As-hyperaccumulator Pteris Vittata Research Articles

Foliar-selenium enhances plant growth and arsenic accumulation in As-hyperaccumulator Pteris vittata: Critical roles of GSH-GSSG cycle and arsenite antiporters PvACR3

It is known that selenium (Se) enhances plant growth and arsenic (As) accumulation in As-hyperaccumulator Pteris vittata, but the associated mechanisms are unclear. In this study, P. vittata was exposed to 50 μM arsenate (AsV) under hydroponics plus 25 or 50 μM foliar selenate. After 3-weeks of growth, the plant biomass, As and Se contents, As speciation, malondialdehyde (MDA) and glutathione (GSH and GSSG) levels, and important genes related to As-metabolism in P. vittata were determined. Foliar-Se increased plant biomass by 17 − 30 %, possibly due to 9.1 − 19 % reduction in MDA content compared to the As control. Further, foliar-Se enhanced the As contents by 1.9–3.5 folds and increased arsenite (AsIII) contents by 64 − 136 % in the fronds. The increased AsV reduction to AsIII was attributed to 60 − 131 % increase in glutathione peroxidase activity, which mediates GSH oxidation to GSSG (8.8 −29 % increase) in the fronds. Further, foliar-Se increased the expression of AsIII antiporters PvACR3;1–3;3 by 1.6 − 2.1 folds but had no impact on phosphate transporters PvPht1 or arsenate reductases PvHAC1/2. Our results indicate that foliar-Se effectively enhances plant growth and arsenic accumulation by promoting the GSH-GSSG cycle and upregulating gene expression of AsIII antiporters, which are responsible for AsIII translocation from the roots to fronds and AsIII sequestration into the fronds. The data indicate that foliar-Se can effectively improve phytoremediation efficiency of P. vittata in As-contaminated soils.

Arsenic-Hyperaccumulator Pteris vittata Effectively Uses Sparingly-Soluble Phosphate Rock: Rhizosphere Solubilization, Nutrient Improvement, and Arsenic Accumulation.

Sparingly-soluble phosphate rock (PR), a raw material for P-fertilizer production, can be effectively utilized by the As-hyperaccumulator Pteris vittata but not most plants. In this study, we investigated the associated mechanisms by measuring dissolved organic carbon (DOC) and acid phosphatase in the rhizosphere, and nutrient uptake and gene expression related to the As metabolism in P. vittata. The plants were grown in a soil containing 200 mg kg-1 As and/or 1.5% PR for 30 days. Compared to the As treatment, the P. vittata biomass was increased by 33% to 4.6 g plant-1 in the As+PR treatment, corresponding to 27% decrease in its frond oxidative stress as measured by malondialdehyde. Due to PR-enhanced DOC production in the rhizosphere, the Ca, P, and As contents in P. vittata fronds were increased by 17% to 9.7 g kg-1, 29% to 5.0 g kg-1, and 57% to 1045 mg kg-1 in the As+PR treatment, thereby supporting its better growth. Besides, PR-induced rhizosphere pH increase from 5.0 to 6.9 promoted greater P uptake by P. vittata probably via upregulating low-affinity P transporters PvPTB1;1/1;2 by 3.7-4.1 folds. Consequently, 29% lower available-P induced the 3.3-fold upregulation of high-affinity P transporter PvPht1;3 in the As+PR treatment, which was probably responsible for the 58% decrease in available-As content in the rhizosphere. Consistent with the enhanced As translocation and sequestration, arsenite antiporters PvACR3/3;3 were upregulated by 1.8-4.4 folds in the As+PR than As treatment. In short, sparingly-soluble PR enhanced the Ca, P, and As availability in P. vittata rhizosphere and improved their uptake via upregulating genes related to As metabolism, suggesting its potential application for improving phytoremediation in As-contaminated soils.

Novel Phosphate Transporter-B PvPTB1;1/1;2 Contribute to Efficient Phosphate Uptake and Arsenic Accumulation in As-Hyperaccumulator Pteris vittata.

Arsenic (As) contamination in soil poses a potential threat to human health via crop uptake. As-hyperaccumulator Pteris vittata serves as a model plant to study As uptake and associated mechanisms. This study focuses on a novel P/AsV transport system mediated by low-affinity phosphate transporter-B 1 family (PTB1) in P. vittata. Here, we identified two plasma-membrane-localized PTB1 genes, PvPTB1;1/1;2, in vascular plants for the first time, which were 4.4-40-fold greater in expression in P. vittata than in other Pteris ferns. Functional complementation of a yeast P-uptake mutant and enhanced P accumulation in transgenic Arabidopsis thaliana confirmed their role in P uptake. Moreover, the expression of PvPTB1;1/1;2 facilitated the transport and accumulation of As in both yeast and A. thaliana shoots, demonstrating a comparable AsV uptake capacity. Microdissection-qPCR analysis and single-cell transcriptome analysis collectively suggest that PvPTB1;1/1;2 are specifically expressed in the epidermal cells of P. vittata roots. PTB1 may play a pivotal role in efficient P recycling during phytate secretion and hydrolysis in P. vittata roots. In summary, the dual P transport mechanisms consisting of high-affinity Pht1 and low-affinity PTB1 may have contributed to the efficient P/As uptake in P. vittata, thereby contributing to efficient phytoremediation for As-contaminated soils.

Arsenic-enhanced plant growth in As-hyperaccumulator Pteris vittata: Metabolomic investigations and molecular mechanisms

The first-known As-hyperaccumulator Pteris vittata is efficient in As uptake and translocation, which can be used for phytoremediation of As-contaminated soils. However, the underlying mechanisms of As-enhanced plant growth are unknown. We used untargeted metabolomics to investigate the potential metabolites and associated metabolic pathways regulating As-enhanced plant growth in P. vittata. After 60 days of growth in an MS-agar medium containing 15 mg kg−1 As, P. vittata biomass was 33–34 % greater than the no-As control. Similarly, the As contents in P. vittata roots and fronds were 272 and 1300 mg kg−1, considerably greater than the no-As control. Univariate and multivariate analyses based on electrospray ionization indicate that As exposure changed the expression of 1604 and 1248 metabolites in positive and negative modes. By comparing with the no-As control, As exposure significantly changed the expression of 14 metabolites including abscisic acid, d-glucose, raffinose, stachyose, chitobiose, xylitol, gibberellic acids, castasterone, citric acid, riboflavin-5-phosphate, ubiquinone, ubiquinol, UDP-glucose, and GDP-glucose. These metabolites are involved in phytohormone synthesis, energy metabolism, and sugar metabolism and may all potentially contribute to regulating As-enhanced plant growth in P. vittata. Our data provide clues to understanding the metabolic regulations of As-enhanced plant growth in P. vittata, which helps to enhance its phytoremediation efficiency of As-contaminated soils.

Insoluble-Phytate Improves Plant Growth and Arsenic Accumulation in As-Hyperaccumulator Pteris vittata: Phytase Activity, Nutrient Uptake, and As-Metabolism.

Phytate, the principal P storage in plant seeds, is also an important organic P in soils, but it is unavailable for plant uptake. However, the As-hyperaccumulator Pteris vittata can effectively utilize soluble Na-phytate, while its ability to utilize insoluble Ca/Fe-phytate is unclear. Here, we investigated phytate uptake and the underlying mechanisms based on the phytase activity, nutrient uptake, and expression of genes involved in As metabolisms. P. vittata plants were cultivated hydroponically in 0.2-strength Hoagland nutrient solution containing 50 μM As and 0.2 mM Na/Ca/Fe-phytate, with 0.2 mM soluble-P as the control. As the sole P source, all three phytates supported P. vittata growth, with its biomass being 3.2-4.1 g plant-1 and Ca/Fe-phytate being 19-29% more effective than Na-phytate. Phytate supplied soluble P to P. vittata probably via phytase hydrolysis, which was supported by 0.4-0.7 nmol P min-1 g-1 root fresh weight day-1 phytase activity in its root exudates, with 29-545 μM phytate-P being released into the growth media. Besides, compared to Na-phytate, Ca/Fe-phytate enhanced the As contents by 102-140% to 657-781 mg kg-1 in P. vittata roots and by 43-86% to 1109-1447 mg kg-1 in the fronds, which was accompanied by 21-108% increase in Ca and Fe uptake. The increased plant As is probably attributed to 1.3-2.6 fold upregulation of P transporters PvPht1;3/4 for root As uptake, and 1.8-4.3 fold upregulation of arsenite antiporters PvACR3/3;1/3;3 for As translocation to and As sequestration into the fronds. This is the first report to show that, besides soluble Na-phytate, P. vittata can also effectively utilize insoluble Ca/Fe-phytate as the sole P source, which sheds light onto improving its application in phytoremediation of As-contaminated sites.

Copper enhanced arsenic-accumulation in As-hyperaccumulator Pteris vittata by upregulating its gene expression for As uptake, translocation, and sequestration

In contaminated soils, arsenic (As) often co-exists with copper (Cu). However, its effects on As accumulation and the related mechanisms in As-hyperaccumulator Pteris vittata remain unclear. In this study, P. vittata plants were exposed to 50 µM As and/or 50 µM Cu under hydroponics to investigate the effects of Cu on plant growth and As accumulation, as well as gene expression related to arsenic uptake (P transporters), reduction (arsenate reductases), and translocation and sequestration (arsenite antiporters). After 14 d of growth and compared to the As treatment, the As concentration in P. vittata fronds increased by 1.4-times from 793 to 1131 mg·kg−1 and its biomass increased by 1.2-fold from 18.0 to 21.1 g·plant−1 in the As+Cu treatment. Copper-enhanced As accumulation was probably due to upregulated gene expressions related to As-metabolisms including As uptake (1.9-fold in P transporter PvPht1;3), translocation (2.1–2.4 fold in arsenite antiporters PvACR3/3;2) and sequestration (1.5–2.0 fold in arsenite antiporters PvACR3;1/3;3). Our results suggest that moderate amount of Cu can help to increase the As accumulation efficiency in P. vittata, which has implication in its application in phytoremedation in As and Cu co-contaminated soils.

Arsenic induced plant growth by increasing its nutrient uptake in As-hyperaccumulator Pteris vittata: Comparison of arsenate and arsenite

Arsenic-hyperaccumulator Pteris vittata is efficient in taking up arsenate (AsV) and arsenite (AsIII), however, their impacts on P. vittata growth and nutrient uptake remain unclear. The uptake of AsV and AsIII, their influences on nutrient uptake and plant biomass, and As speciation were investigated in P. vittata after exposing to 5 or 50 μM AsV or AsIII for 12 d under hydroponics. The results show that AsV uptake in P. vittata was 1.2 times more efficient than AsIII, corresponding to 1.7–2.1 fold greater biomass than the control at 50 μM As. While AsV was dominant in the roots at ∼60%, AsIII was more dominant in the fronds at ∼70% in all treatments. Macronutrients P, K, Ca, and S were increased by 118−185% at 50 μM As, with greater uptake of micronutrients Fe, Mn, Cu, and Zn at 5 μM As. Further, positive correlations between P. vittata biomass and its As contents (r = 0.97), and P. vittata biomass and its S, Mg, P, or Ca contents (r = 0.70–0.98) were observed. Our results suggest that its increased nutrient uptake probably enhanced P. vittata growth under As exposure.

Phytate and Arsenic Enhance Each Other's Uptake in As-hyperaccumulator Pteris vittata: Root Exudation of Phytate and Phytase, and Plant Uptake of Phytate-P.

Phytate as a root exudate is rare in plants as it mainly serves as a P storage in the seeds; however, As-hyperaccumulator Pteris vittata effectively secretes phytate and utilizes phytate-P, especially under As exposure. This study investigated the effects of As on its phytate and phytase exudation and the impacts of As and/or phytate on each other's uptake in P. vittata through two hydroponic experiments. Under 10-100 μM arsenate (AsV), the exudation of phytate and phytase by P. vittata was increased by 50-72% to 20.4-23.4 μmol h-1 g-1 and by 28-104% to 18.6-29.5 nmol h-1 plant-1, but they were undetected in non-hyperaccumulator Pteris ensiformis at 10 μM AsV. Furthermore, compared to 500 μM phytate, the phytate concentration in the growth media was reduced by 69% to 155 μM, whereas the P and As contents in P. vittata fronds and roots were enhanced by 68-134% and 44-81% to 2423-2954 and 82-407 mg kg-1 under 500 μM phytate plus 50 μM AsV. The increased P/As uptake in P. vittata was probably attributed to 3.0-4.5-fold increase in expressions of P transporters PvPht1;3-1;4. Besides, under As exposure, plant P may be converted to phytate in P. vittata roots, thereby increasing phytate's contents by 84% to 840 mg kg-1. Overall, our results suggest that As-induced phytate/phytase exudation and phytate-P uptake stimulate its growth and As hyperaccumulation by P. vittata.

Selenium Increased Arsenic Accumulation by Upregulating the Expression of Genes Responsible for Arsenic Reduction, Translocation, and Sequestration in Arsenic Hyperaccumulator Pteris vittata

Selenate enhances arsenic (As) accumulation in As-hyperaccumulator Pteris vittata, but the associated molecular mechanisms are unclear. Here, we investigated the mechanisms of selenate-induced arsenic accumulation by exposing P. vittata to 50 μM arsenate (AsV50) and 1.25 (Se1.25) or 5 μM (Se5) selenate in hydroponics. After 2 weeks, plant biomass, plant As and Se contents, As speciation in plant and growth media, and important genes related to As detoxification in P. vittata were determined. These genes included P transporters PvPht1;3 and PvPht1;4 (AsV uptake), arsenate reductases PvHAC1 and PvHAC2 (AsV reduction), and arsenite (AsIII) antiporters PvACR3 and PvACR3;2 (AsIII translocation) in the roots, and AsIII antiporters PvACR3;1 and PvACR3;3 (AsIII sequestration) in the fronds. The results show that Se1.25 was more effective than Se5 in increasing As accumulation in both P. vittata roots and fronds, which increased by 27 and 153% to 353 and 506 mg kg-1. The As speciation analyses show that selenate increased the AsIII levels in P. vittata, with 124-282% more AsIII being translocated into the fronds. The qPCR analyses indicate that Se1.25 upregulated the gene expression of PvHAC1 by 1.2-fold, and PvACR3 and PvACR3;2 by 1.0- to 2.5-fold in the roots, and PvACR3;1 and PvACR3;3 by 0.6- to 1.1-fold in the fronds under AsV50 treatment. Though arsenate enhanced gene expression of P transporters PvPht1;3 and PvPht1;4, selenate had little effect. Our results indicate that selenate effectively increased As accumulation in P. vittata, mostly by increasing reduction of AsV to AsIII in the roots, AsIII translocation from the roots to fronds, and AsIII sequestration into the vacuoles in the fronds. The results suggest that selenate may be used to enhance phytoremediation of As-contaminated soils using P. vittata.

Novel Mycorrhiza-Specific P Transporter PvPht1;6 Contributes to As Accumulation at the Symbiotic Interface of As-Hyperaccumulator Pteris vittata.

Arsenic (As) is toxic and ubiquitous in the environment, posing a growing threat to human health. As-hyperaccumulator Pteris vittata has been used for phytoremediation of As-contaminated soil. Symbiosis with arbuscular mycorrhizal fungi (AMF) enhances As accumulation by P. vittata, which is different from As inhibition in typical plants. In this study, P. vittata seedlings inoculated with or without AMF were cultivated in As-contaminated soils for 2 months. AMF-root symbiosis enhanced plant growth, with 64.5% greater As contents in the fronds. After exposure to AsV for 2 h, the arsenate (AsV) and arsenite (AsIII) contents in AMF-roots increased by 1.8- and 3.6-fold, suggesting more efficient As uptake by P. vittata with AMF-roots. Plants take up and transport AsV via phosphate transporters (Phts). Here, for the first time, we identified a novel mycorrhiza-specific Pht transporter, PvPht1;6, from P. vittata. The transcripts of PvPht1;6 were strongly induced in AMF-roots, which were localized to the plasma membrane of arbuscule-containing cells. By complementing a yeast mutant lacking 5-Phts, we confirmed PvPht1;6's transport activity for both P and AsV. In contrast to typical AMF-inducible phosphate transporter LePT4 from tomato, PvPht1;6 showed greater AsV transport capacity. The results suggest that PvPht1;6 is probably critical for AsV transport at the periarbuscular membrane of P. vittata root cells, revealing the underlying mechanism of efficient As accumulation in P. vittata with AMF-roots.

Arsenic-induced up-regulation of P transporters PvPht1;3–1;4 enhances both As and P uptake in As-hyperaccumulator Pteris vittata

Plants often up-regulate gene expression of P transporters under P deficiency, but down-regulate them under arsenic stress. Different from other P transporters, PvPht1;3 and PvPht1;4 expressions in As-hyperaccumulator Pteris vittata are up-regulated under P deficiency and As stress, showing strong transport capacity for both As and P. This study examined the mechanisms behind As-induced up-regulation of P transporters in P. vittata after exposing to 10–50 µM arsenate (AsV) for 14 d under hydroponics, with non-hyperaccumulator P. ensiformis as a control. Under As stress, P. vittata was more efficient in taking up both As and P than P. ensiformis, showing 48–84% more P content in the fronds and roots, leading to 18–79% greater biomass. Though As enhanced the P uptake by P. vittata, the inorganic P was reduced by 25–64% from 140–347 to 65–126 mg kg−1. It is likely that, under As stress, more P was utilized by P. vittata to counter As toxicity, causing reduction in inorganic P. This was supported by As-induced conversion of inorganic P to phytate in P. vittata, with phytate-P being increased by 26–75% from 239–713 to 418–1221 mg kg−1, maintaining internal low P levels. Under As-induced low inorganic-P conditions, the expressions of P transporters PvPht1;3 and PvPht1;4 were up-regulated by 1.4- and 2.7-fold in the roots, helping greater As and P uptake by P. vittata. Clearly, As-induced overexpression of P transporters in P. vittata roots plays a critical role in taking up both As and P, thereby increasing its efficiency in As-hyperaccumulation from contaminated media.

Effects of copper and arsenic on their uptake and distribution in As-hyperaccumulator Pteris vittata

Arsenic (As) and copper (Cu) are common co-contaminates in soils. However, their interactive effects on their accumulation and distribution in As-hyperaccumulator Pteris vittata are poorly understood. A hydroponic experiment was conducted with As being 0, 5, or 50 μM and Cu being 0.32, 3.2, or 32 μM to evaluate their phytotoxicity, accumulation, and distribution in P. vittata. In addition, As and Cu uptake kinetics were examined using the Michaelis-Menten kinetics model. Total As and Cu concentrations in P. vittata were up to 487 and 1355 mg kg−1. About 39–81% of the As was in the fronds compared to 0.6–18% for Cu. At 50 μM As, increasing Cu concentration from 0.32 to 32 μM increased root As while decreasing frond As concentrations, with the translocation factor (ratio of As in fronds to roots) being reduced from 4.0 to 0.31. In contrast, As did not affect Cu accumulation in P. vittata. Michaelis constant Km value for As was higher than that of Cu (6.49–24.9 vs. 0.43–3.36), consistent with higher Cu uptake than As. Besides, Cu reduced root K but increased P levels in the roots, whereas As increased the K and P concentrations in the fronds. Our results suggest that P. vittata accumulated more Cu than As in the roots, contributing to its low As translocation. As such, high levels of Cu are likely to reduce As uptake by P. vittata during phytoremediation of As-contaminated sites.

Arsenic-Induced Up-Regulation of P Transporters Pvpht1;3-1;4 Enhances Both as and P Uptake in As-Hyperaccumulator Pteris Vittata

Arsenic-Induced Up-Regulation of P Transporters Pvpht1;3-1;4 Enhances Both as and P Uptake in As-Hyperaccumulator Pteris Vittata

Selenate increased plant growth and arsenic uptake in As-hyperaccumulator Pteris vittata via glutathione-enhanced arsenic reduction and translocation

The beneficial effects of selenium on As uptake and plant growth in As-hyperaccumulator Pteris vittata are known, but the associated mechanisms remain unclear. Here, we investigated the effects of selenate on arsenic accumulation by P. vittata under two arsenate levels. P. vittata plants were exposed to 13 (As13) or 133 µM (As133) arsenate and 5 µM selenate in 0.2-strength Hoagland solution. After 14 d of growth, plant biomass, Se and As content, As speciation, and malondialdehyde (MDA), glutathione reductase (GR), glutathione peroxidase (GPX), and glutathione (GSH and GSSG) levels were determined. The results show that selenate promoted P. vittata growth and increased As concentrations in the roots and fronds by 256% from 97 to 346 mg kg−1 and 142% from 213 to 514 mg kg−1 under As13 treatment, and by 166% from 500 to 1332 mg kg−1 and 534% from 777 to 4928 mg kg−1 under As133 treatment. In addition, selenate increased the glutathione content in P. vittata roots and fronds by 75–86% under As13 treatment and 44–45% under As133 treatment. Selenate also increased the GPX activity by 161–173%, and GR activity by 72–79% in P. vittata under As13 and As133 treatments. The HPLC-ICP-MS analysis indicated that selenate increased both AsIII and AsV levels in P. vittata, with AsIII/AsV ratio being lower in the roots and higher in the fronds, i.e., more AsIII was being translocated to the fronds. Taken together, our results suggest that, via GPX-GR mediated enhancement of GSH-GSSG cycle, selenate effectively increases plant growth and As uptake in P. vittata by improving AsV reduction in the roots and AsIII translocation from the roots to the fronds.

Novel phytase PvPHY1 from the As-hyperaccumulator Pteris vittata enhances P uptake and phytate hydrolysis, and inhibits As translocation in Plant

Developing P-efficient plants helps improve P uptake from soils with low-available P and reduce environmental damage by P runoff. Here, we investigated a novel root-specific phytase PvPHY1 from As-hyperaccumulator Pteris vittata, which can efficiently utilize phytate, a recalcitrant organic phosphorus in soil. Unlike other plants, expression of PvPHY1 in P. vittata was greater in the roots than the fronds. A pure phytase with considerable activity was obtained via prokaryotic expression. Expressing PvPHY1 in tobacco (PvPHY1-Ex) enhanced its growth (2.8 to 3.5–3.9 g per plant) and increased its P accumulation by 10–50% under low- and adequate-P conditions. Further, PvPHY1-Ex tobacco showed 25–32% lower intracellular phytate and 30–56% higher inorganic P in the roots, likely due to phytase-mediated hydrolysis of phytate. Decrease of phytate levels up-regulated phosphate transporter genes (NbPht1;1, NbPht1;2 and NbPht1;6), leading to greater P and As uptake. However, As translocation to the shoots was low, probably due to competition from increased inorganic P via phytate hydrolysis. As such, PvPHY1 facilitated P uptake from soils and phytate hydrolysis in plants, thereby promoting tobacco growth. Overall, PvPHY1 from P. vittata helps better understand the novel phytase to increase soil P utilization efficiency, thereby reducing P fertilizer requirements for crop production.

Geographical distribution of As-hyperaccumulator Pteris vittata in China: Environmental factors and climate changes

Understanding the distribution of hyperaccumulators helps to implement more efficient phytoremediation strategies of contaminated sites, however, limited information is available. Here, we investigated the geographical distribution of the first-known arsenic-hyperaccumulator Pteris vittata in China and the key factors under two climate change scenarios (SSP 1-2.6 and SSP 5-8.5) at two time points (2030 and 2070). Species distribution model (MaxEnt) was applied to examine P. vittata distribution based on 399 samples from field surveys and existing specimen records. Further, among 23 environmental factors, 11 variables were used in the MaxEnt model, including temperature, precipitation, elevation, soil property, and UV-B radiation. The results show that P. vittata can grow in ~23% of the regions in China. Specifically, it is mainly distributed in 11 provinces of southern China, including Hainan, Guangdong, Guangxi, Yunnan, Guizhou, Hunan, Hubei, Jiangxi, Fujian, Zhejiang, and Jiangsu. Besides, eastern Sichuan, and southern Henan, Shaanxi, and Anhui are suitable for P. vittata growth. Under two climate change scenarios, P. vittata distribution in China would decrease by ~5.76-7.46 × 104 km2 in 2030 and ~3.22-4.68 × 104 km2 in 2070, with southern Henan and most Jiangsu being unsuitable for P. vittata growth. Among the 11 environmental variables, the minimum temperature of coldest month (bio6) and temperature annual range (bio7) are the two key factors limiting P. vittata distribution. At bio6 <-5 °C and/or bio7 >33 °C, the regions are unsuitable for P. vittata growth. Based on the MaxEnt model, precipitation had limited effects, so P. vittata can probably survive under both dry and moist environments. This study helps guide phytoremediation of As-polluted soils using P. vittata and provides an example to evaluate habitat suitability of hyperaccumulators at international scales.

Novel PvACR3;2 and PvACR3;3 genes from arsenic-hyperaccumulator Pteris vittata and their roles in manipulating plant arsenic accumulation

Arsenite (AsIII) antiporter ACR3 is crucial for arsenic (As) translocation and sequestration in As-hyperaccumulator Pteris vittata, which has potential for phytoremediation of As-contaminated soils. In this study, two new ACR3 genes PvACR3;2 and PvACR3;3 were cloned from P. vittata and studied in model organism yeast (Saccharomyces cerevisiae) and model plant tobacco (Nicotiana tabacum). Both ACR3s mediated AsIII efflux in yeast, decreasing its As accumulation and enhancing its As tolerance. In addition, PvACR3;2 and PvACR3;3 were expressed in tobacco plant. Localized on the plasma membrane, PvACR3;2 mediated both AsIII translocation to the shoots and AsIII efflux from the roots in tobacco, resulting in 203 − 258% increase in shoot As after exposing to 5 μM AsIII under hydroponics. In comparison, localized to the vacuolar membrane, PvACR3;3 sequestrated AsIII in tobacco root vacuoles, leading to 18 − 20% higher As in the roots and 15 − 36% lower As in the shoots. Further, based on qRT-PCR, both genes were mainly expressed in P. vittata fronds, indicating PvACR3;2 and PvACR3;3 may play roles in AsIII translocation and sequestration in the fronds. This study provides not only new insights into the functions of new ACR3 genes in P. vittata, but also important gene resources for manipulating As accumulation in plants for phytoremediation and food safety.

Expressing Phosphate Transporter PvPht2;1 Enhances P Transport to the Chloroplasts and Increases Arsenic Tolerance in Arabidopsis thaliana.

Arsenic (As) contamination in soils is of great concerns due to its toxicity to plants. As an analogue, phosphorus plays an important role in protecting plants from As toxicity. In this study, we identified a new phosphate transporter 2 (PHT2), PvPht2;1, from As-hyperaccumulator Pteris vittata and analyzed its functions in As and P transport in a yeast mutant, and model plant Arabidopsis thalian. PvPht2;1 contained 12 transmembrane domains, sharing high identity with PHT2 genes in diverse plants. Further, independent of external P or As levels, PvPht2;1 was mainly expressed in P. vittata fronds with the expression being 3-4 folds higher than that in the roots and rhizomes. Localized to the chloroplasts based on GFP-fused PvPht2;1 in model plant tobacco, PvPht2;1 functioned as a low-affinity P transporter. Under As exposure, PvPht2;1 yeast transformants showed comparable growth with the control while high-affinity P transporter PvPht1;3 transformants showed better growth, suggesting that PvPht2;1 transported P but slower than PvPht1;3 transporter. Expressing PvPht2;1 in A. thaliana increased its shoot P concentration without influencing its As accumulation. Further, the chloroplasts' P content in transgenic A. thaliana increased by 37-59% than wild-type (WT) plants. Under As exposure, the photosynthesis of PvPht2;1-expressing A. thaliana remained stable but that of WT plants decreased. The data indicate that, under As stress, expressing PvPht2;1 in A. thaliana enhanced its P transport to the chloroplasts and protected its photosynthesis. In short, highly expressed in the fronds and not impacted by As exposure, chloroplast-located PvPht2;1 may have protected As-hyperaccumulator P. vittata from As toxicity by efficiently transporting only P to its chloroplasts.

Efficient arsenate reduction in As-hyperaccumulator Pteris vittata are mediated by novel arsenate reductases PvHAC1 and PvHAC2

Arsenic-hyperaccumulator Pteris vittata is efficient in As absorption, reduction, and translocation. But the molecular mechanisms and locations of arsenate (AsV) reduction in P. vittata are still unclear. Here, we identified two new arsenate reductase genes from P. vittata, PvHAC1 and PvHAC2. Two PvHAC genes encoded a rhodanase-like protein, which were localized in the cytoplasm and nucleus. Both recombinant Escherichia coli strains and transgenic Arabidopsis thaliana lines showed arsenate reductase ability after expressing PvHAC genes. Further, expressing PvHAC2 enhanced As tolerance and reduced As accumulation in A. thaliana shoots under AsV exposure. Based on expression pattern analysis, PvHAC1 and PvHAC2 were predominantly expressed in the rhizomes and fronds of P. vittata. Different from those of HAC homologous genes in non-hyperaccumulators, little PvHAC was expressed in the roots. Besides, PvHAC1 expression was strongly upregulated under AsV exposure but not AsIII. The data suggest that arsenate reductase PvHAC1 in the rhizomes coupled with arsenate reductase PvHAC2 in the fronds of P. vittata played a critical role in As-hyperaccumulation by P. vittata, which helps to further improve its utility in phytoremediation of As-contaminated soils.

Pteris vittata coupled with phosphate rock effectively reduced As and Cd uptake by water spinach from contaminated soil

Arsenic (As) and cadmium (Cd) are ubiquitous in the environment and they are both toxic to humans. When present in soils, they can enter food chain, thereby threatening human health. Water spinach (Ipomoea aquatica) is an important leafy vegetable, which is widely consumed in Asian countries. However, it is efficient in taking up As and Cd from soils and accumulating them in the edible parts. Therefore, it is of significance to reduce its As and Cd content, especially in contaminated soil. In this study, pot experiments were conducted to investigate the ability of As-hyperaccumulator Pteris vittata in reducing As and Cd uptake by water spinach under different phosphorus treatments. P. vittata was grown for 60 d in a contaminated-soil amended with P fertilizer (+P) or phosphate rock (+PR), followed by water spinach cultivation for another 30 d. Plant biomass, As and Cd contents in plants and soils, and soil pH were analyzed. We found that, P. vittata coupled with PR decreased the As concentration in water spinach shoots by 42%, probably due to As uptake by P. vittata. Moreover, P. vittata decreased the Cd accumulation in water spinach by 24–44%, probably due to pH increase of 0.47–0.61 after P. vittata cultivation. Taking together, the results showed that P. vittata coupled with PR decreased the As and Cd content in water spinach, which is of significance for improving food safety and protecting human health.