RESUMEN
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
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.
Asunto(s)
Arsénico , Pteris , Contaminantes del Suelo , Humanos , Arsénico/metabolismo , Pteris/metabolismo , Biodegradación Ambiental , Raíces de Plantas/metabolismo , Perfilación de la Expresión Génica , Suelo , Contaminantes del Suelo/metabolismoRESUMEN
Rare earth elements (REEs) are critical for numerous modern technologies, and demand is increasing globally; however, production steps are resource-intensive and environmentally damaging. Some plant species are able to hyperaccumulate REEs, and understanding the biology behind this phenomenon could play a pivotal role in developing more environmentally friendly REE recovery technologies. Here, we identified a REE transporter NRAMP REE Transporter 1 (NREET1) from the REE hyperaccumulator fern Dicranopteris linearis. Although NREET1 belongs to the natural resistance-associated macrophage protein (NRAMP) family, it shares a low similarity with other NRAMP members. When expressed in yeast, NREET1 exhibited REE transport capacity, but it could not transport divalent metals, such as zinc, nickel, manganese, or iron. NREET1 is mainly expressed in D. linearis roots and predominantly localized in the plasma membrane. Expression studies in Arabidopsis thaliana revealed that NREET1 functions as a transporter mediating REE uptake and transfer from root cell walls into the cytoplasm. Moreover, NREET1 has a higher affinity for transporting light REEs compared to heavy REEs, which is consistent to the preferential enrichment of light REEs in field-grown D. linearis. We therefore conclude that NREET1 may play an important role in the uptake and consequently hyperaccumulation of REEs in D. linearis. These findings lay the foundation for the use of synthetic biology techniques to design and produce sustainable, plant-based REE recovery systems.
Asunto(s)
Helechos , Proteínas de Transporte de Membrana , Metales de Tierras Raras , Membrana Celular , Helechos/metabolismo , Zinc/metabolismoRESUMEN
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.
Asunto(s)
Arsénico , Arsenitos , Micorrizas , Pteris , Contaminantes del Suelo , Arseniatos , Arsénico/metabolismo , Arsenitos/metabolismo , Biodegradación Ambiental , Humanos , Micorrizas/metabolismo , Proteínas de Transporte de Fosfato/metabolismo , Raíces de Plantas/metabolismo , Pteris/metabolismo , Suelo , Contaminantes del Suelo/metabolismo , SimbiosisRESUMEN
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
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.
Asunto(s)
Arabidopsis , Arsénico , Pteris , Contaminantes del Suelo , Arabidopsis/genética , Arabidopsis/metabolismo , Arsénico/análisis , Cloroplastos/química , Cloroplastos/metabolismo , Proteínas de Transporte de Fosfato/genética , Raíces de Plantas/metabolismo , Pteris/metabolismo , Contaminantes del Suelo/análisisRESUMEN
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
Arsenic-hyperaccumulator Pteris vittata is efficient in As uptake, probably through phosphate transporters (Pht). Here, for the first time, we cloned a new PvPht1;4 gene from P. vittata and investigated its role in arsenate (AsV) uptake and transport in yeast and transgenic tobacco plants. On the basis of quantitative real-time polymerase chain reaction (qRT-PCR), PvPht1;4 was abundantly expressed in P. vittata fronds and roots, with its transcripts in the roots being induced by both P deficiency and As exposure. PvPht1;4 was localized to the plasma membrane, which complemented a yeast-mutant defective in P uptake and showed higher P transport affinity than PvPht1;3. Under AsV exposure, PvPht1;4 yeast transformants showed comparable tolerance as PvPht1;3, but higher As accumulation than PvPht1;2 transformants, indicating that PvPht1;4 had considerable AsV and P transport activity. However, in soil and hydroponic experiments, PvPht1;4 expressing tobacco lines accumulated 26-44 and 37-55% lower As in the shoots than wild type plants, with lower root-to-shoot As translocation. In the roots of PvPht1;4 lines, higher glutathione (GSH) contents and expression levels of GSH synthetase gene NtGSH2 were observed. In addition, the transcripts of AsIII-GSH transporter NtABCC1 in PvPht1;4 lines were upregulated. The data suggested that PvPht1;4 lines probably detoxified As by reducing AsV to AsIII, which was then complexed with GSH and stored in the root vacuoles, thereby reducing As translocation in transgenic tobacco. Given its strong AsV transport capacity, expression of PvPht1;4 provides a new molecular approach to reduce As accumulation in plant shoots.
Asunto(s)
Arsénico , Pteris , Contaminantes del Suelo , Biodegradación Ambiental , Proteínas de Transporte de Fosfato , Raíces de Plantas , NicotianaRESUMEN
Arsenic-hyperaccumulator Pteris vittata is efficient in As accumulation and has been used in phytoremediation of As-contaminated soils. Arsenate (AsV) is the predominant As species in aerobic soils and is taken up by plants via phosphate transporters (Pht) including P. vittata. In this work, we cloned the PvPht1;3 full length coding sequence from P. vittata and investigated its role in As accumulation by yeast and plants. PvPht1;3 complemented a yeast P uptake mutant strain and showed a stronger affinity and transport capacity to AsV than PvPht1;2. In transgenic tobacco, PvPht1;3 enhanced AsV absorption and translocation, increasing As accumulation in the shoots under both hydroponic and soil experiments. On the basis of the expression patterns via qRT-PCR, PvPht1;3 was strongly induced by P deficiency but not As exposure. To further understand its expression pattern, transgenic Arabidopsis thaliana and soybean expressing the GUS reporter gene, driven by PvPht1;3 promoter, were produced. The GUS staining showed that the reporter gene was mainly expressed in the stele cells, indicating that PvPht1;3 was expressed in stele cells and was likely involved in P/As translocation. Taken together, the data suggested that PvPht1;3 was a high-affinity AsV transporter and was probably responsible for efficient As translocation in P. vittata. Our results suggest that expressing PvPht1;3 enhances As translocation and accumulation in plants, thereby improving phytoremediation of As-contaminated soils.
Asunto(s)
Arsénico , Pteris , Contaminantes del Suelo , Biodegradación Ambiental , Proteínas de Transporte de Fosfato , Raíces de Plantas , NicotianaRESUMEN
While phosphate (P) inhibits arsenic (As) uptake by plants, phytate increases As uptake by As-hyperaccumulator Pteris vittata. Here we tried to understand the underling mechanisms by investigating the roles of phytate in soil As desorption, P transport in P. vittata, short-term As uptake, and plant growth and As accumulation from soils. Sterile soil was used to exclude microbial degradation on phytate. Results showed that inorganic P released 3.3-fold more As than that of phytate from soil. However, P. vittata accumulated 2-2.5 fold more As from soils with phytate than that in control and P treatment. In addition, different from P suppression on As uptake, solution uptake experiment showed that As uptake in phytate treatment was comparable to that of control under 0.1-7.5⯵M As after 1-24â¯h. Moreover, responding to phytate, P. vittata P transporter PvPht1;3 increased by 3-fold while PvPht1;1 decreased by 65%. The data suggested that phytate upregulated PvPht1;3, thereby contributing to As uptake in P. vittata. Our results showed that, though with lower As release from soil compared to P, phytate induced more As uptake and better growth in P. vittata by upregulating P transporters.
Asunto(s)
Arsénico/metabolismo , Fósforo/metabolismo , Ácido Fítico/metabolismo , Pteris/metabolismo , Contaminantes del Suelo/metabolismo , Arsénico/análisis , Biodegradación Ambiental , Fosfatos/metabolismo , Desarrollo de la Planta , Raíces de Plantas/metabolismo , Suelo , Contaminantes del Suelo/análisisRESUMEN
It is known that arsenic (As) promotes growth of As-hyperaccumulator Pteris vittata (PV), however, the associated mechanisms are unclear. Here we examined As-induced nutrient uptake in P. vittata and their potential role to enhance plant growth in sterile agar by excluding microbial effects. As-hyperaccumulator P. multifida (PM) and non-hyperaccumulator P. ensiformis (PE) belonging to the Pteris genus were used as comparisons. The results showed that, after 40â¯d of growth, As induced biomass increase in hyperaccumulators PV and PM by 5.2-9.4 fold whereas it caused 63% decline in PE. The data suggested that As played a beneficial role in promoting hyperaccumulator growth. In addition, hyperaccumulators PV and PM accumulated 7.5-13, 1.4-3.6, and 1.8-4.4 fold more As, Fe, and P than the non-hyperaccumulator PE. In addition, nutrient contents such as K and Zn were also increased while Ca, Mg, and Mn decreased or unaffected under As treatment. This study demonstrated that As promoted growth in hyperaccumulators and enhanced Fe, P, K, and Zn uptake. Different plant growth responses to As among hyperaccumulators PV and PM and non-hyperaccumulator PE may help to better understand why hyperaccumulators grow better under As-stress.