KR100917764B1 - Liquid spawn culture method - Google Patents

Abstract

The present invention relates to a method for cultivating a liquid spawn of mushrooms, and more specifically, protozoa comprising heavy white sugar, starch syrup, MgSo ₄ · 7H ₂ O, KH ₂ PO ₄ , CaCl₂, Yeast, Peptone, Agar, and water. Culture medium preparation step (S110); Prokaryotic culture and harvesting the end point of the secondary growth section instead of the first growth when the mycelium grows while spreading 1 ~ 2cm circular after the passage using the petri dish containing the prokaryotic culture medium and Collecting step (S120); A liquid medium consisting of roasted soy flour, refined white sugar, KH ₂ PO ₄ , MgSO ₄ ㆍ 7H ₂ O, anti form, and distilled water is placed in an Erlenmeyer flask and sterilized at a predetermined temperature for a certain period of time. (S130) inoculating prokaryotic bacteria in the triangular flask; After the infiltration of various bacteria by using a silly stopper, incubating the culture medium until the mycelium grows more than 2 cm for 7 to 10 days at a temperature of 2 to 3 degrees lower than the temperature in order to facilitate the distinction of contamination. S140); Shaking culture for 5 to 7 days at 100 to 120 RPM (S150); After the blade portion of the homogenizer is submerged in the culture medium step of grinding the cultured mycelia mass at 8000 ~ 10000rpm for about 10-15 seconds (S160); It relates to a cultivation method of liquid spawn for mushroom cultivation, characterized in that it comprises a; (S160) containing the liquid spawn in a separate container (capsules).

 Mushrooms, liquid spawn, culture, static culture, shaking culture

Description

Liquid spawn culture method for growing mushrooms

As mushroom cultivation area increases, demand for mushroom spawn increases. The spawn is the mushroom seed, so the production of good spawn is an important factor in determining the success of mushroom cultivation. The first artificial cultivation was manufactured in the form of cultivation using solid wood, and then advanced to the form of crests. Recently, a method of manufacturing sawdust as a main raw material and mixing appropriate additives has been developed. Conventional spawn production method using a conventional sawdust, after mixing sawdust, rice bran, lime, etc., injecting the mixed material into a glass bottle, sterilization, cooling to complete the medium and inoculate the mushroom seed bacteria to seed the mushroom spawn The preparation was completed.

However, the spawn of mushrooms produced by inoculating fungi on a sawdust-based medium is not easy to crush the sawdust spawn to a certain grain size when inoculating them on a medium for mushroom cultivation. In addition, because the nutrient medium is mixed together, the spawn of the mushroom itself when the spawn is stored or has a problem that is contaminated with other various bacteria. In addition, it is not easy to obtain enough sawdust for producing mushroom spawn and the price is high, which is a factor in raising the production cost of spawn, which is a great burden for producing mushroom spawn, and it is difficult for mushroom growers to purchase expensive spawn. It has been.

In the management of spawn, which is the seed of mushrooms, it is not easy to handle except in the case of mass production or professional handling. It became the cause.

The management of the liquid spawn which has been recently carried out by the new technology has not yet been established, however, and the liquid spawn cannot be easily used to grow mushrooms. In addition, the liquid liquid spawn production equipment requires a lot of equipment and requires a professional manpower has not yet been popularized.

An object of the present invention is to reduce the mushroom cultivation period in the production of the most important spawn in mushroom cultivation, to remove contaminants and to produce high-quality mushrooms, bring about an increase in cell mass, and easy storage of spawn It is to provide a method for culturing liquid spawn for mushroom cultivation.

Liquid seed culture method according to the present invention is the step of inoculating the progeny in the culture medium preparation step (S110), the progeny culture and collection step (S120), and the container (triangular flask) containing the liquid medium (S130) and , Static culture step (S140), shaking culture step (S150), the step of crushing the mycelium mass with a homogenizer (S150), and the step of containing the liquid spawn in a separate container (capsule) (S160) .

The first step is the step of preparing a culture medium for prokaryotic culture (S110). The medium for probiotic culture was 30g per medium, 10g starch syrup, 1g MgSo ₄ · 7H ₂ O, KH ₂ PO ₄ Composed of 1g, CaCl2g, Yeast 1g, Peptone 1g, Agar 15g, 1L water. Mycelial growth is visible so that there is a marked difference with the naked eye. After selecting the Petri dishes that are cultivated in a circular shape, the mycelia of the terminal which grows secondly are collected and used.

The second step is the probiotic culture and collection step (S120). There are two methods of culturing prokaryotes using test tubes and Patridish. However, in the present invention, the prokaryotic culture uses a petri dish, which is good for determining the presence of contamination (in the case of test tubes, it is difficult to distinguish the form of mycelial growth). After subculture (cultivation method that increases the generation of spawns, maintains the characteristics and prevents aging and extends the lifespan of spawns), when the mycelium grows about 1-2 cm (proliferation of mycelium is strong, less than this) In this case, it can be referred to as the preparation period of the early stage of growth, and if it grows more than this, the aging of bacteria may come). At this time, the end point of the part where the hyphae grows is not the first growth but the second growing part. (The first part of the progeny that grows rapidly is likely to change due to nutritional bias. A stable part growing up, down, left, and right should be collected to maintain the characteristics of spawn). If you use the existing end part, it does not retain its own characteristics. That is, the progeny culture and harvesting step (S120) is the end point of the second growth part instead of the first growth part when the mycelium grows about 1 to 2 cm after subculture using a petri dish containing the prokaryotic culture medium. Consists of harvesting. In addition, the passage was made of a combination of four medium media made of PDA, 30 g per medium, 10 g starch syrup, 10 g MgSo ₄ · 7H ₂ O 1, KH ₂ PO ₄ 1 g, CaCl ₂ 1 g, Yeast 1 g, Peptone 1 g, Agar 15 g, and 1 L of water. In the medium, primary and secondary growth is good for the naked eye to check the growth status of the hyphae, so one is added and subcultured using a total of five petri dishes. At this time, the rest of the mycelium should be discarded except when the growth of hyphae spreads out in a perfect circle. This can be used as a spawn using the mycelium selected in the progenitor stage.

The third step is the step (S130) of inoculating the bacteria in the container (triangular flask) containing the liquid medium. The petri dish is inoculated into a triangular flask, which is the source of inoculation of liquid spawn, at this time, a cut of 15 mm × 2 mm is firstly carried out using a scalpel, and then cut into a size of 2 mm × 1 mm, and then 10 using a scalpel. Inoculate 15 flasks at a time in the Erlenmeyer flask. If repeated several times, the density of contamination will increase. The liquid medium in the Erlenmeyer flask will be described. The liquid medium is soybean meal (30g per tablet white roasted soybean powder 3g,, 0.5g KH ₂ PO _4, 0.5g MgSO ₄ ㆍ 7H ₂ O 0.5g, Distilled water 1.0L) medium and 300ml culture medium in 1L container of Erlenmeyer flask After sterilization for 20 minutes at 121 degrees (low temperature or short time, sterilization is impossible, because if the temperature is high or time increases, the medium carbonizes and loses nutrients).

The fourth step is the political culture step (S140). After inoculation into the Erlenmeyer flask, use a silly stopper to prevent infiltration of various bacteria (hyphae of mushrooms also require oxygen when breathing, which allows for smooth flow of air and uses a silly stopper, which is a device that cannot infiltrate bacteria). At this time, do not shake so that mycelium does not go away, and incubate at 2 ~ 3 degrees lower than the temperature in the thermostat (even if the temperature is higher than the temperature due to the deviation because the temperature is easily changed. The reason is that the sinking hyphae has less oxygen than the surface, so the respiratory rate is lower. In case of shaking culture from the beginning, it is difficult to distinguish the contamination because the pellet is formed in a round shape. Therefore, the mycelium is largely grown for easy identification. In this way, the mycelia grow more than 2cm on the surface when the cultivation period is 7-10 days (the period of mycelial growth is faster than this and the hyphae is less and the aging progresses later). At this time, the distinction of contamination can be judged whether the intrinsic color of mycelium appears and whether mold is formed on the surface. If it is turbid, it should be discarded because it is bacterial contamination.

The fifth step is a shaking culture step (S150). Shaking cultivation is a task to enable the supply of oxygen in the medium while rotating the triangular flask on the rotating plate to the rotation of the shaking speed is suitable about 100 ~ 120 rpm. One day of shaking culture is turbidity in the soybean meal medium, which is not contaminated, but 5 to 7 days (when rpm is low in the triangle flask, the oxygen supply is low, and when it is higher, the amount of evaporation of the culture solution is increased. After a long time, aging progresses), the mycelium mass and the soybean meal get together and form a mycelium mass, and the medium becomes transparent. Again, pollutants can be found in the same way as in political culture. This also allows you to select bacteria that have better vitality. If the mycelial growth is faster after the mycelial mass is formed, the mycelia separated from the mass will be distributed in the medium in the form of mycelium rather than pellets.

The sixth step is the step of crushing the mycelium mass with a homogenizer (S150). When the incubation of the Erlenmeyer flask is completed, the homogenizer is pulverized using a homogenizer to expand the number of individuals by pulverizing with a homogenizer and pulverizing the mycelium mass at about 8000 to 10,000 rpm for about 10 to 15 seconds. At this time, when using the homogenizer, the blade part should be submerged in the culture solution to prevent the culture solution from splashing. (If the rotation speed is small, the mycelium is not smoothly crushed and crushed. If the amount is small and large, the possibility of mutation is increased by increasing the temperature). When the culture solution is pulverized in this way is not turbid. It appears to float on top of the culture. At the bottom, only a transparent culture layer is visible.

Seventh step is a step (S160) to contain the liquid seed in a separate container (capsules). In this case, the prepared seed is placed in a small capacity individual container. First, the individual container containing the spawn is used as a separate container that can be surely sealed and inoculating the medium into the container (triangular flask) containing the liquid medium (S130). The rod part of the piston is made detachable to allow for role and storage. This individual container is prepared after sterilization. Inject the culture solution into the sterilized individual container and seal it to prevent contamination. The seed capsule thus prepared can be stored for a long time at a temperature of 4 degrees.

According to the present invention, in the production of the most important spawn in mushroom cultivation, the following steps can be used to remove contaminants and produce high-quality mushrooms, which can lead to an increase in cell mass and to facilitate storage of spawn. can do.

Claims (1)

Crude sugar, starch syrup, MgSo ₄ · 7H ₂ O, KH ₂ PO ₄ , CaCl ₂ , Yeast, Peptone, Agar, water culture medium preparation step consisting of water (S110); Prokaryotic culture and harvesting the end point of the secondary growth section instead of the first growth when the mycelium grows while spreading 1 ~ 2cm circular after the passage using the petri dish containing the prokaryotic culture medium and Collecting step (S120); A liquid medium consisting of roasted soy flour, refined white sugar, KH ₂ PO ₄ , MgSO ₄ ㆍ 7H ₂ O, and distilled water is placed in an Erlenmeyer flask and sterilized at a predetermined temperature for a certain period of time. Step of inoculating prokaryotic strains (S130); After the infiltration of various bacteria by using a silly stopper, incubating the culture medium until the mycelium grows more than 2 cm for 7 to 10 days at a temperature of 2 to 3 degrees lower than the temperature in order to facilitate the distinction of contamination. S140); Shaking culture for 5 to 7 days at 100 to 120 RPM (S150); After the blade portion of the homogenizer is submerged in the culture medium step of grinding the cultured mycelia mass at 8000 ~ 10000rpm for about 10-15 seconds (S160); A method for culturing liquid spawn for mushroom cultivation, comprising the step (S160) of holding a liquid spawn in a separate container (capsule).