ISOLATION OF PURE CULTURE OF MEDICINAL Lentinus tigrinus (Buqa) Fr. MACROFUNGUS IN THE TERRITORY OF UZBEKISTAN

ABSTRACT

In this article, the isolation of a pure culture of the medicinal fungus Lentinus tigrinus for the first time in the territory of Uzbekistan is analysed, and the methods and results of its cultivation in potato-glucose agar and Saburo nutrient media are presented. As a result of the research, the work carried out in both nutrient conditions was successfully carried out, and it was possible to obtain a pure culture of the medicinal fungus Lentinus tigrinus in the territory of Kashkadarya region for the first time.

АННОТОЦИЯ

В данной статье впервые на территории Узбекистана выделена чистая культура лекарственного гриба Lentinus tigrinus., представлены методы и результаты ее культивирования на картофельно-глюкозном агаре и питательных средах Сабуро. В результате исследований работа, проведенная на обеих питательных средах, была успешно проведена и впервые на территории Кашкадарьинской области удалось получить чистую культуру лекарственного гриба Lentinus tigrinus.

Keywords: Lentinus tigrinus, fruit body, fungus, medicinal. 

Ключевые слова: Lentinus tigrinus, плодовое тело, гриб, лекарственный.

More than 2,000 eatable and about 700 medicinal species of mushrooms have been registered in the world. (Manzi & Pizzoferrato, 2000) [15]. A lot of research is being done on the study of medicinal mushrooms, as well as on increasing their biomass (Bellettini et. al. 2016) [14].

Isolation of pure cultures from medicinal fungi and their morpho-cultural characteristics are the most important source for selection of promising species.

It is known that Lentinus tigrinus (Bull) Fr. - are saprophytic fungi belonging to the division Basidiomycota, class Agaricomycetes, order Polyporales, family Polyporaceae, genus Lentinus, which grow singly or in groups on the sides of various trees. They are common in tropical regions and grow in a wide range of temperatures  (Karunarathna et al. 2011) [11].

Lentinus tigrinus is reported to be rare in Britain, mainly in central and southern England, to be found mainly in central and southern England, most of Europe, and parts of Asia and North America. This tiger saw mushroom was first scientifically described in 1782 by the French mycologist Jean Baptiste France Pierre Bulliard and given the binomial scientific name Agaricus tigrinus.

Swedish mycologist Elias Magnus Friss redescribed the species in 1825 and transferred it to the genus Lentinus, thus creating its current scientific name, Lentinus tigrinus. https://www.first-nature.com/fungi/lentinus-tigrinus.php. 

It was reported that the raw extract of L. tigrinus fruit body collected in Gaziantep province of Turkey showed antimicrobial and antioxidant effects (Mohammadnejad et al., 2019).

The composition and functionality of Lentinus tigrinus in the Philippines have been investigated. Evaluation of the functionality of this fungus focused on the antibacterial and hypoglycemic activity of the mushroom extract [10]. In addition, a strain of Lentinus tigrinus in the Philippines was evaluated for maximum mycelial growth in different nutrient conditions, pH, and light and temperature  [13].

Acute toxicity of a strain of the macrofungus Lentinus tigrinus was tested in mice in Iran. It was evaluated by the composition of amino acids and the dependence of elements on the substrate. L tigrinus can be considered as a source of macronutrients and micronutrients with anticancer activity, while it is less likely to have harmful effects on humans at low intake levels[12]. 

It is known that a number of works are being carried out in our Republic regarding the study of macrofungi. In 2023, the medicinal mushroom Lentinus tigrinus (Bull) was found in districts of Kamashi, Shahrisabz and Yakkabog during  our scientific research which was aimed to study the macromycetes in the deserts, hills and mountain ranges of the districts of Kashkadarya region.

This macrofungus was first studied in Uzbekistan by A.A. Petrova (1989) [7] in Zaamin Mountainous Forest (Juniper) Reserve, by G.M.Baltaeva (1992) [1] macromycetes belonging to the Polyporaceae family in the conditions of Uzbekistan. Later, in 2005, during the study of macromycetes of the Fergana Valley, it was reported that Juglans regia, Malus domestica, and Cerasus avium are abundant in trees [6 Iminova]. For the first time, a pure culture of the macrofungus Schizophyllum commune was isolated in Uzbekistan by Sherkulova J., Eshonkulov E. [8] and Sherkulova J., Koziboev X. [9]. 

Taking into account that this fungus is a medicinal species, we aimed to isolate a pure culture for growing and multiplying in laboratory conditions in our Republic, and to study its biological properties.

Research source and methods

This research was conducted in the Kashkadarya region of the Republic of Uzbekistan in March 2023, in Langar village of Kamashi district, Kitob district in November, Sharq Star MFY Mingchinor Temirchi district, Kitob district forestry area and in March Yakkabog district Shahrisabz city Khojakhuroson neighborhood. samples were collected from the fruit bodies of the fungus L. tigrinus from poplar trunks. Identification of samples was carried out by preparing temporary preparations in laboratory conditions and using B-382PHiALC DC6V1000 mA digital microscope [5] and mycological analysis of the samples using species determiners [2,3,4]. Experimental mycology and microbiological methods were used to extract a pure culture from the fruit body of the fungus [3,4].

Results and discussion

The convex fruit body of L. tigrinus grows singly or often  as a clump, the fruit body is white and brown in color, reminiscent of tiger skin. It was observed that the center of the cap is hollow and the fruit body is small. The diameter of the cap is 8-17 cm, at first convex, the edge is turned down, dry, fleshy, white or yellowish, fibrous brown, Plates are smooth, narrow white-creamy, uneven, jagged, sometimes split.

The leg is 5-12 cm high, 1.5-3.5 cm wide, cylindrical, narrowed at the bottom and sometimes elongated like a root, white-red-brown.

Figure 1. Fruit body of Lentinus tigrinus fungus

Spores 6 – 10 × 2.5 – 3.5 μm, cylindrical, almond-shaped, slightly unequal, smooth, thin-walled, [1].

Pure culture was obtained using experimental mycology methods.

In this case, two types of optimal nutrient media were selected and prepared for the good growth of the fungus.

Potato-glucose agar and Saburo nutrient conditions were used. In order to isolate a pure culture of the fungus from the fruiting body, an improved, undamaged fruiting body was selected and washed in running water in order to clean it from various foreign substances. The first cleaned fruit was cut into a small piece of 0.5-1.5 cm from the internal tissue of the body using a sterilized scalpel, immersed in 3% hydrogen peroxide liquid for a few seconds, and planted in nutrient media in Petri dishes and test tubes near the spirit lamp . The seeded tubes and Petri dishes were placed in a thermostat at 280C. Within 3 days, the mycelium began to grow and covered the Petri dish within a week (Fig. 2,3).

Figure 2. Growth of Lentinus tigrinus fungus in two different nutrient condition

The color of the mycelium in the petri dishes is white, the growth is radial, i.e. almost the same growth in all directions. Daily growth of mycelium was 0.6-0.8 cm. It was noted that the mycelium sticks to the substrate and grows densely. Fruit body growth was carried out in two different nutrient media. The growth of mycelium in two different nutrient media was almost the same and no significant difference was observed.

Conclusion

Thus, in order to isolate pure cultures of L. tigrinus species, two different nutrient mediums with different contents were prepared, and the formation and development of mycelium was observed in them. The work carried out in both nutrient environments was successfully carried out, and for the first time in the territory of Kashkadarya region, it was achieved to obtain a pure culture of medicinal fungus.

References:

1. Балтаева Г.М. Трутовые грибы (Polyporaceae S. Lato) Узбекистана: Автореф. Дис ... канд. биолог. наук. – Санкт- Петербург: 1992. - 17 с.
2. Билай В.И. Методы экспериментальной микологии. - Киев: Наук думка, 1973. - 238 с.
3. Бухало А.С. Высшие съедобные базидиомицеты в чистой культуре / Киев Наукова думка 1988. – 144 с.
4. Бухало А.С., Бисько Н.А., Соломко Э.Ф., Билай В.Т., Митропольская Н.Ю., Поединок Н.Л., Гродзинская А.А., Михайлова О.Б. Культивирование съедобных и лекарственных грибов. Киев, 2004. – 128 с.
5. Роскин Г.И. Миcросcопиc течниқуе. – М.: Сов. Наука, 1967. –п.447.
6. Иминова М.М. Макромицеты Ферганской долины (в пределах Республики Узбекистан): Автореф. дис. ... канд. биол. наук. – Ташкент, 2009. – 20 с.
7. Петрова А.А. Базидиальные макромецеты Зааминского горно лесного заповодника Уз ССР.: Автореф. Дис…. канд. биол. наук. – Киев, 1989. - 23 с.
8. Еркин Е., Жамила С., Дилмурод М. Schizophyllum sommunasi Фр. он тҳе территорй оф Узбекистан исолатион оф пуре cултуре оф медиcинал фунгус //Универсум: химия и биология. – 2022. – №. 6-4 (96). – С. 4-7.
9. Шерқулова Ж., Қоъзибоев Х. Исолатион оф пуре cултуре оф медиcинал ҳерб маcрофунгус Inonotus hispidus (Bull.) P. Karst. ин Узбекистан. Универсум: химия и биология. – 2023/ № 5 (107) ). – С. 43-46.
10. Dulay, R. M. R., Arenas, M. C., Kalaw, S. P., Rees, R. G. & Cabrera, Ye. C. (2014). Proximate composition and functionality of culinary-medicinal tiger saw gill mus’hroom, Lentinus tigrinus (Higher Basidiomycetes) from the Philippines. International Journal of Medicinal Mus’hrooms 16(1): 85-94.
11. Karunarathna SC, Yang ZL, Zhao RL, Vellinga YeC yet al. 2011 – Three new species of Lentinus from northern Thailand. Mycol Progress 10, 389–398.
12. Hamid R Pourianfar, Safoora Mohammadnejad, S’hadi S’hahtahmasebi, Alireza Madjid Ansari, Saaid Zibaei, Reyhane Ghadirian, S’harareh Rezaeian, Samaneh Attaran Dowom. Toxicity and Nutritional Assessment of Yextracts of Medicinal Tiger Sawgill Mus’hroom, Lentinus tigrinus (Agaricomycetes), a Newly Domesticated in Iran. Med Mus’hrooms. 2020;22(11):1079-1088.
13. Sofronio P. Kalaw, Rich Milton R. Dulay, Yeduardo J. Damaso Jr., Jos’hua C. Ramos, Mark Adrian G. Del Rosario, Marcelino D. Abon, Angeles M. De Leon, and Renato G. Rees Culture conditions for mycelial growth of yeight Philippine wild strains of tiger sawgill mus’hroom, Lentinus tigrinus. CLSU International Journal of Science and Technology. Submitted: January 26, 2023. Accepted: June 15, 2023. Publis’hed Online: August 31, 2023.
14. Bellettini, M.B.; Fiorda, F.A.; Maieves, H.A.; Teixeira, G.L.; Avila, S.; Hornung, P.S.; Junior, A.M. and Ribani, R.H. (2016). Factors affecting mushroom Pleurotus spp. Saudi J. Biol. Sci., 26 (4) : 633-646.
15. Manzi P., Pizzoferrato L. Beta-glucans in yedible mushrooms. Food Chemistry. Volume 68, Issue 3, 15 February 2000, Pages 315-318.