Gymnadenia Conopsea Research Articles

Shifts in bacterial community composition during symbiotic seed germination of a terrestrial orchid and effects on protocorm development.

It is well known that orchid seeds depend on mycorrhizal fungi to supply the necessary nutrients that support germination in natural environments. Apart from fungi, bacteria may also be involved in the germination process of orchid seeds, but so far, their role has not been intensively studied. This research provides evidence that bacterial community composition changes during seed germination of the terrestrial orchid Gymnadenia conopsea. Interestingly, in vitro experiments showed that Pseudomonas spp., which were the most dominant bacteria in the later germination stages, improved protocorm growth. These results suggest that bacteria contribute to the germination of orchid seeds, which may open new perspectives to apply bacteria as a biofertilizer in the introduction and restoration of G. conopsea populations.

×Gymnigritella suaveolens (Orchidaceae), a New Natural Hybrid to the Flora of Serbia

×Gymnigritella suaveolens (Orchidaceae), a natural hybrid between Gymnadenia conopsea and Nigritella rhellicani, was found at Mt Kopaonik for the first time in Serbia, representing its single known locality in the Central Balkans. Morphological characteristics, distribution data, habitat and ecological preferences are indicated. The hybrid was found at 1552 m a.s.l., on sericite schists, as a member of the Festuco-Deschampsietum flexuosae grassland community. Two hybrid specimens were recorded at the site where the two parental species grow in sympatry, and where the population size of G. conopsea was larger than the population size of N. rhellicani.

Genetic differentiation and diversity do not explain variation in heterosis or inbreeding depression: empirical evidence from a long-lived iteroparous plant

AbstractAssisted gene flow can restore genetic diversity when genetic drift has driven deleterious alleles to high frequencies in small, isolated populations. Previous crosses among 20 populations of Gymnadenia conopsea documented the strongest heterosis and the weakest inbreeding depression in sparse and small populations, consistent with fixation of mildly deleterious alleles by genetic drift. We genotyped the populations used for crosses, and used 1200–1728 SNPs to test the following predictions: (1) heterosis increases with genetic differentiation (FST) to donor populations and decreases with genetic diversity in the recipient population, (2) inbreeding depression increases with genetic diversity, and (3) genetic diversity increases, and mean FST to other populations decreases, with population size and density. Pairwise FST ranged from very low to moderate (0.005–0.20) and genetic diversity varied moderately among populations (proportion of polymorphic loci = 0.52–0.75). However, neither FST between populations, nor genetic diversity in the recipient population, were related to the strength of heterosis. There was also no association between genetic diversity and the strength of inbreeding depression. Genetic diversity increased and mean FST decreased with population size, consistent with reduced diversity and increased differentiation of small populations by genetic drift. The results indicate that the loci conferring heterosis are not mirrored by overall population differentiation, and limited additional information on potential source populations for genetic rescue is gained by the genetic data. Instead, the use of controlled crosses can directly reveal positive effects of introducing new genetic material, and is a simple method with high potential in conservation.

Separation and purification of benzylester glucosides and derivatives from tubers of Gymnadenia conopsea (L.) R. Br. by linear gradient counter-current chromatography combined with elution-extrusion mode.

Tubers of Gymnadenia conopsea (L.) R. Br. (Orchidaceae), a traditional medicine and food homologous plant, has a broad application and development prospect in the food and drug industries. Benzylester glucosides, the main effective active components in this plant, are difficult to separate due to their similar structures and high polarity. In this study, linear gradient counter-current chromatography was used to separate benzylester glucosides and derivatives, combined with elution-extrusion mode. The main separation parameters were optimized, including the ratio of mobile phase and sample loading. Finally, seven compounds were successfully separated, including 4-hydroxybenzyl alcohol (1), 4-hydroxybenzaldehyde (2), dactylorhin B (3), loroglossin (4), dactylorhin A (5), 4-(ethoxymethyl) phenol (6), and militarine (7). The structures were analyzed by mass spectrometry and nuclear magnetic resonance spectrometry. According to our findings, the established method was an efficient approach to separate benzylester glucosides and derivatives from tubers of G. conopsea. The established strategy could be applied to purify other similar high-polarity compounds from complex natural products.

Fine-scale genetic structure in the orchid Gymnadenia conopsea is not associated with local density of flowering plants.

Density-dependent pollinator visitation can lead to density-dependent mating patterns and within-population genetic structure. In Gymnadenia conopsea, individuals in low-density patches receive more self pollen than individuals in high-density patches, suggesting higher relatedness at low density. Ongoing fragmentation is also expected to cause more local matings, potentially leading to biparental inbreeding depression. To evaluate whether relatedness decreases with local density, we analyzed 1315 SNP loci in 113 individuals within two large populations. We quantified within-population genetic structure in one of the populations, recorded potential habitat barriers, and visualized gene flow using estimated effective migration surfaces (EEMS). We further estimated the magnitude of biparental inbreeding depression that would result from matings restricted to within 5 m. There was no significant relationship between local density and relatedness in any population. We detected significant fine-scale genetic structure consistent with isolation by distance, with positive kinship coefficients at distances below 10 m. Kinship coefficients were low, and predicted biparental inbreeding depression resulting from matings within the closest 5 m was a modest 1-3%. The EEMS suggested that rocks and bushes may act as barriers to gene flow within a population. The results suggest that increased self-pollen deposition in sparse patches does not necessarily cause higher selfing rates or that inbreeding depression results in low establishment success of inbred individuals. The modest relatedness suggests that biparental inbreeding depression is unlikely to be an immediate problem following fragmentation of large populations. The results further indicate that habitat structure may contribute to governing fine-scale genetic structure in G. conopsea.

Actuarial senescence progresses similarly across sites and species in four boreal orchids

Abstract Whole‐plant senescence, defined as a decrease in individual fitness as an organism grows older, has often been assumed to not occur in plants; however, it has now been detected in a range of plant taxa. Still, reported senescence patterns vary substantially, and it remains unknown how consistent patterns are within phylogenetic groups and how they may be affected by environmental factors. Plants show a high diversity in life‐history traits within phylogenetic groups and environments, but shared traits among related species are also common, making both diverse and similar patterns probable. Here, we explore how mortality changes with advancing age in four closely related species (Dactylorhiza incarnata, D. lapponica, D. maculata and Gymnadenia conopsea) across two sites in Norway: the coastal Nordmarka and inland Sølendet. Using data collected over 34 years, following more than 2500 individual plants, we conduct Bayesian survival trajectory analysis to assess mortality age trajectories. A simple Weibull model, illustrating increasing mortality at a decelerating rate with age, was the best fit for all species at both sites. From these models, we calculate rates of senescence and compare them using Kullback–Leibler divergences, finding no notable differences in rates between species or sites. Synthesis. Our findings suggest that actuarial senescence, an increase in mortality with advancing age, may be common in orchids and show that demographic ageing can proceed similarly in closely related taxa across different environments.

PHYTODIVERSITY OF THE TERRITORY OF THE CHERNIVTSI NATIONAL UNIVERSITY «SHEPIT» BIOBASE AS AN OBJECT OF STUDENTS’ COGNITIVE ACTIVITY

The results of study of grassland habitats of the biobase «Shepit» of Yuriy Fedkovych Chernivtsi National University are presented. 123 species of vascular plants were found in the studied communities, of them, 58,54 % are pratants, 30,89 % are sylvants, and 10,57 % are synanthropants. 7 species from the Red Data Book of Ukraine (Perelik…, 2021) grow on the territory of the biobase, in particular, Botrychium lunaria (L.) Sw., Colchicum autumnale L., Gymnadenia conopsea (L.) R.Br., Listera ovata (L.) R.Br., Neotinea ustulata (L.) R.M. Bateman, Pridgeon et M.W. Chase, Platanthera bifolia (L.) Rich., Traunsteinera globosa (L.) Rchb. This proves the value of this territory as a cell of rare phytodiversity. Grassland habitats of the «Shepit» biobase on a small area well represent the meadows of the region, in particular hay meadows and pastures Arrhenatheretalia elatioris Tx. 1931 order, which belong to two unions: Arrhenatherion elatioris Luquet 1926 і and Cynosurion cristati Tx. 1947 and correspond to the habitats «Т2.3.1 Low and medium altitude hay meadows» and «Т2.2.1 Low and medium altitude pastures» (2018). Bryoflora of the territory of the base is represented by 29 species from 23 genera, 17 families, 6 orders, 3 classes, 2 divisions. In the systematic spectrum, species of the Bryophyta are predominating. In relation to the type of substrate most bryophytes are epigeous. The identified species are typical as for the Bukovyna Carpathians bryoflora and other natural regions of Ukraine, so the ability to recognize them in nature will be useful to students in their further work. It has been established that the territory of the «Shepit» biobase provides a number of ecosystem services classified into four types: Provisioning Services, Regulating Services, Supporting Services, and Cultural and Social Services. The study area is a model environment for learning methodological approaches to the study of phytodiversity, which promotes information about the flora and habitats of the Carpathians and their sozological value, emphasizes the importance of monitoring studies, and serves to form students’ ecological awareness. The methods mastered by students can be applied in their professional activities. Key words: phytodiversity, vascular plants, bryophytes, grasslands, ecosystem services, «Shepit» biobase.

Bioactive steroids from seed germination supporting fungus (Ceratobasidium GS2) of the terrestrial orchid Gymnadenia conopsea

ABSTRACT Almost all orchids rely on mycorrhizal fungus to support their seed germination. To date, the effect of active components in mycorrhizal fungus on orchid seed germination largely remains unknown. In this study, we aimed to investigate the impact of active components found in mycorrhizal fungus on orchid seed germination. Specifically, we focused on a terrestrial orchid Gymnadenia conopsea and its host-specific seed germination supporting fungus Ceratobasidium GS2. In total, several steroids (1–7) were isolated from this fungus. Notably, compounds 1, 2, 4, and 5 exhibited significant enhancements in protocorm volume. Moreover, compounds 1–6 demonstrated strong promotion of protocorm differentiation. These findings suggest that steroids may play a crucial role in the symbiotic germination of G. conopsea seeds. Future studies should continue to explore the specific mechanisms through which these steroids exert their effects, contributing to our understanding of orchid biology and mycorrhizal interaction.

Orchids of Russia and Crimea: herbarium collections provide sufficient data to recover the distributional changes with time

Monitoring the distributional changes of native vascular plants over time is an important task both from the scientific and conservation point. However, numerical data for such studies are often missing, especially for countries lacking national biodiversity datasets, such as Russia. The main aim of this study was to assess whether direct data from herbarium collections are sufficient to recover the distributional changes effectively. The object of this study was the orchid family, a conservationally important group of plants, in the Flora of Russia and Crimea. Primary source of the data were georeferenced herbarium labels from 59 herbarium collections. The distributional changes were assessed by comparisons between the numbers of grid cells with records in different time intervals, taking into account the variation of recording intensity with time.The method proved to be applicable to 108 out of 135 species of orchids occurring within the studied territory. For many of them, reliable changes in the number of localities were confirmed. Among widely distributed orchid species constant negative change was detected for Anacamptis coriophora, Calypso bulbosa, Coeloglossum viride, Cypripedium guttatum, C. macranthos, Gymnadenia conopsea, Herminium monorchis, Neotinea ustulata, Orchis militaris, Platanthera hologlottis; constant positive change was recovered for Dactylorhiza baltica and D. fuchsii only. In many species, trends over time were uneven. The direction of change proved to be generally consistent for a given species across the whole studied territory, suggesting the principal influence of general factors, such as climate change and general eutrophication of the environment. The direction of change is consistent with type of geographic distribution of plants.Although the method used in the current study for recovering distributional changes has limitations, it may be further recommended for use with other taxa.

Gymnadenia conopsea (L.) R. Br.: Comprehensive review of propagation and breeding, traditional uses, chemical composition, pharmacology, quality control, and processing.

Gymnadenia conopsea, a perennial herbaceous flowering plant that belongs to the family of Orchidaceae, sporadic distributed in the altitudes of 200-4700m across northern Europe and, temperate and subtropical Asia region. The dried tubers of G. conopsea have been used to treat cough, asthma, and their syndromes, and also as a tonic in China and surrounding countries for a long history. G. conopsea is often processed deeply processed before use to enhance its efficacy. In recent years, because of its remarkable pharmacological activity and health care function, G. conopsea has been used more and more widely. Due to its extensive application and bad growth environment, the wild distribution of G. conopsea is decreasing and it has been listed as an endangered plant. This review aims to summarize the propagation and breeding, traditional uses, chemical composition, pharmacology, quality control, and processing of G. conopsea. Moreover, it also provides suggestions for the future high-value utilization of G. conopsea. A literature search on Gymnadenia genus and G. conopsea was performed using scientific databases including SciFinder, ACS, Web of Science, Springer, ScienceDirect, PubMed, and CNKI. Information was also collected from classic books of Chinese herbal medicine, official websites, Ph.D. and M.Sc. Dissertations, and so on. Structures of chemical compounds were drawn by ChemDraw software. As of submission date of this manuscript, total 170 natural compounds have been isolated and characterized from G. conopsea, and all of the compounds were isolated from the tubers. The isolated compounds including benzylester glucosides, dihydrostilbenes, phenanthrenes, phenolic compounds, alkaloids, polysaccharide, lignans, flavones, triterpenoids, steroids, and other compounds. Some of these compounds and active extracts exhibited a wide range of pharmacological activities, in which, the tonifying, anti-fatigue, anti-oxidant, anti-viral, sedative and hypnotic activities are consistent with the traditional uses for the treatment of diseases. In addition, a variety of new pharmacological activities, such as preventing and treating gastric ulcers, immunoregulatory, anti-hyperlipidemia, anti-anaphylaxis, anti-silicosis, anti-cancer and neuroprotective activities have also been reported. However, the bioactive compounds responsible for most of the above pharmacological effects have not been well summarised till now. In this manuscript, analysis, speculation and summary of compounds that responsible for pharmacological effects were conducted. The chemical constituents and pharmacological activities studies of G. conopsea extract have been summarised in this context, the isolated compounds responsible for the pharmacological activities were also analyzed and deduced according to the publications, all above led to suggestions for the future high-value utilization of G. conopsea.

Increasing density enhances pollination and fecundity in the rewarding orchid Gymnadenia conopsea

Conspecific density is an important factor in determining the fecundity of pollinator-dependent plants. However, the outcome of increased conspecific density can range from reduced per capita pollination rates and fecundity (competition) to increased per capita pollination rates and fecundity (facilitation). I tested whether local conspecific density influenced pollination and fecundity in the rewarding orchid, Gymnadenia conopsea. I measured the distance to each of the nearest five plants in sixty patches of varying density in a large population in the Italian Apennine mountains. I examined whether pollen removal, pollen deposition, fruit, and seed set varied according to mean distance from conspecifics and plant height. After controlling for plant position in the population, the proportion of pollen removal, pollen deposition, and fruit set decreased with increasing distance from conspecifics, while pollen removal and deposition, but not fruit set, increased with increasing plant height. The proportion of viable seeds per fruit did not vary according to conspecific distance. Decreases in pollen removal, pollen deposition, and fruit set occurred within an average of 1 m distance from conspecific plants, indicating that local aggregation is an important determinant of both pollination success and fecundity. These results suggest that despite being a geographically widespread orchid, G. conopsea may suffer reduced fecundity when it occurs in small, isolated populations. Such field investigations can potentially lead to better predictions for successful re-establishment of populations of orchids under conservation-aided restoration efforts.

Furanoids from the Gymnadenia conopsea (Orchidaceae) seed germination supporting fungus Ceratobasidium sp. (GS2).

Five furanoids including a new analog (S)-1,4-di(furan-2-yl)-2-hydroxybutane-1,4-dione (1) together with four known ones, rhizosolaniol (2), 5-hydroxymethylfurfural (3), 2-furoic acid (4) and (2-furyl) oxoacetamide (5), were isolated from the fungal strain Ceratobasidium sp. (GS2) inducing seed germination of the endangered medicinal plant Gymnadenia conopsea of Orchidaceae. The structure of new furanoid 1 was determined mainly based on HR-ESI-MS and NMR spectral data. Modified Mosher's reactions were used to establish the stereochemistry of the hydroxyl group in 1, which was not stable in Mosher's reagents and transformed into four analogs 6-9. These degraded products (6-9) were elucidated based on UPLC-Q-TOF-MS/MS analysis, and compound 8 was further isolated from the degraded mixture and its structure was characterized through NMR experiments. Therefore, the absolute configuration of compound 1 was determined by electronic circular dichroism combined with quantum-chemical calculations adopting time-dependent density functional theory. Compounds (1-5), and 8 showed weak antioxidant activities, and compounds (2-4) displayed phytotoxicity on punctured detached green foxtail leaves. In addition, compounds 3 and 4 strongly showed inhibition activities on the seed germination of G. conopsea. This was the first chemical investigation of the symbiotic fungus of G. conopsea.

In vitro culture conditions for asymbiotic germination and seedling development of Anacamptis pyramidalis (L.) Rich. and Gymnadenia conopsea (L.) R. Br.

• Positive influence of MM medium on germination of A. pyramidalis and G. conopsea was determined. • Germination was influenced by full darkness and nutrient medium enriched with organic aditives. • In seedling growth, 2iP initiated the formation of the aboveground part of G. conopsea. • In A. pyramidalis, Kin initiated the leaf formation, while Ba promote root formation. Protocols for asymbiotic seed germination and seedling development of two terrestrial orchid species – Gimnadenia conopsea and Anacamptis pyramidalis are presented in this paper. Detail examination covered nutrient media: Knudson C (KC) and Malmgren (MM); organic supplements: coconut water (CW), pineapple juice (PJ), peptone (PE), casein hydrolysate (CA) and glutamine (A); different lighting conditions – 0/24 Light / Dark (L / D) and 16/8 (L / D) as well as cytokinins: 6-benzyladenine (BA), kinetin (Kin), and 6- (γ, γ-dimethylallylamino) purines (2 -iP) aiming to assess asymbiotic seed germination in controlled in vitro conditions, to define the optimal protocol for their mass propagation. The obtained results indicate that the nutrient medium MM and conditions of complete darkness encourage seed germination in both investigated species, while the organic supplements amendment resulted in a shortened period necessary for seed germination as well as a significantly higher percentage of germination. In Gymnadenia conopsea , the highest percentage of germination (73,79%) accompanied by the highest embryo/protocorm morphometric characteristics (1.40 mm average height, 1.21 mm average width, and 0.30 mm average shoot height) was achieved on MM-CW nutrient medium. A similar result was observed for Anacamptis pyramidalis with 69,88% germinated seeds on MM-CW nutrient medium, while the highest morphometric values were 1.10 mm (on MM-CW) for height, 1.00 mm (on MM-PE) for width and 0.57 mm (on MM-PE) for bud height. On the contrary, the results obtained on nutrient medium KC and under the 16/8 L / D lighting conditions indicate the inhibitory effect of these conditions on the seeds germination in examined species. A experiment with chitokinin treatments indicates that the best result in G. conopsea was achieved on the medium with the addition of 0.3 mg L −1 2iP with 5.33 mm for plant height, 1,13 root numbers and 12.26 mm root lenght, while in A. pyramidalis 0.3 mg L −1 Kin emerged as the most suitable for the development of the upper part of the plant (3.33 mm), while the medium enriched with 0.3 mg L −1 BA had a positive effect on the formation of the root system.

Protected species in grassland habitats of Ukraine

Based on the analysis of more than 17,000 vegetation plots (relevés), the participation of 261 protected species (254 vascular plants, six lichens, and one bryophyte) in 30 EUNIS grassland habitat types was revealed. Vegetation plots were assigned to the habitat types using the EUNIS-ESy expert system with further verification. We consider as protected species those listed in the current edition of the Red Data Book of Ukraine, Resolution 6 of the Bern Convention, Annexes II and IV of the Habitat Directive, and the IUCN Red List (only categories VU, EN, CR). The participation of protected species was studied according to the following three criteria: (1) the total number of protected species in the plots assigned to a certain habitat type, (2) the number of plots in which at least one protected species is present, and (3) the mean number of protected species per plots within each habitat type. True steppes (R1B) and meadow steppes (R1A) differed with a significant predominance of the total number of protected species. Arctic alpine calcareous grassland (R44) and Continental dry rocky steppic grassland and dwarf scrub on chalk outcrops (R15) had the largest proportion of plots with protected species and the highest mean numbers of protected species per relevé. Saline habitats, in particular Temperate inland salt marsh (R63) and Semi desert salt pan (R64), were characterized by the smallest number of plots with protected species. Among all species, Gymnadenia conopsea, Stipa capillata, Colchicum autumnale and Gladiolus imbricatus occurred in the largest number of studied habitat types. Based on the results of the analysis, appropriate ways of optimizing the protection of grassland habitats and protected species are proposed.

Addition of fungal inoculum increases seed germination and protocorm formation in a terrestrial orchid

Because orchid mycorrhizal fungi are important for orchid seed germination, addition of germination-supporting fungal inoculum to the soil can be an efficient way to improve in situ seed germination of terrestrial orchids, but empirical evidence is still largely lacking. In this study, we used seed inoculation experiments to test the hypothesis that addition of fungal inoculum increases seed germination of the terrestrial orchid Gymnadenia conopsea under natural field conditions. In China, this species has suffered dramatic declines in abundance due to habitat loss, grazing and over-collection, while observational studies have shown very low recruitment of seedlings into natural populations. Previous research has shown that the fungal strain GS2 (Ceratobasidiaceae) was able to support germination under in vitro conditions. Seed packages were used to bury orchid seeds in the soil with and without fungal inoculum and to assess seed germination. In total, 11.96 ± 2.65 % of seeds had the testa ruptured when fungal inoculum was added to the seed packages, 10.63 ± 3.32 % of seeds germinated to the protocorm formation stage, 5.24 ± 2.71 % of the seeds grew to the protocorm differentiation stage, and 3.16 ± 1.63 % of seeds finally formed early seedlings. In contrast, no seed germination was observed in the control groups without fungal inoculum. We isolated the fungi associating with the germinating seeds and confirmed that seed germination was supported by fungal strain GS2. In addition, several other fungi were isolated from germinating seeds, including four other Ceratobasidiaceae strains that are known to support seed germination in vitro. Our results highlight the potential of fungal inoculum to induce seed germination of terrestrial orchids under field conditions and may be useful when developing more efficient symbiotic seed germination protocols for the restoration and conservation of terrestrial orchids, especially in situations where natural germination is severely limited.

Addition of fungal inoculum increases germination of orchid seeds in restored grasslands

Grasslands restored on arable land often retain high residual nutrients, modified soil biota, and lower plant species diversity. Establishment of rare plant species with complex multitrophic interactions, typical of undisturbed nutrient-poor environments, may be hindered by the absence of interacting organisms. We hypothesised that the addition of a mycorrhizal symbiont improves the seed germination of orchids that crucially depend on fungi. We focused on grasslands restored on arable land 1–15 years ago featuring residual mineral nutrients and low organic matter contents compared to semi-natural grasslands and on four orchid species differing in the level of mycorrhizal specificity: high – Anacamptis pyramidalis and Orchis mascula – and low – Platanthera bifolia and Gymnadenia conopsea. Five fungal isolates obtained from non-green underground mycorrhizal orchid seedlings (protocorms) or adults' roots were tested for orchid-fungus compatibility under conditions in vitro. Orchid seeds inserted in retrievable seed packets were subsequently co-introduced with selected fungal isolates grown either on agar or sterilized hay into the soil of nine restored grasslands and incubated for twelve months. The identity of mycorrhizal fungi in retrieved protocorms was verified by molecular methods. The isolates that supported protocorm establishment in vitro enabled also protocorm formation in situ, but success rates differed among orchid species. While mycorrhizal specialists produced most protocorms after inoculation, the mycorrhizal generalists took advantage of naturally occurring fungi and produced some protocorms both in inoculated and uninoculated treatments. We showed that the addition of mycorrhizal fungi enhanced protocorm formation regardless of the modified soil environment, especially in mycorrhizal specialist orchids. This method may help to restore populations of native orchid species in their former distribution ranges, including farming-altered habitats.

CONSERVATION STATUS OF PLANTS AND HABITATS UNDER TEN YEARS OPERATION PHASE OF A WIND FARM. CASE STUDY: SFANTA ELENA KARST PLATEAU, IRON GATES NATURAL PARK, ROMANIA

Located in the Iron Gates Natural Park, Romania, at the foot of the Locvei Mountains, the Sfanta Elena Karst Plateau is a very picturesque territory, characterized by a karst relief, covered by High Nature Value grasslands. In the last decade this territory has been subjected to increasing anthropogenic pressures, and specific concerns on biodiversity have been raised after the construction of a wind farm in 2011. Starting from a concern expressed by a scientist, our study aims to provide current data on plant species with conservation value and to assess the conservation status of orchid-rich grasslands around wind turbines after ten years of wind farm operation. During the operation period we identified 19 plant taxa with conservation value, eight of which which are orchid species. Four of them are new reports for this territory: Gymnadenia conopsea, Neotinea ustulata, Neottia nidus-avis and Orchis simia, as well other rare taxa such as Cirsium grecescui, Lathyrus sphaericus, Linum hologynum and Rumex thyrsiflorus, are mentioned for the first time. The orchid-rich grasslands in the wind farm area correspond to the Natura 2000 habitat 6210* whose structure and floristic composition are within the acceptable limits of Favourable conservation status. The conservation status of plant species is directly dependent on the quality of their habitat, as well as the variety of pressure and threat factors in the area.

Notes on the distribution, ecology, associated vegetation and conservation status of Gymnadenia (Orchidaceae) in Kosovo

Four species of Gymnadenia are native to Kosovo: G. conopsea, G. frivaldii, G. nigra, and G. odoratissima. In this study, field expedition data, phytosociological relevés, herbarium specimens along with extensive literature sources were used to analyse vegetation and ecological characteristics, habitat types, distributional patterns as well as provide notes on conservation. Gymnadenia conopsea is distributed throughout the country, from lowlands to the alpine belt. It was recorded in various plant communities. Gymnadenia frivaldii grows in the alpine zone of mountains, close to streams and in wet meadows. Its relevés belong to the class Scheuchzerio-Caricetea fuscae. This species is classified as “Near Threatened” (NT) in Kosovo. Gymnadenia nigra grows in subalpine and alpine grassland on preferably calcareous substrate. It has been found in almost all mountains reaching >2000 m a.s.l., and occurs in different plant communities belonging to the class: Elyno-Seslerietea. Gymnadenia odoratissima was recorded from one locality only in Kosovo, on the massif of Maja e Zezë, Sharri Mts. It was growing in degraded beech forest and meadows on silicate bedrock. Its floristically diverse relevés associate with the class: Mulgedio-Aconitetea. Of the four studied species, G. frivaldii deserves more conservation attention because of its fragile populations.

Variability of Population and Individual Characters of Gymnadenia conopsea (L.) R. Br. (Orchidaceae, Liliopsida) under Anthropogenic Load Conditions in Meadow Ecosystems of Central Caucasus

We studied the variability and phytocenotic plasticity of the morphological characters of individuals, and the numbers, density, ontogenetic structure and vitality of 11 coenopopulations of Gymnadenia conopsea (L.) R. Br. within the Kabardino-Balkar Republic in 2015–2019. The variability and plasticity of plants features provide their adaptation to various growing conditions and improve the morphological heterogeneity of coenopopulations. The dynamics of abundance, density, age structure and vitality of coenopopulations in the changing conditions of the environment provides information about life strategies of the species. Age conditions, the variability and plasticity of plants were studied on the basis of morphological parameters of the aboveground organs without digging the plants. The studies were conducted in meadow plant communities under various conditions of anthropogenic load (recreation and grazing). The species is characterized by medium-high phytocenotic plasticity of morphological characters (Ip = 38–68%) and high variability of morphological characters (CVх¯ср = 17.10– 33.63%) under changeable growth conditions. Intensification of growth processes in the individuals with the maximum increase of the coenopopulation vitality (IVC = 0.98– 1.16) was observed in the composition of undisturbed highly wet meadows; the portion of generative individuals increased in their ontogenetic spectra (48.6–56.5%). Under the anthropogenic load intensification, the vitality of coenopopulations (IVC = 0.82– 0.95) and the density of individuals decreased; the portion of virginal individuals increased in their ontogenetic spectra (rubbly meadows) or coenopopulations were clearly aging. The rate of the seed reproduction of coenopopulations, where the density of individuals accounted for 16 ind./m2 , was high on rubbly substrata with low total projective cover (60%). Our studies have identified the mixed type of life strategy for Gymnadenia conopsea with stress-tolerant and ruderal components. The ruderal-stress-tolerant life strategy allows the species to grow for a long time in conditions of high interspecific competition and anthropogenic load, spreading quickly to unoccupied territories.

Genome size variability of the population of Gymnadenia conopsea (Orchidaceae) in the Novosibirsk Region (Russia)

Gymnadenia conopsea (Fragrant orchid) is a widespread model species for the study of polymorphism in the European part of Russia and abroad. Flow cytometry refers to rapid methods effective for assessing the structure of a population by DNA size estimation. However, this method is only available in habitats with close proximity to the place of the experiment, because this indicator is determined only in fresh leaves. A small population of G. conopsea in the industrial area adjacent to the Novosibirsk urban agglomeration in the Novosibirsk Region (NR) was studied. Only diploids were detected in the G. conopsea cenopopulation occurred in the industrial zone of the NR. In this study we pointed on significance of genome size variation, which correlates with the adaptive evolution of the G. conopsea in West Siberia. The revealed absence of minor cytotypes (individuals with a higher level of ploidy) in G. conopsea population reduces intraspecific and intrapopulation diversity of the species.