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

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

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.

Directional Electron Flow in a Selenoviologen-Based Tetracationic Cyclophane for Enhanced Visible-Light-Driven Hydrogen Evolution.

Directional electron flow in the photocatalyst enables efficient charge separation, which is essential for efficient photocatalysis of H2 production. Here, we report a novel class of tetracationic cyclophanes (7) incorporating bipyridine Pt(II) and selenoviologen. X-ray single-crystal structures reveal that 7 not only fixes the distances and spatial positions between its individual units but also exhibits a box-like rigid electron-deficient cavity. Moreover, host-guest recognition phenomena are observed between 7 and ferrocene, forming host-guest complexes with a 1:1 stoichiometry in MeCN. 7 exhibits good redox properties, narrow energy gaps, and strong absorption in the visible range (370-500 nm) due to containing two selenoviologen (SeV2+) units. Meanwhile, the femtosecond transient absorption (fs-TA) reveals that 7 has stabilized dicationic biradical, efficient charge separation, and facilitates directional electron flow to achieve efficient electron transfer due to the formation of rigid cyclophane and electronic architecture. Then, 7 is applied to visible-light-driven hydrogen evolution with high hydrogen production (132 µmol), generation rate (11 µmol/h), turnover number (221), and apparent quantum yield (1.7%), which provides a simplified and efficient photocatalytic strategy for solar energy conversion.

Commercializable Naphthalene Diimide Anolytes for Neutral Aqueous Organic Redox Flow Batteries.

Neutral aqueous organic redox flow batteries (AORFBs) hold the potential to facilitate the transition of renewable energy sources from auxiliary to primary energy, the commercial production of anolyte materials still suffers from insufficient performance of high-concentration and the high cost of the preparation problem. To overcome these challenges, this study provides a hydrothermal synthesis methodology and introduces the charged functional groups into hydrophobic naphthalene diimide cores, and prepares a series of high-performance naphthalene diimide anolytes. Under the synergistic effect of π-π stacking and H-bonding networks, the naphthalene diimide exhibits excellent structural stability and the highest water solubility (1.85 M for dex-NDI) reported to date. By employing the hydrothermal method, low-cost naphthalene diimides are successfully synthesized on a hundred-gram scale of $0.16 g-1 ($2.43 Ah-1), which is also the lowest price reported to date. The constructed full battery achieves a high electron concentration of 2.4 M, a high capacity of 54.4 Ah L-1, and a power density of 318 mW cm-2 with no significant capacity decay observed during long-duration cycling. These findings provide crucial support for the commercialization of AORFBs and pave the way for revolutionary developments in neutral AORFBs.

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.

Transdural circumferential decompression for thoracic spinal stenosis caused by beak-type ossification of the posterior longitudinal ligament: a technical note.

PURPOSE: Thoracic myelopathy caused by ossification of the posterior longitudinal ligament (OPLL) in the thoracic spine is usually progressive and responds poorly to conservative therapy, making surgery the only effective treatment option. A variety of surgical procedures have been developed to treat thoracic OPLL. However, the optimal surgical approach for removal of thoracic OPLL remains unclear. In the present study, we described a newly modified posterior approach for the removal of OPLL: circular decompression via dural approach, and complete removal of OPLL can be achieved under direct vision and without neurological deficit. MATERIALS AND METHODS: Three patients with beak-type thoracic OPLL presented with progressive thoracic myelopathy and leg weakness. Magnetic resonance imaging showed the spinal cord severely compressed. The surgical management of the three patients involved the 'cave-in' circular decompression and transdural resection of OPLL. RESULTS: Transdural circumferential decompression was successfully performed in all three patients. Clinical outcome measures, including pre- and postoperative radiographic parameters, were assessed. All of the patients were followed up for an average of 12 months (ranging from 10 to 15 months), and no surgery-related complications occurred. Weakness relief and neural function recovery were satisfactorily achieved in all patients by the final follow-up. CONCLUSIONS: Transdural circumferential decompression was an effective method for thoracic spinal stenosis caused by concurrent beak-type OPLL, by which OPLL could be safely removed. It is especially useful when there is a severe adhesion between the dura OPLL.

Anti-Sandwich Structured Photo-Electronic Wound Dressing for Highly Efficient Bacterial Infection Therapy.

Photo-electronic devices based on reactive oxygen species (ROS) generation suffer a crucial limitation in wound treatment due to their sandwich structure, which prevents the contact of ROS with wound tissue. In this work, the first anti-sandwich structured visible-light/electricity dual-responsive wound dressing is constructed for treatment of methicillin-resistant Staphylococcus aureus (MRSA), based on selenoviologen-appendant polythiophene (SeV2+ -PT)-containing polyacrylamide hydrogels. The new wound dressing is named an anti-sandwich structured photo-electronic wound dressing (PEWD). The unique structure of PEWD enables its use in synergistic electrodynamic and photodynamic therapy (EDT and PDT), providing rapid, on-demand, and sustained generation of ROS in situ via short-time light irradiation and/or wireless-controlled electrification. The PEWD possesses good flexibility, excellent biocompatibility, and fast response, as well as sustained ROS generation in a physiological environment. Animal experiments demonstrate effective ROS generation in 6 s under irradiation and electrification, inhibiting infection at an early stage, and substantially shortening the healing time of bacterial infection (to within 7 days). This proof-of-concept research holds great promise in developing new flexible PEWD, and novel strategies to improve wound treatment.

Application of open-door laminoplasty with ARCH plate fixation in cervical intraspinal tumors.

BACKGROUND: The open-door laminoplasty is an effective procedure for the treatment of cervical spondylotic myelopathy. However, little information is available about the surgical results of open-door laminoplasty in the treatment of intraspinal tumors. In the present study, we aimed to investigate the clinical effect of open-door laminoplasty with ARCH plate fixation in the treatment of cervical intraspinal tumors. METHODS: This was a retrospective study. From January 2013 to May 2018, 38 patients (13 males and 25 females, the average age of 44 ± 17 years) with cervical intraspinal tumors underwent open-door laminoplasty with ARCH plate fixation in our hospital. The operation time, blood loss, pre- and postoperative visual analog scale (VAS), and Japanese Orthopedic Association (JOA) scores were determined. To determine the radiographic outcomes, cervical X-ray film and magnetic resonance imaging (MRI) were performed before and after the operation, and cervical X-ray sagittal film was used to measure Cobb angle. The clinical data before and after the operation were compared by t-test. RESULTS: A total of 38 patients underwent a successful operation and demonstrated primary healing. The average operation time was 113 ± 12 min. The average blood loss was 120 ± 19 mL. All patients were followed up for 26.1 ± 2.8 months, and the final follow-up time was more than 24 months. VAS scores were much better at 24 months after operation compared with those before the operation, which were decreased from 6.1 ± 1.1 to 1.4 ± 0.7 (t = 32.63, P < 0.01). The JOA score was improved from 9.9 ± 1.5 to 15.5 ± 0.6 (t = - 18.36, P < 0.01), and the mean JOA recovery rate was 79% ± 11% at 24 months after the operation. There was no significant difference in Cobb angle between pre-operation and 24 months after the operation, which was 9.8 ± 2.6 and 10.3 ± 3.1 respectively (t = - 0.61, P > 0.05). Neither spinal malalignment on the coronal plane nor displacement of the laminoplasty flap was observed on postoperative cervical X-ray and MRI examinations at the final follow-up. CONCLUSIONS: Open-door laminoplasty with ARCH plate fixation was a safe and effective surgical approach for the treatment of cervical intraspinal tumors.

Culture-negative versus culture-positive in pyogenic spondylitis and analysis of risk factors for relapse.

OBJECTIVES: This study aims to compare and analyze the clinical features, diagnosis, treatment and prognosis of culture-negative and culture-positive primary pyogenic spondylitis. METHODS: In a retrospective analysis, 202 cases of adult primary pyogenic spondylitis with complete clinical data in our hospital from January 2013 to January 2020 were divided into two groups according to bacterial culture results: culture negative (n = 126) and culture positive (n = 76). We compare the clinical characteristics, diagnosis, treatment and prognosis of patients with different culture results. RESULTS: The culture positive rate was 37.62% (76/202). There were no significant differences in age, gender, affected segment, spinal abscess, diabetes mellitus, course of disease, surgery, recurrence, and follow-up time between the two groups (p>.05). There were statistically significant differences in hospital admission erythrocyte sedimentation rate (ESR), admission C-reactive protein (CRP), admission white blood cell (WBC) count, discharge ESR, discharge CRP, ESR decline rate, CRP (p<.05). There were statistically significant differences in the rate of decline, hospitalization days, and body temperature ≥38 °C (p<.05). Higher CRP levels on admission, antibiotic treatment time <6 weeks, and body temperature ≥ 38 °C are independent risk factors for infection recurrence. CONCLUSIONS: The culture-negative group's admission WBC, admission ESR, admission CRP, discharge ESR, discharge CRP, ESR decline rate, CRP decline rate, and hospital stay were lower than the culture positive group, the difference was statistically significant (p<.05). The independent risk factors for infection recurrence are higher CRP levels in hospital admission, antibiotic treatment time <6 weeks, and body temperature ≥ 38 °C.

Responses of microbial communities and metabolic activities in the rhizosphere during phytoremediation of Cd-contaminated soil.

Phytoremediation is an effective way to repair heavy metal contaminated soil and rhizosphere microorganisms play an important role in plant regulation. Nevertheless, little information is known about the variation of microbial metabolic activities and community structure in rhizosphere during phytoremediation. In this study, the rhizosphere soil microbial metabolic activities and community structure of Trifolium repensL. during Cd-contaminated soil phytoremediation, were analyzed by Biolog EcoPlate™ and high-throughput sequencing. The uptake in the roots of Trifolium repensL. grown in 5.68 and 24.23 mg/kg Cd contaminated soil was 33.51 and 84.69 mg/kg respectively, causing the acid-soluble Cd fractions decreased 7.3% and 5.4%. Phytoremediation significantly influenced microbial community and Trifolium repensL. planting significantly increased the rhizosphere microbial population, diversity, the relative abundance of plant growth promoting bacteria (Kaistobacter and Flavisolibacter), and the utilization of difficultly metabolized compounds. The correlation analysis among substrate utilization and microbial communities revealed that the relative abundance increased microorganisms possessed stronger carbon utilization capacity, which was beneficial to regulate the stability of plant-microbial system. Collectively, the results of this study provide fundamental insights into the microbial metabolic activities and community structure during heavy metal contaminated soil phytoremediation, which may aid in the bioregulation of phytoremediation.

RASGRP1 mutation in autoimmune lymphoproliferative syndrome-like disease.

BACKGROUND: Autoimmune lymphoproliferative syndrome (ALPS) is a genetic disorder of lymphocyte homeostasis due to impaired apoptosis. It was initially regarded as a very rare disease, but recent studies show that it may be more common than previously thought. Defects in a couple of genes have been identified in a proportion of patients with ALPS, but around one-third of such patients remain undefined genetically. OBJECTIVE: We describe 2 siblings presenting with ALPS-like disease. This study aimed to identify the genetic cause responsible for this phenotype. METHODS: Whole-exome sequencing and molecular and functional analyses were used to identify and characterize the genetic defect. Clinical and immunological analysis was also performed and reported. RESULTS: The 2 patients presented with chronic lymphadenopathy, hepatosplenomegaly, autoimmune hemolytic anemia, immune thrombocytopenia, and the presence of antinuclear autoantibody and other autoantibodies, but normal double-negative T cells. They also suffered from recurrent infections. Novel compound heterozygous mutations of RASGRP1 encoding Ras guanyl nucleotide releasing protein 1 were identified in the 2 siblings. The mutations impaired T-cell receptor signaling, leading to defective T-cell activation and proliferation, as well as impaired activation-induced cell death of T cells. CONCLUSIONS: This study shows for the first time that RASGRP1 mutation should be considered in patients with ALPS-like disease. We also propose to investigate the intracellular proteins involved in the T-cell receptor signaling pathway in similar patients but with unknown genetic cause.

Investigation on microbial community in remediation of lead-contaminated soil by Trifolium repensL.

Trifolium repensL. is a plant with strong adaptability and large biomass, which possess great potential for phytoremediation. However, little is known concerning its remediation effects and changes in rhizosphere microbial activity and community structure under heavy metal pressure. The aims of this study were to evaluate lead accumulation of Trifolium repensL., study microbial lead resistance, metabolism and community structure characteristics in rhizosphere soils. The accumulated Pb concentration of Trifolium repensL. was observed in 100 and 500 mg/kg Pb contained soil at 55.81 and 90.3 mg/kg, respectively, which cause the decrease of acid-soluble fractions in rhizosphere soil. In the progress of lead-contaminated soil phytoremediation by Trifolium repensL., Pb resistance and metabolic activities of microorganisms have been prompted gradually. In addition, the microbial community composition and abundance were investigated using Illumina sequencing and quantitative PCR. The result showed that after phytoremediation, beneficial microorganisms, such as Flavisolibacter, Kaistobacter, and Pseudomonas, increased, becoming the dominant genera. This study has provided insight into the distribution and activity of the microbial community.

3D printed polycaprolactone/ß-tricalcium phosphate/carbon nanotube composite - Physical properties and biocompatibility.

Three-dimensional (3D) printing is a bio-fabrication technique used to process tissue-engineered scaffolds for bone repair and remodeling. Polycaprolactone (PCL)/ß-tricalcium phosphate (TCP) has been used as a base and osteoconductive biomaterial for bone tissue engineering in the past decades. The current study reveals the fabrication of a polycaprolactone (PCL)/ß-tricalcium phosphate (TCP) scaffold by incorporating carbon nanotubes (CNT) via 3D printing. The physical properties and cytocompatibility of a new type of tissue engineering composite from polycaprolactone/ß-tri-calcium phosphate/carbon nanotubes were investigated, and it was an absorbable scaffold prepared via furnace deposition 3D printing technology. The scaffold was designed with CAD software, and the composite material was fabricated via 3D printing. The printed composite material was tested for mechanical strength, scanning electron microscope (SEM) analysis, porosity calculation, systemic toxicity test, hemolysis rate determination, and effect on the proliferation of rat adipose-derived stem cells cultured in vitro. A composite scaffold with a length of 15 mm, width of 10 mm, and height of 5 mm was manufactured through CAD software drawing and 3D printing technology. Scanning electron microscopy measurements and analysis of the internal pore size of the stent are appropriate; the pores are interconnected, and the mechanical strength matches the strength of human cancellous bone. The calculated porosity of the stent was >60%, non-toxic, and non-hemolytic. The proliferation activity of the ADSC co-cultured with different scaffold materials was as follows: polycaprolactone/ß-tricalcium phosphate/0.2% carbon nanotube scaffolds > polycaprolactone/ß-tricalcium phosphate/0.1% carbon nanotube scaffolds > polycaprolactone/ß-tricalcium phosphate/0.3% carbon nanotube scaffolds > polycaprolactone/ß-tricalcium phosphate scaffolds (P < 0.05). The results showed that polycaprolactone/ß-tricalcium phosphate/0.2% carbon nanotube scaffolds promoted the adhesion and proliferation of ADSC. The combination of 3D printing technology and CAD software can be used to print personalized composite stents, which have the characteristics of repeatability, high precision, and low cost. Through 3D printing technology, combining a variety of materials with each other can provide the greatest advantages of materials. The waste of resources was avoided. The prepared polycaprolactone/ß-tri-calcium phosphate/0.2% carbon nanotube scaffold has a good pore structure and mechanical properties that mimic human cancellous bone, is non-toxic and non-hemolytic, and is effective in promoting ADSC proliferation in vitro. Given this correspondence, 3D printed scaffold shows good biocompatibility and strength, and the fabrication method provides a proof of concept for developing scaffolds for bone tissue engineering applications.

Selenium increases antimony uptake in As-hyperaccumulators Pteris vittata and Pteris cretica by promoting antimonate reduction: GSH-GSSG cycle and arsenate reductases HAC1/ACR2.

Selenium-enhanced arsenic uptake by As-hyperaccumulators Pteris vittata and Pteris cretica is known, but how it impacts antimony (Sb) uptake and associated mechanisms are unclear. Here, we investigated the effects of 2.5 µM selenate (Se2.5) on Sb uptake by two plants after growing for 10 days under hydroponics containing 10 or 50 µM antimonate (SbV) (Sb10 or Sb50). Both plants were efficient in taking up SbV, which was reduced to SbIII (17-40 %) and mainly accumulated in the roots (86-97 %). The addition of Se increased the Sb contents by 78-97 and 29-33 % to 242-1358 and 132-697 mg kg-1 in P. vittata and P. cretica roots. Compared with the Sb10 and Sb50 treatments, addition of Se increased the SbV reduction, with more increase in P. vittata than P. cretica roots (181-273 % vs. 17-29 %). Enhanced GSH-GSSG cycle mediated by glutathione peroxidase (GPX) and glutathione reductase (GR) may play an important role in SbV reduction in the roots. Compared with the Sb treatments, addition of Se increased the GPX and GR activity by 71-97 and 2-50 % in P. vittata roots, and 59-153 and 22-63 % in P. cretica roots. Besides, Se upregulated the expression of arsenate reductases PvHAC1 and PvACR2 in P. vittata roots by 1.7-3.4 folds but not in P. cretica. Se-enhanced SbV reduction in P. vittata explains why it was more effective in Sb accumulation than P. cretica. Taken together, Se is effective in increasing the Sb uptake in both plants probably by promoting SbV reduction via GSH-GSSG cycle and/or PvHAC1/PvACR2, suggesting that Se may be used to enhance phytostabilization of Sb-contaminated soils.

High geological background concentrations of As and Cd in karstic soils may not contribute to greater risks to human health via rice consumption.

High geological background concentrations of toxic metal(loid)s arsenic (As) and cadmium (Cd) from natural enrichment in soils of karst regions have attracted much attention. In this study, paired soil-rice samples were collected from karst and non-karst regions in Guangxi, China to assess the potential risks of metal(loid) transfer from soil to rice grains, and rice grains to humans. Our results indicate that the karstic soils had greater As (25.7 vs. 12.4 mg·kg-1) and Cd (2.12 vs. 1.04 mg·kg-1) contents than those in non-karstic soils. However, metal(loid) transfer from soil to rice grains (ratio of rice grains to soil content) of As and Cd was 40 % and 49 % lower in karst regions, which may relate to their 42 % and 61 % lower HNO3-extractable As and CaCl2-extractable Cd, resulting in similar As/Cd contents in karstic and non-karstic rice grains. In vitro assay using a modified physiologically-based extraction test shows that karstic rice grains had a lower As/Cd bioaccessibility than non-karstic grains, which can be attributed to their ∼50 % greater P content, which negatively correlated with As/Cd bioaccessibility. Additionally, karstic rice grains had 39 % greater phytate and exhibited 45 % and 9.4 % lower As and Cd bioaccessibility in the gastric phase with phytate supplement at 0.6 %. Our work indicates that despite the greater As/Cd contents in karstic soils, the risks of As/Cd transfer from soil to rice grains as well as their exposure risks to humans via rice consumption may not be greater than non-karst regions.

[Production of adeno-associated virus in insect cells using multiple gene deleted baculovirus].

Adeno-associated virus (AAV) is one of the most frequently used viral vectors in the field of gene therapy. However, the industrial production of AAV is facing key bottlenecks such as low yield and high-cost. The aim of this study was to establish a technology system for production of AAV in the double virus infected insects by using multiple-gene deleted baculovirus. First, a multiple gene deleted baculovirus for AAV production was constructed, and the baculovirus titer and its effect on infected cells was examined. Subsequently, the insect cells were co-infected with the double baculovirus and the infection conditions were optimized. At the final stage, we performed AAV production based on optimized conditions, and evaluated relevant parameters including production titer and quality. The results showed that the titer of AAV produced in the multiple gene deleted baculovirus was not different from that of the wild type, but the rate of cell death was significantly slower upon infection. Using the double virus route for optimized production of AAV, the genome titers were 1.63×1011 VG/mL for Bac4.0-1 and 1.02×1011 VG/mL for Bac5.0-2, which were elevated 240% and 110%, respectively, compared with that of the wild-type. Electron microscopy observations revealed that all three groups exhibited normal AAV viral morphology and they showed similar transduction activity. Taken together, we developed an AAV production system based on the infection of insect cells using multiple-gene deleted baculovirus, which significantly improved the virus yield and showed application potential.

Selenium alleviates chromium stress and promotes chromium uptake in As-hyperaccumulator Pteris vittata: Cr reduction and cellar distribution.

Arsenic-hyperaccumulator Pteris vittata exhibits remarkable absorption ability for chromium (Cr) while beneficial element selenium (Se) helps to reduce Cr-induced stress in plants. However, the effects of Se on the Cr uptake and the associated mechanisms in P. vittata are unclear, which were investigated in this study. P. vittata plants were grown for 14 days in 0.2-strength Hoagland solution containing 10 (Cr10) or 100 µM (Cr100) chromate (CrVI) and 1 µM selenate (Se1). The plant biomass, malondialdehyde contents, total Cr and Se contents, Cr speciation, expression of genes associated with Cr uptake, and Cr subcellular distribution in P. vittata were determined. P. vittata effectively accumulated Cr by concentrating 96-99% in the roots under Cr100 treatment. Further, Se substantially increased its Cr contents by 98% to 11,596 mg kg-1 in the roots, which may result from Se's role in reducing its oxidative stress as supported by 27-62% reduction in the malondialdehyde contents. Though supplied with CrVI, up to 98% of the Cr in the roots was reduced to insoluble chromite (CrIII), with 83-89% being distributed on root cell walls. Neither Cr nor Se upregulated the expression of sulfate transporters PvSultr1;1-1;2 or phosphate transporter PvPht1;4, indicating their limited role in Cr uptake. P. vittata effectively accumulates Cr in the roots mainly as CrIII on cell walls and Se effectively enhances its Cr uptake by reducing its oxidative stress. Our study suggests that Se can be used to enhance P. vittata Cr uptake and reduce its oxidative stress, which may have application in phytostabilization of Cr-contaminated soils.

Novel phosphatase PvPAP1 from the As-hyperaccumulator Pteris vittata promotes organic P utilization and plant growth: Extracellular exudation and phytate hydrolysis.

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 ∼8 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.

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

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

Arsenic-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.