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.

Arbuscular mycorrhizal fungi promote arsenic accumulation in Pteris vittata L. through arsenic solubilization in rhizosphere soil and arsenic uptake by hyphae.

The introduction of arbuscular mycorrhizal fungi (AMF) is considered an effective strategy for improving the arsenic phytoremediation efficiency of Pteris vittata L. (P. vittata). However, how hyphae take up arsenic and translocate it to the root cells of P. vittata in the symbiotic mycorrhizal structure is currently unclear. In this study, the role of hyphae in arsenic enrichment in P. vittata and the mechanism of arsenic species transformation in the rhizosphere were studied via a compartmented cultivation setup. After Claroidoglomus etunicatum (C. etunicatum) colonization, the arsenic content of P. vittata increased by 234%. Hyphae contributed 32% to the accumulation of arsenic in symbionts. C. etunicatum promoted the conversion of iron and aluminum oxides to crystalline states in rhizosphere soil, promoted the desorption of arsenic bound to iron and aluminum oxides, and increased the content of available arsenic in rhizosphere soil by 116%. The transfer of arsenic from arbuscular structures to root cells was confirmed by transmission electron microscopy (TEM)/scanning electron microscopy- energy dispersive X-ray spectroscopy (SEMEDS) analysis. This study demonstrated that C. etunicatum inoculation enhances the phytoremediation efficiency of P. vittata in arsenic-contaminated soils through hyphal uptake, plant growth promotion, and alteration of the rhizosphere environment.

In Vitro Modulation of Genotoxicity and Oxidative Stress by Polyphenol-Rich Fraction of Chinese Ladder Brake (Pteris vittata L.).

Pteris vittata L. is a terrestrial genus growing in moist, shady forests and on hillsides. The plant has considerable ethnomedicinal importance. Investigations have been carried out on chemical profiling and antioxidant compounds from some genera of pteridophytes but studies on the biological properties of P. vittata are lacking. Therefore, the present study investigates antioxidant, antigenotoxic, and antiproliferative potential of the aqueous fraction of P. vittata (PWE). A battery of assays were carried out to assess the antioxidant potential of the PWE. SOS chromotest and DNA nicking assay were used to evaluate the antigenotoxicity of the fraction. The cytotoxic effect of PWE was analyzed using MTT and Neutral Single Cell Gel Electrophoresis comet assay. EC50 of 90.188 µg/ml, 80.13 µg/ml, 142.836 µg/ml, and 12.274 µg/ml was obtained in DPPH, superoxide anion scavenging, reducing power and lipid peroxidation assays, respectively. PWE was potent in inhibiting Fenton's reagent-induced nicking of pBR322 plasmid. The fraction significantly inhibited hydrogen peroxide (H2O2) and 4-nitroquinoline-N-oxide (4NQO) induced mutagenicity and a reduction in induction factor was found with increased PWE concentration. GI50 of 147.16 µg/ml was obtained in MTT assay in human MCF-7 breast cancer cell line. PWE induced apoptosis as confirmed from confocal microscopy studies. The protective effects can be attributed to the presence of the phytochemicals in PWE. These results will be helpful in the development of functional food characteristics, as well as unravel the benefits of pteridophytes as promoters of health.

Arsenic in the hyperaccumulator Pteris vittata: A review of benefits, toxicity, and metabolism.

Arsenic (As) is a toxic metalloid, elevated levels of which in soils are becoming a major global environmental issue that poses potential health risks to humans. Pteris vittata, the first known As hyperaccumulator, has been successfully used to remediate As-polluted soils. Understanding why and how P. vittata hyperaccumulates As is the core theoretical basis of As phytoremediation technology. In this review, we highlight the beneficial effects of As in P. vittata, including growth promotion, elemental defense, and other potential benefits. The stimulated growth of P. vittata induced by As can be defined as As hormesis, but differs from that in non-hyperaccumulators in some aspects. Furthermore, the As coping mechanisms of P. vittata, including As uptake, reduction, efflux, translocation, and sequestration/detoxification are discussed. We hypothesize that P. vittata has evolved strong As uptake and translocation capacities to obtain beneficial effects from As, which gradually leads to As accumulation. During this process, P. vittata has developed a strong As vacuolar sequestration ability to detoxify overloaded As, which enables it to accumulate extremely high As concentrations in its fronds. This review also provides insights into several important research gaps that need to be addressed to advance our understanding of As hyperaccumulation in P. vittata from the perspective of the benefits of As.

As-hyperaccumulator Pteris vittata and non-hyperaccumulator Pteris ensiformis under low As-exposure: Transcriptome analysis and implication for As hyperaccumulation.

Soil contamination by arsenic (As) poses potential health risks to humans. As-hyperaccumulator P. vittata has been used in As-contaminated soils for phytoremediation. Clarifying the mechanisms of its As-hyperaccumulation is critical to enhance its efficiency in phytoremediation. Here, based on transcriptome analysis, we determined the concentration-dependent patterns of As-related gene families by comparing As-hyperaccumulator P. vittata and non-hyperaccumulator P. ensiformis after exposing to 20 µM arsenate (AsV). As expected, arsenic induced more stress in P. ensiformis than P. vittata. Based on gene ontology, differences in transporter activity are probably responsible for their differential As accumulation. Though As exposure induced expression of phosphate transporter PvPht1;4 for AsV absorption in both plants, stronger AsV reduction, AsIII transport, and AsIII-GSH complexation were found in P. ensiformis roots. Unlike P. ensiformis, As metabolism processes occurred mainly in P. vittata fronds. Notably, tonoplast-localized ACR3s were only present in P. vittata, making it more effective in sequestrating AsIII into frond vacuoles. Further, vesicle As transformation via PvGAPC1 (glyceraldehyde 3-phosphate dehydrogenase), PvOCT4 (organic cation transporter 4), and PvGSTF1 (glutathione S-transferase) contributed little to As-hyperaccumulation. This study provides information on critical genes responsible for As-hyperaccumulation by P. vittata, which can be applied to construct As-hyperaccumulating plants by genetic engineering to enhance their phytoremediation efficiency in As-contaminated soils.

Metal uptake and translocation by Chinese brake fern (Pteris vittata) and diversity of rhizosphere microbial communities under single and combined arsenic and cadmium stress.

Chinese brake fern (Pteris vittata) can increase tolerance to arsenic (As) and cadmium (Cd) toxicity by regulating rhizosphere microbial diversity. However, effects of combined As-Cd stress on microbial diversity and plant uptake and transport remain poorly understood. Therefore, effects of different concentrations of As and Cd on Pteris vittata (P. vittata) metal uptake and translocation and rhizosphere microbial diversity were examined in a pot experiment. The results indicated that As primarily accumulated aboveground in P. vittata (bioconcentration factor (BCF) ≤ 51.3; translocation factor (TF) ≈ 4), whereas Cd primarily accumulated belowground (BCF ≤ 39.1; TF < 1). Under single As, single Cd, and As-Cd combined stress, the most dominant bacteria and fungi were Burkholderia-Caballeronia-P (6.62-27.92%) and Boeremia (4.61-30.42%), Massilia (8.07-11.51%) and Trichoderma (4.47-22.20%), and Bradyrhizobium (2.24-10.38%) and Boeremia (3.16-45.69%), respectively, and their abundance ratios had a significant impact on the efficiency of P. vittata for As and Cd accumulation. However, with increasing As and Cd concentrations, abundances of plant pathogenic bacteria such as Fusarium and Chaetomium (the highest abundances were 18.08% and 23.72%, respectively) increased, indicating that As and Cd concentrations reduced P. vittata resistance to pathogens. At high soil concentrations of As-Cd, although plant As and Cd contents increased and microbial diversity was highest, enrichment efficiency and transportability of As and Cd decreased substantially. Therefore, pollution intensity should be considered when evaluating P. vittata suitability for phytoremediation of combined As-Cd contaminated soils.

Arsenic (As) oxidation by core endosphere microbiome mediates As speciation in Pteris vittata roots.

Pteris vittata is an arsenic(As)-hyperaccumulator that may be employed in phytoremediation of As-contaminated soils. P. vittata-associated microbiome are adapted to elevated As and may be important for host survival under stresses. Although P. vittata root endophytes could be critical for As biotransformation in planta, their compositions and metabolisms remain elusive. The current study aims to characterize the root endophytic community composition and As-metabolizing potentials in P. vittata. High As(III) oxidase gene abundances and rapid As(III) oxidation activity indicated that As(III) oxidation was the dominant microbial As-biotransformation processes compared to As reduction and methylization in P. vittata roots. Members of Rhizobiales were the core microbiome and the dominant As(III) oxidizers in P. vittata roots. Acquasition of As-metabolising genes, including both As(III) oxidase and As(V) detoxification reductase genes, through horizontal gene transfer was identified in a Saccharimonadaceae genomic assembly, which was another abundant population residing in P. vittata roots. Acquisition of these genes might improve the fitness of Saccharimonadaceae population to elevated As concentrations in P. vittata. Diverse plant growth promoting traits were encoded by the core root microbiome populations Rhizobiales. We propose that microbial As(III) oxidation and plant growth promotion are critical traits for P. vittata survival in hostile As-contaiminated sites.

Cytotoxic n-Hexane Fraction of the Egyptian Pteris vittata Functions as Anti-breast Cancer Through Coordinated Actions on Apoptotic and Autophagic Pathways.

We investigated the possible anticancer mechanisms of Pteris vittata [PV] n-hexane extract on MCF-7 [breast cancer cell line]. Cultured cell lines were treated with various concentrations of this extract ± Baf-A1 [autophagic inhibitor]. Cells' viability, apoptotic markers [caspase-7, Bax, and Bcl-2], autophagic markers [light chain 3 [LC-3] and P62/SQSTM1]], and the tumor suppressor P53 and its mRNA were checked by their corresponding methods. Treated cell lines showed significant concentration and time-dependent reductions in cell viability in response to PV-n-hexane extract and also exhibited a concomitant induction of apoptosis [increased chromatin condensation, nuclear fragmentation, and pro-apoptotic Bax, and cleaved caspase-7 levels while decreased Bcl-2 levels] and autophagy [increased autophagosomes vacuoles, and LC3B II levels while decreased P62/SQSTM1 levels]. Moreover, PV-n-hexane extract-treated cells showed significant increases in the P53 and its mRNA levels. The addition of Baf-A1 reversed the PV-n-hexane extract autophagic effects and increased apoptotic cell percentage with a much increase in the cleaved caspase-7 and P53 protein and its mRNA levels. We concluded that the PV-n-hexane extract exhibits cytotoxic effects on the MCF-7 cell line with significant reductions in cell viability and concomitant autophagy and apoptosis induction. Inhibition of autophagy in the PV-treated MCF-7 cells enhances apoptosis via a p35-dependent pathway.

Pteris laeta Wall. and Its New Phytochemical, Pterosinsade A, Promote Hippocampal Neurogenesis via Activating the Wnt Signaling Pathway.

Pteris laeta Wall., as a traditional tea, is popular in Southwest China, but its role in preventing cognitive impairment is unclear. In this study, Pteris laeta Wall. extracts (PW) and its active compounds were evaluated for preventive effects on Alzheimer's disease (AD) in vivo and in vitro. The results showed that PW diminished oxidative stress damage and apoptosis of Aß-induced HT22 cells and also rescued cognitive deficits, and ameliorated pathological injury and inflammatory response in APP/PS1 mice. Besides, a new pterosin sesquiterpene, named pterosinsade A (PA), and nine known compounds were discovered from the EtOAc extract that possessed the best neuroprotective activity. PA reduced apoptosis of APP-overexpressing neural stem cells and promoted their proliferation and neuronal differentiation. Meanwhile, PW and PA promoted hippocampal neurogenesis, which proved to be associated with activating the Wnt signaling pathway. These findings suggest that PW and PA are candidates for AD prevention.

Pterosin sesquiterpenoids from Pteris laeta Wall. ex Ettingsh. protect cells from glutamate excitotoxicity by modulating mitochondrial signals.

ETHNOPHARMACOLOGICAL RELEVANCE: The genus Pteris (Pteridaceae) has been used as a traditional herb for a long time. In particular, Pteris laeta Wall. ex Ettingsh. has been widely used in traditional Chinese medicine to treat nervous system diseases and some pterosin sesquiterpenes from Pteris show neuroprotective activity, but their underlying molecular mechanisms remain elusive. Therefore, to investigate the neuroprotective activity and working mechanism of pterosin sesquiterpenes from P. laeta Wall. ex Ettingsh. will provide a better understanding and guidance in using P. laeta Wall. ex Ettingsh. as a traditional Chinese medicine. AIM OF THE STUDY: We aim to develop effective treatments for neurodegenerative diseases from pterosin sesquiterpenes by evaluating their neuroprotective activity and investigating their working mechanisms. MATERIALS AND METHODS: Primary screening on the glutamate-induced excitotoxicity cell model was assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay. Fluorescent-activated cell sorting (FACS) was used to analyze the activation level of glutamate receptors and mitochondria membrane potential after treatment. Transcriptomics and proteomics analysis was performed to identify possible targets of pterosin B. The key pathways were enriched by the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis through the Database for Annotation, Visualization, and Integrated Discovery (DAVID). The core targets were visualized by a protein-protein interaction network using STRING. The mRNA and protein expressions were evaluated using real-time quantitative polymerase chain reaction (Q-PCR) and western blot, respectively. Immunocytochemistry was performed to monitor mitochondrial and apoptotic proteins. Cellular reactive oxygen species (ROS) were measured by ROS assay, and Ca2+ was stained with Fluo-4 AM to quantify intracellular Ca2+ levels. RESULTS: We found pterosin B from Pteris laeta Wall. ex Ettingsh. showed significant neuroprotective activity against glutamate excitotoxicity, enhancing cell viability from 43.8% to 105% (p-value: <0.0001). We demonstrated that pterosin B worked on the downstream signaling pathways of glutamate excitotoxicity rather than directly blocking the activation of glutamate receptors. Pterosin B restored mitochondria membrane potentials, alleviated intracellular calcium overload from 107.4% to 95.47% (p-value: 0.0006), eliminated cellular ROS by 36.55% (p-value: 0.0143), and partially secured cells from LPS-induced inflammation by increasing cell survival from 46.75% to 58.5% (p-value: 0.0114). Notably, pterosin B enhanced the expression of nuclear factor-erythroid factor 2-related factor 2 (NRF2) and heme oxygenase-1 (HO-1) by 2.86-fold (p-value: 0.0006) and 4.24-fold (p-value: 0.0012), and down-regulated Kelch-like ECH-associated protein 1 (KEAP1) expression by 2.5-fold (p-value: 0.0107), indicating that it possibly promotes mitochondrial biogenesis and mitophagy to maintain mitochondria quality control and homeostasis, and ultimately inhibits apoptotic cell death. CONCLUSIONS: Our work revealed that pterosin B protected cells from glutamate excitotoxicity by targeting the downstream mitochondrial signals, making it a valuable candidate for developing potential therapeutic agents in treating neurodegenerative diseases.

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.

Comparative Analysis of Arsenic Transport and Tolerance Mechanisms: Evolution from Prokaryote to Higher Plants.

Arsenic (As) is a toxic metalloid for all living organisms and can cause serious harm to humans. Arsenic is also toxic to plants. To alleviate As toxicity, all living organisms (from prokaryotes to higher plants) have evolved comprehensive mechanisms to reduce cytosolic As concentration through the set of As transporters localized at the plasma and tonoplast membranes, which operate either in arsenite As(III) extrusion out of cells (via ArsB, ACR3, and aquaporins) or by sequestering arsenic into vacuoles (by ABC transporters). In addition, a special arsenate resistance mechanism found in some bacterial systems has evolved in an As hyperaccumulating fern Pteris vittata, which involves transforming arsenate As(V) to an As(V) phosphoglycerate derivative by a glyceraldehyde 3-phosphate dehydrogenase and transporting this complex by an efflux transporter. In the present review, we summarize the evolution of these arsenic resistance mechanisms from prokaryotes to eukaryotes and discuss future approaches that could be utilized to better understand and improve As resistance mechanisms in plants.

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.

Luteolin-7-O-rutinoside from Pteris cretica L. var. nervosa attenuates LPS/D-gal-induced acute liver injury by inhibiting PI3K/AKT/AMPK/NF-κB signaling pathway.

Pteris cretica L. var. nervosa is one of the most well-known Chinese medicines. Although it is widely used to treat jaundice hepatitis, the main ingredient for its treatment was not thoroughly explored until recently. Essentially, the purpose of this study is to find the monomer compound in Pteris cretica L. var. nervosa, which is most likely to be effective in treating liver injury. Through the model of LPS/D-gal-induced liver injury in mice, the best therapeutic site of the total extract was explored, the chemical components of the parts with the best therapeutic effect were separated, a total of 10 flavonoids were isolated, and the RAW264.7 cells induced by LPS were used as the experimental model to explore the preliminary anti-inflammatory activity of NO production in vitro. Finally, the anti-inflammatory activity and the highest content in this plant Luteolin-7-O-rutinoside (LUT) were selected, as the object of study in vivo. It was found that LUT could not only reduce alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, but also significantly reduce the release of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß), and inhibit PI3K/AKT/AMPK/NF-κB pathway. In addition, LUT can increase levels of SOD and GSH to reduce oxidative stress. It has an obvious therapeutic effect on acute liver injury induced by LPS/D-gal in mice. Therefore, infer LUT is a functional substance in Pteris cretica L. var. nervosa.

Arsenic accumulation in Pteris vittata: Time course, distribution, and arsenic-related gene expression in fronds and whole plantlets.

In this work, arsenic (As) accumulation and distribution over time in Pteris vittata young fronds from adult plants and in whole plantlets, grown on a highly contaminated As-soil, was determined by µ-XRF. A linear increase in As content up to 60 days was found in young fronds at different times, and a progressive distribution from the apex to the base of the fronds was observed. In whole plantlets, As signal was detectable from 9 to 20 days in the apex of a few fronds and fiddleheads. Later, up to 60 days, As was localized in all fronds, in the rhizome and in basal part of the roots. The dynamics of expression of As-related genes revealed a good correlation between As content and the level of the As (III)-antiporter PvACR3 transcript in plantlets roots and fronds and in young fronds. Moreover, the transcription of As (V)-related gametophytic genes PvGAPC1, PvOCT4 increases over time during As accumulation while PvGSTF1 is expressed only in roots. Here, we demonstrate the suitability of the µ-XRF technique to monitor As accumulation, which allowed us to propose that As is initially directly transported to fiddleheads and apex of fronds, is later distributed to the whole fronds and simultaneously accumulated in the rhizome and roots. We also provide indications on the expression of candidate genes possibly involved in As (hyper)accumulation.

Ionomics and metabolomics analysis reveal the molecular mechanism of metal tolerance of Pteris vittata L. dominating in a mining site in Thai Nguyen province, Vietnam.

This study aims to find the interaction between ionome and metabolome profiles of Pteris vittata L., an arsenic hyperaccumulator plant, to reveal its metal tolerance mechanism. Therefore, at the Pb-Zn mining sites located in Thai Nguyen province, Vietnam, where these species dominate, soil and plant samples were collected. Their multi-element compositions were analyzed using inductively coupled plasma mass spectrometry (ICP-MS) and thus referred to as the "ionomics" approach. In parallel, the widely targeted metabolomics profiles of these plant samples were performed using liquid chromatography-tandem mass spectrometry (UPLC-QqQ-MS). Nineteen elements, including both metals and nonmetals, were detected and quantified in both tissues of thirty-five plant individuals. A comparison of these elements' levels in two tissues showed that above-ground parts accumulated more As and inorganic P, whereas Zn, Pb, and Sb were raised mostly in the under-ground samples. The partial least squares regression (PLSR) model predicting the level of each element by the whole metabolome indicated that the enhancement of flavonoids content plays an essential contribution in adaptation with the higher levels of Pb, Ag, and Ni accumulated in the aerial part, and Mn, Pb in subterranean part. Moreover, the models also highlighted the effect of Mn and Pb on the metabolic induction of adenosine derivatives in subterranean parts. At the same time, the model presented the most contribution of As to the metabolisms of the amino acids of this tissue. On those accounts, the developed integration approach linking the ionomics and metabolomics data of P. vittata improved the understanding of the molecular mechanism of hyperaccumulation characteristics and provided markers that could be targeted in future investigations.

Uptake, speciation and detoxification of antimonate and antimonite in As-hyperaccumulator Pteris Cretica L.

Antimony (Sb) and arsenic (As) are chemical analogs, but their behaviors in plants are different. To investigate the Sb uptake, translocation and speciation in As-hyperaccumulator P. cretica, a hydroponic experiment was conducted. In this study, P. cretica was exposed to 0.2-strength Hoagland nutrient solution, which contained 0.5 or 5 mg/L antimonite (SbIII) or antimonate (SbV). After 14 d exposure, P. cretica took up 1.4-2.8 times more SbIII than SbV. Since P. cretica was unable to translocate Sb, its roots accumulated >97% Sb with the highest at 7965 mg/kg. In both SbIII and SbV treatments, SbIII was the predominant species in P. cretica, with 90-100% and 46-100% SbIII in the roots. As the first barrier against Sb to enter plant cells, more Sb was accumulated in cell wall than cytosol or organelles. The results suggest that P. cretica may detoxify Sb by reducing SbV to SbIII and immobilizing it in root cell walls. Besides, the presence of SbIII significantly reduced the concentrations of dissolved organic C including organic acids in P. cretica root exudates. Further, increasing Sb levels promoted P accumulation in the plant, especially in the fronds, which may help P. cretica growth. The information from this study shed light on metabolic transformation of Sb in As-hyperaccumulators P. cretica, which helps to better understand Sb uptake and detoxification by plants.