Antiviral Properties of Water Extracts of Mycelium Inonotus rheades (Pers.) P. Karst. (1882) against the Virus of Tick-Borne Encephalitis Virus in vitro

Background. Tick-borne encephalitis virus is dangerous and widespread pathogen that is transmitted to humans through the bites of hard ticks. Wild fungi, such as xylotrophic basidiomycetes, are widely used in traditional medicine to treat the infectious diseases and are promising natural sources of new antiviral agents. It was previously shown that aqueous extracts from the mycelium of the Inonotus rheades (Pers.) P. Karst. (1882) fungus exhibit significant antiviral activity against tick-borne encephalitis virus, however, the mechanisms of this activity remain unclear.Aim. To analyze the relationship between the virucidal properties of I. rheades extract and the substrate on which the cultivation was carried out.Materials and methods. The mycelium was grown either in a standard liquid medium with wort or on wooden disks from birch. Extracts of water-soluble polysaccharides were prepared from both mycelium samples. The concentration of infectious tick-borne encephalitis virus was determined using the method of titration of plaque-forming components (PFU). Approximately 30 000 PFU of tick-borne encephalitis virus was mixed with an equal volume of corresponding I. rheades extract at concentration of 8 mg/mL and incubated for 30 min at 37 °C. Afterwards, the residual infectivity of tick-borne encephalitis virus was determined using the identical virus sample incubated with sterile water as a reference.Results. It was found that treatment of tick-borne encephalitis virus with extracts from I. rheades mycelium resulted in inhibition of the infectivity of the virus in the cell culture. However, the same strain of I. rheades, grown on medium with wort, did not exhibit antiviral properties.Conclusions. Virucidal substances are likely to be not the main metabolites of the mycelium of I. rheades, but are rather metabolized wood polysaccharides. Further research is needed to more accurately identify the active ingredients and assess their antiviral activity.

Peculiarities of successional changes of xylotrophic basidiomycetes during decomposition of dominant tree species of forest formations of the Pitsunda-Myussera Reserve of the Republic of Abkhazia

The article is about the biodiversity of xylotrophic fungi of oak and hornbeam forests of Abkhazia and related destructive processes of the main forest-forming species (oak and hornbeam). The species composition of fungi subjected to phytopathogenic and saprotrophic methods of decomposition was determined. The species composition of mycobiota, causing wood xylolysis practically up to the last stages of decomposition, has been determined. Key words: biodiversity, xylotrophic fungi, succession, Republic of Abkhazia.

BIOTECHNOLOGICAL POTENTIAL OF THE SIBERIAN STRAINS OF BASIDIOMYCETES – PRODUCERS OF LIG-NOLYTIC AND CELLULOLYTIC ENZYMES

The results of a study of the wood-destroying properties of Siberian strains of xylotrophic basidiomycetes (Armillaria, Ganoderma, Fomitopsis, Heterobasidion and Porodaedalea) are presented. The growth parameters and enzymatic activity of the strains were determined during solid-phase and deep cultivation. Fomitopsis pinicola and Ganoderma lucidum are fast-growing fungi on cellulose-, tannin-containing nutrient media, coniferous and deciduous plant substrates (source and hydrodynamically activated). The growth coefficient on media with tannin and Na-carboxymethyl cellulose is more than 45; on plant substrates - more than 30. The strains exhibit a different growth reaction to the preliminary activation of birch sawdust (growth acceleration / growth slowdown / indifference). The maximum wood-destroying activity on wood of A. sibirica noted for the fungi A. borealis, G. tsugae, G. lucidum, F. pinicola, and P. niemelaei. The decrease in substrate mass was 8–11%; the amount of polysaccharides decreased on average 1.4 times mainly due to the fermentolysis of hard-hydrolyzable polysaccharides. High enzymatic activity of fungi observed during solid-phase and deep cultivation with an inducer. The maximum activity of phenol oxidase is characteristic of G. tsugae (1.21 units/g·s); carboxymethyl cellulase – for F.pinicola and G. lucidum strains (11.8 and 10.3 units/ml, respectively); xylanases – for H. abietinum (3.8 u/ml). The maximum accumulation of extracellular protein observed in F. pinicola (0.89 mg/ml). According to the totality of rapid test indicators, quantitative determination of enzymatic activity, growth parameters on lignocellulosic substrates and the degree of wood bioconversion, the most promising producer of lignolytic enzymes in vitro is the Gl4-16A Ganoderma lucidum strain; cellulolytic enzymes – strain Fp6-17 Fomitopsis pinicola.

XYLOTROPHIC BASIDIOMYCETES OF SMALL-LEAVED FORESTS OF THE KOSTROMA REGION

This paper provides an overview of the biology and species diversity of xylotrophic basidiomycetes, as well as the biotopic relationships of xylotroph mycobiota with phytocenoses. Ecological groups of these organisms are considered in connection with substrate specialization and their species similarity in birch and aspen small-leaved forests of different pharmacy. A brief assessment of the influence of the anthropogenic factor on the biological diversity of the represented group of organisms is given. For the first time, the Kostroma region is divided into southern, central, and northern parts to assess species diversity depending on weather, climate, and phytocenotic factors. The data on the study of changes in the degree of integration of mycocenoses of birch and aspen forests within the studied areas of the region are presented. In total, over 60 species of xylotrophic basidiomycetes were found in the small-leaved forests in the region, among which 38 species primarily affected birch forests and 40 species of pathogenic xylotrophic basidiomycetes have tropism for P. tremula. It was established that species belonging to the Coriolaseae family predominate in the species composition of wood-destroying mycobiota; they comprise about 27% of the total species composition in birch forests and about 24% of the species composition in aspen forests.

Cultivation of mycelium and the study of the phytopathogenicity of certain xylotrophic basidiomycetes under in vitro conditions

The paper discusses the cultivation of the mycelium of some economically significant xylotrophic basidiomycetes using methods of clonal micropropagation. For cultivation, Chapek, Murashige-Skoog synthetic media are used, a comparative analysis of mycelium growth in these media is carried out. Particular attention is paid to the study of the phytopathogenic properties of mycelium obtained in vitro. Two variants of phytopathogenicity are being studied: the cultural one, which is associated with the nature and intensity of mycelium growth on a nutrient medium, and phytopathogenicity for anatomical structures during the joint cultivation of xylotrophic basidiomycetes mycelium with parts of shoots from B. pendula and P. tremula. Significant variability was shown both in the expansivity of mycelial growth and in the manifestation of the degree of phytopathogenicity in different types of xylotrophs. Microscopy of the in vitro mycelium obtained was also carried out in order to more accurately identify the organism under study and to study the anatomical and morphological features. In general, this study will make it possible to better understand the ecology of these organisms in vivo, to model the interaction of the host para-site, and to more quickly and accurately conduct a specific determination of the phytopathogen, which can be used when conducting sanitary-protective and other economic activities in the forest industry.

Xylotrophic Basidiomycetes of protected species of woody plants of the Crimea

The species composition of xylotrophic basidiomycetes on woody plants listed in Red Book of the Crimea was studied. Xylotrophic basidiomycetes were detected on 10 species of protected woody plants of the Crimea: Arbutus andrachne - 3 species, Crataegus pojarkovae - 13, Juniperus deltoides - 2, Juniperus excelsa - 7, Malus sylvestris - 9, Pinus brutia - 3, Pistacia mutica - 15, Taxus baccata - 5, Tilia dasystyla - 2, Vitex agnus-castus - 1 species. A total of 46 species were identified. All discovered species of fungi belong to 8 orders, 20 families and 39 genera of Basidiomycota division. The order Polyporales dominates by the number of species of fungi - 18 species. There are 9 species in the orders Agaricales and Hymenochaetales. Xylotrophic basidiomycetes of the orders Hymenochaetales and Polyporales (27 species) are the most dangerous to protected species of woody plants. The greatest danger to growing trees are 12 species of fungi from the orders Hymenochaetales and Polyporales. Among the xylotrophic macromycete polyphagans, the most dangerous are Phellinus torulosus and Ganoderma applanatum , with a range of 100 and 82 host plants, respectively. Among the specialized xylotrophic basidiomycetes, Pyrophomes demidofii on Juniperus excelsa is the most common in park and forest plantations of the Southern Coast of the Crimea.

ИЗУЧЕНИЕ ВОЗМОЖНОСТИ ИСПОЛЬЗОВАНИЯ ОТХОДОВ ГРИБОВОДСТВА В БИОРЕМЕДИАЦИИ ПОЧВ СЕЛЬСКОХОЗЯЙСТВЕННОГО НАЗНАЧЕНИЯ

В работе рассматриваются возможности использования органических субстратов после культивирования на них мицелиальных культур ксилотрофных базидиомицетов для биоремедиации почв, загрязненных полифенольными соединениями и тяжелыми металлами. Показано снижение в 0,9-3 раза показателя интегральной токсичности почв под действием мицелиально-субстратных комплексов (отходов грибоводства и биотехнологий). Изучена динамика содержания гумуса в почвенных образцах в результате внесения в качестве удобрения отходов производства вешенки устричной (Pleurotus ostreatus). Установлено, что содержание гумуса в опытных вариантах увеличилось на 0,08-0,14. При этом использование отходов грибоводства в качестве удобрений позволило повысить урожайность яровой пшеницы Тулайковская 10 на 10,7-29,5.The paper considers the possibilities of using organic substrates after cultivation mycelial cultures of xylotrophic basidiomycetes for bioremediation of soils contaminated with polyphenolic compounds and heavy metals. A 0.9-3-fold decrease in the integral soil toxicity indicator under the influence of mycelial-substrate complexes (mushroom cultivation and biotechnology waste) was shown. The dynamics of the humus content in soil samples as a result of introducing oyster mushrooms (Pleurotus ostreatus) as a fertilizer was studied. It was found that the content of humus in the experimental variants increased by 0.08-0.14. At the same time, the use of mushroom cultivation waste as fertilizers made it possible to increase the yield of spring wheat Tulaykovskaya 10 by 10.7-29.5.