RESUMEN
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.
Asunto(s)
Arsénico , Arsenitos , Glutatión , Hojas de la Planta , Pteris , Selenio , Arseniatos/metabolismo , Arsénico/metabolismo , Arsenitos/metabolismo , Biodegradación Ambiental , Glutatión/metabolismo , Malondialdehído/metabolismo , Hojas de la Planta/metabolismo , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/efectos de los fármacos , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Pteris/metabolismo , Pteris/crecimiento & desarrollo , Pteris/genética , Pteris/efectos de los fármacos , Selenio/metabolismo , Selenio/farmacología , Contaminantes del Suelo/metabolismoRESUMEN
Organic phosphorus (Po) is a large component of soil P, but it is often unavailable for plant uptake. Purple acid phosphatases (PAP) can hydrolyze a wide range of Po, playing an important role in Po utilization by plants. In this study, we investigated a novel secretary PvPAP1 from the As-hyperaccumulator Pteris vittata, which can effectively utilize exogenous Po, including adenosine triphosphate (ATP) and phytate. Unlike other PAP, PvPAP1 was abundantly-expressed in P. vittata roots, which was upregulated 3.5-folds under P-deprivation than P-sufficient conditions. When expressed in tobacco, its activity in the roots of PvPAP1-Ex lines was â¼8 folds greater than that in wild-type (WT) plants. Besides, PvPAP1 exhibited its secretory ability as evidenced by the sapphire-blue color on the root surface after treating with 5-bromo-4-chloro-3-indolyl phosphate. In a long-term experiment using sand media, PvPAP1-expressing tobacco plants showed 25-30 % greater root biomass than WT plants when using ATP as the sole P source. This is because PvPAP1-expression enhanced its phosphatase activity by 6.5-9.2 folds in transgenic tobacco, thereby increasing the P contents by 39-41 % in its roots under ATP treatment and 9.4-30 % under phytate treatment. The results highlight PvPAP1 as a novel secreted phosphatase crucial for external Po utilization in P. vittata, suggesting that PvPAP1 has the potential to serve as a valuable gene resource for enhancing Po utilization by crop plants.
Asunto(s)
Nicotiana , Fósforo , Ácido Fítico , Raíces de Plantas , Pteris , Ácido Fítico/metabolismo , Nicotiana/metabolismo , Nicotiana/genética , Nicotiana/crecimiento & desarrollo , Fósforo/metabolismo , Pteris/metabolismo , Pteris/genética , Pteris/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Raíces de Plantas/crecimiento & desarrollo , Hidrólisis , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Fosfatasa Ácida/metabolismo , Fosfatasa Ácida/genética , Arsénico/metabolismo , Regulación de la Expresión Génica de las PlantasRESUMEN
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.
Asunto(s)
Arsénico , Proteínas de Transporte de Fosfato , Fosfatos , Pteris , Pteris/metabolismo , Pteris/genética , Arsénico/metabolismo , Fosfatos/metabolismo , Proteínas de Transporte de Fosfato/metabolismo , Proteínas de Transporte de Fosfato/genética , Arabidopsis/metabolismo , Arabidopsis/genética , Contaminantes del Suelo/metabolismo , Transporte BiológicoRESUMEN
BACKGROUND AND AIMS: Niche differentiation is a crucial issue in speciation. Although it has a well-known role in adaptive processes of hybrid angiosperms, it is less understood in hybrid ferns. Here, we investigate whether an intermediate ecological niche of a fern hybrid is a novel adaptation that provides insights into fern hybrid speciation. METHODS: Pteris fauriei (Pteridaceae) is a natural hybrid fern, occurring in environments between its parent species. The maternal Pteris minor is found in sunny areas, but the habitat of the paternal Pteris latipinna is shady. We combined data from morphology, leaf anatomy and photosynthetic traits to explore adaptation and differentiation, along with measuring the environmental features of their niches. We also performed experiments in a common garden to understand ecological plasticity. KEY RESULTS: The hybrid P. fauriei was intermediate between the parent species in stomatal density, leaf anatomical features and photosynthetic characteristics in both natural habitats and a common garden. Interestingly, the maternal P. minor showed significant environmental plasticity and was more similar to the hybrid P. fauriei in the common garden, suggesting that the maternal species experiences stress in its natural habitats but thrives in environments similar to those of the hybrid. CONCLUSIONS: Based on the similar niche preferences of the hybrid and parents, we propose hybrid superiority. Our results indicate that the hybrid P. fauriei exhibits greater fitness and can compete with and occupy the initial niches of the maternal P. minor. Consequently, we suggest that the maternal P. minor has experienced a niche shift, elucidating the pattern of niche differentiation in this hybrid group. These findings offer a potential explanation for the frequent occurrence of hybridization in ferns and provide new insights into fern hybrid speciation, enhancing our understanding of fern diversity.
Asunto(s)
Ecosistema , Hibridación Genética , Fenotipo , Pteris , Pteris/fisiología , Pteris/anatomía & histología , Pteris/genética , Hojas de la Planta/anatomía & histología , Hojas de la Planta/fisiología , Hojas de la Planta/crecimiento & desarrollo , Especiación Genética , Fotosíntesis/fisiología , Helechos/fisiología , Helechos/anatomía & histología , Adaptación FisiológicaRESUMEN
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.
Asunto(s)
Arsénico , Arsenitos , Pteris , Cobre , Arsénico/toxicidad , Pteris/genética , Proteínas de Transporte de Membrana , Antiportadores , Expresión Génica , SueloRESUMEN
Pteris vittata, as the firstly discovered arsenic (As) hyperaccumulator, has great application value in As-contaminated soil remediation. Currently, the genes involved in As hyperaccumulation in P. vittata have been mined continuously, while they have not been used in practice to enhance phytoremediation efficiency. Aiming to better assist the practice of phytoremediation, this review collects 130 studies to clarify the progress in research into the As hyperaccumulation process in P. vittata from multiple perspectives. Antioxidant defense, rhizosphere activities, vacuolar sequestration, and As efflux are important physiological activities involved in As hyperaccumulation in P. vittata. Among related 19 genes, PHT, TIP, ACR3, ACR2 and HAC family genes play essential roles in arsenate (Asâ ¤) transport, arsenite (Asâ ¢) transport, vacuole sequestration of Asâ ¢, and the reduction of Asâ ¤ to Asâ ¢, respectively. Gene ontology enrichment analysis indicated it is necessary to further explore genes that can bind to related ions, with transport activity, or with function of transmembrane transport. Phylogeny analysis results implied ACR2, HAC and ACR3 family genes with rapid evolutionary rate may be the decisive factors for P. vittata as an As hyperaccumulator. A deeper understanding of the As hyperaccumulation network and key gene components could provide useful tools for further bio-engineered phytoremediation.
Asunto(s)
Arsénico , Pteris , Filogenia , Pteris/genética , Biología Molecular , Fenómenos Fisiológicos de las PlantasRESUMEN
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.
Asunto(s)
Arsénico , Arsenitos , Pteris , Selenio , Contaminantes del Suelo , Antiportadores/metabolismo , Antiportadores/farmacología , Arseniato Reductasas/genética , Arseniato Reductasas/metabolismo , Arseniatos , Arsénico/metabolismo , Arsenitos/metabolismo , Biodegradación Ambiental , Raíces de Plantas/metabolismo , Pteris/genética , Pteris/metabolismo , Ácido Selénico , Selenio/metabolismo , Suelo , Contaminantes del Suelo/metabolismoRESUMEN
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.
Asunto(s)
Arsénico , Pteris , Contaminantes del Suelo , Arsénico/análisis , Biodegradación Ambiental , Expresión Génica , Raíces de Plantas/metabolismo , Pteris/genética , Pteris/metabolismo , Contaminantes del Suelo/análisisRESUMEN
Arsenic (As) is a toxic semi-metallic element that is ubiquitous in the environment and poses serious human health risks. Phytoextraction by Pteris vittata is considered a low-cost and environmentally friendly approach to treat As-contaminated soil. P. vittata mainly absorbs arsenate thus the bioavailability of As to P. vittata depends on the chemical form of As. Microbial redox of As contributes to the biogeochemical cycling of As, and rhizobacterium-assisted phytoextraction by P. vittata was proposed. In this study, this microbe-assisted phytoextraction was applied to two fields, and the effectiveness of phytoextraction was evaluated. The results revealed that P. vittata was able to grow in temperate and subarctic climate zones. The biomass was influenced by the weather, and the As concentration in plants was dependent on the As content in the soil. The ratio of arsenite oxidase genes (aioA-like genes) to 16S rRNA genes was employed to evaluate the effect of As phytoextraction, and the results exhibited that the ratio was related to the As concentration in P. vittata. Our results showed that arsenite oxidation in the rhizosphere might not be achieved by single-strain inoculation, while this study provided empirical evidence that the rhizospheric aioA-like genes could be an indicator for evaluating the effectiveness of As phytoextraction.
Asunto(s)
Arsénico , Pteris , Contaminantes del Suelo , Arsénico/análisis , Biodegradación Ambiental , Oxidorreductasas/genética , Pteris/enzimología , Pteris/genética , ARN Ribosómico 16S , Contaminantes del Suelo/análisisRESUMEN
Pteris vittata is an arsenic (As) hyperaccumulator that can accumulate several thousand mg As kg-1 DW in aboveground biomass. A key factor for its hyperaccumulation ability is its highly efficient As long-distance translocation system. However, the underlying molecular mechanisms remain unknown. We isolated PvAsE1 through the full-length cDNA over-expression library of P. vittata and characterized it through a yeast system, RNAi gametophytes and sporophytes, subcellular-location and in situ hybridization. Phylogenomic analysis was conducted to estimate the appearance time of PvAsE1. PvAsE1 was a plasma membrane-oriented arsenite (AsIII) effluxer. The silencing of PvAsE1 reduced AsIII long-distance translocation in P. vittata sporophytes. PvAsE1 was structurally similar to solute carrier (SLC)13 proteins. Its transcripts could be observed in parenchyma cells surrounding the xylem of roots. The appearance time was estimated at c. 52.7 Ma. PvAsE1 was a previously uncharacterized SLC13-like AsIII effluxer, which may contribute to AsIII long-distance translocation via xylem loading. PvAsE1 appeared late in fern evolution and might be an adaptive subject to the selection pressure at the Cretaceaou-Paleogene boundary. The identification of PvAsE1 provides clues for revealing the special As hyperaccumulation characteristics of P. vittata.
Asunto(s)
Arsénico , Arsenitos , Helechos , Pteris , Contaminantes del Suelo , Arsénico/metabolismo , Arsenitos/metabolismo , Biodegradación Ambiental , Helechos/metabolismo , Raíces de Plantas/metabolismo , Pteris/genética , Contaminantes del Suelo/análisis , Contaminantes del Suelo/metabolismoRESUMEN
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.
Asunto(s)
6-Fitasa , Arsénico , Pteris , Contaminantes del Suelo , 6-Fitasa/genética , Arsénico/análisis , Biodegradación Ambiental , Hidrólisis , Ácido Fítico , Raíces de Plantas/química , Pteris/genética , Contaminantes del Suelo/análisisRESUMEN
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.
Asunto(s)
Arsénico , Pteris , Contaminantes del Suelo , Arsénico/toxicidad , Biodegradación Ambiental , Raíces de Plantas/genética , Pteris/genética , Nicotiana/genéticaRESUMEN
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.
Asunto(s)
Arsénico , Pteris , Contaminantes del Suelo , Arseniato Reductasas/genética , Arseniatos , Biodegradación Ambiental , Raíces de Plantas/química , Pteris/genética , Contaminantes del Suelo/análisisRESUMEN
Acute and chronic arsenic (As) toxicity is a global health issue affecting millions of people, which leads to inactivation of over 200 enzymes, particularly those involved in cellular energy pathways and DNA synthesis and repair. The fern Pteris vittata acts as a hyperaccumulator of As and may be useful for phytoremediation to reduce disposal risks by utilizing metal-enriched plant biomass in energy and metal recovery. However, these ferns grow in limited environments and its transplantation and transport can be challenging. Therefore, we generated a transgenic Arabidopsis plant as a seed plant model, capable of accumulating As in their vacuole lumen. This was achieved by transforming the As-resistant bacterial As transporter, ArsB, via fusion with a organelle-targeting signal to the vacuolar membrane, N-ethyl-maleimide-sensitive factor attachment protein receptors (SNAREs) protein, VAMP711. In this study, we developed the iVenus assay as a method for detecting whether the N- or C-terminus of a membrane protein is located on the cytoplasmic or exoplasmic side, and from the result of the iVenus assay, we generated the transgenic plant introduced N-terminal end of ArsB with VAMP711, localized to the central vacuolar membrane to accumulate As in the shoot and differentiation zone of root.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Arsénico/metabolismo , Biodegradación Ambiental , Proteínas SNARE/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Brotes de la Planta/genética , Brotes de la Planta/metabolismo , Pteris/genética , Pteris/metabolismo , Proteínas SNARE/genética , Vacuolas/metabolismoRESUMEN
Arsenic accumulation in soil is a global problem typically addressed using phytoremediation methods. Pteris vittata, a model arsenic hyperaccumulator, has great potential as a genetically engineered plant for phytoremediation. However, the lack of omic information on this species has severely limited the identification and application of its arsenic hyperaccumulation and regulation components. In this study, we used an optimized single-molecular real-time (SMRT) strategy to create a de novo full-length transcriptomic-tonoplast proteomic database for this unsequenced fern and to determine the genetic components underlying its arsenic hyperaccumulation-regulation mechanisms. We established a comprehensive network consisting of six major transporter families, two novel resistance pathways, and a regulatory system by examining alternative splicing (AS) and long non-coding RNA (lncRNA) in different tissues following As(III) and As(V) treatment. The database and network established in this study will deepen our understanding of the unique hyperaccumulation and regulation mechanisms of P. vittata, ultimately providing a valuable resource for futher research on phytoremediation of arsenic-contaminated soil.
Asunto(s)
Arsénico/toxicidad , Pteris/efectos de los fármacos , Contaminantes del Suelo/toxicidad , Biodegradación Ambiental , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Secuenciación de Nucleótidos de Alto Rendimiento , Proteínas de Transporte de Membrana/genética , Proteínas de Plantas/genética , Pteris/genética , Pteris/metabolismo , ARN Largo no Codificante , ARN de Planta , TranscriptomaRESUMEN
Hyperaccumulation of arsenic (As) by brake fern Pteris vittata has been described as an important genetic trait that provides an option for development of a sustainable phytoremediation process for As mitigation. Accumulation of very high concentration of arsenic in above-ground tissues may be the result of arsenic vacuole compartmentalization, but the mechanism(s) of arsenic uptake and transport by underground tissues are largely unknown. In this study, we made an attempt towards understanding the molecular mechanism of As hyperaccumulation in this plant. A time-dependent As accumulation study indicates an exponential accumulation of As from 7 to 30 days of arsenic exposure in fronds, and day 3-7 in roots. Root transcriptome analysis identified 554,973 transcripts. Further, subsets of 824 transcripts were differentially expressed between treated and control samples. Many of the genes of critical As-stress response, transcription factors and metal transporters, biosynthesis of chelating compounds involved in uptake and accumulation mechanisms were identified. The genes that were highly expressed such as cysteine-rich RLK, and ABC transporter G family member 26 needs further studies along with arsenite transmembrane transporter. The analysis of generated transcriptome dataset has provided valuable information and platform for further functional studies.
Asunto(s)
Arsénico/metabolismo , ATPasas Transportadoras de Arsenitos/genética , Proteínas de Plantas/genética , Raíces de Plantas/genética , Pteris/genética , Contaminantes del Suelo/metabolismo , Transcriptoma , Arsénico/aislamiento & purificación , ATPasas Transportadoras de Arsenitos/metabolismo , Biodegradación Ambiental , Transporte Biológico , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Ontología de Genes , Anotación de Secuencia Molecular , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Proteínas de Plantas/metabolismo , Raíces de Plantas/metabolismo , Pteris/metabolismo , Contaminantes del Suelo/aislamiento & purificaciónRESUMEN
Arsenic (As) is an important environmental and food-chain toxin. We investigated the key components controlling As accumulation and tolerance in Arabidopsis thaliana. We tested the effects of different combinations of gene knockout, including arsenate reductase (HAC1), γ-glutamyl-cysteine synthetase (γ-ECS), phytochelatin synthase (PCS1) and phosphate effluxer (PHO1), and the heterologous expression of the As-hyperaccumulator Pteris vittata arsenite efflux (PvACR3), on As tolerance, accumulation, translocation and speciation in A. thaliana. Heterologous expression of PvACR3 markedly increased As tolerance and root-to-shoot As translocation in A. thaliana, with PvACR3 being localized to the plasma membrane. Combining PvACR3 expression with HAC1 mutation led to As hyperaccumulation in the shoots, whereas combining HAC1 and PHO1 mutation decreased As accumulation. Mutants of γ-ECS and PCS1 were hypersensitive to As and had higher root-to-shoot As translocation. Combining γ-ECS or PCS1 with HAC1 mutation did not alter As tolerance or accumulation beyond the levels observed in the single mutants. PvACR3 and HAC1 have large effects on root-to-shoot As translocation. Arsenic hyperaccumulation can be engineered in A. thaliana by knocking out the HAC1 gene and expressing PvACR3. PvACR3 and HAC1 also affect As tolerance, but not to the extent of γ-ECS and PCS1.
Asunto(s)
Arabidopsis/genética , Arsénico/metabolismo , Proteínas de Plantas/metabolismo , Pteris/genética , Aminoaciltransferasas/genética , Aminoaciltransferasas/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arseniato Reductasas/genética , Arseniato Reductasas/metabolismo , Transporte Biológico , Técnicas de Inactivación de Genes , Mutación , Proteínas de Plantas/genética , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Brotes de la Planta/genética , Brotes de la Planta/metabolismoRESUMEN
A hydroponic experiment was conducted to investigate the effects of indole-3-acetic acid (IAA) on arsenic (As) uptake and antioxidative enzymes in fronds of Pteris cretica var. nervosa (As hyperaccumulator) and Pteris ensiformis (non-hyperaccumulator). Plants were exposed to 2 mg L-1 As(III), As(V) or dimethylarsinic acid (DMA) and IAA concentrations for 14 d. The biomass and total As in the plants significantly increased at 30 mg L-1 IAA. Superoxide dismutase (SOD) activities significantly increased with IAA addition. Catalase (CAT) activities showed a significant increase in P. ensiformis exposed to three As species at 30 or 50 mg L-1 IAA but varied in P. cretica var. nervosa. Peroxidase (POD) activities were unchanged in P. ensiformis except for a significant decrease at 50 mg L-1 IAA under As(III) treatment. However, a significant increase was observed in P. cretica var. nervosa at 10 mg L-1 IAA under As(III) or DMA treatment and at 50 mg L-1 IAA under As(V) treatment. Under DMA stress, malondialdehyde contents in fronds of P. cretica var. nervosa showed a significant decrease at 10 mg L-1 IAA but remained unchanged in P. ensiformis. Therefore, IAA enhanced As uptake and frond POD activity in P. cretica var. nervosa under As stress.
Asunto(s)
Arsénico/metabolismo , Ácidos Indolacéticos/farmacología , Reguladores del Crecimiento de las Plantas/farmacología , Pteris/efectos de los fármacos , Contaminantes del Suelo/metabolismo , Antioxidantes/metabolismo , Biomasa , Hidroponía , Ácidos Indolacéticos/administración & dosificación , Malondialdehído/metabolismo , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/enzimología , Hojas de la Planta/metabolismo , Proteínas de Plantas/metabolismo , Pteris/enzimología , Pteris/genética , Pteris/metabolismo , Especificidad de la EspecieRESUMEN
To date, the response of the fern gametophyte to its environment has received considerable attention. However, studies on the influence of plant invasion on the fern gametophyte are fewer. Allelopathy has been hypothesized to play an important role in biological invasion. Hence, it is necessary to study the allelopathy of invasive plant species to the fern gametophyte and elucidate the mechanisms by which invasive plants cause phytotoxicity. As one of the main invasive plants in China, Bidens pilosa exhibits allelopathic effects on the gametophytic growth of Pteris multifida. The root exudate plays an important role among various allelochemical delivery mechanisms in B. pilosa. The effect invasive plant species has on photosynthesis in native species is poorly understood. To elucidate this effect, the changes in photosynthesis in the gametophytes of P. multifida are analyzed to examine the mechanisms of the root exudates of B. pilosa. Meanwhile, a non-invasive plant, Coreopsis basalis, was also applied to investigate the effects on fluorescence and pigments in P. multifida gametophytes. We found that gametophytes exposed to both B. pilosa and C. basalis had decreased fluorescence parameters in comparison with the control, except for non-photochemical quenching. Furthermore, it was found that these parameters were markedly affected from day 2 to day 10 in the presence of both exudates at a concentration of 25% or above. B. pilosa exudate had a negative dose-dependent effect on chlorophyll a, chlorophyll b, carotenoid, and the total chlorophyll in the gametophyte. The inhibitory effects increased with increasing exudate concentrations of both species, exhibiting the greatest inhibition at day 10. In conclusion, B. pilosa irreversibly affected the photosynthesis of P. multifida on both PS I and PS II. Root exudates caused the primary damage with respect to the decrease of the acceptors and donors of photon and electron in photosynthetic units and the production and the relative yield of photochemical quantum in PS II. With the effects of exudates, part of the energy is released as heat in chloroplasts. The comparison of invasive and non-invasive plants in allelopathic experiments demonstrated that invasive plants were responsible for the critical damage to the photosynthetic process in local species.
Asunto(s)
Células Germinativas de las Plantas/metabolismo , Especies Introducidas , Fotosíntesis , Pteris/metabolismo , Transporte de Electrón , Pigmentos Biológicos/metabolismo , Pteris/genética , Espectrometría de FluorescenciaRESUMEN
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.