KR100232710B1 - A cultivating method for cordyceps militaris - Google Patents

Abstract

The present invention relates to a method for cultivating and cultivating milliliter cordyceps using silkworm chrysalis, and to cultivate seedlings of militus cordyceps using only silkworm chrysalis as a medium, and to cultivate millyless cordyceps with only milletless cordyceps. Militerless Cordyceps fungi cultured by the invention can be used as a biological pesticide to combat pests, and the cultivated Millyless Cordyceps fungi can be used as a raw material for pharmaceuticals such as nutrient tonics, anticancer drugs, drug addiction drugs, and inflammation inhibitors. It works.

Description

Cultivation Method and Cultivation Method of Milletless Cordyceps Sinensis Using Silkworm Pupa

The present invention relates to a method for culturing and cultivating seedlings of Millyless cordyceps using silkworm chrysalis, and more particularly, to a method for culturing and cultivating seedlings of Millyless cordyceps using only silkworm chrysalis itself as a medium.

Cordyceps militaris (L. Ex Fr.) Link,] is a mushroom that forms fruiting bodies in bundles based on the chrysalis of lepidoptera insects in the natural state. The fruiting bodies have the shape of clubs and are 17 ~ 28mm in size. It consists of a dark orange head and a sack supporting it, with clear boundaries. The lobules are projected in the shape of eggs, densely distributed, and the size is 490 ~ 550㎛ × 280 ~ 350㎛. Asylum is 400 ~ 420㎛ × 3 ~ 4㎛, arranged in a line, and there are thread-shaped vesicles. Aspergillus consists of cells separated by a clear and clear diaphragm and develops as a round secondary spore and begins to germinate. Spores of an insecure generation coincide with the form of Verticillium.

Cordyceps sinensis has been handed down in China since ancient times as the "Emperor's Bulwark Elixir", and it is also recorded in the herbaceous tree as "the obvious effect of strengthening weakness and strengthening immunity". Produces mannitol for medicinal use and contains other substances. In addition, a component called cordycepin, which is found to be an isomer of quinic acid in the fruiting body, is known as an active substance that acts as a toxin in insects and has antibacterial, antifungal, antiviral and anticancer effects. Currently, the Chinese medical community is using Cordyceps sinensis to delay the death of a patient or promote the recovery of senile patients, and to treat chronic cough, asthma, pulmonary tuberculosis, male sexual dysfunction, hypertension and diabetes from old age.

In addition, it has been recently proved that Cordyceps sinensis has a very powerful immune function, and nutrient extract extracted from Cordyceps sinensis significantly enhances the immune function of the organism and promotes humoral and cellular immunity. Increase the resistance of the organism In particular, recent studies have found that there is a very high anti-cancer component with a tumor suppression rate of 83%. In this regard, Professor Kondo of the Faculty of Pharmacy, Tofuku University, Japan, found that some of the components of Cordyceps sinensis have an anti-cancer component that strongly attacks cancer called "ergosterol and paroxide." These ingredients have no side effects and have been shown to be effective in terminal cancer patients.

Cordyceps sinensis is highly effective as a drug detoxifying agent, in addition to the above-described effects, and because of its parasitic properties to insects, it is also effective as a biological pesticide capable of preventing environmental pollution while providing effective pest control.

The native Cordyceps is usually collected between May and September, and the harvested Cordyceps can be dried and consumed by month or by other means.

However, since the Cordyceps sinensis having such a wonderful effect is difficult to find, the price is very high, so there are many difficulties for the general public to use, and a method for mass production of Cordyceps sinensis has been requested.

It is known to mainly use sawdust medium as a spawn culture method of common mushrooms except for Cordyceps sinensis. In the case of culturing millilis cordyceps fungi using sawdust as described above, mycelia are formed or fruiting bodies are not formed. Cordyceps cultivation was not possible. In addition, during the spawn culture of common mushrooms, excessive light has been known to adversely affect spawn culture. In nature, miterless cordyceps were formed mainly from the pupa of lepidoptera insects in the ground.

It is an object of the present invention to provide a method for cultivating a mass of miterless Cordyceps spp. Having a mysterious effect using silkworm pupa as a medium, and a method for cultivating Millyless Cordyceps sp.

Therefore, the present invention was made to solve the above problems by adjusting the silkworm pupae to 65 ~ 70% moisture content, put in a spawn bottle and then cooled by autoclaving in the medium cultured in the sterile chamber By using a silkworm pupa, the method of culturing milliliters Cordyceps sinensis seedlings, characterized in that the silkworm pupa and the milliliters Cordyceps sp. The medium selected from the media such as sawdust, soil, and sand for supply is adjusted to have a water content of 90 to 95%, and then placed in a cultivation container and cooled by autoclaving, and a suitable amount of milletless cordyceps is placed on the medium. Silkworms characterized in that they are grown without directly supplying moisture Provides a method for cultivating miterous Cordyceps sinensis using pupa.

Hereinafter, the present invention will be described in more detail.

In the case of cultivating the seedlings of the milliliter Cordyceps sinensis according to the present invention, the dried silkworm pupa or wet pupa should first be adjusted to have a water content of 65 to 70%. At this time, the dried silkworm pupa is soaked in water for about 24 hours in water temperature of about 20 ℃ and take out to adjust the moisture. Here, the hyphae growth is poor when the moisture content is below the above range, and the hyphae growth is very good on the outer surface of the silkworm pupa, but the hyphae growth is caused by the excessive moisture content in the silkworm pupa. Because of this, it is not possible to obtain a good millless Cordyceps sinensis fruiting body during cultivation. Therefore, it is important to control the water content within the above range in order to obtain a good seed.

Subsequently, the silkworm pupa with controlled moisture content was placed in a spawn bottle, autoclaved at 121 ° C. for 90 minutes, cooled to 20 ° C., and then incubated with a medium-cultured millisless Cordyceps fungus seedling in a clean room, followed by stirring to mix the silkworm pupa with the spawn well. While stirring three more days. At this time, because the miterous Cordyceps Bacillus very much needs water, if it is not shaken to mix well the silkworm pupa and the Milletless Cordyceps Bacillus, it will be active while absorbing the water at the bottom of the spawn bottle when the spawn sticks at the top of the spawn bottle. Since only the top part of the bottle is well cultured and the bottom part lacks water, it is not possible to obtain high quality spawn and the seed spawn takes more than 50 days. Therefore, in the present invention, it is essential to shake the seedlings after inoculating the miterous cordyceps fungi in the silkworm pupa medium, and for this purpose, it is preferable to put about 2/3 of the silkworm pupa whose moisture content is controlled without filling the spawn disease.

Roughness when culturing milliliters Cordyceps spp. On silkworm cultivation medium is preferably 200 to 500 Lx, and seedlings cultured at roughness beyond the above range are seedlings cultivated within the range of roughness in milliliters Cordyceps spp. About 10 days later the formation is slow. Therefore, in the present invention, it is essential to administer a suitable amount of light after inoculating the miterous Cordyceps sinensis in the silkworm pupa medium.

When culturing milletless Cordyceps sinensis on silkworm pupa medium, the temperature is preferably 21-23 ° C. If it is less than 21 ° C, the culture rate is slow and uneconomical. On the contrary, when the temperature exceeds 23 ° C, the culture rate is faster. Due to the weakening of the activity, the formation of fruiting bodies slowed down during the cultivation of militaris Cordyceps sinensis, resulting in a problem of failing to obtain good fruiting bodies. Therefore, in the present invention, the culturing temperature of the milliliter cordyceps fungi is required to be within the above range.

In addition, when cultivating the bacterium of the miterless Cordyceps sinensis on silkworm pupa medium, humidity and air are required so that the culture chamber is not dried. To maintain 65-70% humidity and to prevent the mycelial growth due to lack of oxygen, It is necessary to perform approximately three times of ventilation.

In the case of cultivating miteris Cordyceps sinensis according to the present invention, first, it should be adjusted to 90 to 95% of the moisture content of the medium selected from a medium capable of absorbing moisture such as sawdust, soil, sand, sponge, waste paper, etc. . At this time, since the mycelial Cordyceps mycelium requires a large amount of water to form the fruiting body, it is important to adjust the water content within the above range because the complete fruiting body is not formed when the water content is below the above range.

Subsequently, about 10 mm of the water supply medium with the moisture content adjusted in the cultivation container was autoclaved at 121 ° C. for 60 minutes, cooled to 20 ° C., and then supplied with the method of cultivating the bacterium of the cell cultured by the above method in a clean room. After proper amount is added to the medium, it is grown in a growing room.

When cultivation of milletless Cordyceps sinensis using silkworm chrysalis is preferably 200 ~ 500Lx as in the spawn culture, when grown in the roughness outside the above range has been shown to adversely affect the growth of fruiting bodies. In addition, when there was little light, the fruiting body did not form. Therefore, the roughness at the time of cultivating milllessless Cordyceps sinensis in this invention needs to be in the said range.

In addition, the cultivation of milletless Cordyceps sinensis using silkworm chrysalis requires more humidity and oxygen. Therefore, the cultivation room always maintains 85 ~ 95% humidity and prevents the fruiting of fruiting bodies due to lack of oxygen. It is necessary to provide about 7 ventilations, but it is not necessary to supply water directly to the medium.

Although the action of the present invention is not precisely known, the miteris Cordyceps sinensis is a kind of mushrooms mainly parasitic to the lepidoptera pupa, so when the silkworm chrysalis is used as in the present invention, the active metabolic material contained in the body is the growth of the miteris cordyceps fruiting body. Is supposed to help.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto.

Example 1

Soak 150g of dried silkworm pupa for 24 hours in water with a temperature of 20 ℃ and take it out to adjust the water content to 65 ~ 70%, then put about 2/3 in 850cc translucent polypropylene bottle and pressurize at 121 ℃ for 90 minutes. After sterilization and cooling to 20 ° C, the medium cultured Cordyceps militaris spawn was added in a clean room, and the silkworm chrysalis and the injected millilis cordyceps spawn were mixed well and shaken two to three times a day. After incubation in a culture room of roughness 200 ~ 500Lx, humidity 65 ~ 70%, temperature 21 ~ 23 ℃ three times after shaking once more and continued incubation. As a result, in just 20 days, a miterless Cordyceps spp. Using the silkworm pupa was obtained.

It was cultivated in two ways as follows using the miterless Cordyceps sinensis seed obtained by the above method.

First, 2 g of agar and 100 g of agar were put in a 850 cc semi-transparent polypropylene bottle, autoclaved at 121 ° C. for 60 minutes, cooled to 20 ° C., and a medium for water supply was prepared. After putting the appropriate amount in the clean room, cover the bottle cap, and ventilate 6 ~ 7 times a day without supplying water directly to the medium, with the illuminance of 200 ~ 500Lx, the humidity of 85 ~ 95%, and the temperature of 18 ~ 20 ℃. Cultivated by the grower. As a result, in the input Millilis cordyceps fungi itself, a millisuls fungus Cordyceps fungus formed in a bundle having a length of about 15 to 35 mm in 35 days was obtained.

Second, adjust the moisture content of the soil mixed with 100g of sand and ocher soil to 90 ~ 95%, put about 10mm in 850cc translucent polypropylene bottle, put it in 121 ℃ and autoclave for 60 minutes, cool it to 20 ℃ Put the appropriate amount of the above-mentioned milletless Cordyceps sinensis seedlings on the soil medium prepared for supply in a clean room, close the bottle cap, and illuminate 6 ~ 7 times a day without supplying water directly to the medium. 200-500Lx, humidity was 85-95%, temperature was grown in the cultivator 18 ~ 20 ℃. As a result, in the militia cordyceps fungi seedling itself, the militia fungus cordyceps fungus formed in a bundle having a length of about 15 to 35 mm in 35 days was obtained.

Example 2

After adjusting 200g of wet silkworm pupa to 65 ~ 70% water content, put about 2/3 in 850cc translucent polypropylene bottle, autoclave at 121 ℃ for 90 minutes, cool to 20 ℃, and then cultured mediumless cordyceps. (Cordyceps militaris) Inoculate the spawn in a clean room, shake the silkworm pupa and the millilis cordyceps fungus spawn well to mix well, and ventilate 2 ~ 3 times a day with 200 ~ 500Lx light intensity, 65 ~ 70% humidity, and temperature After culturing in a culture room at 21-23 ° C., the solution was shaken once more and then cultured. As a result, in just 20 days, a miterless Cordyceps spp. Using the silkworm pupae was obtained.

It was cultivated by the following two methods using the miteris Cordyceps sinensis seed obtained by the above method.

First, after supplying water to 150g of sawdust to adjust the water content to 90 ~ 95%, put about 10mm in 850cc semi-transparent polypropylene bottle and autoclaved at 121 ℃ 60 minutes and cooled to 20 ℃, obtained above Inject the appropriate amount of milliliter Cordyceps sinensis seedlings into a sawdust medium made for hydration in a clean room, close the bottle cap, and ventilate 6-7 times a day without directly supplying water to the medium. ~ 500Lx, humidity was 85 ~ 95%, temperature was grown in the cultivator 18 ~ 20 ℃. As a result, in the militia cordyceps fungi seedling itself, the militia fungus cordyceps fungus formed in a bundle having a length of about 15 to 35 mm in 35 days was obtained.

Second, after adjusting the water content to 90 ~ 95% of soil mixed with 100g of sand and ocher soil, put about 10mm in a 850cc translucent polypropylene bottle, autoclave at 121 ℃ for 60 minutes and cool it to 20 ℃. , Put the appropriate amount of the above-mentioned milletless Cordyceps sinensis seed on the soil medium for water supply in a clean room, close the bottle cap, and ventilate 6-7 times a day without directly supplying water to the medium. It was cultivated in a cultivator with roughness 200 ~ 500Lx, humidity 85 ~ 95%, temperature 18 ~ 20 ℃. As a result, in the militia cordyceps fungi seedling itself, the militia fungus cordyceps fungus formed in a bundle having a length of about 15 to 35 mm in 35 days was obtained.

Comparative Example 1

After spraying water to 500g of sawdust according to the conventional method, the water content is adjusted to 60-63%, and filled into a 850cc translucent polypropylene bottle, autoclaved at 121 ° C for 90 minutes, cooled to 20 ° C, and then incubated in medium. In the sterile chamber was added to the militialess Cordyceps sinensis seedling without shaking, the culture was incubated at 200-500 Lx, 60-63% humidity, 21-23 ° C. with roughness of 2 ~ 3 times a day. As a result, after 50 days, the miterless Cordyceps spp. Using the sawdust having low mycelial activity was obtained.

The cultivation was attempted using the miterless Cordyceps spp. Seed obtained by the above method, but fruiting body was not obtained.

Comparative Example 2

150g of dried silkworm pupa was soaked in water at 20 ℃ for 24 hours, then taken out, adjusted to water content of 60 ~ 63%, filled with 850cc translucent polypropylene bottle, autoclaved at 121 ℃ for 90 minutes After cooling to ℃, the medium culture 돤 milllessless Cordyceps sinensis seedlings are put in a clean room without shaking, and the ventilation room is 50Lx, humidity 60 ~ 63%, temperature 21 ~ 23 ℃ while ventilating 2 ~ 3 times a day. Incubated at. As a result, in 50 days, the mycelial Cordyceps spp. Did not form as a whole, and in part, the mycelium was not formed due to lack of water.

It was cultivated as follows using the miterless Cordyceps sinensis seed obtained by the above method.

2g of agar and 100g of water in a 850cc semi-transparent polypropylene bottle were autoclaved at 121 ° C. for 90 minutes, cooled to 20 ° C. to make a medium for water supply, and the appropriate amount of millilis cordyceps fungi obtained above was laid on the medium. It was grown in a cultivator with roughness of 50Lx, humidity of 85 ~ 95%, and temperature of 18 ~ 20 ℃ with ventilation of 6 ~ 7 times a day without directly supplying water to the medium after putting it in a clean room. . As a result, only about 5 to 9 mm of millless cordyceps was obtained after only 50 days in the injected milletless alumni seedling itself.

The milliliter cordyceps fungi cultured according to the present invention can be used as a biopesticide to combat pests, and the cultivated millilis cordyceps fungus can be used as a pharmaceutical raw material such as nutrient tonic, anticancer drug, drug addiction treatment and inflammation inhibitor. .

According to the method of the present invention, it is possible to mass-produce miterless Cordyceps sinensis which has an excellent effect which has been considered to be difficult to grow artificially.

Claims (4)

In the method of cultivating Cordyceps spp. By injecting the Cordyceps spp. Into the medium, the medium is a silkworm chrysalis medium consisting only of silkworm chrysalis in which the moisture content of the silkworm chrysalis is adjusted to 65 to 70%, and the silkworm chrysalis medium Put about 2/3 of the autoclave to cool, and then put the medium cultured militia Cordyceps sinensis strain in the sterile chamber to shake the silkworm pupa medium and the militia cordyceps strain well to cultivate in the light of 200 ~ 500Lx A method of culturing the millilis cordyceps fungi using silkworm pupa characterized by the above-mentioned. After adjusting the water content medium to 90 ~ 95%, put 10mm into the cultivation container, cool it by autoclaving, and then put in the sterile cultivated Cordyceps sinensis seedling cultured using silkworm pupae in the aseptic chamber, and the temperature of 18 ~ 20 ℃. Method of cultivating millilis Cordyceps sinensis, characterized in that the fruiting body is formed from the seedlings of the Millilis Cordyceps sinensis cultured by the culture method of claim 1 by supplying light of 200 ~ 500Lx. The method of claim 1, wherein the silkworm chrysalis medium and the medium-cultured militia cordyceps fungi are cultured by stirring twice. The method of claim 2, wherein the medium for supplying water is a method of cultivating wheatless cordyceps, characterized in that it is selected from sawdust, soil and sand, sponge, waste paper and the like.