TECHNOLOGY OF ISOLATING A PURE CULTURE OF THE MEDICINAL F. fomentarius FUNGUS IN THE TERRITORY OF UZBEKISTAN

ABSTRACT

This article provides information about the methods and results of isolating a pure culture of the medicinal fungus Fomes fomentarius for the first time in Uzbekistan and growing it in potato-glucose agar and wort-agar nutrient condition.

АННОТОЦИЯ

В данной статье приведены сведения о методах и результатах выделения впервые в Узбекистане чистой культуры лекарственного гриба Fomes fomentarius и выращивания ее на картофельно-глюкозном агаре и сусло-агаровых питательных средах.

Keywords: intensive, fruit body, antioxidant, fungal, medicinal.

Ключевые слова: интенсивный, плодовое тело, антиоксидантный, грибковый, лекарственный.

Introduction

Nowadays, the demand for intensive breeding of medicinal fungi and the development of technologies for extracting their pure cultures and preserving them for a long time is increasing year by year. Fomes fomentarius is one of the important medicinal fungi in the pharmaceutical industry. The fruit body of this medicinal fungus is perennial, the base of the fruit body is wide, the diameter is 5-40 cm, the thickness is up to 5-20 cm, and the lower surface has a flat, hard appearance [1].

F. fomentarius fungus grows on the trunks of almost all trees. It can be found most often in the trunk of a birch tree broken by the wind. The fruit body is easily separated from the substrate. The upper surface of the fruit has a hard shell up to 2 mm thick. Its color is gray, light red, and the overwintered fruit body is black in early spring [2].

F. fomentarius fungus lives mainly as a parasite on trees. The fungus first begins to form in the form of a light yellow swelling on the trunk of the tree. It slowly causes the tree to rot. Rotten wood produces black lines that distinguish it from healthy wood. These strips form layers up to 2 mm thick and 50 mm or more in length [3].

F. fomentarius fungus contains up to 70% natural antioxidants. Medicines obtained from the fungus F. fomentarius are used to treat lung diseases, bronchial inflammation, purulent wounds [4]. In cosmetology, creams that rejuvenate the body and improve the condition of the face skin are prepared. Many medicines are produced from this fungus in Japan. The body of the fruit itself is used as a means of stopping bleeding when injured [5].

This article presents the results of research on selection of optimal nutrient condition for isolating of pure culture of medicinal fungus F. fomentarius.

Materials and method

To determine the composition of the fungus species, temporary or permanent preparations were prepared and observed first with small objective lens, then with 20 x, 40 x lenses [6].

To obtain a pure culture of the medicinal fungus F. fomentarius, nutrient condition with the following contents are used:

1. Potato - glucose agar (potato 200, glucose 100, agar 20 g);
2. Wort - agar: water up to 700C is added to beer wort. 1.5-2% agar-agar was added to the resulting wort and heated.

Wort - agar: water up to 700C is added to beer wort. 1.5-2% agar-agar was added to the resulting wort and heated.

Mycological and microbiological methods were used to obtain pure cultures of the fungus in laboratory conditions [7].

Results and discussion

Medicinal fungus F. fomentarius was found in the trunks of walnut (Juglans regia L.) and plum (Prunus domestica L.) trees growing in the mountainous regions of Yakkabog and Kitob districts during scientific trips and expeditions conducted throughout the region of Kashkadarya of the Republic of Uzbekistan in 2020-2023.

Figure 1. The fruit body of the medicinal F.fomentarius fungus

The sample taken from the fruit skin of the medicinal fungus F. fomentarius was thoroughly cleaned of iodine bodies, cut into small pieces using a scalpel and disinfected in a 3% solution of hydrogen peroxide.

Small pieces of the mushroom fruit body were planted 3 times on potato-glucose agar and suslo-agar mediums. The cultivated cultures of the fungus F. fomentarius were incubated in a TC-80 thermostat at a temperature of 28±1°C.

Mycelium of F. fomentarius fungus began to grow on potato-glucose agar in 4 days and formed white colonies. The fungus began to grow in the wort-agar nutrient medium from day 3 and formed a white mycelium (Fig. 2).

Figure 2. Appearance of mycelia of F. fomentarius fungus in a) Potato-glucose agar and b) Wort-agar nutrient condition

A pure culture of the medicinal fungus F. fomentarius was obtained by repeated inoculation and completely covered the petri dish in the suslo-agar nutrient medium on the day 14 of the growth cycle. The fungus grew well on potato-glucose agar for up to 9 days, but could not completely cover the petri dish. In both media, the fungal colony has a unique appearance, and the growth point of the fungus stopped completely in 15 days (Fig. 3).

Figure 3. Growth days of F. fomentarius fungus in potato-glucose agar and wort-agar nutrient condition

As a conclusion, a pure culture of the medicinal fungus F. fomentarius was isolated for the first time in Uzbekistan. The fungus completely covered the petri dish in 14 days in the wort-agar nutrient condition, while it grew in the potato-glucose agar for 9 days. In both nutrient mediums, the fungal colony formed a characteristic white mycelium. It is possible to show the most optimal wort - agar nutrient condition for obtaining a pure culture of the medicinal fungus F. fomentarius.

A pure culture of the medicinal fungus F. fomentarius is currently kept in the collection under fat oil in the laboratory of Microbiology and virology under the authority Department of Microbiology and Biotechnology of Karshi State University.

References:

1. Wasser S.P., Weis A.L. Medical properties of substances occurring in higher basidiomycetes mushrooms: current perspectives (review) // Int. J. of Medicinal Mushrooms. — 1999. — Vol.l — PP.31-62.
2. Mizuno T. The extraction and development of antitumor-active polysaccharides from medicinal mushrooms in Japan (review) // Int. J. of Medicinal Mushrooms. — 1999. — Vol.l — PP.195-206.
3. Kalitukha L, Galiano A, Harrison F. Medicinal Potential of the Insoluble Extracted Fibers Isolated from the Fomes fomentarius (Agaricomycetes) Fruiting Bodies: A Review. Int J Med Mushrooms. 2023;25(3):21-35. doi: 10.1615/IntJMedMushrooms.2022047222. PMID: 37017659.
4. Kalitukha L. An approach to change the basic polymer composition of the milled Fomes fomentarius fruiting bodies. Fungal Biol Biotechnol. 2021 Apr 15;8(1):5. doi: 10.1186/s40694-021-00112-9. PMID: 33858513; PMCID: PMC8051122.
5. Dudka I.A., Vasser S.P. Ellanskaya I.A. and others. Methods of experimental mycology: Handbook / Under. ed. IN AND. Bilay. – Kyiv: Naukova Dumka, 1982. – P. 549.
6. Pidoplichko N.P. Fungi parasites of cultivated plants key. T. II. Mushrooms Imperfect. – Kiev, 1977. – PP. 251-258.
7. Bukhalo A.S. Higher edible basidiomycetes in pure culture. - Kyiv: Naukova Duma, 1988.- P. 143.