Fungus-like Organisms Research Articles

Systematics and Phylogeny of Myxomycetes: Yesterday, Today, Tomorrow

Myxomycetes are amoeboid fungus-like organisms (Amoebozoa) with a unique life cycle characterized by a great morphological diversity of fruiting bodies. Due to the similarity of these structures to the fruiting bodies of some representatives of Ascomycota and Basidiomycota, myxomycetes have been classified as fungi since the first known scientific description in 1654. Only in the XIX century, when their life cycle was studied, the difference of this group from fungi became clear. During the same period, microscopic structures of fruiting bodies, as well as ornamentation of the spore surface, began to be considered as diagnostic features. Due to this, in the period from the end of XIX to the middle of XX century, a rather stable system was formed. However, as further studies have shown, both macro- and micromorphological characters are often quite variable, depend on environmental conditions, and often result from a convergent evolution, which causes difficulties in defining species and taxonomic units of higher ranks. Since the first decade of the 21st century, thanks to the development of molecular genetic methods and accumulation of data on nucleotide sequences of marker genes together with the improvement of microscopic studies, it has been possible to obtain data on the evolutionary relationships of different groups of myxomycetes. A milestone in this process was the publication of the first phylogenetic system of myxomycetes in 2019. This work was the starting point for a number of studies on the relationships of different groups of myxomycetes at a lower taxonomic level. Thus, there has been a surge in the number of studies that bring us closer to constructing a natural system. The latest iteration of the myxomycete system, incorporating all modifications and enhancements as of June 2024, is presented.

The Systematics and Phylogeny of Myxomycetes: Yesterday, Today, and Tomorrow.

Abstract-Myxomycetes are amoeboid fungus-like organisms (Amoebozoa) with a unique life cycle characterized by a great morphological diversity of fruiting bodies. Due to the similarity of these structures to the fruiting bodies of some representatives of Ascomycota and Basidiomycota, myxomycetes have been classified as fungi since the first known scientific description in 1654. Only in the 19th century, when their life cycle was studied, did the difference of this group from fungi become clear. During the same period, microscopic structures of fruiting bodies, as well as ornamentation of the spore surface, began to be considered as diagnostic features. Due to this, in the period from the end of the 19th to the middle of the 20th century, a rather stable system was formed. However, as further studies have shown, both macro- and micromorphological characters are often quite variable, depend on environmental conditions, and often result from convergent evolution, which causes difficulties in defining species and taxonomic units of higher ranks. Since the first decade of the 21st century, due to the development of molecular genetic methods and the accumulation of data on nucleotide sequences of marker genes together with the improvement of microscopic studies, it has been possible to obtain data on the evolutionary relationships of different groups of myxomycetes. A milestone in this process was the publication of the first phylogenetic system of myxomycetes in 2019. This work was the starting point for a number of studies on the relationships between different groups of myxomycetes at a lower taxonomic level. Thus, there has been a surge in the number of studies that bring us closer to constructing a natural system.

Visualizing liquid distribution across hyphal networks with cellular resolution.

Filamentous fungi and fungal-like organisms contribute to a wide range of important ecosystem functions. Evidence has shown the movement of liquid across mycelial networks in unsaturated environments, such as soil. However, tools to investigate liquid movement along hyphae at the level of the single cell are still lacking. Microfluidic devices permit the study of fungal and fungal-like organisms with cellular resolution as they can confine hyphae to a single optical plane, which is compatible with microscopy imaging over longer timescales and allows for precise control of the microchannel environment. The aim of this study was to develop a method that enables the visualization and quantification of liquid movement on hyphae of fungal and fungal-like microorganisms. For this, the fungal-fungal interaction microfluidic device was modified to allow for the maintenance of unsaturated microchannel conditions. Fluorescein-containing growth medium solidified with agar was used to track liquid transported by hyphae via fluorescence microscopy. Our key findings highlight the suitability of this novel methodology for the visualization of liquid movement by hyphae over varying time scales and the ability to quantify the movement of liquid along hyphae. Furthermore, we showed that at the cellular level, extracellular movement of liquid along hyphae can be bidirectional and highly dynamic, uncovering a possible link between liquid movement and hyphal growth characteristics. We envisage that this method can be applied to facilitate future research probing the parameters contributing to hyphal liquid movement and is an essential step for studying the phenomenon of fungal highways.

The Disease Triangle of Chestnut: A review of host, pathogen, and environmental interactions of chestnuts cultivated in the eastern United States

Chestnuts, the edible seeds of the genus Castanea, are a perennial food crop closely tied to the global migration of humans throughout history and have recently been gaining popularity in agriculture and forest restoration in eastern North America. Cultivation of chestnuts yields nutritionally balanced food while fostering economic development, food security, and environmental health. However, diseases and insect pests threaten successful ecological restoration and food production. In this review we explore conditions affecting chestnuts in the eastern United States through the lens of the disease triangle. A “host” in the disease triangle is not merely a single tree, but a tree including its constituent population of fungal endophytes. Chestnut trees are rich with microbial life, and the sustainability of chestnuts in forest and cultivated settings may lie in understanding and manipulating microbial communities to improve plant health and control destructive diseases. To benefit from the ecological and economic potential of chestnuts on the landscape, it may be necessary to select locally adapted chestnut trees, regardless of pedigree, that are resilient against cosmopolitan pathogens. With transport of plants and pathogens throughout the globe, and with landscape level environmental changes over the last century, chestnut trees in the eastern United States (U.S.) are in a unique disease landscape compared to their ancestors. Diseases of economic concern from fungi and fungal-like organisms include chestnut blight (Cryphonectria parasitica) and ink disease (Phytophthora cinnamomi) on American and European chestnuts, oak wilt (Bretziella fagacearum) on all chestnut species, and the emerging diseases of brown rot (Gnomoniopsis smithogilvyi) and chestnut anthracnose (Colletotrichum henanense). The eastern U.S. has experienced profound environmental changes over the 20th century and into the early 21st century. These changes happen to coincide with the demise of the American chestnut in the eastern forest, efforts to re-establish chestnut as a forest species, and the rise in cultivation of multiple chestnut species and hybrids as a culinary crop. Chestnut trees growing in the early 21st century face different environmental circumstances than the American chestnuts of pre-colonial times, including changes in forest composition, rainfall changes and acidification, industrialized agriculture’s increased chemical inputs, rising global temperatures, and increased levels of carbon dioxide in the atmosphere. We conclude that chestnut tree species for both forestry and agriculture should be considered based on current fitness, adaptability, and economic and ecological value considering continued dynamics in host and pathogens distributions and anthropogenically driven climatic and edaphic conditions.

Fungal and Fungal-like Diseases of Halophytes in the Mediterranean Basin: A State-of-the-Art Review

Halophytes are salt-tolerant plants growing in saline ecosystems and are spread throughout the Mediterranean area. Recently, there has been a renewed interest in agricultural exploitation of halophytes, but poor attention has been given to pest and disease management of these species. The objective of this review is to assess the extent and distribution of pathogenic fungal and fungal-like (Oomycota) organisms on major Mediterranean halophytes. We retrieved 92 references spanning over 100 years, with more than half published since 2000. Ascomycota is the best-represented phylum, and the order Pleosporales has the highest species diversity. The order Pucciniales prevails in Basidiomycota, whereas Peronosporales is the richest order within Oomycota. Most of the pathogenic species have been isolated from aboveground organs, especially from leaves. Portulaca oleracea is the species with the most associated pathogens (16) and records (28). Leveillula powdery mildew, caused by Leveillula taurica, is the most common disease among the selected species. Cakile maritima was found to be vulnerable to different mycotoxigenic Alternaria species. Strengthening the research on diseases of halophytes is essential to successfully grow these species and to evaluate the risks related to the presence of mycotoxigenic species, which is crucial for the effective exploitation of halophytes as crops.

A Survey of Fungi and Fungus-Like Organisms at the Albany Pine Bush Preserve, a Globally Rare Inland Pine Barren System

A Survey of Fungi and Fungus-Like Organisms at the Albany Pine Bush Preserve, a Globally Rare Inland Pine Barren System

Current state of the species composition of mycobiota in South-East Kazakhstan

To date, no information has been available on the species composition of fungi in southeastern Kazakhstan, the center of agrobiodiversity but also home to rare and endemic species. Therefore, our objective was to investigate the diversity of fungi, fungus-like organisms, and lichens in southeastern Kazakhstan. The scientific and practical significance of the work is to obtain data on the species composition of mycobiota, which will allow planning measures for its monitoring, protection and restoration. The material for the article served as the authors’ own collections, conducted in 2021-2023. The study and identification of fungi were carried out according to the standard methodology. In the study area 2864 species of fungi, fungus-like organisms and lichens from 3 kingdoms, 22 classes, 84 orders, 260 families and 770 genera were found. Among them, 36 species were new to southeastern Kazakhstan, and 8 species were new to Kazakhstan. It is worth noting that the Karasay region had the largest number of species (1547) when analyzing the distribution of fungi, fungus-like organisms, and lichens. The study’s value and practical significance lies in the data obtained on the species composition of fungi, fungus-like organisms, and lichens, including their distribution and confinement to different types of substrates.

Micromycetes associated with barley and their significance as pathogens in Russia

Despite the great attention paid to the study of barley diseases, inaccuracies in the names of diseases, their practical significance, and incorrect use of the scientific names of causal agents can be commonly found in the scientific literature. This may lead to confusion and misidentification of the pathogens that can affect this crop especially as relates to phytosanitary requirements. This review continues the series started with a publication devoted to wheat diseases. This review includes information about the main barley diseases and pathogenic organisms causing them, as well as the species emerging as a potential threat to barley. The current taxonomic status of fungal species and fungal-like organisms associated with various organs of barley is given, and the breadth of their distribution and degree of impact on the crop are summarized. The micromycetes were divided into two groups according to their phytosanitary importance. The first group is represented by fungi of great importance as the pathogens causing the 29 economically important common barley diseases. The second group consists of fungi causing 20 minor and poorly studied diseases with unconfirmed harmfulness, or potential endophytic fungi. Perceptions of their ability to cause disease remain controversial and the available data require confirmation. This dataset can be used as a reference for a more accurate description of the phytosanitary situation. The review will also be helpful for more targeted studies using molecular techniques to clarify taxonomy and areals of fungi associated with barley and to provide more detailed data on disease damage in this crop.

ГРИБЫ, АССОЦИИРОВАННЫЕ С NUPHAR LUTEA (L.) SMITH, В р. КОТОРОСЛЬ (ЯРОСЛАВЛЬ, РОССИЯ)

Studies of mycobiota of Nuphar lutea leaves in the estuary part of the Kotorosl River, located in the center of a large city, showed that phylloplane fungal complexes in 2006, 2007 and 2021 years form and consist of a small number of more or less constant species and a large number are temporary. Fungi and fungal-like organisms in all cases become participants in the primary stages of destructions of dead leaves. These are some inhabitants of phylloplane complexes, spotable to switch to feeding on leaf tissues, and new colonizers, the life cycle of which completely occurs in water in 2006 and 2007 years. In 2021 we found only poorly developed mycelium. Antropogenic impact on the river affects the species composition, and most importantly – the intensity of grows and the passage of the full life cycle of micodestructors.

Conservation of putative transcription factor binding sites of co-expressed Plasmopara halstedii genes in two Phytophthora species

Oomycetes, a large group of fungus-like organisms, include some destructive plant pathogens causing enormous economic damage. Phylogenetically, oomycetes belong to the kingdom Straminipila and have diverse lifestyles, including saprotrophs and both general and specialized pathogens of various eukaryotic supergroups. A rapid increase in genomic studies and next-generation sequencing technologies have led to significant progress in understanding oomycete lifestyles. However, their genetics, including transcriptional regulation, have been studied to a much lesser extent. Here, we provide a cross-species analysis of oomycete promoter for providing a first step towards elucidating gene regulation networks related to pathogenicity and life cycle stages. The clustered DNA sequences of Plasmopara halstedii transcriptome time-series expression level dataset from a preliminary study have been used as a core reference for cross-species comparisons. Using a computational pipeline, 46 potential transcription factor binding site (TFBS) motifs in 25 clusters with functionally conserved downstream genes of downy mildew and two Phytophthora species, regardless of the gene expression levels of Phytophthora transcriptomes, were found. This can now be followed up by knock-out experiments in oomycete species amenable for genetic modification.

Marine Labyrinthulomycetes

Labyrinthulomycetes are a small but very important group of marine fungi-like organisms. Labyrinthulomycetes are ubiquitous, can be associated with living plants, algae, and animals and can decompose a variety of organic remains. These organisms are able to synthesize and abundantly accumulate lipids, in particular, polyunsaturated fatty acids, some of which are of great interest for biotechnology. The scientific information about labyrinthulomycetes is currently accumulating very quickly. Our study is devoted to reviewing presently available data on the structure, biology and taxonomy of these organisms. The review examines the problems of methodological approaches to the study of these organisms, as well as the diversity, distribution and significance of Labyrinthulomycetes in marine ecosystems.

Adding a missing piece to the puzzle of oomycete phylogeny: the placement of Rhipidium interruptum (Rhipidiaceae).

Oomycetes are a group of fungus-like organisms, which phylogenetically comprise early diverging lineages that are mostly holocarpic, and two crown classes, the Peronosporomycetes and Saprolegniomycetes, including many well-investigated pathogens of plants and animals. However, there is a poorly studied group, the Rhipidiales, which placement amongst the crown oomycetes is ambiguous. It accommodates several taxa with a sophisticated vegetative and reproductive cycle, as well as structural organisation, that is arguably the most complex in the oomycete lineage. Despite the remarkable morphological complexity and their notable perseverance in the face of faster-growing saprotrophic oomycetes and fungi, the knowledge on Rhipidiales is limited to date, as the most complex members are not easily cultured, even by targeted approaches. This also leads to inadequate sequence data for the order, which was sourced from only the two least complex out of seven introduced genera, i.e. Sapromyces and Salispina. In the present study, ex-situ baiting was done using various fruit substrates, and naturally-shed twigs or fruits acquired from water bodies were examined. As a result of these efforts, the species Rhipidium interruptum was obtained and gross cultivation was accomplished using poplar (Populus nigra) twigs as substrate, which allowed further documentation of both asexual and sexual reproduction. This enabled phylogenetic and detailed morphological study, as well as an epitypification of the species. Phylogenetic analyses based on cox2 and nrLSU sequences revealed Rhipidium as the sister genus of Sapromyces. The morphological studies done support a conspecificity of R. interruptum and R. continuum, which might in turn be conspecific with R. americanum. Though several further studies will be required to fit the scattered missing pieces of knowledge on Rhipidiales together revealing a more complete picture of oomycete evolution, we hope that the current study can serve as a cornerstone for future investigations in the group. Citation: Tsai I, Thines M (2023). Adding a missing piece to the puzzle of oomycete phylogeny: the placement of Rhipidium interruptum (Rhipidiaceae). Fungal Systematics and Evolution 11: 95-108. doi: 10.3114/fuse.2023.11.08.

The Volyn biota (Ukraine) – indications of 1.5 Gyr old eukaryotes in 3D preservation, a spotlight on the “boring billion”

Abstract. The Volyn biota, fossilized organisms with a minimum age of 1.5 Ga, were found in cavities in granitic pegmatites from the Korosten Pluton, NW Ukrainian shield. Fossilization was due to an influx of hydrothermal fluorine-rich waters, which silicified the outermost part of the organisms, thus preserving the 3D morphology. Details of the morphology (investigated by scanning electron microscopy) show that the majority of the specimens are filamentous, of a large variety with diameters ranging from ∼ 10 to ∼ 200 µm, thin filaments with typical branching and thick filaments with ball-shaped outgrowths and dented surface. Filaments can be straight or conical, curvilinear, or strongly curved, up to millimeters in length, some with a central channel. Some filaments show indications of segmentation and are grown as sessile organisms onto substrate; others show both intact ends, indicating a non-sessile, free-living lifestyle. Objects with flaky morphology and agglutinating filaments are interpreted as fossil biofilms. Other objects are hollow and show a large variety of forms; spherical objects are scarce. Infrared spectroscopy indicates the presence of chitosan in one filament type, electron microprobe analysis of nanometer-sized inclusions in filaments identified the presence of Bi(Te,S) minerals, and both observations are compatible with the interpretation as fungi-like organisms. Stable C- and N-isotope data of bulk samples are in the range of −31 ‰ to −47 ‰ δ13C and of +3 ‰ to +10 ‰ δ15N, indicating possible methanogens as part of the subsurface microecosystem. The Volyn biota indicate that at 1.5 Ga complex forms of life existed in the continental deep biosphere, well above the microscopic level, including fungi-like organisms resembling eukaryotes.

How Does the Sample Preparation of Phytophthora infestans Mycelium Affect the Quality of Isolated RNA?

RNA isolation from fungi and fungus-like organisms is not an easy task. Active endogenous RNases quickly hydrolyze RNA after the sample collection, and the thick cell wall prevents inhibitors from penetrating the cells. Therefore, the initial collection and grinding steps may be crucial for the total RNA isolation from the mycelium. When isolating RNA from Phytophthora infestans, we varied the grinding time of the Tissue Lyser and used TRIzol and beta-mercaptoethanol to inhibit the RNase. In addition, we tested the mortar and pestle grinding of mycelium in liquid nitrogen, with this method showing the most consistent results. During the sample grinding with the Tissue Lyser device, adding an RNase inhibitor proved to be a prerequisite, and the best results were achieved using TRIzol. We considered ten different combinations of grinding conditions and isolation methods. The classical combination of a mortar and pestle, followed by TRIzol, has proved to be the most efficient.

Copper Can Be Elevated in Hydroponics and Peat-based Media for Potential Disease Suppression: Concentration Thresholds for Lettuce and Tomato

Copper (Cu) is typically adequate at 0.5 μM (0.03 ppm) in hydroponics and at 2 μM (0.125 ppm) in soilless media, but elevated levels can be used to inhibit pathogenic fungal growth. We studied the effect of elevated Cu on the growth of lettuce and tomato in peat-based media and deep-flow hydroponics. Lettuce growth in hydroponics was not hindered until a concentration greater than 4 μM (0.25 ppm) Cu was used, which is eight times greater than the adequate level. Tomato was more tolerant of elevated Cu, with no growth suppression up to 8 μM (0.5 ppm) in hydroponics. Organic matter tightly binds Cu, and bioavailability is thus determined by organic components in soilless media. We confirmed an adsorption capacity of 19 mg Cu per g of peat, which explains why there was no inhibition of lettuce or tomato growth up to 1000 μM (64 ppm) Cu in peat-based media. When chelated with ethylenediaminetetraacetic acid, Cu binding to organic matter was reduced and growth was decreased in lettuce but not tomato. Both species tolerated a 100-fold greater concentration of Cu in peat-based media than in deep-flow hydroponics. Elevated Cu in solution increased concentrations 20 times greater in root tissue than in leaves. These solution and tissue concentrations are greater than identified toxicity thresholds of pathogenic fungal and fungal-like organisms, and could thus be used to suppress root-borne fungal and fungal-like diseases.

The history of research of fungi and fungi-like organisms on the territory of the Hutsulshchyna NNP and the role of security, protection and the environmental measures by in situ, ex situ, re situ in conserving their diversity

The chronology of researches the fungi and fungi-like organisms on the territory of the Hutsulshchyna National Nature Park is described. Information is bases on the Chronicles of Nature of the Park since the time of it creation. The contribution of researchers in the study of the fungal biodiversion of the Park is noted. 1.01.2022 List of fungi of the Park counts 1214 species, of which 23 species are included in the Red Book of Ukraine. The information about in situ, ex situ and re situ methods are represented. In situ method aims the preservation of fungi and their localities. Ex situ method is expressed by introduction into artificial culture of fungi and maintenance of viability of fungal mycelium in collections of pure cultures. Re situ method is expressed by reproduction of aboriginal strains species of fungi in their natural conditions, involving in situ and ex situ practice. The environmental effect of using in situ, ex situ and re situ measures on the territory of Hutsulshchyna National Park is described. The development of growing methods for rare species of fungi in laboratory conditions is pointed. Reducing the threat of extinction of rare fungi due to re-situ technology and systematic fruiting of fungi at the mycological reproductive areas is explained. Certain legal aspects of protection of fungi and mushroom-like organisms are described within the "Program of protection, preservation and reproduction of rare species of fungi", approved by the Scientific and Technical Council of NNP "Hutsulshchyna". The question of creating the protection zones for rare species of fungi is outlined. The collections of macromycetes of Hutsulshchyna National Park are briefly described. The collection of pure cultures numbers 7 species of 13 strains, the collection of herbarium and wet preparations numbers 175 specimens. The important scientific and educational role of the collections is noted, the main purpose of their further operation is marked, as well as the methods and results of popularization of scientific researches among children, youth and adults. The connection of scientific achievements with the creation of a favorable economic climate for the community is indicated. Probably positive results of introduction of new recreational services - fungi therapy routes and commercial cultivation of new species of edible, rare, pharmaceutically popular species of fungi are marked.

Fungi and fungus-like organisms of the Kivertsi National Nature Park “Tsumanska Pushcha”

As a result of mycological surveys of the territory of Kivertsi National Nature Park “Tsumanska Pushcha” during 2004–2021, 203 species of fungi and fungus-like organisms were detected and registered. A comparative analysis of the species composition of the mycoflora of the park was carried out. The most diversified and rich was the phylum Basidiomycota, which comprises 172 species, belonging to 48 genera of 49 families, ten orders of the class Agaricomycetes. Many rare macromycetes that are rare or relatively rare for Ukraine (i.e., Cortinarius bolaris, Craterellus cornucopioides, Entoloma bloxamii, Fistulina hepatica, and Gyroporus castaneus) and listed in the Red Book of Ukraine (Grifola frondosa, Phaeolepiota aurea, Pseudoboletus parasiticus, and Sparassis crispa) have been discovered. For two species listed in the Red Book of Ukraine (Grifola frondosa and Pseudoboletus parasiticus), new localities have been discovered. As of today, the conducted studies expand information about the mycoflora of the Kivertsi National Nature Park “Tsumanska Pushcha”. However, the mycoflora of the park still requires further, more detailed research and inventory.

Pythium spp. – rosnący problem w uprawie ogórka pod osłonami

In 2022 and 2023, a record number of cucumber samples came to the Plant Diseases Clinic in Institute of Plant Protection – National Research Institute in Poznan. The dominant problem (more than 90% of the samples) was the sudden wilting of healthy-looking plants, despite adequate water availability. The yield decreased drastically, and the plants in the entire rows dried up and died prematurely. Until 2019, similar symptoms appeared sporadically on cucumbers delivered to the Clinic. On the basis of microscopic observation, confirmed by isolation on artificial substrates from fragments of infected cucumber, the cause of the disease was identified – fungus-like organisms (Oomycetes) of the genus Pythium. In soilless cultivation systems, they cause soft rot of the roots and the shoot base of cucumber and the so-called sudden wilting of a cucumber. Typical symptoms of infection are stunted plant growth, brown discoloration of the root tips and wilting of the plant during the warmer times of the day (they regain turgor during the night). Infected plants turn yellow and die. Infection can occur all year round. In summer, thermophilic species are the most serious problem, while in winter and early spring, spe¬cies preferring lower temperatures. The samples delivered to the Clinic were dominated by the species Globisporangium ultimum (Trow) Uzuhashi, Tojo & Kakish commonly known as Pythium ultimum Trow. The importance of Pythium spp. in the cultivation of cucumber under cover increases year by year, taking into account the frequency of occurrence in the samples delivered to the Clinic.

Mycobiota of Dead Reed Fragments Immersed in Water (Yaroslavl Region, Russia)

The mycobiota on dead submerged plant parts of Phragmites australis reed (leaves, stems, leaf sheaths) was studied in lakes – Pleshcheyevo, Mostetskoye, a lake in the Zavolzhsky district of Yaroslavl and the Kotorosl river (Yaroslavl region, Russia). The kingdom Fungi is represented by 53 species belonging to 33 genera, 25 families, 12 orders, and 7 classes in 2 taxonomic phyla: Mucoromycota (4 species), Ascomycota (48). The kingdom Chromista (fungi-like organisms) includes 8 species belonging to 3 genera, 2 families, 2 orders, and 1 class under phylum Oomycota. For the first time, 23 species of micromycetes were found on reeds. Representatives of the orders Pleosporales (16 species), Helotiales (11), Saprolegniales (7), Hypocreales (5) prevailed. The number of species of fungi on fragments of plants in different aquatic bodies varied from 16 to 39, the statistically significant difference in the species composition of communities of dead reeds was found between a lake in the Zavolzhsky district of Yaroslavl and the Kotorosl River (R = 0.646, significance level of sample statistic 0.1%).

Біотичне різноманіття урочища «Сатанівська дача» — об’єкта Всесвітньої спадщини ЮНЕСКО (Україна)

Abstract. The Satanivska Dacha tract in Khmelnytskyi Oblast, Ukraine, is unique in that Fagus sylvatica L. grows here at the edge of its geographic range. In 2017, the forest community was included in the trans-European World Heritage site ‘Primeval beech forests and ancient forests of the Carpathians and other regions of Europe" by UNESCO under the name Site No. 61 ‘Satan’s Dacha.’ In the territory of the tract, 163 species of vertebrates have been recorded, including 12 species of amphibians, 6 species of reptiles, 96 species of birds, and 50 species of mammals, which are representatives of 24 orders. Within the world heritage site, 72 species of vertebrates have been recorded: 7 species of amphibians, 3 species of reptiles, 39 species of birds, and 23 species of mammals. There are 135 species of vertebrates that have protection statuses and included in the Red Data Book of Ukraine (33 species), Regional Red List (23 species), Bern (134 species), Bonn (43 species), and Washington (17 species) conventions. Of higher plants, 234 species have been recorded, including 169 species within the world heritage site, and 7 species are included in the Red Data Book of Ukraine (2009, 2021) and 5 species are included in the CITES list. The List of Regionally Rare Species in Need of Protection in Khmelnytskyi Oblast includes 24 species that were found in the Satanivska Dacha tract, of which 17 species grow in the territory of the world heritage site. The bryoflora of the tract consists of 98 bryophytes. The mycoflora, according to preliminary studies, includes 61 species of fungi and fungus-like organisms. According to the EUNIS classification, rare biotopes represent the following type—G: Woodland, forest and other wooded land; G 1.6: Beech forests (Fagus woodland). Three rare associations have been identified that have a synsozological status according to the Directive of the Council of Europe 92/43/EEC. The population status of Allium ursinum L., Galanthus nivalis L., Epipactis purpura Smith, Epipactis helleborine (L.) Crantz, and Barbastella barbastellus Schreber was monitored. The territory is characterised by a high potential as an environment for the self-regeneration of the biota on the whole. Continuation of monitoring programmes of this type is extremely important.