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.

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.

Response of cytokinins and nitrogen metabolism in the fronds of Pteris sp. under arsenic stress.

Given the close relationship between cytokinins (CKs), photosynthesis and nitrogen metabolism, this study assessed the effect of arsenic (As) contamination on these metabolic components in the As-hyperaccumulators Pteris cretica L. var. Albo-lineata (Pc-A) and var. Parkerii (Pc-P) as well as the As-non-hyperaccumulator Pteris straminea Mett. ex Baker (Ps). The ferns were cultivated in a pot experiment for 23 weeks in soil spiked with As at the levels 20 and 100 mg·kg-1. For the purpose of this study, the CKs were placed into five functionally different groups according to their structure and physiological roles: bioactive forms (bCKs; CK free bases); inactive or weakly active forms (dCKs; CK N-glucosides); transport forms (tCKs; CK ribosides); storage forms (sCKs; O-glucosides); and primary products of CK biosynthesis (ppbCKs; CK nucleotides). An important finding was higher CKs total content, accumulation of sCKs and reduction of dCKs in As-hyperaccumulators in contrast to non-hyperaccumulator ferns. A significant depletion of C resources was confirmed in ferns, especially Ps, which was determined by measuring the photosynthetic rate and chlorophyll fluorescence. A fluorescence decrease signified a reduction in the C/N ratio, inducing an increase of bioactive CKs forms in Pc-P and Ps. The impact of As on N utilization was significant in As-hyperaccumulators. The glutamic acid/glutamine ratio, an indicator of primary N assimilation, diminished in all ferns with increased As level in the soil. In conclusion, the results indicate a large phenotypic diversity of Pteris species to As and suggest that the CKs composition and the glutamic acid/glutamine ratio can be used as a tool to diagnose As stress in plants.

Pteridophytes in phytoremediation.

Soil contamination by heavy metals and metalloids is a serious problem which needs to be addressed. There are several methods for removal of contaminants, but they are costly, while the method of phytoremediation is eco-friendly and cost-effective. Pteridophytes have been found to remediate heavy metal-contaminated soil. Pteridophytes are non-flowering plant that reproduces by spores. Pteris vittata has been reported as the first fern plant to hyperaccumulate arsenic. The Pteris species belongs to the order Pteridales. Other ferns that are known phytoremediators are, for example, Nephrolepis cordifolia and Hypolepis muelleri (identified as phytostabilisers of Cu, Pb, Zn and Ni); similarly Pteris umbrosa and Pteris cretica accumulate arsenic in leaves. So, pteridophytes have a number of species that accumulate contaminants. Many of them have been identified, while various other are being explored. The present review article describes the phytoremediation potential of pteridophytes plants and suggests as a potential asset for phytoremediation programs.

Interactive effects of As, Cd and Zn on their uptake and oxidative stress in As-hyperaccumulator Pteris vittata.

The effects of arsenic (As), cadmium (Cd) and zinc (Zn) on each other's uptake and oxidative stress in As-hyperaccumulator Pteris vittata were investigated. P. vittata plants were exposed to 50 µM As, Cd and/or Zn for 15 d in 0.2-strength Hoagland solution. When applied alone, P. vittata accumulated 185 mg kg-1 As, 164 mg kg-1 Cd and 327 mg kg-1 Zn in the fronds. While Cd and Zn did not impact each other's uptake, As affected Cd and Zn uptake. Whereas As decreased Zn uptake, Zn affected As speciation in P. vittata fronds, with more arsenate (AsV) than arsenite (AsIII) being present. At 50 µM As, 75 µM Zn increased As accumulation in P. vittata fronds by 10 folds to 2363 mg kg-1 compared to 50 µM Zn. Although AsV was the predominant As species in all tissues, Cd enhanced AsIII levels in the fronds but increased AsV in the roots. Co-exposure of Cd + Zn elevated oxidative stress basing on thiobarbituric acid reactive substances, H2O2 content, Evans blue dye uptake, membrane injury index and reactive oxygen species (ROS) relative to single metal. By lowering Cd and Zn concentrations in P. vittata fronds, As reduced the associated stress comparative to Cd or Zn treatment. The results enhance our understanding of the mechanisms underlying the interactions between As, Cd and Zn in As-hyperaccumulator P. vittata.

Potential use of the Pteris vittata arsenic hyperaccumulation-regulation network for phytoremediation.

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.

Metal tolerance of arsenic-resistant bacteria and their ability to promote plant growth of Pteris vittata in Pb-contaminated soil.

Soils contaminated with Pb and As are difficult to remediate. In this study, the utility of coupling As-hyperaccumulator Pteris vittata with metal-resistant rhizobacteria was explored. Siderophore-producing and P-solubilizing As-resistant bacteria from the P. vittata rhizosphere were screened for resistance to multiple metals. Results indicated Pseudomonas spp. strain PG-12 was most efficient in resisting multiple metals, i.e., up to 0.6 mM Cd and 10 mM Pb. Amplification of gene fragments encoding various metal efflux transporters (PbrA and CadA2) from genomic DNA of PG-12 suggested that metal efflux might play a role in its metal resistance and detoxification. In addition, PG-12 produced significant levels of plant growth hormones including 17.4 µg mL-1 indole acetic acid and 3.54 µg mL-1 gibberellin. P. vittata sporophytes inoculated with PG-12 were grown in Pb-contaminated medium and exhibited improved growth, increased P uptake, and reduced Pb uptake into plant tissue compared to the control. Results demonstrated that viable PG-12 cells were responsible for Pb immobilization and plant growth enhancement in P. vittata. The ability of PG-12 cells to solubilize P and display resistance to multiple metals combined with the production of plant hormones indole acetic acid and gibberellin make PG-12 a suitable candidate for plant growth promotion in metal-contaminated soil.

Effect of arsenate on endogenous levels of cytokinins with different existing forms in two Pteris species.

Our previous results showed that content of trans-zeatin (tZ) increases in leaves of heavy metal hyperaccumulators but decreases in non-hyperaccumulators growing in multiple heavy metal polluted soils. However, the relationship between arsenic (As) accumulation and endogenous forms of cytokinins (CTKs) in As hyperaccumulators remains unknown. Here a hydroponic experiment was conducted to compare the CTK forms in the As hyperaccumulator Pteris cretica var. nervosa and non-hyperaccumulator Pteris ensiformis under arsenate stress (0, 2, 5, and 10 mg L-1). A simple and cost-effective procedure for the determination of CTK forms in plants was established, and a stepwise regression analysis was used to study the relationship among total As contents and different forms of endogenous CTKs in fronds of two plants. The results showed that the optimized chromatographic parameters were Zobax SB-C18 column (5 µm × 4.6 mm × 250 mm), UV detection detector at 269 nm, a flow rate of 0.6 mL min-1, constant temperature of 45 °C and gradient elution with methanol-acetonitrile-1% acetic acid. Contents of chlorophylls in the fronds of P. ensiformis were significantly decreased with addition of As compared to P. cretica var. nervosa. Furthermore, the total As content in fronds of P. cretica var. nervosa was positively correlated to the contents of N6-(2-isopentenyl) adenine-7-ß-D-glucoside (iP7G) and N6-(2-isopentenyl) adenosine (iPR). However, the total As content in fronds of P. ensiformis was negatively correlated to its trans-zeatin riboside (ZR) content. Therefore, iP7G and iPR could positively improve As accumulation by P. cretica var. nervosa.

Effects of kinetin on plant growth and chloroplast ultrastructure of two Pteris species under arsenate stress.

Cytokinins (CTKs) are effective in alleviating abiotic stresses on plants, but little information is available regarding the effects of CTKs on arsenic (As) accumulation and changes of chloroplast ultrastructure in plants with different As-accumulating ability. Here a hydroponic experiment was designed to evaluate the effects of different concentration of kinetin (KT, 0-40 mg/L) on growth and chloroplast ultrastructure of As hyperaccumulator Pteris cretica var. nervosa and non-hyperaccumulator Pteris ensiformis treated by 5 mg/L arsenate for 14 days. The growth parameters, As accumulation, contents of photosynthetic pigments and chloroplast ultrastructure were examined. The results showed that KT promoted the growth of two plants, and significantly increased As accumulation and translocation in P. cretica var. nervosa and P. ensiformis at 5 and 20 mg/L, respectively. Additionally, the contents of chlorophyll a and carotenoid in two plants showed no significant difference at 20 mg/L KT compared to the control. Chloroplast ultrastructure of P. cretica var. nervosa was integral with KT application. Comparatively, the swollen chloroplasts were increased, plasmolysis appeared, and chloroplast grana slice layers and stroma lamellas were clearly separated or distorted at 5 mg/L KT in P. ensiformis. The length and width of chloroplasts in P. cretica var. nervosa were significantly increased with KT addition compared to the control. However, the length of chloroplasts in P. ensiformis was significantly decreased but their width showed no significant change. Furthermore, the deterioration of chloroplast ultrastructure in P. ensiformis was ameliorated by 40 mg/L KT. These results suggested that KT increased As accumulation and was beneficial to maintain the photosynthetic pigments for a good growth of plants. Therefore, KT could maintain and reorganize the ultrastructure integrality of As-stressed chloroplasts to some extent for the two plants, especially at high concentration.

Arsenic-induced nutrient uptake in As-hyperaccumulator Pteris vittata and their potential role to enhance plant growth.

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 40 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.

Interactions between Pteris vittata L. genotypes and a polycyclic aromatic hydrocarbon (PAH)-degrading bacterium (Alcaligenes sp.) in arsenic uptake and PAH-dissipation.

The effects of two Pteris vittata L. accessions and a polycyclic aromatic hydrocarbon (PAH)-degrading bacterium (Alcaligenes sp.) on arsenic (As) uptake and phenanthrene dissipation were studied. The Alcaligenes sp. survived in the rhizosphere and improved soil As bioavailability with co-exposure. However, bacterial inoculation altered Pteris vittata L. stress tolerance, and substantially affected the As distribution in the rhizosphere of the two P. vittata accessions. Bacterial inoculation was beneficial to protect the Guangxi accession against the toxic effects, and significantly increased plant As and phenanthrene removal ratios by 27.8% and 2.89%, respectively. In contrast, As removal was reduced by 29.8% in the Hunan accession, when compared with corresponding non-inoculated treatments. We conclude that plant genotype selection is critically important for successful microorganism-assisted phytoremediation of soil co-contaminated with As and PAHs, and appropriate genotype selection may enhance remediation efficiency.

Arsenate and fluoride enhanced each other's uptake in As-sensitive plant Pteris ensiformis.

We investigated the effects of arsenate (AsV) and fluoride (F) on each other's uptake in an As-sensitive plant Pteris ensiformis. Plants were exposed to 1) 0.1 × Hoagland solution control, 2) 3.75 mg L-1 As and 1.9, 3.8, or 7.6 mg L-1 F, or 3) 1 mg L-1 F and 3.75 mg L-1 or 7.5 mg L-1 As for 7 d in hydroponics. P. ensiformis accumulated 14.7-32.6 mg kg-1 As at 3.75 mg L-1 AsV, and 99-145 mg kg-1 F at 1 mg L-1 F. Our study revealed that AsV and F increased each other's uptake when co-present. At 1.9 mg L-1, F increased frond As uptake from 14.7 to 40.3 mg kg-1, while 7.5 mg L-1 As increased frond F uptake from 99 to 371 mg kg-1. Although, AsV was the predominant As species in all tissues, F enhanced AsIII levels in the rhizomes and fronds, while the reverse was observed in the roots. Increasing As concentrations also enhanced TBARS and H2O2 in tissues, indicating oxidative stress. However, F alleviated As stress by lowering their levels in the fronds. Frond and root membrane leakage were also evident due to As or F exposure. The results may facilitate better understanding of the mechanisms underlying the co-uptake of As and F in plants. However, the mechanisms of how they enhance each other's uptake in P. ensiformis need further investigation.

Effects of indole-3-acetic acid on arsenic uptake and antioxidative enzymes in Pteris cretica var. nervosa and Pteris ensiformis.

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.

Arsenic-enrichment enhanced root exudates and altered rhizosphere microbial communities and activities in hyperaccumulator Pteris vittata.

Phytoremediation of arsenic (As)-contaminated soil by hyperaccumulator Pteris vittata is promising. A better understanding of the rhizosphere microbial dynamics that regulate As availability and plant growth is important to optimize the phytoremediation process. In this study, Illumina sequencing of 16S rRNA genes was applied to assess the rhizosphere microbial community structure of P. vittata. Microbial functionality was monitored by soil enzyme activities and MPN-PCR targeting genes of interest. Arsenic (100mgkg-1 AsV) addition to soil significantly increased DOC, root exudates, As and P uptake and the frond biomass of P. vittata. Moreover, As-enrichment significantly increased soil enzyme activities involved in N, P and S cycling and the gene abundance of As transforming bacteria, Fe- and S-reducing bacteria and N and C fixing bacteria in the rhizosphere of P. vittata. Together, the results revealed that the combined selective pressure of As and rhizosphere resulted in stimulation of microbial community, which most likely has a role in reductive dissolution of Fe and S, As and P mobilization, C degradation and fixation, and N fixation. These changes appeared to have a role in mitigation of As toxicity and to promote growth and the As uptake ability of P. vittata under As-enriched conditions.

Arsenic-induced plant growth of arsenic-hyperaccumulator Pteris vittata: Impact of arsenic and phosphate rock.

Phosphate rock (PR) has been shown to promote plant growth and arsenic (As) uptake by As-hyperaccumulator Pteris vittata (PV). However, little is known about its behaviors in agricultural soils. In this study, impact of 50 mg kg(-1) As and/or 1.5% PR amendment on plant As accumulation and growth was investigated by growing PV for 90 d in three agricultural soils. While As amendment significantly increased plant As uptake and substantially promoted PV growth, the opposite was observed with PR amendment. Arsenic amendment increased plant frond As from 16.9-265 to 961-6017 mg kg(-1),whereas PR amendment lowered frond As to 10.2-216 mg kg(-1). The As-induced plant growth stimulation was 69-71%. While PR amendment increased plant Ca and P uptake, As amendment showed opposite results. The PV biomass was highly correlated with plant As at r = 0.82, but with weak correlations with plant Ca or P at r < 0.30. This study confirmed that 1) As significantly promoted PV growth, probably independent of Ca or P uptake, 2) PR amendment didn't enhance plant growth or As uptake by PV in agricultural soils with adequate available P, and 3) PV effluxed arsenite (AsIII) growing in agricultural soils.

A member of the Phosphate transporter 1 (Pht1) family from the arsenic-hyperaccumulating fern Pteris vittata is a high-affinity arsenate transporter.

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.

Arsenic uptake, arsenite efflux and plant growth in hyperaccumulator Pteris vittata: Role of arsenic-resistant bacteria.

Bacteria-mediated arsenic (As) transformation and their impacts on As and P uptake and plant growth in As-hyperaccumulator Pteris vittata (PV) were investigated under sterile condition. All As-resistant bacteria (9 endophytic and 6 rhizospheric) were As-reducers except one As-oxidizer. After growing two months in media with 37.5 mg kg(-1) AsV, As concentrations in the fronds and roots were 3655-5389 (89-91% AsIII) and 971-1467 mg kg(-1) (41-73% AsIII), corresponding to 22-52% decrease in the As in the media. Bacterial inoculation enhanced As and P uptake by up to 47 and 69%, and PV growth by 20-74%, which may be related to elevated As and P in plants (r = 0.88-0.97, p < 0.05). Though AsV was supplied, 95% of the As in the bacteria-free media was AsIII, suggesting efficient efflux of AsIII by PV roots (120 µg g(-1) root fw). This was supported by the fact that no AsV was detected in media inoculated with As-reducers while 95% of AsV was detected with As-oxidizer. Our data showed that, under As-stress, PV reduced As toxicity by efficient AsIII efflux into media and AsIII translocation to the fronds, and bacteria benefited PV growth probably via enhanced As and P uptake.

Arsenic transformation and plant growth promotion characteristics of As-resistant endophytic bacteria from As-hyperaccumulator Pteris vittata.

The ability of As-resistant endophytic bacteria in As transformation and plant growth promotion was determined. The endophytes were isolated from As-hyperaccumulator Pteris vittata (PV) after growing for 60 d in a soil containing 200 mg kg(-1) arsenate (AsV). They were isolated in presence of 10 mM AsV from PV roots, stems, and leaflets, representing 4 phyla and 17 genera. All endophytes showed at least one plant growth promoting characteristics including IAA synthesis, siderophore production and P solubilization. The root endophytes had higher P solubilization ability than the leaflet (60.0 vs. 18.3 mg L(-1)). In presence of 10 mM AsV, 6 endophytes had greater growth than the control, suggesting As-stimulated growth. Furthermore, root endophytes were more resistant to AsV while the leaflet endophytes were more tolerant to arsenite (AsIII), which corresponded to the dominant As species in PV tissues. Bacterial As resistance was positively correlated to their ability in AsV reduction but not AsIII oxidation. The roles of those endophytes in promoting plant growth and As resistance in P. vittata warrant further investigation.

Changes in gametophyte physiology of Pteris multifida induced by the leaf leachate treatment of the invasive Bidens pilosa.

In recent years, the response of fern gametophytes to environment has raised much attention. However, studies on the influence of plant invasion to fern gametophytes are scarce. Allelopathy plays an important role in biological invasion. Hence, it is necessary to study the allelopathic effects of invasive plants on fern gametophytes 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 spermatophyte growth. Field investigation shows that many ferns are threatened by the invasion of B. pilosa. The distribution of Pteris multifida overlaps with that of B. pilosa in China. To examine the potential involvement of allelopathic mechanisms of B. pilosa leaves, changes in the physiology in P. multifida gametophytes are analyzed. We found that cell membrane and antioxidant enzyme activities as well as photosynthesis pigment contents of the gametophytes were affected by B. pilosa leachates. Gametophytes of P. multifida exposed to B. pilosa had increased damages to cell membranes, expressed in thiobarbituric acid reacting substance (TBARS) concentrations, malondialdehyde (MDA), electrolyte leakage (membrane permeability), and degree of injury. Enzyme activities, assessed by superoxide dismutase (SOD) and catalase (CAT) as well as guaiacol peroxidase (GPX) enhanced with the increase in leachate concentration after 2-day exposure. Meanwhile, lower chlorophyll a (Chl a), chlorophyll b (Chl b), carotenoid (Car), and the total chlorophyll were measured as leachate concentrations increased. At day 10, leaf leachates of B. pilosa exhibited the greatest inhibition. These results suggest that the observed inhibitory or stimulatory effects on the physiology studied can have an adverse effect on P. multifida and that allelopathic interference seems to have involved in this process.

[Influence of Arsenate and Phenanthrene on Carbon-groups of Pteris vittata L. Roots].

To ascertain absorption of arsenate and phenanthrene as well as their influence on carbon groups in excised roots of Pteris vittata L., the chemical structure of the carbon groups in excised roots was characterized by solid state13C-Nuclear Magnetic Resonance (13C-NMR). The results showed that the excised roots could effectively absorb As and PHE without transpiration, and PHE promoted As accumulation in the roots. Similarly, arsenate increased the adsorption of PHE by the excised roots, the concentration of PHE was increased by 15%-53% compared with CK. The carbon groups of the excised roots were dominated by O-alkyl C, the percentage of carboxyl C was the lowest, mainly composed of carboxylic acids, esters and amides. With the addition of As and PHE, the percentage of carboxyl C increased significantly. The more stable and complex aromatic organic matter was formed to improve the resistance and adaptability in excised roots of Pteris vittata L. under As and PHE stress.