Hart's Tongue Research Articles

Phytotoxicity, cytogenotoxicity and antimicrobial potential of extracts with gold-silver bimetallic nanoparticles obtained from pteridophyte spores

Investigating the toxicity of naturally occurring or synthesized nanoparticles for various applications is absolutely necessary for environmental protection and safety use. The aim of these research was to investigated the phytotoxicity, cytogenotoxicity and antibacterial potential of the extracts with gold-silver bimetallic nanoparticles (Au-Ag NPs) obtained from green synthesis in Asplenium scolopendrium L. and Dryopteris filix-mas (L.) Schott spores extracts. To our knowledge, this is the first report of the Au-Ag NPs phytosynthesis based on extracts obtained from fern spores. UV-Vis spectroscopy analysis of the samples revealed the maximum absorbance, characteristic of samples with bimetallic nanoparticles, which varied depending on the Au:Ag ratio. Energy-dispersive X-ray spectroscopy confirmed the presence and distribution of Au, Ag and other chemical elements. The presence of specific secondary metabolites in the extracts that helped in NPs biosynthesis stimulated growth processes. Good results were recorded for some Dryopteris filix-mas samples, correlated with a significantly increased mitotic index. Cell viability decreased significantly in three of the nanoformulations. Only extracts with Au-Ag NPs showed antimicrobial effect against Staphylococcus aureus ATCC 25923, Bacillus subtilis ATCC 6633 and Escherichia coli ATCC 8739. The testing of the antibacterial potential of these extracts must be extended to other bacterial strains and other microorganisms, the search of new antimicrobial resources being an urgent necessity nowadays.

Impacts of Invasive Earthworms on Early Life Stages of the Threatened American Hart's-Tongue Fern

Impacts of Invasive Earthworms on Early Life Stages of the Threatened American Hart's-Tongue Fern

Utilizing Novel Lignocellulosic Material from Hart's-Tongue Fern (Asplenium scolopendrium) Leaves for Crystal Violet Adsorption: Characterization, Application, and Optimization.

In this work, a new lignocellulosic adsorbent was obtained and tested for crystal violet dye removal from water. The material was obtained from hart's-tongue fern (Asplenium scolopendrium) leaves after minimal processing, without chemical or thermal treatment. The surface of the material was characterized using a variety of techniques, including FTIR, SEM, and color analysis. The effect of various factors on the adsorption capacity was then investigated and discussed. The kinetic and equilibrium studies showed that the general-order kinetic model and the Sips isotherm are the most suitable to describe the adsorption process. The equilibrium time was reached after 20 min and the maximum calculated value of the adsorption capacity was 224.2 (mg g-1). The determined values for the thermodynamic parameters indicated physical adsorption as the main mechanism involved in the process. The Taguchi method was used to optimize the adsorption conditions and identify the most influential controllable factor, which was pH. ANOVA (general linear model) was used to calculate the percentage contribution of each controllable factor to dye removal efficiency. Analysis of all the results shows that hart's-tongue fern (Asplenium scolopendrium) leaves are a very inexpensive, readily available, and effective adsorbent for removing crystal violet dye from aqueous solutions.

Molecular phylogenetic assessment of three major taxa in the Asplenium scolopendrium complex (Aspleniaceae)

AbstractTaxonomic ambiguities of the Asplenium scolopendrium complex arise from multiple synonyms and reclassifications, subtle phenotypic variations, and a lack of information on phylogenetic relationships. This study thus aimed to resolve this taxonomic uncertainty and provide insight into identifying evolutionarily significant units (ESUs). We first utilized genomic approaches to resolve the phylogenetic position of the East Asian taxon, A. komarovii, which is currently treated as a separate species. Phylogenetic trees based on whole plastomes suggested that A. komarovii is a variant of A. scolopendrium most closely related to A. scolopendrium var. americanum, with A. scolopendrium var. scolopendrium as the sister to these in the clade. This three‐lineage relationship was also validated with the nuclear marker gapCp and newly developed infraspecific plastid markers. Asplenium komarovii should therefore be subsumed into A. scolopendrium, rather than remaining a distinct species. In addition, our phylogenetic analyses further revealed that A. scolopendrium var. americanum consisted of subclades with potential to be treated as distinct ESUs. Our results also grouped a newly discovered population from New Mexico (U.S.A.) as a member of A. scolopendrium var. americanum and identified a genetically admixed population in New York (U.S.A.) containing putative hybrids. Well‐defined taxonomy and ESUs can greatly improve the implementation of tailored conservation actions by adequately reflecting the underlying evolutionary potentials of the A. scolopendrium complex.

Ecotoxicological Risk Assessment of Actellic 50 EC Insecticide on Non-Target Organisms in Parallel with the Application of Standardized Tests.

This paper contributes to the ecotoxicological risk assessment of the Actellic 50 EC insecticide (with 50% pirimiphos-methyl as the active substance) tested on non-target organisms. The insecticide concentrations tested were the same for all organisms (0.1, 0.01, and 0.001 mg L-1 of Actellic 50 EC), with an exposure of 3-5-21 days for plants and 4-5-14 days for animals. The non-target organisms tested were both plants (wheat and two ferns) and animals (the Prussian carp and marsh frog tadpoles). The tested insecticide significantly inhibited the growth of roots in wheat, a result that was also confirmed by a microbiotest application (62% root growth inhibition in sorghum and 100% germination inhibition in white mustard and garden cress). In ferns, even for the lowest concentration, the percentage of germinated spores was inhibited by 40% for Asplenium scolopendrium. The recorded toxicological effects of Actellic 50 EC upon the Prussian carp included a decrease in the respiratory rate and oxygen consumption, an increase in the number of erythrocytes and leukocytes, and an increase in blood glucose levels. The highest concentration (0.1 mg L-1 of Actellic 50 EC) caused a 50% decrease in the survival rate of marsh frog tadpoles after 5 days of exposure, negatively affecting body volume and length. Given the high degree of toxicity of the insecticide Actellic 50 EC, we recommend continuing investigations on non-target species, including both plants and animals, as the sub-chronic effects are quite little known in the scientific literature.

Global and regional drivers of abundance patterns in the hart's tongue fern complex (Aspleniaceae).

The hart's tongue fern (HTF) complex is a monophyletic group composed of five geographically segregated members with divergent abundance patterns across its broad geographic range. We postulated hierarchical systems of environmental controls in which climatic and land-use change drive abundance patterns at the global scale, while various ecological conditions function as finer scale determinants that further increase geographic disparities at regional to local scales. After quantifying the abundance patterns of the HTF complex, we estimated their correlations with global climate and land-use dynamics. Regional determinants were assessed using boosted regression tree models with 18 potential ecological variables. Moreover, we investigated long-term population trends in the USA to understand the interplay of climate change and anthropogenic activities on a temporal scale. Latitudinal climate shifts drove latitudinal abundance gradients, and regionally different levels of land-use change resulted in global geographic disparities in population abundance. At a regional scale, population isolation, which accounts for rescue effects, played an important role, particularly in Europe and East Asia where several hot spots occurred. Furthermore, the variables most strongly influencing abundance patterns greatly differed by region: precipitation seasonality in Europe; spatial heterogeneity of temperature and precipitation in East Asia; and magnitudes of past climate change, temperature seasonality and edaphic conditions in North America. In the USA, protected populations showed increasing trends compared with unprotected populations at the same latitude, highlighting the critical role of habitat protection in conservation measures. Geographic disparities in the abundance patterns of the HTF complex were determined by hierarchical systems of environmental controls, wherein climatic and land-use dynamics act globally but are modulated by various regional and local determinants operating at increasingly finer scales. We highlighted that fern conservation must be tailored to particular geographic contexts and environmental conditions by incorporating a better understanding of the dynamics acting at different spatiotemporal scales.

Phytochemical Study and Biological Activity of Three Fern Species of the Asplenium Genus Growing in Bulgaria

Background: Ferns are underestimated as medicinal plants and their use in traditional medicine is limited despite their diversity. Most of the research on their healing properties and phytochemical composition related to their biological activity has been focused on the secondary metabolites synthesized by ferns. Objective: In this study, we aimed to make an in-depth chemical characterization of three ferns widely spread in Bulgaria - Asplenium ceterach L., Asplenium scolopendrium L. and Asplenium trichomanes L. Methods: Micro elemental analysis was carried out using ICP-MS. Standard laboratory methods were used to determine the content of proteins, fats and fatty acids. Chromatographic methods were applied to quantify some secondary metabolites. Two antioxidant methods and two antimicrobial tests were used to evaluate the biological properties of the ferns tested. Results: Micro elemental analysis showed that these ferns could be useful source of zinc and iron. Protein content slightly varied among the species (13.6% - 18.4% of the dried plant weight). Negligible was the variation in oil content (3.1% - 4.0%, dry weight base); oil composition was dominated by saturated fatty acids. Some principal classes secondary metabolites, individual phenolic acids and flavonoids were quantified. The extract obtained from Asplenium ceterach L. demonstrated the highest antioxidant capacity. Fern extracts showed moderate antimicrobial activity against six of the 14 microorganisms tested. Conclusion: The results obtained may reveal new areas of application for ferns and are a solid basis for comparison with the same species growing in other geographical and climatic conditions, which may affect their healing properties.

Significance of Lipid Fatty Acid Composition for Resistance to Winter Conditions in Asplenium scolopendrium.

Ferns are one of the oldest land plants. Among them, there are species that, during the course of evolution, have adapted to living in temperate climates and under winter conditions. Asplenium scolopendrium is one such species whose fronds are able to tolerate low subzero temperatures in winter. It is known that the resistance of ferns to freezing is associated with their prevention of desiccation via unique properties of the xylem and effective photoprotective mechanisms. In this work, the composition of A. scolopendrium lipid fatty acids (FAs) at different times of the year was studied by gas-liquid chromatography with mass spectrometry to determine their role in the resistance of this species to low temperatures. During the growing season, the polyunsaturated FA content increased significantly. This led to increases in the unsaturation and double-bond indices by winter. In addition, after emergence from snow, medium-chain FAs were found in the fronds. Thus, it can be speculated that the FA composition plays an important role in the adaptation of A. scolopendrium to growing conditions and preparation for successful wintering.

CHARACTERIZATION IN TERMS OF COMPOSITION AND PHYTOTOXICITY OF AQUEOUS SPORES EXTRACT

In this study, we used aqueous extracts of fern spores and solution of AgNO3 and HAuCl4 for the synthesis of bimetallic nanoparticles Au:Ag in different proportions: 1:1 and 1:10. The spores used come from 2 species of ferns: Asplenium scolopendrium and Dryopteris filix-mas. For the characterization of the extracts with or without bimetallic nanoparticles we applied Fourier transform infrared spectroscopy (FT-IR). Phytotoxicity was tested using Pisum sativum seeds. Each extract was tested in 2 dilutions: 1:10 (D10) and 1:100 (D100). The parameters, which we determinate were the root and stem growth and fresh biomass. Root growth was stimulated in variants with Asplenium scolopendrium extract: without nanoparticles both dilution and with Au:Ag nanoparticles 1:10 D10. The highest values obtained for the stem were at D10 at the variants with Asplenium scolopendrium extract with or without bimetallic nanoparticles. The influence of extracts on fresh biomass was smaller than on the growth of root and stem.

Inventory of vascular spore plants and gymnosperms of the Kabardino-Balkar State Highland Reserve

The Kabardino-Balkar State Highland Reserve (KBGWZ) was established in 1976 to protect the alpinefloristic and faunal complexes of the Central Caucasus. The flora of the reserve includes about 1000 species of vascularplants. The inventory showed the presence in its flora of 40 species of vascular spore plants belonging to the divisionsLycopodiophyta (5 species, 2 families), Polypodiophyta (35 species, 12 families). Gymnosperms are represented by thedivisions Pinophyta, containing 3 families and 7 taxa of the species rank and below, as well as the division Gnetophytawith one representative of the genus Ephedra. Among the protected plants in the Red Book of the Russian Federation(2008), Taxus baccata is included. Asplenium scolopendrium (sub nom. Phyllitis scolopendrium), Ceterach officinarum andPolystichum lonchitis are in the Red Data Book of Kabardino-Balkar (2018). Among gymnosperms, Abies nordmannianaand Picea orientalis are also regionally protected species.

The complete chloroplast genome sequence of Asplenium komarovii Akasawa, a rare fern in South Korea

We sequenced the complete chloroplast genome of Asplenium komarovii Akasawa (syn: Asplenium scolopendrium L. subsp. japonicum (Komarov) Rasbach, Reichstein & Viane), which is designated as a rare species in South Korea. The complete chloroplast genome is 149,393 bp in total length and comprised of the following regions: large single copy (82,464 bp), small single copy (21,345 bp), and a pair of inverted repeats (22,792 bp). The overall GC content is 40.9% and the genome encoded a total of 115 genes, including 84 protein-coding, 27 transfer RNA, and 4 ribosomal RNA genes. Phylogenetic analysis based on 21 representative chloroplast genomes of the suborder Aspleniineae (and one outgroup) indicates that Aspleniaceae is monophyletic and sister to Diplaziopsidaceae, with Rhadidosoraceae as the basal group in this three family clade. Asplenium komarovii is sister to A. nidus and A. prolongatum with strong bootstrap support. The chloroplast genome of A. komarovii will be useful in establishing its relationships within the A. scolopendrium complex, which is currently unresolved.

Relicts of the forest cenoflora in the eastern Russian Caucasus

The article is devoted to the analysis of relics of the forest cenoflora occurred in the eastern part of the North Caucasus. Using the methods of morphological-ecological-geographical, florogenetic analysis and synthesis, etc., we analyzed the results of field observations and herbarium funds available in the region, and also scientific publications. The species composition of relics was revealed, their territorial distribution in forest communities, their origin, and florogenetic relationships were determined. The purpose of the analysis is to establish the geographical origin of relict species. In the studied flora, two types of relics were identified-tertiary and glacial, their importance for solving florogenetic issues is noted. The presence of relics of different geographic origin in forest cenoflora is explained by the consequences of migration processes that have taken place since the middle of the Tertiary period, which led to the movement of floristic complexes from the northern territories, from Transcaucasia and the adjacent parts of Asia Minor and Western Asia. Many relict species (Huperzia selago (L.) Bernh.ex Schrank & Mart., Lycopodium annotinum L, Selaginella helvetica (L.) Spring, Equisetum hyemale L., Polypodium vulgare L., Phyllitis scolopendrium (L.) Newman, Polystichym braunii (Spenn.) Fee, Taxus baccata L., Actaea spicata L) have rather extensive ranges in the Caucasus, and in general, their relictness is determined by the insular position of forests in this area.

Selective Anticancer Properties, Proapoptotic and Antibacterial Potential of Three Asplenium Species

The ferns Asplenium ceterach L., Asplenium scolopendrium L. and Asplenium trichomanes L. have wide application in traditional medicine worldwide. However, the scientific research on their anticancer and antibacterial properties is insufficient. The present article aims to provide more information on this topic. Extracts derived from the aerial parts of A. ceterach, A. scolopendrium and A. trichomanes were examined using a panel of in vitro assays with different bacterial and mammalian cells. The cytotoxicity and anticancer activity of the samples were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Trypan blue assays with three human (A549, FL, HeLa) and three murine (3T3, TIB-71, LS48) cell lines. Inhibitory effects on the growth of Gram-positive (Bacillus cereus) and Gram-negative (Pseudomonas aeruginosa) bacteria were determined by the agar diffusion assay. Apoptosis-inducing properties of the extracts were analyzed by flow cytometry. Superoxide dismutase (SOD) activity in extract-treated cells was investigated by ELISA. The obtained results demonstrate selective anticancer activity of all three Asplenium species. The extract from A. ceterach displayed the strongest inhibitory properties against human cervical cancer cells and bacterial cells. It induced a lower level of cytotoxicity against mouse cell lines, indicating a species-specific effect. The extract from A. trichomanes demonstrated better anticancer and antibacterial properties than the sample from A. scolopendrium. Further experiments linked the mechanism of action of A. ceterach extract with oxidative stress-inducing potential and strong proapoptotic potential against the cervical cancer cell line HeLa. A. trichomanes and A. scolopendrium extracts appeared to be potent inducers of necrotic cell death.

Preparation of stable nanosuspensions from Asplenium scolopendrium leaves via rapid expansion of supercritical solution into aqueous solutions (RESSAS)

The present study intended to stabilize the nanosuspension of powerful antioxidants such as genistein, quercetin, as well as caffeic acid from the leaves related to Asplenium scolopendrium by utilizing the RESSAS process. These compounds offer low efficacy and bioavailability since they are barely soluble in aqueous media. Thus, producing stable nanosuspensions can resolve this problem by reducing the size of the particle. In addition, central composite design (CCD) was used for analyzing the impact of oven temperature, pressure, CO2 flow rate, and modifier volume on the antioxidant activity index (AAI) in the outcome of RESSAS process. Further, DLS, FE-SEM, and LC-MS techniques were implemented for evaluating the nanosuspension features. Based on the results, the behavior of the particles after forming particle was emphasized, which indicated that the RESSAS process results in diminished agglomeration of the particle and enhanced AAI for the extract. Thus, the bioavailability of the products related to herbal medicinal can be dramatically increased in biological media.

Receptor-mediated biological effects of extracts obtained from three Asplenium species.

This study reports the effects of aqueous extracts obtained from three fern species of Bulgarian origin: Asplenium ceterach L., Asplenium scolopendrium L., and Asplenium trichomanes L. on the contractility and bioelectrogenesis of rat gastric smooth muscle tissues. In the concentration range 0.015-0.150mg/mL the three extracts contracted smooth muscle tissues in a concentration-dependent manner. The contractions caused by A.ceterach L. and A.scolopendrium L. extracts (0.150mg/mL) were reduced by ketanserin (5×10-7 and 5×10-6mol/L), an antagonist of serotonin 5-HT2 receptor. The contraction evoked by A.trichomanes L. (0.150mg/mL) was significantly reduced by 1×10-6mol/L atropine, an antagonist of muscarinic receptors, and turned into relaxation against the background of 3×10-7mol/L galantamine. After combined pretreatment with galantamine and l-arginine (5×10-4mol/L), this relaxation become more pronounced. The study demonstrates that constituents of A.ceterach L. and A.scolopendrium L. extracts act as agonists of 5-HT2 receptors and cause contraction by activating serotonergic signaling system. A.trichomanes L.-induced reaction is an additive result of two opposite-in-character effects. Thedominant contraction is initiated by inhibition of acetylcholinesterase activity. The relaxation develops with pre-inhibited acetylcholinesterase, it is significantly potentiated by l-arginine, and therefore associated with nitrergic signaling pathway.

Influence of gamma irradiation on the biological properties of Asplenium scolopendrium L. hydroalcoholic extracts

Gamma irradiation represents a method extensively used for the sterilization of natural products, thus leading to the extension of the products’ shelf life. However, a limited number of studies presented the effects of irradiation on the biological properties of the extracts. The present work aims to evaluate the effects of gamma irradiation on the biological properties (antimicrobial, cytotoxic and phytotoxic) of two types of hydroalcoholic fern extracts (obtained from leaves of Asplenium scolopendrium L.). The results obtained revealed that the extracts are not negatively affected by the irradiation procedure, the antimicrobial properties being enhanced, especially at a radiation dose of 0.6 kGy. The same observations were made for the cytotoxic and phytotoxic properties.

Frond Optical Properties of the Fern Phyllitis scolopendrium Depend on Light Conditions in the Habitat.

Ferns display an elevated degree of phenotypic plasticity to changes in irradiance levels; however, only a few reports deal with their response to different light conditions. To get an insight into the extent of phenotypic plasticity of the fern Phyllitis scolopendrium, thriving in a forested area along a radiation gradient at the entrance of a cave, we examined selected biochemical, morphological, and physiological frond traits of the ferns from three different habitats. Sampling was performed two times during the vegetation season, in April and June. We also measured frond optical properties to point out the differences in leaf/light interactions between different plant samples. According to frond size, the middle habitat, receiving 125 µmol m−2s−1 of photosynthetically active radiation at both sampling times, appeared to be the most favourable. The production of UV-absorbing substances was highest in the habitat with the lowest radiation level. At the beginning of the season, the level of photosynthetic pigments in this habitat was the same as in the other habitats, while it was significantly lower in June when the tree canopy was closed. Frond reflectance was similar when comparing habitats and different sampling times. The most significant differences were obtained in the UV-A and near-infrared regions. The reflectance spectra depended mainly on frond biochemical properties, which altogether explained 54% (p ≤ 0.05) of the spectra variability. Frond transmittance depended on both, morphological parameters, explaining 51% (p ≤ 0.05), and frond biochemistry, explaining 73% (p ≤ 0.05) of the spectra variability. P. scolopendrium was revealed to be highly plastic regarding light conditions. The shapes of the frond reflectance and transmittance optical curves were similar to those typical of leaves of seed plants. The fronds exhibited high morphological plasticity when comparing different habitats. However, their biochemical and optical traits differed more between the two sampling times than between the habitats.

Epiphyllous bryophytes in Arboretum Mlyňany (Slovakia)

Epiphyllous bryophytes in Arboretum Mlyňany (Slovakia)

Morphology and anatomy of three species of <i>Asplenium L.</i> at Obafemi Awolowo University Ile-Ife, Nigeria

Comparative morphological and foliar anatomical studies were carried out on three species of Asplenium: Asplenium nidus L., Asplenium scolopendrium (L.) Newn and Asplenium barterii Hook with a view to identifying important morphological and anatomical characters that can be employed in the separation of the species. Mature healthy plants of each species were collected from Erin Ijesa Waterfall and nursed in the FernGarden of Natural History Museum at Obafemi Awolowo University, Ile-Ife. Morphological observations and measurements were documented for each species. The transverse sections of the petiole and stem foliar sections were prepared by cutting at 10 μm thickness using Reichert Sliding Microtome and microphotographs were taken. Quantitative data obtained from this study were subjected to one-way Analysis of Variance with Duncan Multiple Range Test for mean separation. Findings revealed that the predominantly irregular epidermal cells on the adaxial surface of A. barterii is diagnostic of the species. The presence of tetracytic stomata in addition to anisocytic type also separates A. nidus from the other taxa. Similarly, A. scolopendrium had a significantly higher guard cell area compared to the other taxa. Morphological and foliar anatomical features in the genus can be used to delimit the three species while the possession of two vascular bundles broadly admit the species into the genus Asplenium.
 Keywords: Asplenium, Taxonomic, Foliar anatomy, Morphology.

ВІДДІЛ POLYPODIOPHYTA У СТРУКТУРІ ФЛОРИ НАЦІОНАЛЬНОГО ПРИРОДНОГО ПАРКУ “ГУЦУЛЬЩИНА” (УКРАЇНСЬКІ КАРПАТИ)

National Natural Park “Hutsulshchyna” is situated on the territory of the Kosiv district of Ivano-Frankivsk region on the area of 322.71 km 2 . In the borders of NNP we have found 27 species of ferns, which belong to 16 generas and 9 families. Here we are representing a list of the flora of Polypodiophyta plants section, biomorphological, ecological, regional and geographical characteristic of each species of higher spore plants on the territory of NNP “Hutsulshchyna”. According to zonal distribution the largest group among ferns are plurizonal species – 33.3 %, boreal-submeridional – 22.2 %, temperate-meridional – 14.8%, boreal-meridional – 11.1 %, arctic-meridional and boreal-subtropic – 7.4 % respectively and boreal-temperate – by 3.7 %. An altitude analysis of pteridophytes shows that among seven groups of flora’s distribution the biggest is the group of montana-plain species – 85.1 %. The climatic analysis of Polypodiophyta plants section shows, that dominative is the group of oceanists – 81.5 %. The character of distribution of ferns across the globe designate the regional distribution of species. The dominative group, which includes more than half of all species composition are circumpolar species – 59.3 %, they are occurring in the entire of all Northern hemisphere. The other groups make a small part of the total numbers of ferns. The result of analyze of the life forms of Polypodiophyta by K. Raunkier was next: they are represented by two groups, hemicryptophytes dominate – 70.4 %, geophytes are less then one third of the species – 29.6 %. The ratio of the ferns according to the level of moisture content of the soil (substrate) is as follows: the largest group is mesophytes – 70.4 %, mesogigrophytes – 22.2 %, while hihrophytes and xeromesophytes includes only one species – 3.7 % of the total numbers of ferns. A distribution of the ferns on the territory of NNP “Hutsulshchyna” as a measure of trophy of the substrate is: mesotrophs are 55.6 %, mesoeutrophs – 40.7 % of the total number of species, shows a moderate curiosity to the richness of the soil. In the researching group of the flora of NNP “Hutsulshchyna” there is significant advantage of sciophytes, which life cycle undergo in the conditions of rather weak light (55.6 %), they are confined to forest phytocenoses. Sciogeliophytes which can existing on the open areas and withstand a certain degree of shading make up 33.3 % of the total number of species. Among the 27 species of ferns which are founded on the on the territory of the National park two species ( Botrychium lunaria , Cystopteris sudetica ) are included in the Red Book of Ukraine. The list of endemic, relict, rare and endangered plant species known from the territory of the Ivano-Frankivsk region includes Ophioglossum vulgatum , Phyllitis scolopendrium , Polystichum braunii , Polystichum aculeatum.