Armillaria Research Articles

5'-Methoxyarmillane, a Bioactive Sesquiterpenoid Aryl Ester from the Fungus Armillaria ostoyae.

Higher fungi of the genus Armillaria belonging to the phylum Basidiomycota produce bioactive sesquiterpenoid aryl esters called melleolides. A bioactivity-guided discovery process led to the identification of the new melleolide 5'-methoxyarmillane (1) in organic extracts from the mycelium of Armillaria ostoyae. Remarkably, supplementation of rapeseed oil to the culture medium potato dextrose broth increased the production of 1 by a factor of six during the course of the 35 days fermentation. Compound 1 was isolated and its structure elucidated by UHPLC-QTOF-HR-MS/MS and NMR spectroscopy. It showed toxicity against Madin-Darby canine kidney II (MDCK II, IC50 19.2 μg/mL, 44.1 μM) and human lung cancer Calu-3 cells (IC50 15.2 μg/mL, 34.9 μM) as well as moderate bioactivity against Mycobacterium tuberculosis (MIC 8 mg/mL, 18.4 μM) and Mycobacterium smegmatis (MIC 16 mg/mL, 36.8 μM), but not against Staphylococcus aureus, Escherichia coli, Candida albicans, and Septoria tritici. No inhibitory effects of 1 against the influenza viruses H3N2, H1N1pdm, B/Malaysia, and B/Massachusetts were observed.

Molecular identification of wood-decaying fungi of <i>Armillaria</i> genus widespread in Eastern Siberia and the Far East of Russia using ITS, IGS-1-1 and <i>Tef-1α</i> genetic markers

The genus Armillaria is an essential component of forest ecosystems playing very important ecological role in dead wood decomposition, but it often becomes a serious pathogen causing white root rot in trees. It is also known that Armillaria species significantly differ in the level of pathogenicity. Thus, accurate identification of Armillaria is critical for assessing the risk of tree disease. In this study we analyzed 28 Armillaria isolates from Siberia and the Far East using nucleotide sequences of ITS, IGS-1-1 and TEF-1α gene regions and generated phylogenetic trees based on maximum likelihood method. In total, four Armillaria species were identified: A borealis, A. cepistipes, A. ostoyae and A. gallica. A. borealis was the most frequent among collected isolates (18 out of 28 isolates). A. gallica, A. cepistipes and A. ostoyae were much less frequent with two, five and three isolates out of 28, respectively. Thedistribution of Armillaria species in Siberia and the Far East was described for the first time. It is concluded that further studies are necessary to determine the role of Armillaria in trees pathological dieback, and A. borealis should be a key focus.

Characterization of Mycoviruses in Armillaria ostoyae and A. cepistipes in the Czech Republic.

Members of the genus Armillaria are widespread forest pathogens against which effective protection has not yet been developed. Due to their longevity and the creation of large-scale cloning of Armillaria individuals, the use of mycoviruses as biocontrol agents (BCAs) against these pathogens could be an effective alternative. This work describes the detection and characterization of viruses in Armillaria spp. collected in the Czech Republic through the application of stranded total RNA sequencing. A total of five single-stranded RNA viruses were detected in Armillaria ostoyae and A. cepistipes, including viruses of the family Tymoviridae and four viruses belonging to the recently described "ambivirus" group with a circular ambisense genome arrangement. Both hammerhead (HHRz) and hairpin (HpRz) ribozymes were detected in all the ambiviricot sequences. Armillaria viruses were compared through phylogenetic analysis and confirmed their specific host by direct RT-PCR. One virus appears to infect both Armillaria species, suggesting the occurrence of interspecies transmission in nature.

Armillaria korhonenii, the sixteenth biological species of Armillaria from China

Armillaria korhonenii, designated as Chinese biological species Q (CBSQ) here, is described as a new species based on phylogenetic and morphological evidence as well as the results of mating tests. This species is the sixteenth biological species of Armillaria reported from China following the discovery of 15 previously known biological species. Phylogenetically, A. korhonenii is the earliest divergent lineage of the genus Armillaria as inferred from the nucleotide sequences of the Elongation Factor 1-Alpha (EF1a). Morphologically, it is characterized by its ellipsoid to elongate basidiospores, yellowish orange to honey yellow pileus with fibrillose squamules. Mating tests indicated that A. korhonenii is a tetrapolar heterogeneous species and is inter-sterile with all the tested species except partial compatibility with CBSH.

Decoding the RNA virome of the tree parasite Armillaria provides new insights into the viral community of soil-borne fungi.

The globally distributed basidiomycete genus Armillaria includes wood decomposers that can act as opportunistic parasites, causing deadly root rot on woody plants. To test whether RNA viruses are involved in this opportunistic behaviour, a large isolate collection of five Armillaria species collected over 40 years in Switzerland from trees, dead wood and soil was analysed. De novo assembly of RNA-Seq data revealed 21 viruses, 14 of which belong to putative new species. Two dsRNA viruses and an unclassified Tymovirales are formally described for the first time for Armillaria. One mitovirus occurred with a high prevalence of 71.1%, while all other viruses were much less prevalent (0.6%-16.9%). About half of all viruses were found only in one fungal species, others occurred in 2-6 fungal species. Co-infections of 2-7 viruses per isolate were not uncommon (34.9%), and most viruses persisted circulating within fungal populations for decades. Some viruses were related to viruses associated with other Armillaria species, supporting the hypothesis that virus transmission can occur between different fungal species. Although no specific correlation between viruses and the fungal trophic strategy was found, this study opens new insights into viral diversity hidden in the soil microbiome.

An introduction to edible Armillaria novae-zelandiae from Indonesia

The majority study of Armillaria globally had been focused on A. mellea. The Indonesian data of genus Armillaria remains poor. During our fungus foray in West Java (Indonesia), some basidiomata of supa jambu (local name) consumed by the local people were collected. In the field, the specimens were morphologically identified as Armillaria. The present study was to confirm the species identity of our specimens based on morphological and molecular evidence. The fresh basidiomata were used for the examination. The combination of morphological and molecular analyses confirmed our specimen as Armillaria novae-zelandiae. Armillaria novae-zelandiae FIPIA-DEP50 was identified by its viscid, thin, striate, honey-yellow pileus, semi-bulbous to bulbous stipe which darken to black with age towards the base. Spore moderately thick walled, ellipsoid, to elongate-ellipsoid, smooth with some internal structure. Armillaria novae-zelandiae FIPIA-DEP50 is generally smaller in stature than the other closely related species (A. sparrei and A. hinnulea). The BLAST result revealed that our specimen exhibits 98% similarity to Armillaria novae-zelandiae as the top hit. The phylogenetic tree nested our specimens with A. novae-zelandiae clade (BS 100%) and A. mellea as its sister clade. The current study provides the first edibility information and herbarium collection of A. novae-zelandiae for Indonesia.

The Armillaria response to Gastrodia elata is partially mediated by strigolactone-induced changes in reactive oxygen species

Armillaria root diseases, caused by Armillaria spp., pose a significant threat to woody plants worldwide and result in substantial economic losses. However, certain species in the genus Armillaria can establish a unique symbiotic relationship with Gastrodia elata, which is the only known example of a plant benefiting from Armillaria. Although various plant signals that play a role in this interaction have been identified, the mechanism remains largely unknown from the Armillaria’s perspective. In this study, we performed whole-genome sequencing of an Armillaria gallica strain named NRC001 isolated from G. elata. Comparative genomic analysis showed it is low-pathogenic Armillaria spp., which possesses 169 expanded gene families compared to high-pathogenic Armillaria spp. Among these expanded families, transcriptomic analysis revealed a significant increase in expression levels of four reactive oxygen species (ROS)-related gene families in A. gallica on G. elata compared to A. gallica on wood. Thus, a systematic survey of ROS-related gene families was carried out, and a total of 218 genes belonging to 44 ROS-related gene families in A. gallica were identified. Physiological experiments and transcriptome analysis showed that strigolactones (SLs) released by G. elata have a mediation impact on ROS, particularly enhancing the ROS scavenging activities by increasing the expression level and activity of several enzymes, such as catalase and glutathione reductase. Among the ROS-related genes, the aquaporin (AQP) is crucial as it is responsible for transporting hydrogen peroxide (H2O2) across the cell membrane. Five orthologs of AQP genes in A. gallica were identified and overexpressed in yeast. Only AgAQPA from the so-called ‘other aquaglyceroporin’ subfamily was demonstrated to be capable of mediating H2O2 transport in A. gallica. To our best knowledge, this is the first ‘other aquaglyceroporins’ gene in fungi to be identified as having transporter capacity. This study not only provides new insights into the mechanisms by which SL signaling regulates interactions between Armillaria and G. elata, but also sheds light on the function of fungal AQPs.

Honey Mushroom, Armillaria mellea (Agaricomycetes) and Its Fermentation Products Target Regulation of OAT1/OAT3 Proteins to Reduce Hyperuricemia in Mice.

Disorders of purine metabolism are the main cause of hyperuricemia. Current drugs for the treatment of hyperuricemia usually cause a degree of cardiovascular damage. This study aimed to investigate the therapeutic effects of Armillaria mellea fruiting body (AFB), Armillaria rhizomorph (AR) and Armillaria mellea fermentation product (after rhizomorphs removal) (AFP) on hyperuricemic mice. The hyperuricemia mouse model was established by oral administration of potassium oxonate 0.9 g⋅kg-1 and hypoxanthine 0.5 g⋅kg-1 for two weeks. Starting from the third week, the intragastric administration of the intervention drug group was as follows: Allopurinol 0.013 g⋅kg-1, AFB (3.9 and 7.8 g⋅kg-1), AR (3.9 and 7.8 g⋅kg-1), AFP (1.95 and 3.9 g⋅kg-1) once daily for 14 days. Results showed that AFB, AR, and AFP reduced the contents of serum uric acid, serum creatinine, and blood urea nitrogen in hyperuricemic mice and the mechanism of action might be through up-regulation of the expression levels of organic anion transporter 1/organic anion transporter 3 proteins in kidney tissue. AR and AFP both exhibited better uric acid-lowering effects than AFB, which may be due to the higher purine content of AFB. Armillaria mellea and its fermentation products can treat hyperuricemia by up-regulating OAT1 protein and OAT3 protein, reducing uric acid content in mice.

5′-O-methyl-14-hydroxyarmillane, a new armillane-type sesquiterpene from cultures of Guyanagaster necrorhiza

Protoilludene-type sesquiterpene aryl esters are a unique and very diverse compound class that were exclusively isolated from members of the genus Armillaria (Agaricomycetes, Physalacriaceae) up to this point. Herein, we describe the isolation and structural characterization of 5′-O-methyl-14-hydroxyarmillane (1), a new armillane-type derivative, that was obtained from submerged cultures of Guyanagaster necrorhiza (CBS 138623) together with the known congeners melleolide G (2), melleolide B (3), and 10-dehydroxy-melleolide B (4). ROESY data and coupling constants assigned the relative configurations of 1, while common absolute configurations were confirmed from comparison of its ECD spectrum to the one of 10-hydroxy-5′-methoxy-6′-chloroarmillane (5). Additionally, the configuration of melleolide G (2) was revised based on observed ROESY correlations. It is the first time that protoilludene-type sesquiterpene aryl esters were isolated from another genus, namely, Guyanagaster, that is closely related to Armillaria. 1–4 were evaluated for their biological activities in a serial dilution assay against several yeast, fungi, and bacteria, as well as in a cytotoxicity assay against different cell lines. Compound 4 was moderately active against Bacillus subtilis, Staphylococcus aureus, and Mucor hiemalis. Furthermore, 1, 3, and 4 showed weak cytotoxic effects against the mouse fibroblast cell line L929 and the cervix carcinoma cell line KB3.1.

Belarusian Oak Forests Afflicted with Armillaria Root Rot during Their Mass Desiccation

During the period of the oak forests’ mass desiccation, Armillaria root rot became widespread, exacerbating the state of depression. The infection of stands by white sapwood rot caused by fungi of the genus Armillaria was more often detected in Bug-Polesia and Berezina-Pre-Polesia forest growth areas. The incidence of the disease in the oak forests of Belarus increases with the raising of the stands age and the decrease in their density. More often, the affliction with Armillaria root rot occurs in floodplain oak forests, as well as in the dry lands forest types with the most fertile soils. The oak’s proportion in the stand’s composition does not significantly affect the occurrence of the infection. Under the conditions of massive weakening of the Belarussian oak forests, facultative parasites from the Armillaria genus can occur as dangerous secondary pathogens that accelerate the death of weakened oak trees. Their pathogenicity persists in the north of the republic, where the negative role of other pathological factors in oak forests decreases. After the end of the depression period, the transition of Armillaria from a parasitic to a predominantly saprotrophic strategy was recorded. The affliction of oak forests by Armillaria root rot can act as one of the oak formation’s condition indicators, marking the depression phase in oak stands. Therefore, the spread of foci of Armillaria root rot should be monitored while conducting forest pathology surveys and monitoring the condition of oak forests.

Vertical and horizontal gene transfer shaped plant colonization and biomass degradation in the fungal genus Armillaria.

The fungal genus Armillaria contains necrotrophic pathogens and some of the largest terrestrial organisms that cause tremendous losses in diverse ecosystems, yet how they evolved pathogenicity in a clade of dominantly non-pathogenic wood degraders remains elusive. Here we show that Armillaria species, in addition to gene duplications and de novo gene origins, acquired at least 1,025 genes via 124 horizontal gene transfer events, primarily from Ascomycota. Horizontal gene transfer might have affected plant biomass degrading and virulence abilities of Armillaria, and provides an explanation for their unusual, soft rot-like wood decay strategy. Combined multi-species expression data revealed extensive regulation of horizontally acquired and wood-decay related genes, putative virulence factors and two novel conserved pathogenicity-induced small secreted proteins, which induced necrosis in planta. Overall, this study details how evolution knitted together horizontally and vertically inherited genes in complex adaptive traits of plant biomass degradation and pathogenicity in important fungal pathogens.

Vliv mykorhizního přípravku a hnojiva na růst a napadení sazenic smrku ztepilého václavkami

On Norway spruce (Picea abies L. [H.] Karst.) seedlings, influence of three treatments – (i) inoculation with Ectovit® preparation containing ectomycorrhizal fungi (INOCULATION), (ii) Ectovit® + Conavit® fertilizer (INOCULATION + FERTILIZATION), and (iii) untreated group (CONTROL) – were evaluated in this study. In autumn 2021 (3.5 years after inoculation), 100 seedlings/treatment (300 seedlings in total) were sampled to evaluate seedlings survival percentage and growth characteristics, and 10 live seedlings/treatment (30 seedlings in total) to evaluate dry weight and the number of seedlings with Armillaria rhizomorphs. By relating these values to the previous evaluation (autumn 2019), values for period autumn 2019 – autumn 2021 were obtained for statistical analyses. Increment of the seedling height, increment of the root collar diameter and increment of the seedling aboveground dry weight were significantly higher in INOCULATION compared to INOCULATION + FERTILIZATION, the first and second named variables were also significantly higher in INOCULATION compared to CONTROL. There were no significant differences in seedlings survival percentage nor number of seedlings with Armillaria rhizomorphs (all identified as Armillaria ostoyae [Romagn.] Herink). Contrary to the previous evaluation (1.5 years after inoculation), the protective effect of artificial ectomycorrhizal inoculation against infection of seedling by honey fungus was not confirmed.

Metabolomics analysis of mycelial exudates provides insights into fungal antagonists of Armillaria

ABSTRACT The genus Armillaria has high edible and medical values, with zones of antagonism often occurring when different species are paired in culture on agar media, while the antagonism-induced metabolic alteration remains unclear. Here, the metabolome of mycelial exudates of two Chinese Armillaria biological species, C and G, co-cultured or cultured separately was analysed to discover the candidate biomarkers and the key metabolic pathways involved in Armillaria antagonists. A total of 2,377 metabolites were identified, mainly organic acids and derivatives, lipids and lipid-like molecules, and organoheterocyclic compounds. There were 248 and 142 differentially expressed metabolites between group C-G and C, C-G, and G, respectively, and fourteen common differentially expressed metabolites including malate, uracil, Leu-Gln-Arg, etc. Metabolic pathways like TCA cycle and pyrimidine metabolism were significantly affected by C-G co-culture. Additionally, 156 new metabolites (largely organic acids and derivatives) including 32 potential antifungal compounds, primarily enriched into biosynthesis of secondary metabolites pathways were identified in C-G co-culture mode. We concluded that malate and uracil could be used as the candidate biomarkers, and TCA cycle and pyrimidine metabolism were the key metabolic pathways involved in Armillaria antagonists. The metabolic changes revealed in this study provide insights into the mechanisms underlying fungal antagonists.

Enzymatic Degradation of Aflatoxin Accumulation in Dry Maize Kernels as A Promising Post-Harvest Mitigation Strategy

Background: Aflatoxin is one of the most problematic fungal-produced toxins as it is responsible for massive global agricultural losses and is deleterious to both human and animal health. Contamination of crops by certain strains of Aspergillus fungi accumulate aflatoxin in both post- and pre-harvest conditions. Methods: In this report, we tested the aflatoxin-degradation efficiency of an endogenously expressed enzyme in harvested maize kernels. In post-harvest conditions, equivalent loads of A. flavus were used to infect harvested maize kernels previously engineered to express an aflatoxin-degrading enzyme from the Honey mushroom fungus. Results: No measurable, or significantly reduced, levels of aflatoxin were detected in all the enzyme-expressing harvested kernels initially and then 3 days post –harvest the transgenic kernels amassed aflatoxin. Conclusions: This is the first report of an enzyme degradation of aflatoxin in a crop in harvested kernels. This demonstrates the potential of this strategy to aid in the mitigation of aflatoxin in post-harvest conditions.

Desirability Assessment with the Aid of Satellite Imagery and Chorographic Data in the Areas with Agricultural Land use in Esfahan; Geo-Spatial Analysis and Fuzzy Approach Nexus

Background: Aflatoxin is one of the most problematic fungal-produced toxins as it is responsible for massive global agricultural losses and is deleterious to both human and animal health. Contamination of crops by certain strains of Aspergillus fungi accumulate aflatoxin in both post- and pre-harvest conditions. Methods: In this report, we tested the aflatoxin-degradation efficiency of an endogenously expressed enzyme in harvested maize kernels. In post-harvest conditions, equivalent loads of A. flavus were used to infect harvested maize kernels previously engineered to express an aflatoxin-degrading enzyme from the Honey mushroom fungus. Results: No measurable, or significantly reduced, levels of aflatoxin were detected in all the enzyme-expressing harvested kernels initially and then 3 days post –harvest the transgenic kernels amassed aflatoxin. Conclusions: This is the first report of an enzyme degradation of aflatoxin in a crop in harvested kernels. This demonstrates the potential of this strategy to aid in the mitigation of aflatoxin in post-harvest conditions.

Dual RNA-Seq Profiling Unveils Mycoparasitic Activities of Trichoderma atroviride against Haploid Armillaria ostoyae in Antagonistic Interaction Assays.

Armillaria ostoyae, a species among the destructive forest pathogens from the genus Armillaria, causes root rot disease on woody plants worldwide. Efficient control measures to limit the growth and impact of this severe underground pathogen are under investigation. In a previous study, a new soilborne fungal isolate, Trichoderma atroviride SZMC 24276 (TA), exhibited high antagonistic efficacy, which suggested that it could be utilized as a biocontrol agent. The dual culture assay results indicated that the haploid A. ostoyae-derivative SZMC 23085 (AO) (C18/9) is highly susceptible to the mycelial invasion of TA. In the present study, we analyzed the transcriptome of AO and that of TA in in vitro dual culture assays to test the molecular arsenal of Trichoderma antagonism and the defense mechanisms of Armillaria. We conducted time-course analysis and functional annotation and analyzed enriched pathways and differentially expressed genes including biocontrol-related candidate genes from TA and defense-related candidate genes from AO. The results indicated that TA deployed several biocontrol mechanisms when confronted with AO. In response, AO initiated multiple defense mechanisms to protect against the fungal attack. To our knowledge, the present study offers the first transcriptome analysis of a biocontrol fungus attacking AO. Overall, this study provides insights that aid the further exploration of plant pathogen-biocontrol agent interaction mechanisms. IMPORTANCE Armillaria species can survive for decades in the soil on dead woody debris, develop rapidly under favorable conditions, and harmfully infect newly planted forests. Our previous study found Trichoderma atroviride to be highly effective in controlling Armillaria growth; therefore, our current work explored the molecular mechanisms that might play a key role in Trichoderma-Armillaria interactions. Direct confrontation assays combined with time course-based dual transcriptome analysis provided a reliable system for uncovering the interactive molecular dynamics between the fungal plant pathogen and its mycoparasitic partner. Furthermore, using a haploid Armillaria isolate allowed us to survey the deadly prey-invading activities of the mycoparasite and the ultimate defensive strategies of its prey. Our current study provides detailed insights into the essential genes and mechanisms involved in Armillaria defense against Trichoderma and the genes potentially involved in the efficiency of Trichoderma to control Armillaria. In addition, using a sensitive haploid Armillaria strain (C18/9), with its complete genome data already available, also offers the opportunity to test possible variable molecular responses of Armillaria ostoyae toward diverse Trichoderma isolates with various biocontrol abilities. Initial molecular tests of the dual interactions may soon help to develop a targeted biocontrol intervention with mycoparasites against plant pathogens.

Chemical Characterization and Bioactive Properties of the Edible and Medicinal Honey Mushroom Armillaria mellea (Agaricomycetes) from Serbia

Honey mushroom, Armillaria mellea, is an edible and medicinal lignicolous basidiomycete. In this study, we investigated the chemical composition and bioactive properties of its methanolic and acetonic extracts. The chemical characterization of extracts was done with the HPLC-DAD-MS/MS method. The results showed that potassium was the most abundant mineral; chlorogenic acid was the most abundant polyphenol; malic acid was the most abundant organic acid; and among carbohydrates, the most abundant were sorbitol, glucose, fructose, and saccharose. Antioxidative activity was assessed by DPPH (IC50 of the methanolic extract was 608.32 μg/mL and of the acetonic extract 595.71 μg/mL) and reducing power assays (the results ranged between 0.034 and 0.102 μg/mL). Total phenolic content was determined as gallic acid equivalent (methanolic extract: 4.74 mg GAE/g; acetonic extract: 5.68 mg GAE/g). The microdilution assay was used to evaluate the antimicrobial activity of the extracts, and the results ranged from 1.25 to 20 mg/mL. The antidiabetic effect of the extracts was tested by the α-amylase (the results ranged from 34.90% to 41.98%) and α-glucosidase assays (the results were in the range of 0.55-2.79%). The neuroprotective activity was explored by the acetylcholinesterase inhibition assay (the results were in the range of 1.94-7.76%). The microtetrazolium assay was used to investigate the cytotoxic activity of the extracts (the IC50 values ranged from 212.06 to > 400 μg/mL). Although some results suggest that some activities of the extracts are relatively moderate, the honey mushroom can still be considered an excellent source of food and bioactive compounds with medicinal value.

ОСОБЛИВОСТІ ЕТИЧНОГО ПРОЯВУ МОРАЛІ КИЯН ХІХ СТОЛІТТЯ (ЗА П'ЄСАМИ І. НЕЧУЯ-ЛЕВИЦЬКОГО "НА КОЖУМ'ЯКАХ", "ГОЛОДНОМУ Й ОПЕНЬКИ – М'ЯСО")

The article analyzes the binary position of folk and bourgeois morality, the ethical features of the behavior of Kyivan residents in the 19th century, using the example of the dramatic works "On the Kozhumyaki" and "For hungry person honey mushrooms like meat", by classic of Ukrainian literature I. Nechuy-Levytsky. Kyiv city loci in the comedies of I. Nechuy-Levytsky are a means of historical, realistic and everyday depiction of the city landscape. The plays combined tendencies towards objectivity, locality and concreteness - social, ethnographic, geographical (places of Kyiv: Kozhumyaki, Hlybochytsia, Podil, Lypky, etc.) and moral with the author's desire for a realistic description of various aspects of Kyiv life, a depiction of Kyiv urban world in relief-plastic images of drama, ridiculing the arrogance, cunning and ambition of individual representatives of Kyiv society. The article examines the way in which the classic of Ukrainian prose conveyed the atmosphere of the city of Kyiv in the 19th century, showed the customs and ethics of Kyivans through stage means. Thanks to the humorous description of some characters, the author showed moral flaws in the life of the city. With realistic completeness, I. Nechuy-Levytsky recreated the Ukrainian types of Kyiv burghers, who have a secure financial position compared to petty officials, who, in turn, neglect the commandments of morality and try to arrange their lives at the expense of the burghers or pay off their debts, like Svyrid Hostrohvosty. Like the representatives of the "common people" – Kyivans without ranks and large fortunes, the townspeople and petty officials of the city do not shy away from simple folk proverbs, but are ambitious and demonstrate their own pride. We interpret two plays by I. Nechuy-Levytsky, in which, through the actions and lines of the characters, the facets of morality and ethical positions of various representatives of Kyiv city society are reflected: officials, townspeople, and ordinary people in Kyiv. These two dramatic works have not yet been studied in Ukrainian literary studies from the standpoint of describing the life of the city at that time.

Spread and Harmfulness of Infectious Diseases of the Main Forest-Forming Species in Zhytomyr Polissia of Ukraine

Given the multifunctional role of forests, there is an urgent need in forming biologically stable and productive tree stands to obtain the maximum ecological and economic effect for Ukraine in present-day conditions. Therefore, studies of the species composition, spread, pathogenic action, and harmful effects of pathogens of infectious diseases of the main forestforming species in Zhytomyr Polissia of Ukraine allow designing a real phytosanitary situation that will take place in the forests in the coming years and developing timely measures to limit the spread of pathological phenomena, which is the relevant research vector. The purpose of this study is to assess the current phytosanitary state of the forests of the surveyed region and identify abiotic and biotic factors that most contribute to weakening and degradation of forest tree species. This paper uses classical forest inventory and phytopathological methods to establish the general phytosanitary condition of the surveyed forests. Special mycological and microbiological methods were also involved to investigate the aetiology and pathogenesis of infectious diseases pathogens. Currently, the comprehensive sanitary condition of the forests of Zhytomyr Oblast can be described as satisfactory. However, every year there is a slow but steady increase in the dieback of forest areas, specifically the main forest-forming species – pine, oak, ash, and birch. During the examinations, the authors noted typical symptoms of bacteriosis (bacterial dropsy), necrosis (dieback), vascular (graphiosis, tracheomycosis) and other diseases of contradictory aetiology (transverse cancer, dieback), as well as fruit bodies of aphilophoroid macromycetes, which are the causes of stem (pine polypore, birch polypore, false oak polypore, etc.) and root (heterobasidion perennial, honey mushroom) rot were noted. Monitoring of the phytosanitary condition with the allocation of predictors of degradation of forest stands will further allow forming a “global map of plant resilience and sensitivity” for analysis of phytosanitary risk and rapid and rational decision-making on forest protection measures

Effects of an Armillaria mellea Polysaccharide on Learning and Memory of D-Galactose-Induced Aging Mice.

Arm illaria mellea has been known and used in traditional medicine in East Asia for hundreds of years. It has already been reported that A. mellea extracts have various pharmacological effects, and the polysaccharides of A. mellea exhibit antioxidant and anti-apoptotic activities. In this study, a water-soluble polysaccharide (AMP-N-a-1), with an average molecular weight of 17 kD, was isolated and purified from the water extract of A. mellea using DEAE-52, Sepharose CL-4B, and Sephadex G-100 column chromatography. AMP-N-a-1 was mainly composed of Man (1.65%), Glca (1.64%), Rha (1.82%), Gala (2.49%), Glc (90.48%), Gal (0.89%), Xyl (0.42%), and Ara (0.61%). AMP-N-a-1 was used to study the effect on the learning and memory of mice and its underlying mechanisms. The results showed that AMP-N-a-1 could significantly increase the activities of catalase (CAT) and superoxide dismutase (SOD) and reduce the content of nitric oxide (NO) in mouse brain tissue. Meanwhile, AMP-N-a-1 could reduce the contents of norepinephrine (NE) and dopamine (DA) but could increase the content of 5-hydroxytryptamine (5-HT) in mouse brain tissue. In addition, the immunofluorescence experiment showed that AMP-N-a-1 could promote the proliferation of hippocampal dentate gyrus neurons. The above results indicate that AMP-N-a-1 can significantly improve the learning and memory of mice, and the mechanism may be that AMP-N-a-1 can participate in the regulation of learning and memory through a variety of ways.