RU2446206C1 - METHOD OF GROWING Ganoderma lucidum MYCELIUM - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: method of growing Ganoderma lucidum mycelium involves preparation of inoculum on agarised culture medium in the presence of a growth stimulant. The growth stimulant used is selective light with wavelength 620-680 nm, intensity 60-100 mcmol/cm²·s for 30-120 minutes. Mycelium is grown at temperature 24-26°C or 5-7 days. With exposure time of 30 minutes, the output of air-dry mass of mycelium is approximately 160 mg.

EFFECT: method increases output of the air-dry mass of mycelium.

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Description

The invention relates to biotechnology and agricultural production, in particular to mushroom growing.

A known method of producing seed mycelium of edible mushrooms (see RF Patent RU 2249614, published March 21, 2003), including growing seed mycelium on a potato-wheat nutrient medium of the following composition (weight parts): flour - 20 g, potato broth - based on 200 g of potatoes per 1 liter of water. The culture was grown at a temperature of 26 ° C for three days. A suspension of bacteria Azospirillum brasilense Sp 7 grown on a synthetic liquid medium of the following composition (g / l) was used as a stimulator of mycelial growth: KH ₂ PO ₄ - 13.61; NaOH - 1.7; FeSO ₄ · 7H ₂ O — 0.01; CaCl ₂ - 0.026; Na ₂ MoO ₄ - 0.002; MgSO ₄ 0.2; KNO ₃ - 0.05; biotin - 10 ^-4 ; fructose - 1.44; pH 6.8. Cultivation was carried out at a temperature of 30-35 ° C. The disadvantage of this method is the complexity of obtaining mycelium due to the need to grow crops on a potato-wheat culture medium using a bacterial growth stimulator Azospirillum brasilense Sp 7, as well as an increase in material costs. A known method of growing mycelium of higher fungi, including sowing basidiospores on a nutrient medium and incubating them until the formation of mycelium. A pre-pasteurized or sterilized substrate is mixed with mycelium and introduced into a rigid culturing container lined with a flexible pad with waterproof properties inside. Germination of mycelium occurs inside a closed flexible laying space in conditions of high humidity and microaerophilic conditions, which is optimal for the development of substrate mycelium. (RU, patent 2101913, class A01G 1/04, 1998). The disadvantage of this method is its significant complexity and technological complexity of the stages of this method, which significantly increases the duration of the method and increases material costs. The closest is a method of producing seed mycelium of edible mushrooms (see RF Patent No. 2183056 according to IPC A01G 1/04, published June 10, 2002), including the preparation of seed on an agarized nutrient medium in the presence of a growth stimulator, which is used as an immunocytophyte in a concentration of 5.2 · 10 ^-5 mg / ml of the active substance, moreover, the stimulator is introduced at the initial stage of obtaining seed, namely in an agarized nutrient medium. The duration of cultivation of seed is 6 days. The disadvantage of this method is the need to use immunocytophyte as a growth promoter of the chemical substance, which is based on arachidonic acid ethyl ester, which does not provide an environmentally friendly food product.

The objective of the present invention is to develop a method for growing Ganoderma lucidum mycelium with the aim of reducing the time of growing primary mycelium, as well as increasing the air-dry mass of mycelium without any chemical additives, which allows you to grow environmentally friendly product with competitive properties.

The object of our research was one of the famous basidiomycetes Ganoderma lucidum, used for medicinal purposes for more than two thousand years by the peoples of Southeast Asia. It was used for a variety of diseases, including bronchial asthma, neurasthenia, gastritis, and liver diseases. The varnished polypore has many names. In China and Korea it is known as “ling-ji” (mushroom / herb of immortality), in Japan this mushroom is best known by the names “reishi” (mushroom of spiritual strength).

Intensive studies of G. lucidum over the past decades have shown that biologically active substances isolated from this fungus have immunoregulatory, antitumor, antiviral, antibiotic, lipid-lowering, hypoglycemic, hepatoprotective, gene-protective, anti-inflammatory, anti-allergenic, antioxidant, and antioxidant effects. vascular, respiratory and nervous systems.

The fruiting bodies and mycelium of G. lucidum contain carbohydrates (reducing sugars and polysaccharides), amino acids, peptides, proteins, triterpenes, including steroids, lipids, alkaloids, glycosides, volatile essential oils, vitamins, trace elements such as magnesium, manganese, molybdenum, calcium , zinc, potassium, sodium, iron, copper, sulfur, germanium. The proposed method of growing mycelium is implemented as follows. The claimed invention uses a standard, agarized nutrient medium (Malt Extract Agar from Mycomedia), which is prepared by adding the required amount of distilled water to it and autoclaving for 15-30 minutes at 1-1.5 atm. Glass Petri dishes are used to grow mycelium, after preliminary sterilization in a dry heat oven for 2 h 20 min at 160 ° C, they are cooled and the medium is poured so that the bottom of the cup is completely covered by 1-2 mm, this and all further procedures are carried out in sterile conditions (laminar boxing). Cups are left for a while in a laminar box (for 10-15 minutes), so that the medium freezes. On the frozen nutrient medium, a mycelium of a certain strain of Ganoderma lucidum is planted in the center of a Petri dish.

After sowing, the Petri dishes are closed with parafilm (a film to seal open vessels) and exposed for 24 hours in the dark at a temperature of 24-26 ° C. The next step is to stimulate growth with selective light with a wavelength of 620-680 nm, for 30-120 minutes daily, at a temperature of 24-26 ° C, while the cups are placed on special lighting installations (the lighting installation is a rack with shelves adjustable in height , each of which has a backlight (the lamps are located in a horizontal plane, on top, the installation is covered with a dark, impermeable fabric on all sides, thereby excluding the penetration of daylight or any other light during the experiment), the rest of the time shki are in the dark at the same temperature. This procedure is continued until complete fouling of the substrate surface (5-7 days).

Figure 1 shows the difference in the readings of the air-dry mass of the Ganoderma lucidum mycelium grown under 30-minute illumination with red selective light. Figure 2 shows the data of a 5-day experiment with Ganoderma lucidum mycelium grown under daily illumination with red selective light, which differs from the control data grown in the dark.

The following are examples of specific implementation.

Example 1

A method for growing varnished Tinder fungus mycelium (Ganoderma lucidum), including the preparation of seed on an agarized nutrient medium (Malt Extract Agar from Mycomedia), at a temperature of 24-26 ° C. As a seed, a piece of agar medium with a culture of 5 × 5 mm is used, which is placed in the center of a Petri dish with medium. The culture is incubated in the dark, daily illuminating for 30 min with red selective light (620 nm), intensity 60 μmol / cm ² · s, at a temperature of 24-26 ° C until the surface of the substrate is completely overgrown. Duration of cultivation is 5-7 days.

Example 2

A method for growing varnished Tinder fungus mycelium (Ganoderma lucidum), including the preparation of seed on an agarized nutrient medium (Malt Extract Agar from Mycomedia), at a temperature of 24-26 ° C. As a seed, a piece of agar medium with a culture of 5 × 5 mm is used, which is placed in the center of a Petri dish with medium. The culture is incubated in the dark, daily illuminating for 2 hours (120 min) with selective red light (680 nm), intensity 100 μmol / cm ² · s at a temperature of 24-26 ° C until the surface of the substrate is completely overgrown. Duration of cultivation is 5-7 days.

Using this method of growing can significantly accelerate the release of mycelium, without damaging it at the stage of growth by lower fungi, due to the complete sterility of the process, as well as the absence of chemical additives for cultivation, it can ensure its ecological purity and also increase the yield of air-dry mass of mycelium.

Claims (1)

A method of growing Ganoderma lucidum mycelium, including the preparation of seed on an agarized nutrient medium in the presence of a growth stimulator, characterized in that selective light with a wavelength of 620-680 nm, an intensity of 60-100 μmol / (cm ² · s) is used as a growth stimulator within 30-120 minutes at a temperature of 24-26 ° C, 5-7 days.

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