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Azolla is a floating aquatic fern, having amazing capacity for concentrating toxic heavy metals. Metallothioneins (MTs) and phytochelatins (PCs) are well-defined heavy metal-binding ligands in plants. Bioaccumulation potential of different Azolla species varies according to their heavy metal ions. Therefore, the accumulation of Ni, Zn, Cu, and Cd was studied in A. pinnata, A. filiculoides, and a sample taken from Anzali wetland. Moreover, the expression of metallothionein and phytochelatin synthase encoding genes was examined at different metal concentrations. The highest level of Cu and Cd absorption was detected in A. pinnata, while the maximum amount of Ni and Zn absorption was observed in A. filiculoides and the sample taken from Anzali, respectively. The MT2 and PCS1 gene expression patterns were significantly induced by the heavy metal treatments, confirming their roles in phytoremediation potential of Azolla. However, as the results concerning heavy metal accumulation and gene expression vary in different species, only specific species of Azolla can be used for special purposes. It can be concluded that the Azolla is a good candidate for phytoremediation purposes, and the formation of phytochelatin-heavy metal complexes and their sequestration in vacuole are the main processes influencing susceptibility of Azolla to heavy metals., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Arsenic is a carcinogenic contaminant of water and food and a growing threat to human health in many regions of the world. This study focuses on the fern Pteris vittata (Pteridaceae), which is extraordinary in its ability to tolerate and hyperaccumulate very high levels of arsenic that would kill any other plant or animal outside the Pteridaceae. Here, we use RNA-seq to identify three genes (GLYCERALDEHYDE 3-PHOSPHATE DEHYDROGENASE (PvGAPC1), ORGANIC CATION TRANSPORTER 4 (PvOCT4), and GLUTATHIONE S-TRANSFERASE (PvGSTF1) that are highly upregulated by arsenic and are necessary for arsenic tolerance, as demonstrated by RNAi. The proteins encoded by these genes have unexpected properties: PvGAPC1 has an unusual active site and a much greater affinity for arsenate than phosphate; PvGSTF1 has arsenate reductase activity; and PvOCT4 localizes as puncta in the cytoplasm. Surprisingly, PvGAPC1, PvGSTF1, and arsenate localize in a similar pattern. These results are consistent with a model that describes the fate of arsenate once it enters the cell. It involves the conversion of arsenate into 1-arseno-3-phosphoglycerate by PvGAPC1. This "chemically trapped" arsenate is pumped into specific arsenic metabolizing vesicles by the PvOCT4 protein. Once inside these vesicles, 1-arseno-3-phosphoglycerate hydrolyses to release arsenate, which is then reduced by PvGSTF1 to arsenite, the form of arsenic stored in the vacuoles of this fern. This mechanism is strikingly similar to one recently described Pseudomonas aeruginosa, whose tolerance to arsenic also involves the biosynthesis and transport of 1-arseno-3-phosphoglycerate, indicating that P. vittata has evolved a simple, bacterial-like mechanism for arsenic tolerance., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Lead (Pb) removal from water column was evaluated in batch experiments using naturally occurring Salvinia biloba Raddi (S. biloba) specimens collected from Middle Paraná River and exposed every 24 h to a fresh discharge of water contaminated with 2.65 ± 0.07, 12.62 ± 0.02 or 30.57 ± 0.01 mg L -1 Pb, during 10 consecutive days. S. biloba demonstrated a great ability for metal concentration-dependent Pb removal under these stressful conditions. Additionally, Pb toxicity in plants was assessed by the quantification of physiological parameters in root-like modified fronds (named "roots"), and its aerial leaf-like fronds (named "leaves") of submerged S. biloba. Photosynthetic (carotenoids, chlorophyll a, b, and total) and antioxidant pigments (anthocyanins and flavonoids), soluble carbohydrate content, and membrane stability index of both roots and leaves were affected as the metal concentration increased. In general, root deterioration was more pronounced than that in leaves, suggesting a greater implication of the former organs in Pb removal by S. biloba. All of these deleterious effects were well correlated with qualitative changes observed at plant phenotype during the assay. In conclusion, S. biloba may be considered as a water fern useful in phytoremediation strategies towards management of residual water bodies contaminated with Pb. In addition, these macrophytes could also be valuable for water biomonitoring contributing to improve risk assessments related to metal presence in wastewaters., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Fern spores are unicellular structures produced by the sporophyte generation that give rise to the haploid gametophyte. When released from the sporangium, spores are desiccation tolerant (DT) in the royal fern (Osmunda regalis) and contain fully developed chloroplasts. As a consequence, this type of spores is called chlorophyllous spores (CS). Upon transfer to germination conditions, CS initiate a process of imbibition that suppresses DT in 72 h, before the germination starts. In parallel to such change in DT, thylakoids undergo a profound remodelling in composition and function. Firstly, sustained quenching of chlorophyll fluorescence is relaxed, giving rise to photochemically active CS, while lipid composition shifts from that of a resting structure to a metabolically active cell. Basically trigalactolipids decreased in favour of monogalactolipids, with a parallel desaturation of fatty acids. Storage lipids such as triacylglycerol were quickly depleted. These results highlight the importance of the structure of thylakoids lipid as a key to protect membrane integrity during desiccation, together with the saturation of fatty acids and the constitutive chlorophyll quenching to prevent oxidative damage. The CS used here, in which the same cell shifts from DT to sensitive strategy in 72 h, reveal their potential as unicellular models for future studies on DT., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The present work highlights some preliminary observations on metabolism in rice (Oryza sativa Linn.) when an aquatic fern Azolla pinnata R.Br. was co-cultured under 2,4-Dichlorophenoxy acetic acid toxicity. We have observed the effects of Azolla in both fresh and dried forms. This work signifies the possible physiological changes of a crop plant by using Azolla as a bioremediator. In brief the herbicide 2,4-D is considered as stressor to rice plants and by applying the fresh and dried Azolla we investigate the changes occurred. The activities of different nitrogen metabolizing enzymes and reactive oxygen species were observed. On the other hand chlorophyll and carotenoids synthesis were retrieved by addition of fresh and dried Azolla mass over 2,4-D toxicity. Thus, the efficiency of fresh and dried Azolla mass was evaluated under herbicidal toxicity in rice. We evaluate the bio remediating role of Azolla plants against 2,4-D stress and conclude this species would also be supporting in supplementation of major nutrients to rice plants.

It has been claimed that the green synthesized NPs possess no toxicity in comparison to the NPs fabricated via conventional protocols like reduction by sodium borohydride. Therefore, it is necessary to test the toxic effects of NPs on environment. In the current study, we report the binding of Fe 3 O 4 NPs to galate ions containing biomaterial namely "galate bio-capping agent". The bio-capping agent is simply mixed with the Fe 3+ cations at pH 8 to produce negatively charged bio-capped Fe 3 O 4 NPs. Finally, the toxic effects of the Fe 3 O 4 NPs were investigated on some growth and developmental indices of the aquatic plant species Azolla filiculoides. The relative frond number and relative growth rate were calculated after treatment of plants with different concentrations of bio-capped Fe 3 O 4 NPs. In addition, the content of phenolics as well as antioxidant enzymes' activity including superoxide dismutase and peroxidase were assessed. The Fe 3 O 4 NPs led to growth reduction and significant changes in total phenol and flavonoid content as well as in antioxidant enzymes' activity. All these findings confirm reactive oxygen species formation due to the nanoparticle toxicity. In consequence, the enzymatic and non-enzymatic antioxidant defense systems of plant were stimulated against oxidative stress.

All structure-based information on chlorophyll (Chl) breakdown in the higher plants relies on studies with angiosperms. Herein, the first investigation of a fern is reported, revealing a novel type of Chl catabolites (phyllobilins) in leaves of this large division of the vascular plants, and providing structural insights into an astounding metabolic process of the higher plants that appears to have played a role even in early phases of plant evolution. The tetrapyrrolic Chl catabolites in the cosmopolitan bracken fern were discovered to be phyllobilin isomers with an unprecedented skeleton, proposed to be the striking result of a rearrangement of a hypothetical phyllobilin precursor., (© 2018 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.)

Plants sense and respond to microbes utilizing a multilayered signalling cascade. In seed plants, the phytohormones jasmonic and salicylic acid (JA and SA) are key denominators of how plants respond to certain microbes. Their interplay is especially well-known for tipping the scales in plants' strategies of dealing with phytopathogens. In non-angiosperm lineages, the interplay is less well understood, but current data indicate that it is intertwined to a lesser extent and the canonical JA/SA antagonism appears to be absent. Here, we used the water fern Azolla filiculoides to gain insights into the fern's JA/SA signalling and the molecular communication with its unique nitrogen fixing cyanobiont Nostoc azollae, which the fern inherits both during sexual and vegetative reproduction. By mining large-scale sequencing data, we demonstrate that Azolla has most of the genetic repertoire to produce and sense JA and SA. Using qRT-PCR on the identified biosynthesis and signalling marker genes, we show that Azolla is responsive to exogenously applied SA. Furthermore, exogenous SA application influenced the abundance and gene expression of Azolla's cyanobiont. Our data provide a framework for JA/SA signalling in ferns and suggest that SA might be involved in Azolla's communication with its vertically inherited cyanobiont., (© 2018 John Wiley & Sons Ltd.)

The epiphytic fern Pleopeltis polypodioides can tolerate repeated drying and rehydration events without conspicuous damage. To understand the biochemical principles of drought-tolerance, we analyzed the effect of dehydration and rehydration at 25 °C on hydroperoxide and lipid hydroperoxide, the activities of antioxidative (catalase and glutathione-oxidizing) enzymes and evaluated changes in fatty acid composition and saturation levels. Dehydration increased peroxide concentration and the activity of glutathione oxidases, but reduced catalase activity. During dehydration, the biosynthesis of palmitic (C16:0), linoleic (C18:2), linolenic (C18:3) and stearic acid (C18:0) increased 18, 12, 20, and 8-fold, respectively. In contrast, rehydration lowered levels of peroxides, the activity of glutathione-oxidizing enzymes, and fatty acids but increased catalase activity. The coordinated changes during de- and rehydration suggest that lipids and oxidative and antioxidative enzymes are components of the drought-resistance system., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

We studied how high light causes photoinhibition of photosystem I (PSI) in the shade-demanding fern Nephrolepis falciformis, in an attempt to understand the mechanism of PSI photoinhibition under natural field conditions. Intact leaves were treated with constant high light and fluctuating light. Detached leaves were treated with constant high light in the presence and absence of methyl viologen (MV). Chlorophyll fluorescence and P700 signal were determined to estimate photoinhibition. PSI was highly oxidized under high light before treatments. N. falciformis showed significantly stronger photoinhibition of PSI and PSII under constant high light than fluctuating light. These results suggest that high levels of P700 oxidation ratio cannot prevent PSI photoinhibition under high light in N. falciformis. Furthermore, photoinhibition of PSI in N. falciformis was largely accelerated in the presence of MV that promotes the production of superoxide anion radicals in the chloroplast stroma by accepting electrons from PSI. From these results, we propose that photoinhibition of PSI in N. falciformis is mainly caused by superoxide radicals generated in the chloroplast stroma, which is different from the mechanism of PSI photoinhibition in Arabidopsis thaliana and spinach. Here, we provide some new insights into the PSI photoinhibition under natural field conditions.

Recent studies have revealed that some responses of fern stomata to environmental signals differ from those of their relatives in seed plants. However, it is unknown whether the biophysical properties of guard cells differ fundamentally between species of both clades. Intracellular micro-electrodes and the fluorescent Ca 2+ reporter FURA2 were used to study voltage-dependent cation channels and Ca 2+ signals in guard cells of the ferns Polypodium vulgare and Asplenium scolopendrium. Voltage clamp experiments with fern guard cells revealed similar properties of voltage-dependent K + channels as found in seed plants. However, fluorescent dyes moved within the fern stomata, from one guard cell to the other, which does not occur in most seed plants. Despite the presence of plasmodesmata, which interconnect fern guard cells, Ca 2+ signals could be elicited in each of the cells individually. Based on the common properties of voltage-dependent channels in ferns and seed plants, it is likely that these key transport proteins are conserved in vascular plants. However, the symplastic connections between fern guard cells in mature stomata indicate that the biophysical mechanisms that control stomatal movements differ between ferns and seed plants., (© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.)

Exposure to recurrent desiccation cycles carries a risk of accumulation of reactive oxygen species that can impair leaf physiological activity upon rehydration, but changes in filmy fern stress status through desiccation and rewatering cycles have been poorly studied. We studied foliage photosynthetic rate and volatile marker compounds characterizing cell wall modifications (methanol) and stress development (lipoxygenase [LOX] pathway volatiles and methanol) through desiccation-rewatering cycles in lower-canopy species Hymenoglossum cruentum and Hymenophyllum caudiculatum, lower- to upper-canopy species Hymenophyllum plicatum and upper-canopy species Hymenophyllum dentatum sampled from a common environment and hypothesized that lower canopy species respond more strongly to desiccation and rewatering. In all species, rates of photosynthesis and LOX volatile emission decreased with progression of desiccation, but LOX emission decreased with a slower rate than photosynthesis. Rewatering first led to an emission burst of LOX volatiles followed by methanol, indicating that the oxidative burst was elicited in the symplast and further propagated to cell walls. Changes in LOX emissions were more pronounced in the upper-canopy species that had a greater photosynthetic activity and likely a greater rate of production of photooxidants. We conclude that rewatering induces the most severe stress in filmy ferns, especially in the upper canopy species., (© 2018 John Wiley & Sons Ltd.)

Azolla spp., a water fern often used for phytoremediation, is a strong phosphorus (P) accumulator due to its high growth rate and N 2 fixing symbionts (diazotrophs). It is known that plant growth is stimulated by P, but the nature of the interactive response of both symbionts along a P gradient, and related changes in growth-limiting factors, are unclear. We determined growth, and N and P sequestration rates of Azolla filiculoides in N-free water at different P concentrations. The growth response appeared to be biphasic and highest at levels ≥10 P µmol l -1 . Diazotrophic N sequestration increased upon P addition, and rates were three times higher at high P than at low P. At 10 µmol P l -1 , N sequestration rates reached its maximum and A. filiculoides growth became saturated. Due to luxury consumption, P sequestration rates increased until 50 µmol P l -1 . At higher P concentrations (≥50 µmol l -1 ), however, chlorosis occurred that seems to be caused by iron- (Fe-), and not by N-deficiency. We demonstrate that traits of the complete symbiosis in relation to P and Fe availability determine plant performance, stressing the role of nutrient stoichiometry. The results are discussed regarding Azolla's potential use in a bio-based economy.

This study investigated the occurrence of toxicity, expressed as damage to the photosynthetic apparatus, in the aquatic fern Azolla filiculoides and the lichen Xanthoria parietina following treatments with diclofenac at different concentrations (0.1, 1, 10 and 100 mg/L) and different exposure times (24, 48, 72 and 240 h). Measurements of photosynthetic efficiency, chlorophyll content and chlorophyll degradation indicated dose- and time-dependent toxicity, since significant differences with control samples as well as among treatments, emerged mainly for the highest concentration (100 mg/L) and the longest time (240 h). In addition, also the mycobiont of the lichen X. parietina showed similar toxic effects, expressed as ergosterol content. The absence of relevant alterations at the lowest concentration (0.1 mg/L) suggested a very limited susceptibility of these species to environmentally relevant levels of this pharmaceutical.

Main Conclusion: Unlike most plant cell walls, the five consecutive walls laid down during spermatogenesis in the model fern Ceratopteris contain sparse cellulose, lack pectin and are enriched with callose and hemicelluloses. Seed-free plants like bryophytes and pteridophytes produce swimming male gametes for sexual reproduction. During spermatogenesis, unique walls are formed that are essential to the appropriate development and maturation of the motile gametes. Other than the detection of callose and general wall polysaccharides in scattered groups, little is known about the sequence of wall formation and the composition of these walls during sperm cell differentiation in plants that produce swimming sperm. Using histochemistry and immunogold localizations, we examined the distribution of callose, cellulose, mannan and xylan-containing hemicelluloses, and homogalacturonan (HG) pectins in the special walls deposited during spermatogenesis in Ceratopteris. Five walls are produced in sequence and each has a unique fate. The first wall (W1) contains callose and sparse xylan-containing hemicelluloses. Wall two (W2) is thin and composed of cellulose crosslinked by xylan-containing hemicelluloses. The third wall (W3) is thick and composed entirely of callose, and the fourth wall (W4) is built of cellulose heavily crosslinked by galactoxyloglucan hemicelluloses. Wall five (W5) is an arabinogalactan protein (AGP)-rich matrix in which the gamete changes shape and multiple flagella elongate. We detected no esterified or unesterified HG pectins in any of the walls laid down during spermatogenesis. To consider evolutionary modifications in cell walls associated with motile gametes, comparisons are presented with male gametophyte and spermatogenous cell walls across plant groups.

Salvinia minima was assessed for its ability to accumulate lead (Pb) by exposing it to concentrations of 40µM Pb(NO 3 ) 2 during 24h. At the same time, the expression levels were quantified, of four genes coding for transporters: SmABCC (ABCC-MRP), SmATPase (ATPase-P3A), SmNhaD (Type-Na + /H + ) and SmABCG (ABCG-WBC). In the absence of lead, S. minima had very low expression of those genes, when plants were exposed to the metal however, those genes showed a rapid (in just three hours or less) and sharp increase (up to 60 times) in their expression, particularly the SmNhaD (Type-Na + /H + ) gene. This sharp increase in expression levels of the genes studied, occurred at the same time that the plant accumulated the highest content of lead in its tissues. The first two genes, are apparently implicated in detoxification and lead accumulation mechanisms, while the other two genes are apparently involved in maintaining cell balance (homeostatic control) and membrane integrity. Our results confirmed that S. minima is efficient for phytoremediation of water bodies contaminated by lead, as it is efficient in accumulating this metal in its tissues (bioconcentration factor; BCF) values greater than 1000, in short times of exposure. More importantly, our data on the expression profiles of four genes coding for transporters, represent a first sight scenario of the molecular basis for understanding the different mechanism of detoxification, apparently present in this aquatic fern., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Under conditions of Azolla imbricata restoration, the high-throughput sequencing technology was employed to determine change trends of microbial community structures in the soil that had undergone long-term application of pesticides. The relationship between the content of pesticide residues in the soil and the microbial community structure was analyzed. The results indicated that the microbial diversity was strongly negatively correlated with the contents of pesticide residues in the soil. At a suitable dosage of 5 kg fresh A. imbricata per square meter of soil area, the soil microbial diversity increased by 12.0%, and the contents of pesticide residues decreased by 26.8-72.1%. Sphingobacterium, Sphingopyxis, Thermincola, Sphingobium, Acaryochloris, Megasphaera, Ralstonia, Pseudobutyrivibrio, Desulfitobacterium, Nostoc, Oscillochloris, and Aciditerrimonas may play major roles in the degradation of pesticide residues. Thauera, Levilinea, Geothrix, Thiobacillus, Thioalkalispira, Desulfobulbus, Polycyclovorans, Fluviicola, Deferrisoma, Erysipelothrix, Desulfovibrio, Cytophaga, Vogesella, Zoogloea, Azovibrio, Halomonas, Paludibacter, Crocinitomix, Haliscomenobacter, Hirschia, Silanimonas, Alkalibacter, Woodsholea, Peredibacter, Leptolinea, Chitinivorax, Candidatus_Lumbricincola, Anaerovorax, Propionivibrio, Parasegetibacter, Byssovorax, Runella, Leptospira, and Nitrosomonas may be indicators to evaluate the contents of pesticide residues.

Early-warning biomarkers, such as mitochondrial activity, have become a key tool in ecosystem assessment. This study aims to evaluate the response of mitochondrial activity in spores of the autochthonous fern Cyathea costaricensis as a bioassessment tool concurrently with land use and physicochemical evaluation in 11 sites along Bobos River, Veracruz, Mexico, to assess river water quality. Bobos River is located in the Nautla basin, northeastern Veracruz (Mexico); the upper river runs through a protected natural area (Filobobos River and adjacent areas). The study involved three monitoring periods: February, June and September 2014. In each study site, physicochemical water quality parameters were recorded to calculate the Water Quality Index (WQI); also, study sites were characterized in terms of land use. Water samples were collected to perform bioassays where spores of C. costaricensis were exposed to samples to assess mitochondrial activity; a positive control exposure test was run under controlled conditions to maximize mitochondrial activity. A Principal Component Analysis was performed to correlate land-use attributes with environmental variables and mitochondrial activity. Three river sections were identified: the upper portion was characterized by the dominance of native vegetation, the highest WQI (in September), and the lowest mitochondrial activity (63.87%-77.47%), related to the geological nature of the basin and high hardness levels. Mitochondrial activity peaked in September (98.32% ± 9.01), likely resulting from nutrient enrichment in the rainy season, and was lowest in February (74.54% ± 1.60) (p < 0.05). Mitochondrial activity was found to be a good benchmark for the assessment of water quality, reflecting the effects of physicochemical characteristics. Mitochondrial activity showed changes along the river and between seasons, associated with environmental characteristics such as land use and the geological nature of the basin, as well as with those related to human impacts., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Lead (Pb) is one of the most serious environmental pollutants. The aquatic fern Salvinia minima Baker is capable to hyper-accumulate Pb in their tissues. However, the molecular mechanisms involved in its Pb accumulation and tolerance capacity are not fully understood. In order to investigate the molecular mechanisms that are activated by S. minima in response to Pb, we constructed a suppression subtractive hybridization library (SSH) in response to an exposure to 40μM of Pb(NO 3 ) 2 for 12h. 365 lead-related differentially expressed sequences tags (ESTs) were isolated and sequenced. Among these ESTs, 143 unique cDNA (97 were registered at the GenBank and 46 ESTs were not registered, because they did not meet the GenBank conditions). Those ESTs were identified and classified into 3 groups according to Blast2GO. In terms of metabolic pathways, they were grouped into 29 KEGG pathways. Among the ESTs, we identified some that might be part of the mechanism that this fern may have to deal with this metal, including abiotic-stress-related transcription factors, some that might be involved in tolerance mechanisms such as ROS scavenging, membrane protection, and those of cell homeostasis recovery. To validate the SSH library, 4 genes were randomly selected from the library and analyzed by qRT-PCR. These 4 genes were transcriptionally up-regulated in response to lead in at least one of the two tested tissues (roots and leaves). The present library is one of the few genomics approaches to study the response to metal stress in an aquatic fern, representing novel molecular information and tools to understand the molecular physiology of its Pb tolerance and hyperaccumulation capacity. Further research is required to elucidate the functions of the lead-induced genes that remain classified as unknown, to perhaps reveal novel molecular mechanisms of Pb tolerance and accumulation capacity in aquatic plants., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Cyrtomium macrophyllum naturally grown in 225.73 mg kg -1 of soil mercury in mining area was found to be a potential mercury accumulator plant with the translocation factor of 2.62 and the high mercury concentration of 36.44 mg kg -1 accumulated in its aerial parts. Pot experiments indicated that Cyrtomium macrophyllum could even grow in 500 mg kg -1 of soil mercury with observed inhibition on growth but no obvious toxic effects, and showed excellent mercury accumulation and translocation abilities with both translocation and bioconcentration factors greater than 1 when exposed to 200 mg kg -1 and lower soil mercury, indicating that it could be considered as a great mercury accumulating species. Furthermore, the leaf tissue of Cyrtomium macrophyllum showed high resistance to mercury stress because of both the increased superoxide dismutase activity and the accumulation of glutathione and proline induced by mercury stress, which favorited mercury translocation from the roots to the aerial parts, revealing the possible reason for Cyrtomium macrophyllum to tolerate high concentration of soil mercury. In sum, due to its excellent mercury accumulation and translocation abilities as well as its high resistance to mercury stress, the use of Cyrtomium macrophyllum should be a promising approach to remediating mercury polluted soils., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The spores of Lygodium japonicum (Thunb.) Sw. (L. japonicum) have been used in traditional Chinese medicine for the treatment of various inflammatory diseases. However, the molecular mechanisms underlying their anti‑inflammatory effects have yet to be elucidated. In the present study, we investigated the anti‑inflammatory effects of ethanol extracts of L. japonicum spores (ELJ) by measuring the production of inflammatory mediators, and explored the molecular mechanisms underlying the effects of ELJ in murine macrophages in vitro using immunoblotting analyses. At non‑cytotoxic concentrations of (50‑300 µg/ml), ELJ was revealed to significantly suppress the production of nitric oxide (NO) and tumor necrosis factor (TNF)‑α in lipopolysaccharide (LPS)‑stimulated murine RAW 264.7 macrophages; ELJ repressed the production of interleukin (IL)‑6 only at high concentrations (≥200 µg/ml). The ELJ‑mediated decrease in NO production was demonstrated to depend on the downregulation of inducible NO synthase mRNA and protein expression. Conversely, the mRNA and protein expression of cyclooxygenase‑2 were not affected by ELJ. In addition, ELJ was revealed to inhibit the mRNA expression of IL‑6, IL‑1β, and TNF‑α in LPS‑stimulated RAW 264.7 macrophages. The effects of ELJ on proinflammatory mediators may have been due to the stabilization of inhibitor of κBα and the inhibition of p38 mitogen‑activated protein kinase (MAPK). These results suggested that ELJ may suppress LPS‑induced inflammatory responses in murine macrophages in vitro, through the negative regulation of p38 MAPK and nuclear factor (NF)‑κB. Therefore, ELJ may have potential as a novel candidate for the development of therapeutic strategies aimed at alleviating inflammation.

A pot experiment was conducted to evaluate the effect of urea on nitrogen metabolism and membrane lipid peroxidation in Azolla pinnata. Compared to controls, the application of urea to A. pinnata resulted in a 44% decrease in nitrogenase activity, no significant change in glutamine synthetase activity, 660% higher glutamic-pyruvic transaminase, 39% increase in free amino acid levels, 22% increase in malondialdehyde levels, 21% increase in Na+/K+- levels, 16% increase in Ca2+/Mg2+-ATPase levels, and 11% decrease in superoxide dismutase activity. In terms of H2O2 detoxifying enzymes, peroxidase activity did not change and catalase activity increased by 64% in urea-treated A. pinnata. These findings suggest that urea application promotes amino acid metabolism and membrane lipid peroxidation in A. pinnata.

Nuclear-encoded plant FtsZ genes are derived from endosymbiotic gene transfer of cyanobacteria-like genes. The green lineage (Chloroplastida) and red lineage (Rhodophyta) feature FtsZ1 and FtsZ2 or FtsZB and FtsZA, respectively, which are involved in plastid division. These two proteins show slight differences and seem to heteropolymerize to build the essential inner plastid division ring. A third gene, encoding FtsZ3, is present in glaucophyte and charophyte algae, as well as in land plants except ferns and angiosperms. This gene was probably present in the last common ancestor of the organisms united by having a primary plastid (Archaeplastida) and was lost during vascular plant evolution as well as in the red and green algae. The presence/absence pattern of FtsZ3 mirrors that of a full set of Mur genes and the peptidoglycan wall encoded by them. Based on these findings, we discuss a role for FtsZ3 in the establishment or maintenance of plastid peptidoglycan shells.

In higher plants, the electron-sink capacity of photorespiration contributes to alleviation of photoinhibition by dissipating excess energy under conditions when photosynthesis is limited. We addressed the question at which point in the evolution of photosynthetic organisms photorespiration began to function as electron sink and replaced the flavodiiron proteins which catalyze the reduction of O 2 at photosystem I in cyanobacteria. Algae do not have a higher activity of photorespiration when CO 2 assimilation is limited, and it can therefore not act as an electron sink. Using land plants (liverworts, ferns, gymnosperms, and angiosperms) we compared photorespiration activity and estimated the electron flux driven by photorespiration to evaluate its electron-sink capacity at CO 2 -compensation point. In vivo photorespiration activity was estimated by the simultaneous measurement of O 2 -exchange rate and chlorophyll fluorescence yield. All C3-plants leaves showed transient O 2 -uptake after actinic light illumination (post-illumination transient O 2 -uptake), which reflects photorespiration activity. Post-illumination transient O 2 -uptake rates increased in the order from liverworts to angiosperms through ferns and gymnosperms. Furthermore, photorespiration-dependent electron flux in photosynthetic linear electron flow was estimated from post-illumination transient O 2 -uptake rate and compared with the electron flux in photosynthetic linear electron flow in order to evaluate the electron-sink capacity of photorespiration. The electron-sink capacity at the CO 2 -compensation point also increased in the above order. In gymnosperms photorespiration was determined to be the main electron-sink. C3-C4 intermediate species of Flaveria plants showed photorespiration activity, which intermediate between that of C3- and C4-flaveria species. These results indicate that in the first land plants, liverworts, photorespiration started to function as electron sink. According to our hypothesis, the dramatic increase in partial pressure of O 2 in the atmosphere about 0.4 billion years ago made it possible to drive photorespiration with higher activity in liverworts., (© 2017 Scandinavian Plant Physiology Society.)

Effects of seasonality and increasing Cr(VI) concentrations on leaf starch-sucrose partitioning, sucrose- and starch-related enzyme activities, and carbon allocation toward leaf development were analyzed in fronds (floating leaves) of the floating fern Salvinia minima. Carbohydrates and enzyme activities of Cr-exposed fronds showed different patterns in winter and summer. Total soluble sugars, starch, glucose and fructose increased in winter fronds, while sucrose was higher in summer ones. Starch and soluble carbohydrates, except glucose, increased under increasing Cr(VI) concentrations in winter fronds, while in summer ones only sucrose increased under Cr(VI) treatment. In summer fronds starch, total soluble sugars, fructose and glucose practically stayed without changes in all assayed Cr(VI) concentrations. Enzyme activities related to starch and sucrose metabolisms (e.g. ADPGase, SPS, SS and AI) were higher in winter fronds than in summer ones. Total amylase and cFBPase activities were higher in summer fronds. Cr(VI) treatment increased enzyme activities, except ADPGase, in both winter and summer fronds but no clear pattern changes were observed. Data of this study show clearly that carbohydrate metabolism is differently perturbed by both seasonality and Cr(VI) treatment in summer and winter fronds, which affects leaf starch-sucrose partitioning and specific leaf area (SLA) in terms of carbon investment., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

Abscisic acid (ABA)-driven stomatal regulation reportedly evolved after the divergence of ferns, during the early evolution of seed plants approximately 360 million years ago. This hypothesis is based on the observation that the stomata of certain fern species are unresponsive to ABA, but exhibit passive hydraulic control. However, ABA-induced stomatal closure was detected in some mosses and lycophytes. Here, we observed that a number of ABA signaling and membrane transporter protein families diversified over the evolutionary history of land plants. The aquatic ferns Azolla filiculoides and Salvinia cucullata have representatives of 23 families of proteins orthologous to those of Arabidopsis ( Arabidopsis thaliana ) and all other land plant species studied. Phylogenetic analysis of the key ABA signaling proteins indicates an evolutionarily conserved stomatal response to ABA. Moreover, comparative transcriptomic analysis has identified a suite of ABA-responsive genes that differentially expressed in a terrestrial fern species, Polystichum proliferum These genes encode proteins associated with ABA biosynthesis, transport, reception, transcription, signaling, and ion and sugar transport, which fit the general ABA signaling pathway constructed from Arabidopsis and Hordeum vulgare The retention of these key ABA-responsive genes could have had a profound effect on the adaptation of ferns to dry conditions. Furthermore, stomatal assays have shown the primary evidence for ABA-induced closure of stomata in two terrestrial fern species P proliferum and Nephrolepis exaltata In summary, we report, to our knowledge, new molecular and physiological evidence for the presence of active stomatal control in ferns., (© 2017 American Society of Plant Biologists. All Rights Reserved.)

Premise of the Study: This paper investigates the occurrence and evolution of aluminum (Al) accumulation within ferns and lycophytes, which is characterized by Al concentrations above 1000 mg·kg -1 in aboveground plant tissues. We hypothesize that this feature is more common in ferns than in angiosperms, and potentially correlated with growth form and other chemical elements., Methods: Aluminum concentrations were obtained from novel analyses and literature for a total of 354 specimens and 307 species. Moreover, a semiquantitative aluminon test was applied for a subset of 105 species and validated against exact Al measurements., Key Results: Molecular phylogenetic analyses showed that the major Al-accumulating groups were primarily found in the Gleicheniales and Cyatheales, and largely absent in the Polypodiales. At the species and generic level, Al accumulation was typically either absent or present, and mixed results within a single species and genus were limited to less than 30% of the species and genera tested. Epiphytic ferns had significantly lower Al levels than terrestrial ferns, although this finding was not significant after phylogenetic correction. In addition, a significant, positive correlation was found between Al and iron, while Al was negatively correlated with phosphorus and potassium concentrations., Conclusions: Aluminum accumulation is most common outside of the Polypodiales and occurs in 38% of the species studied, indicating that this trait is indeed common within subtropical and tropical ferns, a finding that could be in line with their role as pioneer species on landslides and soils with high levels of soluble Al., (© 2017 Botanical Society of America.)

Removal of Cr(VI) species by dried biomass of the aquatic macrophyte Salvinia auriculata was studied in order to understand the involved sorption mechanisms. Kinetic tests were carried out under the conditions such as concentration range of Cr(VI) from 50 to 250 mg L -1 and a temperature of 30 °C. Modification of the biosorbent by the presence of Cr(VI) species was assessed by analysis of its porosity, density and infrared molecular absorption spectrum. A series of experimental approaches involving directed chemical modifications on the biosorbent surface was performed. The main functional groups involved in the sorption mechanisms were identified. The gas sorption analyser was applied and proved that a strong chemical effect of Cr(VI) species on the surface took place, resulting in a leaching organic matter with an obvious and significant increase in the porosity parameters. The intra-particle diffusion model revealed different mass transfer zones into the adsorbent during Cr(VI) removal. New combined Langmuir and Dubinin-Radushkevich isotherm was the best to fit the equilibrium data of Cr(VI) species removal. Finally, Cr(VI) removal was mainly mediated by a redox process where Cr(III) species were formed., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Asexual reproduction is widespread in land plants, including ferns where 10% of all species are obligate asexuals. In these ferns, apogamous sporophytes are generated directly from gametophytes, bypassing fertilization. In the model fern Ceratopteris richardii, a sexual species, apogamy can be induced by culture on high sugar media. BABY BOOM (BBM) genes in angiosperms are known to promote somatic embryogenesis, which like apogamy produce sporophytes without fertilization. Here, a Brassica napus BBM (BnBBM) was used to investigate genetic similarity between apogamy in ferns and somatic embryogenesis in angiosperms. A C. richardii transcriptome was constructed from which one AINTEGUMENTA-LIKE unigene, CrANT, was identified. Whole mount in situ hybridization showed that CrANT is expressed in sperm and fertilized eggs. Phylogenetic analysis grouped CrANT with other non-seed-plant ANT genes to the euANT clade but in a branch separate from BBM genes. Overexpression of CrANT or BnBBM promotes apogamy in C. richardii without sugar supplement. CrANT knockdown gametophytes responded weakly to sugar for apogamy promotion. Theses results suggest some genetic conservation between apogamy and somatic embryogenesis and that such asexual reproduction may be ancient., (© 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.)

This report describes the development of lab-on-a-chip device designed to measure changes in cellular ion gradients that are induced by changes in gravitational (g) forces. The bioCD presented here detects differential calcium ion concentrations outside of individual cells. The device includes sufficient replicates for statistical analysis of the gradients around multiple single cells and around control wells that are empty or include dead cells. In the data presented, the degree of the cellular response correlates with the magnitude of the g-force applied via rotation of the bioCD. The experiments recorded the longest continuous observation of a cellular response to hypergravity made to date, and they demonstrate the potential utility of this device for assaying the threshold of cells' g-force responses in spaceflight conditions.

Fern spores were traditionally classified into chlorophyllous (green) and nonchlorophyllous (nongreen) types based on the color visible to the naked eye. Recently, a third type, "cryptochlorophyllous spores", is recognized, and these spores are nongreen under white light but contain chlorophylls. Epifluorescence microscopy was previously used to detect chlorophylls in cryptochlorophyllous spores. In addition to epifluorescence microscopy, current study performed some other approaches, including spore-squash epifluorescence, absorption spectra, laser-induced fluorescence emission spectra, thin layer chromatography (TLC), and ultra-high performance liquid chromatography with ultraviolet and mass spectrometric detection (UHPLC-UV-MS) in order to detect chlorophylls of spores of seven ferns (Sphaeropteris lepifera, Ceratopteris thalictroides, Leptochilus wrightii, Leptochilus pothifolius, Lepidomicrosorum buergerianum, Osmunda banksiifolia, and Platycerium grande). Destructive methods, such as TLC and UHPLC-UV-MS, successfully detected chlorophylls inside the spores when their signals of red fluorescence under epifluorescence microscope were masked by spore wall. Although UHPLC-UV-MS analysis was the most sensitive and reliable for determining the chlorophylls of spores, spore-squash epifluorescence is not only reliable but also cost- and time-effective one among our study methods. In addition, we first confirmed that Lepidomicrosorium buergerianum, Leptochilus pothifolius, Leptochilus wrightii, and Platycerium grande, produce cryptochlorophyllous spores.

Mosquitoes act as vectors of devastating pathogens and parasites, representing a key threat for millions of humans and animals worldwide. Eco-friendly control tools are urgently required. We proposed a novel method of fern-mediated biosynthesis of silver nanoparticles (AgNP) using Dicranopteris linearis, acting as a reducing and capping agent. AgNP were characterized by UV-vis spectroscopy, Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), zeta potential and particle size analysis. In mosquitocidal assays, the LC 50 of D. linearis extract against Aedes aegypti ranged from 165.213 (larva I) to 255.055ppm (pupa). LC 50 of D. linearis-synthesized AgNP ranged from 18.905 (larva I) to 29.328ppm (pupa). In the field, the application of D. linearis extract and AgNP (10×LC 50 ) led to 100% larval reduction after 72h. Smoke toxicity experiments conducted against A. aegypti adults showed that D. linearis leaf-, stem- and root-based coils evoked mortality rates comparable to the permethrin-based positive control (58%, 47%, 34%, and 48% respectively). In ovicidal experiments, egg hatchability was reduced by 100% after treatment with 25ppm of AgNP and 300ppm of D. linearis extract. Interestingly, oviposition deterrent assays highlighted that 100ppm of fern extract reduced oviposition rates of more than 65%, while 10ppm of fern-fabricated AgNP reduced oviposition rates of more than 70% in A. aegypti (OAI were -0.52 and -0.55, respectively). Overall, our results highlighted that D. linearis-synthesized AgNP could be useful candidates to develop nano-formulated oviposition deterrents effective against dengue vectors., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

We examined the potential of Azolla japonica as a remediating plant for leachate channels and post-accumulation use as fertilizer for landfill slope. The harvested biomass of Azolla after one month grown in leachate was 254% that of the initial biomass and the predicted annual harvestable biomass of Azolla using a growth model was 32 times that of the initial biomass. Na, Fe, Mn, Mg, and P were accumulated in Azolla at very high concentrations. Such rapid increase of biomass and high accumulation rates suggest that this plant could be an excellent remediating plant. The post-harvest use of Azolla as compost was studied for the management and use of phytoaccumulating Azolla. Metal contents of Azolla compost were below permissible limits for co-composting material. Nitrogen, organic matter, P, and Mg content of the Azolla compost improved the soil condition of the landfill and enhanced ecophysiological responses of the plants. The application of Azolla compost can improve management of sanitary landfills, including the restoration of vegetation. Considering its ease of harvesting, high accumulation rates, harvestable biomass and suitability for composting, Azolla can provide a suitable solution for sustainable management of leachate channels and landfill slopes., (Copyright © 2016. Published by Elsevier Ltd.)

Despite the extraordinary significance leaves have for life on Earth, their origin and development remain vigorously debated. More than a century of paleobotanical, morphological, and phylogenetic research has still not resolved fundamental questions about leaves. Developmental genetic data are sparse in ferns, and comparative studies of lycophytes and seed plants have reached opposing conclusions on the conservation of a leaf developmental program. We performed phylogenetic and expression analyses of a leaf developmental regulator (Class III HD-Zip genes; C3HDZs) spanning lycophytes and ferns. We show that a duplication and neofunctionalization of C3HDZs probably occurred in the ancestor of euphyllophytes, and that there is a common leaf developmental mechanism conserved between ferns and seed plants. We show C3HDZ expression in lycophyte and fern sporangia and show that C3HDZs have conserved expression patterns during initiation of lateral primordia (leaves or sporangia). This expression is maintained throughout sporangium development in lycophytes and ferns and indicates an ancestral role of C3HDZs in sporangium development. We hypothesize that there is a deep homology of all leaves and that a sporangium-specific developmental program was coopted independently for the development of lycophyte and euphyllophyte leaves. This provides molecular genetic support for a paradigm shift in theories of lycophyte leaf evolution., (© 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.)

Whitefly (Bemisia tabaci) damages field crops by sucking sap and transmitting viral diseases. None of the insecticidal proteins used in genetically modified (GM) crop plants to date are effective against whitefly. We report the identification of a protein (Tma12) from an edible fern, Tectaria macrodonta (Fee) C. Chr., that is insecticidal to whitefly (median lethal concentration = 1.49 μg/ml in in vitro feeding assays) and interferes with its life cycle at sublethal doses. Transgenic cotton lines that express Tma12 at ∼0.01% of total soluble leaf protein were resistant to whitefly infestation in contained field trials, with no detectable yield penalty. The transgenic cotton lines were also protected from whitefly-borne cotton leaf curl viral disease. Rats fed Tma12 showed no detectable histological or biochemical changes, and this, together with the predicted absence of allergenic domains in Tma12, indicates that Tma12 might be well suited for deployment in GM crops to control whitefly and the viruses it carries.

Salvinia molesta, an aquatic fern was observed to have a potential of degrading azo dye Rubine GFL up to 97% at a concentration of 100mg/L within 72h using 60±2g of root biomass. Both root as well as stem tissues showed induction in activities of the enzymes such as lignin peroxidase, veratryl alcohol oxidase, laccase, tyrosinase, catalase, DCIP reductase and superoxide dismutase during decolorization of Rubine GFL. FTIR, GC-MS, HPLC and UV-visible spectrophotometric analysis confirmed phytotransformation of the model dye into smaller molecules. Analysis of metabolites revealed breakdown of an azo bond of Rubine GFL by the action of lignin peroxidase and laccase and formation of 2-methyl-4-nitroaniline and N-methylbenzene-1, 4-diamine. Anatomical tracing of dye in the stem of S. molesta confirmed the presence of dye in tissues and subsequent removal after 48h of treatment. The concentration of chlorophyll pigments like chlorophyll a, chlorophyll b and carotenoid was observed during the treatment. Toxicity analysis on seeds of Triticum aestivum and Phaseolus mungo revealed the decreased toxicity of dye metabolites. In situ treatment of a real textile effluent was further monitored in a constructed lagoon of the dimensions of 7m×5m×2m (total surface area 35m(2)) using S. molesta for 192h. This large scale treatment was found to significantly reduce the values of COD, BOD5 and ADMI by 76%, 82% and 81% considering initial values 1185, 1440mg/L and 950 units, respectively., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Nutrients can determine the outcome of the competition between different floating plant species. The response of floating plants to current phosphorus levels may be affected by previously experienced phosphorus concentrations because some species have the ability to store excess phosphorus for later use. This might have an impact on their competition. Here, we investigate the effect of previous and actual phosphorus concentrations on the growth rate of free-floating plant species (Azolla filiculoides, Lemna minor/gibba and Ricciocarpus natans)and the effect of phosphorus history on the competition between L. minor/gibba and A. filiculoides and between L. minor/gibba and R. natans. As expected, plant growth was lower when previously kept at low instead of high phosphorus concentrations. Growth of L. minor/gibba and A. filiculoides with a phosphorus rich history was comparable for low and high actual phosphorus concentrations, however, internal phosphorus concentrations were significantly lower with low actual phosphorus concentration. This indicates that both species perform luxury phosphorus uptake. Furthermore, internal P concentration in Azolla and Lemna increased within two weeks after a period of P deficit without a strong increase in growth. A. filiculoides in a mixture with L. minor/gibba grew faster than its monoculture. Morphological differences may explain why A. filiculoides outcompeted L. minor/gibba and these differences may be induced by phosphorus concentrations in the past. Growth of L. minor/gibba was only reduced by the presence of A. filiculoides with a high phosphorus history. Growth of L. minor/gibba and R. natans in mixtures was positively affected only when they had a high phosphorus history themselves and their competitor a low phosphorus history. These observations clearly indicate that phosphorus history of competing plants is important for understanding the outcome of the competition. Therefore, actual and previously experienced phosphorus concentrations should be taken into account in future studies dealing with competition between plants., Competing Interests: The authors have declared that no competing interests exist.

Premise of the Study: California experienced severe drought between 2012 and 2016. During this period, we compared seasonal changes in tissue-water relations among eight fern species in the Santa Monica Mountains of southern California to elucidate differential mechanisms of drought survival and physiological performance during extreme water deficits., Methods: We monitored seasonal changes in water potential (Ψmd) and dark-adapted chlorophyll fluorescence (Fv/Fm), assessed tissue-water relations including osmotic potential at saturation and the turgor loss point (Ψπ, sat and Ψπ, tlp), and measured, for two evergreen species, xylem-specific and leaf-specific hydraulic conductivity (Ks and Kl) and vulnerability of stem xylem to water stress-induced embolism (water potential at 50% loss hydraulic conductivity, Ψ50)., Key Results: Species grew in either riparian or chaparral understory. The five chaparral species had a wider range of seasonal water potentials, root depths, and frond phenological traits, including one evergreen, two summer-deciduous, and two desiccation-tolerant (resurrection) species. Evergreen species were especially diverse, with an evergreen riparian species maintaining seasonal water potentials above -1.3 MPa, while an evergreen chaparral species had seasonal water potentials below -8 MPa. In those two species the Ψ50 values were -2.5 MPa and -4.3 MPa, respectively., Conclusions: Observed differences in physiological performance among eight fern species reflected niche partitioning in water utilization and habitat preference associated with distinct phenological traits. We predict differential survival among fern species as future drought events in California intensify, with desiccation-tolerant resurrection ferns being the most resistant., (© 2016 Botanical Society of America.)

Natural products are important sources of chemical diversity leading to unique scaffolds that can be exploited in the discovery of new drug candidates or chemical probes. In this context, chemical and biological investigation of ferns and lycophytes occurring in Brazil is an approach adopted by our research group aiming at discovering bioactive molecules acting on neurodegeneration targets. In the present study, rosmarinic acid (RA) isolated from Blechnum brasiliense showed an in vitro multifunctional profile characterized by antioxidant effects, and monoamine oxidases (MAO-A and MAO-B) and catechol-O-methyl transferase (COMT) inhibition. RA showed antioxidant effects against hydroxyl (HO(•)) and nitric oxide (NO) radicals (IC50 of 29.4 and 140 μM, respectively), and inhibition of lipid peroxidation (IC50 of 19.6 μM). In addition, RA inhibited MAO-A, MAO-B and COMT enzymes with IC50 values of 50.1, 184.6 and 26.7 μM, respectively. The MAO-A modulation showed a non-time-dependent profile, suggesting a reversible mechanism of inhibition. Structural insights on RA interactions with MAO-A and COMT were investigated by molecular docking. Finally, RA (up to 5 mM) demonstrated no cytotoxicity on polymorphonuclear rat cells. Taken together, our results suggest that RA may be exploited as a template for the development of new antioxidant molecules possessing additional MAO and COMT inhibition effects to be further investigated on in vitro and in vivo models of neurodegenerative diseases., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

In plants of Salvinia rotundifolia and Salvinia minima the effect of two Cr(VI) concentrations (5 and 20mgL(-1)) applied for 7days was assessed by measuring changes in biomass, photosynthetic pigments, Cr accumulation, malondialdehyde (MDA), membrane stability index (MSI), thiols (TT, NPT and PBT), and phenolics (SP and IP). Biomass in S. minima was decreased at highest Cr(VI) concentration, but there were no changes in S. rotundifolia. Metal accumulation was different in both species. S. minima accumulates more metal in fronds, but S. rotundifolia accumulates more metal in lacinias. Results also showed that S. minima translocates more Cr to fronds than S. rotundifolia, but at the whole plant level higher accumulation occurred in this last. Tolerance index (Ti) was higher in S. rotundifolia. Chl b and carotenoids were decreased only upon exposure to high Cr(VI) concentration in both species. Cr(VI) treatment did not enhance MDA accumulation. Cr exposure had no impact on MSI values when comparing with Cr-untreated values. Thiols in fronds and lacinias showed different distribution patterns between species. IP and NPT were higher in S. rotundifolia lacinias that accumulate more Cr than S. minima lacinias. Whilst SP and NPT were higher in S. minima fronds compared with S. rotundifolia ones. This may indicate that these species can cope with Cr(VI) toxicity, either through metal complexation and/or metal reduction or by the scavenging of ROS derived from Cr-induced oxidative stress. Based on Cr accumulation and biomass production, S. rotundifolia seems more suitable to remove Cr(VI) from polluted waters., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Hymenophyllaceae is a desiccation tolerant family of Pteridophytes which are poikilohydric epiphytes. Their fronds are composed by a single layer of cells and lack true mesophyll cells and stomata. Although they are associated with humid and shady environments, their vertical distribution varies along the trunk of the host plant with some species inhabiting the drier sides with a higher irradiance. The aim of this work was to compare the structure and function of the photosynthetic apparatus during desiccation and rehydration in two species, Hymenophyllum dentatum and Hymenoglossum cruentum, isolated from a contrasting vertical distribution along the trunk of their hosts. Both species were subjected to desiccation and rehydration kinetics to analyze frond phenotypic plasticity, as well as the structure, composition and function of the photosynthetic apparatus. Minimal differences in photosynthetic pigments were observed upon dehydration. Measurements of ϕPSII (effective quantum yield of PSII), ϕNPQ (quantum yield of the regulated energy dissipation of PSII), ϕNO (quantum yield of non-regulated energy dissipation of PSII), and TL (thermoluminescence) indicate that both species convert a functional photochemical apparatus into a structure which exhibits maximum quenching capacity in the dehydrated state with minimal changes in photosynthetic pigments and polypeptide compositions. This dehydration-induced conversion in the photosynthetic apparatus is completely reversible upon rehydration. We conclude that H. dentatum and H. cruentum are homoiochlorophyllous with respect to desiccation stress and exhibited no correlation between inherent desiccation tolerance and the vertical distribution along the host tree trunk., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Current study presents an economic, ecofriendly and simple photo-catalytic green route for the swift biosynthesis of silver nanoparticles (AgNPs) within 20s, devoid of any instrumental support or chemical reductant. Aqueous leaf-extract of an aquatic fern, Salvinia molesta (AES), was used as a bioreductant as well as a stabilizing agent. Rapid change in color of reaction mixture from yellowish green to reddish brown within 20s in direct sun light exposure was considered as the primary visual indication of AgNPs biosynthesis. The biosynthesis of AgNPs was confirmed by UV-visible spectroscopy through the presence of a characteristic surface plasmon resonance (SPR) band for AgNPs at λmax of 425 nm. The process parameters were optimized through one factor at a time approach. Optimal values of different process parameters for the current biosynthetic system were found as; 35 min of reaction time under sun light, 8.0mM AgNO3 concentration and 5.0% (v/v) AES inoculum dose. Field emission scanning electron microscopy (FESEM), energy dispersive X-Ray spectroscopy (EDX), high resolution transmission electron microscopy (HRTEM), atomic force microscopy (AFM) and X-ray diffraction (XRD) analysis showed that most of AgNPs were spherical in shape with average size distribution of 12.46 nm having face centered cubic (fcc) crystal lattice. IR analysis of AES and synthesized AgNPs indicated the involvement of both hydroxyl and amino groups in the biosynthesis and stabilization of AgNPs. The synthesized AgNPs were found to be an effective antibacterial agent against both Gram positive and Gram negative bacteria. On the basis of results and facts, a probable mechanism has also been proposed to explore the possible route of biosynthesis of AgNPs through AES., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Two constructed wetlands, one with Azolla pinnata plant (CW1) and the other without (CW2) for treating domestic wastewaters were developed. Fifteen water parameters which include: Dissolved Oxygen (DO), Biochemical Oxygen Demand (BOD5), Chemical Oxygen Demand (COD), Total Suspended Solid (TSS), Total Phosphorus (TP), Total Nitrogen (TN), Ammoniacal Nitrogen (NH3N), Turbidity, pH, Electrical Conductivity (EC), Iron (Fe), Magnesium (Mg), Manganese (Mn), and heavy metals such as Lead (Pb) and Zinc (Zn) were analyzed using standard laboratory procedures. The experiments were conducted in two (dry and wet) seasons simultaneously. Results showed considerable reductions in all parameters and metals including Zn in CW1 compared with CW2 in the two seasons considered while Pb and Mn were not detected throughout the study. Zn concentration levels reduced significantly in both seasons just as removal efficiencies of 70.03% and 64.51% were recorded for CW1 while 35.17% and 33.45% were recorded for CW2 in both seasons. There were no significant differences in the removal efficiencies of Fe in both seasons as 99.55%, 59.09%, 88.89%, and 53.56% were recorded in CW1 and CW2 respectively. Azolla pinnata has proved effective in domestic wastewater phytoremediation studies.

The aim of this work was to compare Cr(III) and Cr(VI) removal kinetics from water by Pistia stratiotes and Salvinia herzogii. The accumulation in plant tissues and the effects of both Cr forms on plant growth were also evaluated. Plants were exposed to 2 and 6 mg L(-1) of Cr(III) or Cr(VI) during 30 days. At the end of the experiment, Cr(VI) removal percentages were significantly lower than those obtained for Cr(III) for both macrophytes. Cr(III) removal kinetics involved a fast and a slow component. The fast component was primarily responsible for Cr(III) removal while Cr(VI) removal kinetics involved only a slow process. Cr accumulated principally in the roots. In the Cr(VI) treatments a higher translocation from roots to aerial parts than in Cr(III) treatments was observed. Both macrophytes demonstrated a high ability to remove Cr(III) but not Cr(VI). Cr(III) inhibited the growth at the highest studied concentration of both macrophytes while Cr(VI) caused senescence. These results have important implications in the use of constructed wetlands for secondary industrial wastewater treatment. Common primary treatments of effluents containing Cr(VI) consists in its reduction to Cr(III). Cr(III) concentrations in these effluents are normally below the highest studied concentrations in this work.

In this study, the phytoremediation capacity of Azolla filiculoides Lam. for the water resources contaminated with petroleum hydrocarbons was investigated. The plants were grown in nitrogen-free Hoagland nutrient solution containing 0.005%, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, and 0.5% crude oil under greenhouse conditions for 15 days. Although the growth rate of the plants were not negatively influenced by the presence of crude oil in the media for the concentration of 0.005% and 0.01% v/v, a gradual impeding effect of crude oil in the growth media has been observed at concentrations 0.05-0.1%. More than 0.1% crude oil in the growth medium ostensibly retarded the growth. For example, 0.2% oil in the media reduced growth approximately 50% relative to the control, and the presence of crude oil at concentrations 0.3% or more were lethal. The data about the percentage of plant growth, fresh weight increase and root growth clearly indicated that the tolerance level of A. filiculoides plants to crude oil ranges between 0.1% and 0.2%. In comparison to control samples, the biodegradation rate of total aliphatic and aromatic (phenathrene) hydrocarbons at 0.05-0.2% oil concentrations, was 94-73% and 81-77%, respectively. On the other hand, in case of further increases in oil concentration in media, i.e.; 0.3-0.5%, the biodegradation rate was still higher in the experimental samples, respectively 71-63% and 75-71%. The high biodegradation rates of petroleum hydrocarbons in the experimental samples suggested that A. filiculoides plants could be a promising candidate to be used for the phytoremediation of low crude oil contaminated precious freshwater resources.

Physiological and biochemical effects of cylindrospermopsin (CYN), a cyanobacterial toxin that inhibits protein synthesis and released during a harmful cyanobacterial bloom, has been overlooked in plants. Therefore, at the present research, the toxic effects (physiological and biochemical) of a crude extract containing CYN were assessed in the aquatic fern Azolla filiculoides exposed to three concentrations (0.05, 0.5 and 5 μg CYN mL(-1)). At 5 μg CYN mL(-1), fern growth rate has showed a drastic decrease (0.001 g g(-1) day(-1)) corresponding to a 99.8% inhibition, but at the concentrations of 0.05 and 0.5 μg CYN mL(-1) the growth rate was similar to the control plants. Growth rate also indicated a IC50 of 2.9 μg CYN mL(-1). Those data point to the presence of other compounds in the crude extract may stimulate the fern growth and/or the fern is tolerant to CYN. Chlorophyll (a and b), carotenoids and protein content as well as the activities of glutathione reductase (GR) and glutathione-S-transferase (GST) has increased at 5 μg CYN mL(-1) which may indicate that photosynthesis and protein synthesis are not affected by CYN and the probable activation of defense and detoxifying mechanisms to overcome the effects induced by the presence of CYN. Low uptake of cylindrospermopsin (1.314 μg CYN g(-1) FW) and low bioconcentration factor (0.401) point towards to a safe use of A. filiculoides as biofertilizer and as food source, but also indicate that the fern is not suitable for CYN phytoremediation.

We investigated stomatal conductance (g(s)) and mesophyll conductance (g(m)) in response to atmospheric CO2 concentration [CO2] in two primitive land plants, the fern species Pteridium aquilinum and Thelypteris dentata, using the concurrent measurement of leaf gas exchange and carbon isotope discrimination. [CO2] was initially decreased from 400 to 200 μmol mol(-1), and then increased from 200 to 700 μmol mol(-1), and finally decreased from 700 to 400 μmol mol(-1). Analysis by tunable diode laser absorption spectroscopy (TDLAS) revealed a rapid and continuous response in g m within a few minutes. In most cases, both ferns showed rapid and significant responses of g m to changes in [CO2]. The largest changes (quote % decrease) were obtained when [CO2] was decreased from 400 to 200 μmol mol(-1). This is in contrast to angiosperms where an increase in g(m) is commonly observed at low [CO2]. Similarly, fern species observed little or no response of g(s) to changes in [CO2] whereas, a concomitant decline of g(m) and g(s) with [CO2] is often reported in angiosperms. Together, these results suggest that regulation of g(m) to [CO2] may differ between angiosperms and ferns.

Azolla, a small water fern, abscises its roots and branches within 30 min upon treatment with various stresses. This study was conducted to test whether, in the rapid abscission that occurs in Azolla, breakdown of wall components of abscission zone cells by (●) OH is involved. Experimentally generated (●) OH caused the rapid separation of abscission zone cells from detached roots and the rapid shedding of roots from whole plants. Electron microscopic observations revealed that (●) OH rapidly and selectively dissolved a well-developed middle lamella between abscission zone cells and resultantly caused rapid cell separation and shedding. Treatment of abscission zones of Impatiens leaf petiole with (●) OH also accelerated the separation of abscission zone cells. However, compared with that of Azolla roots, accelerative effects in Impatiens were weak. A large amount of (●) OH was cytochemically detected in abscission zone cells both of Azolla roots and of Impatiens leaf petioles. These results suggest that (●) OH is involved in the cell separation process not only in the rapid abscission in Azolla but also in the abscission of Impatiens. However, for rapid abscission to occur, a well-developed middle lamella, a unique structure, which is sensitive to the attack of (●) OH, might be needed., (© 2015 John Wiley & Sons Ltd.)

Athyrium wardii is one of the dominant plant species flourishing on the Pb-Zn mine tailings in Sichuan Province, China. A greenhouse pot experiment was conducted to evaluate the chemical forms, subcellular distribution, and thiol compounds in A. wardii under different Pb treatments. The results showed that plants of the mining ecotype (ME) of A. wardii were more tolerant to Pb than those of the non-mining ecotype (NME) in spite of accumulation of higher Pb concentrations. The Pb concentrations in shoots and roots of the ME were 3.2∼8.6 times and 3.0∼24.6 times higher than those of the NME, respectively. The ME was more efficient in Pb uptake than the NME. Moreover, 27.8∼39.0% of the total Pb in ME was sodium chloride (NaCl) extractable and 38.0∼48.5% was acetic acid (HAc) extractable, whereas only a minority of total Pb was in ethanol and H2O extractable. In subcellular level, 77.4∼88.8% of total Pb was stored in the cell walls of ME and 9.0∼18.9% in soluble fractions. Increasing Pb concentrations enhanced sequestration of Pb into the cell walls and soluble fractions of ME tissues to protect organelles against Pb. Synthesis of non-protein thiols (NP-SH) and phytochelatins (PCs) in roots of ME significantly enhanced in response to Pb stress, and significant increases in glutathione (GSH) were observed in shoots of ME. Higher levels of NP-SH, GSH, and PCs were observed in roots of the ME comparing with NME, especially under high Pb treatments. The results indicated that Pb was localized mainly in cell wall and soluble fraction of ME plants with low biological activity by cell wall deposition and vacuolar compartmentalization, which might be the important adapted Pb detoxification mechanisms of ME.