans. Plants including rice take up arsenate (AsV) by phosphate transporters. In this study, rice phosphate transporter OsPT4 (OsPht1;4) was investigated based on two independent T-DNA insertion mutants of OsPT4 (M1 and M2), which displayed stronger AsV resistance than wild types WT1 and WT2. When cultivated in medium (+P or -P) with AsV, ospt4 mutants accumulated 16-32% lower As in plants, suggesting that OsPT4 mediates AsV uptake. Analysis of the xylem sap showed that AsV concentrations in ospt4 mutants was 20-40% lower than WT controls under -P condition, indicating OsPT4 may also mediate AsV translocation. Moreover, kinetics analysis showed that ospt4 mutants had lower AsV uptake rates than the WT controls, further proving that OsPT4 functions as an AsV transporter in rice. When grown in flooded soils with As, AsV concentrations in rice grains of ospt4 mutants decreased by 50-55%. More importantly, knocking out OsPT4 in M1 and M2 reduced inorganic As accumulation in rice grains by 20-44%, significant for controlling As exposure risk from rice. Taken together, our findings revealed a critical role of OsPT4 in AsV uptake and translocation in rice. Knocking out OsPT4 effectively decreased inorganic As accumulation in rice grains, shedding light on engineering low-As rice to enhance food safety.

Thioarsenate Toxicity and Tolerance in the Model System Arabidopsis thaliana.

Thioarsenates form from arsenite under sulfate-reducing conditions, e.g., in rice paddy soils, and are structural analogues of arsenate. Even though rice is one of the most important sources of human arsenic intake, nothing is published about uptake, toxicity, or tolerance of thioarsenates in plants. Experiments using the model system Arabidopsis thaliana showed that monothioarsenate is less toxic than arsenite, but more toxic than arsenate at concentrations ≥25 µM As, reflected in stronger seedling growth inhibition on agar plates. Despite higher toxicity, total As accumulation in roots was lower upon exposure to monothioarsenate compared to arsenate, and a higher root efflux was confirmed. Root-shoot translocation was higher for monothioarsenate than for arsenate. Compared to the wild type (Col-0), both arsenate and monothioarsenate induced higher toxicity in phytochelatin (PC)-deficient mutants (cad1-3) as well as in glutathione biosynthesis (cad2) and PC transport (abcc12) mutants, demonstrating the important role of the PC pathway, not only for arsenate, but also for monothioarsenate detoxification. In Col-0, monothioarsenate induced relatively higher accumulation of PCs than arsenate. The observed differences in plant uptake, toxicity, and tolerance of thioarsenate vs oxyarsenate show that studying the effects of As on plants should include experiments with thiolated As species.

Phytochelatins play key roles for the difference in root arsenic accumulation of different Triticum aestivum cultivars in comparison with arsenate uptake kinetics and reduction.

In the previous studies, we have found that arsenic (As) accumulation in roots of bread wheat (Triticum aestivum L.) seedlings were significantly different among different wheat cultivars, and As(V) tolerant wheat cultivars have much higher capacities of root As accumulation. However, the reason for the difference remains unclear. Four wheat cultivars with high (MM45 and FM8) or low (QF1 and HM29) levels of arsenic (As) accumulation were selected to investigate the relationship between root As(V) uptake kinetics and root As accumulation. MM45 and HM29 were also used to examine As(V) reduction ability and non-protein thiol (cysteine [Cys], glutathione [GSH], and phytochelatins [PCs]) concentrations in wheat seedlings. MM45 had the lowest Michaelis-Menten constant (Km) and maximum influx rate (Vmax). No difference in the Km values was found among the three other cultivars. No difference in As(V) reduction capacity was observed between MM45 and HM29. GSH and PC2 were significantly induced by 10 µM As(V) in roots of wheat seedlings, particularly in MM45. Synthesis of GSH and PCs was completely suppressed in the presence of l-buthionine sulfoximine (BSO), a specific inhibitor of γ-glutamylcysteine synthetase. BSO markedly decreased the As tolerance of wheat seedlings and decreased the accumulation of As in roots, but increased As accumulation in shoots. No significant difference in As concentrations was found between MM45 and HM29 under the BSO treatment. GSH and PCs are the reason why As accumulation and As(V) tolerance differ in roots of different wheat cultivars.

Phosphorus deficiency modifies As translocation in the halophyte plant species Atriplex atacamensis.

Most arsenic in surface soil and water exists primarily in its oxidized form, as arsenate (As(V); AsO43-), which is an analog of phosphate (PO43-). Arsenate can be taken up by phosphate transporters. Atriplex atacamensis Phil. is native to northern Chile (Atacama Desert), and this species can cope with high As concentrations and low P availability in its natural environment. To determine the impact of P on As accumulation and tolerance in A. atacamensis, the plants were cultivated in a hydroponic system under four treatments: no As(V) addition with 323µM phosphate (control); 1000µM As(V) addition with 323µM phosphate; no As(V) and no phosphate; 1000µM As(V) addition and no phosphate. Phosphate starvation decreased shoot fresh weight, while As(V) addition reduced stem and root fresh weights. Arsenate addition decreased the P concentrations in both roots and leaves, but to a lesser extent than for P starvation. Phosphorus starvation increased the As concentrations in roots, but decreased it in shoots, which suggests that P deficiency reduced As translocation from roots to shoots. Arsenate addition increased total glutathione, but P deficiency decreased oxidized and reduced glutathione in As(V)-treated plants. Arsenate also induced an increase in S accumulation and nonprotein thiol and ethylene synthesis, and a decrease in K concentrations, effects that were similar for the P-supplied and P-starved plants. In contrast, in As(V)-treated plants, P starvation dramatically decreased total soluble protein content and increased lipid peroxidation, compared to plants supplied with P. Phosphorus nutrition thus appears to be an important component of A. atacamensis response to As toxicity.

Effects of phosphate binders on the gastrointestinal absorption of arsenate and of an SGLT2 inhibitor drug on the urinary excretion of arsenite in mice.

Arsenate (AsV) and arsenite (AsIII) are typical sources of acute and chronic arsenic poisoning. Therefore, reducing inner exposure to these arsenicals is a rational objective. Because AsV mimics phosphate, phosphate binder drugs may decrease the intestinal AsV absorption. Indeed, lanthanum and aluminium salts and sevelamer removed AsV from solution in vitro, especially at acidic pH. In mice gavaged with AsV, lanthanum chloride, lanthanum carbonate and aluminium hydroxide given orally also lowered the urinary excretion and tissue levels of AsV and its metabolites, indicating that they decreased the gastrointestinal AsV absorption. As some glucose transporters may carry AsIII, the effect of the SGLT2 inhibitor dapagliflozin was investigated in AsIII-injected mice. While producing extreme glucosuria, dapagliflozin barely affected the urinary excretion and tissue concentrations of AsIII and its metabolites. Thus, phosphate binders (especially lanthanum compounds) can reduce the gastrointestinal absorption of AsV; however, SGLT2 inhibition cannot diminish the renal reabsorption of AsIII.

Micro-distribution of arsenic species in tissues of hyperaccumulator Pteris vittata L.

Arsenic (As) contamination and its harmful consequences have gained increasing attention in research. Phytoextraction, which uses the As hyperaccumulator Pteris vittata L., is a well-established technology adopted in many countries. However, the hyperaccumulation mechanisms of this plant remain controversial. This study investigated the species and the micro-distribution of As species in three P. vittata L. ecotypes after exposure to arsenite (AsIII) and arsenate (AsV) for 7d. Arsenic-accumulating abilities and preferences to As species varied among different ecotypes. The reduction of AsV into AsIII, oxidation of AsIII into AsV, and chelation of AsIII with thiols were all observed in P. vittata. The reduction of As mainly occurred in the rhizoid, whereas oxidation and chelation mainly occurred in the aboveground parts. Correlation analyses showed that the As concentration in pinna was significantly correlated with the AsV percentage in paraxial and abaxial epidermis (positive), AsIII-GSH percentage in paraxial epidermis (positive), and AsIII percentage in paraxial and abaxial epidermis (negative). Results indicated that oxidation and chelation reactions contributed to the accumulation of As in P. vittata.

Toxicological characterisation of a thio-arsenosugar-glycerol in human cells.

Arsenosugars are water-soluble arsenic species predominant in marine algae and other seafood including mussels and oysters. They typically occur at levels ranging from 2 to 50mg arsenic/kg dry weight. Most of the arsenosugars contain arsenic as a dimethylarsinoyl group (Me2As(O)-), commonly referred to as the oxo forms, but thio analogues have also been identified in marine organisms and as metabolic products of oxo-arsenosugars. So far, no data regarding toxicity and toxicokinetics of thio-arsenosugars are available. This in vitro-based study indicates that thio-dimethylarsenosugar-glycerol exerts neither pronounced cytotoxicity nor genotoxicity even though this arsenical was bioavailable to human hepatic (HepG2) and urothelial (UROtsa) cells. Experiments with the Caco-2 intestinal barrier model mimicking human absorption indicate for the thio-arsenosugar-glycerol higher intestinal bioavailability as compared to the oxo-arsenosugars. Nevertheless, absorption estimates were much lower in comparison to other arsenicals including arsenite and arsenic-containing hydrocarbons. Arsenic speciation in cell lysates revealed that HepG2 cells are able to metabolise the thio-arsenosugar-glycerol to some extent to dimethylarsinic acid (DMA). These first in vitro data cannot fully exclude risks to human health related to the presence of thio-arsenosugars in food.

Effect of arsenate As (V) on the biomarkers of Myriophyllum alterniflorum in oligotrophic and eutrophic conditions.

Alternate watermilfoil, Myriophyllum alterniflorum is an aquatic macrophyte found in the Limousin rivers (France) whose potential for biomonitoring of metal pollution has been demonstrated. The objective of the present study carried out in vitro was to identify biomarkers for an early detection of the pollution by a metalloid As (V) in eutrophic and oligotrophic conditions. A synthetic medium of similar composition to the waters of the River Vienne was prepared. The morphological development of watermilfoil was monitored for 30 days, with or without contamination by 100 µg L(-1) As (V). In addition, the mineralization of plants and the analysis of biomarkers (chlorophylls, photosynthetic and respiratory intensities …) were investigated after 21 days. Our results indicated that eutrophic medium, induced a decrease in chlorophyll pigments, in growth and an increase in H2O2 compared to the oligotrophic medium. While, the presence of As (V), led to a decrease in the osmotic potential, pigment content, photosynthesis and respiration rates and an inhibition of shoot branching of plants in both conditions. However, a significant increase in H2O2 content was noted in the eutrophic medium. Finally, As (V) was found to be more accumulated in roots than shoots in both conditions but was more accumulated in oligotrophic one. Therefore, we can conclude that the water trophic level modifies the response of M. alterniflorum in presence of arsenate. Thus, M. alterniflorum shows a great promise in water-quality biomonitoring.

A member of the Phosphate transporter 1 (Pht1) family from the arsenic-hyperaccumulating fern Pteris vittata is a high-affinity arsenate transporter.

Pteris vittata exhibits enhanced arsenic uptake, but the corresponding mechanisms are not well known. The prevalent form of arsenic in most soils is arsenate, which is a phosphate analog and a substrate for Phosphate transporter 1 (Pht1) transporters. Herein we identify and characterize three P. vittata Pht1 transporters. Pteris vittata Pht1 cDNAs were isolated and characterized via heterologous expression in Saccharomyces cerevisiae (yeast) and Nicotiana benthamiana leaves. Expression of the PvPht1 loci in P. vittata gametophytes was also examined in response to phosphate deficiency and arsenate exposure. Expression of each of the PvPht1 cDNAs complemented the phosphate uptake defect of a yeast mutant. Compared with yeast cells expressing Arabidopsis thaliana Pht1;5, cells expressing PvPht1;3 were more sensitive to arsenate, and accumulated more arsenic. Uptake assays with yeast cells and radiolabeled (32)P revealed that PvPht1;3 and AtPht1;5 have similar affinities for phosphate, but the affinity of PvPht1;3 for arsenate is much greater. In P. vittata gametophytes, PvPht1;3 transcript levels increased in response to phosphate (Pi) deficiency and arsenate exposure. PvPht1;3 is induced by Pi deficiency and arsenate, and encodes a phosphate transporter that has a high affinity for arsenate. PvPht1;3 probably contributes to the enhanced arsenate uptake capacity and affinity exhibited by P. vittata.

Arsenic uptake, arsenite efflux and plant growth in hyperaccumulator Pteris vittata: Role of arsenic-resistant bacteria.

Bacteria-mediated arsenic (As) transformation and their impacts on As and P uptake and plant growth in As-hyperaccumulator Pteris vittata (PV) were investigated under sterile condition. All As-resistant bacteria (9 endophytic and 6 rhizospheric) were As-reducers except one As-oxidizer. After growing two months in media with 37.5 mg kg(-1) AsV, As concentrations in the fronds and roots were 3655-5389 (89-91% AsIII) and 971-1467 mg kg(-1) (41-73% AsIII), corresponding to 22-52% decrease in the As in the media. Bacterial inoculation enhanced As and P uptake by up to 47 and 69%, and PV growth by 20-74%, which may be related to elevated As and P in plants (r = 0.88-0.97, p < 0.05). Though AsV was supplied, 95% of the As in the bacteria-free media was AsIII, suggesting efficient efflux of AsIII by PV roots (120 µg g(-1) root fw). This was supported by the fact that no AsV was detected in media inoculated with As-reducers while 95% of AsV was detected with As-oxidizer. Our data showed that, under As-stress, PV reduced As toxicity by efficient AsIII efflux into media and AsIII translocation to the fronds, and bacteria benefited PV growth probably via enhanced As and P uptake.