KR100823681B1 - Composition of culture medium for Tremella fuciformis and culturing method of the same - Google Patents

Abstract

The present invention relates to a medium composition for mushroom cultivation of mushrooms comprising elvan or zeolite, and a method for growing the same in medium, wherein the medium composition of the present invention promotes the mycelial growth of mushrooms and promotes the production of fruiting bodies. Production is possible, and also due to the bottled cultivation method of the present invention, the white wood without contamination concerns by the external environment enables mass production of mushrooms.

Elvanite, Zeolite, Albino Mushroom, Bottle Cultivation

Description

Composition of medium for cultivating white-eyed mushroom and its cultivation method {Composition of culture medium for Tremella fuciformis and culturing method of the same}

1 is a photograph after the bottleneck cultivation of white mushrooms for 15 days.

Figure 2 is a photograph of the incision after the bottle white cultivated mushrooms for 15 days in a bottle.

The present invention relates to a medium composition for mushroom cultivation of mushrooms comprising elvan or zeolite, and a method for growing the same in medium, wherein the medium composition of the present invention promotes the mycelial growth of mushrooms and promotes the production of fruiting bodies. Production is possible, and also due to the bottled cultivation method of the present invention, the white wood without contamination concerns by the external environment enables mass production of mushrooms.

It was not studied in Korea because it was not artificial cultivation, but attempted artificial cultivation in China in 1914. In the early 1950s, scientists at universities and research institutes used spore suspension Successful isolation of pure strains from yeast-like conidia. Around 1959, MP Chan first isolated white mycelium and inoculated the logs as a feather-like mycelium mixed medium. In early 1962, scientists from Shanghai, Zhejiang and Fujian provinces used their own mixed media to grow white wood. In 1965, PR Hsu proved that pure white mycelium from the feather-like mycelium cultures grown on artificial media had a complete life history. In 1968, large-scale tree artificial cultivation began using mixed culture spawn in Fujian, Guangdong and Hubei, China. In 1974, SS Yau of Gutian, China, tried to grow bottlenecks, but it was unsuccessful, and WH Dai of the same province tried to grow plastic bags. Huwang et al. [3] have used the knowledge of previous researchers to conduct various studies on the various fields of white wood cultivation [Huang, NL 1986. Cultivation of Tremella (in chinese), promotion of science press, Beijing. p 31-104]. Early white cultivation was done by placing the logs to be used for cultivation next to the logs with fruiting bodies, by a simple method in which spores scattered from the fruiting bodies fell and inoculated. Chen and Hu then began to inoculate by tissue suspension without relying on natural scattering [Chen, P. C and Hou, HH 1978. Tremella fuciformis in the biology and cultivating of edible mushrooms. Chang, ST and Hays, WA Eds, Academic press, New York. 6: 25]. This spawn was used after grinding with water and fruiting bodies. However, for the short-term mass production of mushrooms, a systematic study was needed to develop special media and make it convenient for cultivation management in containers.

The present invention provides a medium composition for accelerating mushroom mycelial growth and fruiting body production, characterized in that the whites include elvan or zeolite for promoting short-term mass production by promoting mycelial growth of the mushroom and promoting fruiting body production.

In another aspect, the present invention provides a bottle growing method for mass production of mushrooms without affecting the external environment.

The present invention comprises at least one selected from oak sawdust, cottonwood sawdust, sugar cane watermelon as a basic medium component, white rice comprising a rice bran, bran, beer gourd, soybeans as an additive as an additive to the medium for mushroom cultivation In the solid content of the medium composition, it comprises 5 to 11% by weight of gannetite or 0.5 to 2% by weight of zeolite, or white mucus mushroom hyphae, which comprises 5 to 11% by weight and 0.5 to 2% by weight of gannetite and zeolite, respectively. It is characterized by a medium composition for promoting growth and fruiting production.

In another aspect, the present invention is a method of cultivating a white-necked mushroom, which is composed of the separation of white fungus and symbiotic bacteria, cultivation of seedlings, culture medium preparation, production of primary inoculum, culture seed production, inoculation, mycelial culture, fruiting body growth, harvesting. The white wood fills 30 to 70% of the growth volume of the mushroom hyphae growth and fruiting body production, and closes the cap of the growing bottle, and the white rice grows the fruiting body of the mushroom in the remaining empty space of the growing bottle. It is characterized by the method of growing bottled mushrooms.

Hereinafter will be described in more detail the step-by-step detailed process of the white wood mushroom cultivation method according to the present invention.

One. White fungus Symbiosis  detach

Pure cultures of Tremella fuciformis and H. archeri , which are streptococci , are required. Pure white cultivars can be separated by spore separation, tissue separation, native log separation and the like. White-eyed is homozygous when mushroom spores are mononuclear, and fruiting bodies are formed when binuclear conjugates. Fruiting bodies are similar to gelatin and are easily contaminated by bacteria, etc. on the surface of the fruiting bodies. Therefore, in order to receive spores from mushroom fruiting bodies, contaminants on the surface of fruiting bodies must be carefully removed before receiving spores. And these pollutants cause difficulties in pure tissue separation from fruiting bodies. Tissue separation involves separating the tissue from the base of the fruiting body before the tissue changes like gelatin. Unfortunately, the hyphae isolated from the bottom of the fruiting body cannot be guaranteed to succeed in the separation of good pure hyphae because of their very slow growth and low vitality.

The isolation of pure bacteria from mushroom alleys requires close attention. First, wipe off the base of the bark and mushrooms and the surface of the logs with 70% alcohol. Then, cut off the logs of the area where the mushrooms have been generated, remove the bottom pieces, put them in the test tube containing the medium, and incubate in the incubator at 20 ~ 28 ℃. A few days later, mycelia begin to grow from the pieces of wood, and the test tube is carefully examined daily for pure separation of combina- tions, which are albino and aseptic bacteria.

1) Properties of Binary Nucleus Mycelium

Color tack is white to yellow, upright aerial mycelium, and both surfaces exist and some mycelia penetrate into the medium. The diameter of the hyphae is 1.5∼3.0 ㎛ and there is a clamp connector at the hyphae diaphragm. Mycelial growth of white fungus is much slower than other edible fungi.

2) Symbiotic Properties of Bacillus Bacillus

White, feather-like, long, thin main hyphae with branch-like hyphae. Old hyphae are light yellow or light brown, and the culture medium gradually changes from light brown to brown or dark green. The aerial hyphae are grey-white with a thin, velvety surface. Usually there is no conidia, but if produced, the conidia are yellow to green to yellowish green, and are semi-elliptical in shape and approximately 3 to 5 μm in size. Aspergillus was formed on the surface of wood and its classification was confirmed to be asymptomatic genus Hypoxylon . Once the pure culture is separated, the parent strain is mixed and cultured in the test tube in the following procedure.

2. Fungus  culture

When several white fungus are cultured in a test tube and colony diameter is 1 cm, each test tube is inoculated with symbiotic bacteria and incubated at 25 ° C. for 7 days. Except for the fungi used for inoculum preparation, it is kept at 10 ° C for subculture every month.

3. Preparation of Badge

1) Addition of basic medium component

Sawdust or sugar cane watermelon are the basic medium components necessary for white mushrooms to grow mushrooms, and mycelial growth and white wood have the necessary moisture for mushroom growth and are used as nutrients. Sawdust contains few resins and water-soluble growth inhibitors and antibacterial agents. And no chemical is used. Among the solids of the medium composition, it is advantageous to use 25 to 35% by weight of oak sawdust, 16 to 24% by weight of cottonwood sawdust and 16 to 24% by weight of watermelon watermelon.

2) Additive

The basic medium component is oak sawdust, cottonwood sawdust, sugar cane watermelon or a mixture of one or more thereof, but when grown only with the basic medium component, the yield is significantly reduced due to lack of nutrients. When used as an additive, rice bran, bran, beer foil, bean curd, etc. may be mixed with one or more whites to provide the nitrogen needed to grow mushrooms. The amount of such additives is preferably 10 to 30% by weight of the total medium solids, less than this becomes malnutrition, and in many cases the white wood can make the generation of mushrooms in excess of the nitrogen component.

3) Addition of other nutritional ingredients

Depending on the composition of the basic medium component and additives, the medium may be prepared by including one or more of soy flour, sugar, calcium carbonate, lime, selenium, etc.

4) Addition of Elvanite and Zeolite

The medium composition for cultivating agaric mushrooms of the present invention is characterized by using elvan and zeolite.

Elvan is a rock belonging to quartz stool in igneous rocks and is porous and weathered and fragile. These pulsars have a slightly higher pH of 8.7 and have a CEC (cationic substitution capacity) of 9.0 me / 100g, which is significantly lower than other mineral modifiers such as zeolite and bentonite. As it has properties that can inhibit the growth of bacteria, it is advantageous to shorten the period of mycelial culture by using 5 to 11% by weight of the total medium solids.

Zeolite is a porous material having a size of a narrow rice, and has a physical adsorption power to nitrogen, potassium phosphate, and the like, and is water adsorbed. It is advantageous to shorten the period of mycelial culture by using 0.5 to 2% by weight of the total medium solids.

5) Control of moisture content

Moisture content in the medium composition mixed by adding the components 1) to 4) is adjusted to 60 to 75%, preferably 65 to 70%, thereby preparing a medium composition for growing white mushrooms. If the water content is less than 60%, the mycelial growth is good, but the mycelial density tends to be low, which is disadvantageous for mass production, especially when the water content exceeds 75%, inhibiting the growth of commensal bacteria, and water mushrooms are generated. there is a problem.

In addition, the pH of the water content-adjusted media composition is preferably in the range of 5-7. The mycelial density was good in the pH range of 5-9, but in the mycelial growth, it tends to decrease when it is below pH 5 or exceeds pH 7.

6) admission

In the case of encapsulation, the culture composition is placed in a culture bottle, and a culture bottle of 500 to 5000 cc, preferably 1000 to 3000 cc, is used. If the bottle is too small, the yield per unit bottle is small, the harvesting operation is inconvenient, and if the bottle is too large, there is a problem such as sterilization of the medium. The form of the cultivation bottle is gentle to the shoulder line, which can be easily fed into the medium and harvesting of the fruiting body, and can be either transparent or translucent, and the lid has a vent to prevent the air from being completely blocked. Lids with blocking filters, such as nonwovens, cotton, sponges, paper filters, etc., which can be incorporated into the container, are used.

In the present invention, unlike the method of growing the mushroom composition at the inlet of the bottle generally by filling the medium composition just below the inlet of the culture bottle, only 30-70%, preferably 35-60% of the capacity of the culture bottle is filled, In the remaining empty space of the cultivation bottle, the white wood provides a space for the mushroom fruiting body to grow, and the lid of the bottle is closed to provide a method of preventing contamination of airborne bacteria regardless of the growing environment.

7) Sterilize Medium Composition

After introducing the medium composition into the cultivation bottle, the other microorganisms in the medium composition are killed and the material is softened by high pressure steam so that only white fungi grow only mushrooms and symbiotic bacteria.

The sterilization method is not particularly limited, but after high-pressure steam sterilization, alignment is performed at 100 ° C. for 60 minutes, sterilization at 118 ° C. for 90 minutes, and then alignment for 90 minutes is performed by slowly opening the exhaust valve to lower the temperature to 100 ° C. and removing the medium from the sterilization pot. It is available.

8) Cooling Medium Composition

The cooling chamber is maintained aseptic state by ultraviolet sterilization and disinfection, and after the sterilization is completed and the steam pressure is removed, when the internal temperature of the cooling chamber drops to about 50 ° C, the cooling chamber is cooled down to rapidly lower the medium to 20 ° C.

4. Primary Inoculator  production

The parent strain cultured in the above 2. is inoculated into the medium in the cultivation bottle. In vitro strains can be inoculated from 1 to 4 bottles into culture bottles. This cultivation bottle is incubated at 25 ° C. until the formation of the original, and after completion of the cultivation, the first seed is called the inoculation source.

5. Culture spawn production

The amount of medium that can be inoculated with the primary seed is determined by the growth activity of the inoculator (primary seed) and the degree of growth of the white fungus in the medium. If the white fungus and symbiotic bacteria were well mixed in the primary culture mycelia, 100-200 bottles of culture spawn could be made with one bottle of primary spawn. However, in order to guarantee the quality of the spawn, the number of spawn bottles to be transplanted per first spawn bottle should be 40 to 60 bottles.

6. Inoculation

If the temperature of the medium drops below 20 ℃ after sterilization, the seed should be inoculated as soon as possible. The inoculation should be carried out aseptically in the same manner as the bottle mushroom inoculation method, and 0.2-5 g, preferably 0.5-1 g, of the culture spawn is inoculated into the medium composition.

7. Mycelial culture

The inoculated medium is initially incubated in a culture room at 25 to 28 ° C., and after a few days, the growth of mycelia begins to be seen from the inoculation site of the medium, and the temperature is lowered to 18 to 25 ° C., which is the optimum temperature for mycelial growth.

8. Fruiting Body Growth

The growth room only needs to maintain the temperature, not the humidity. The development of the fruiting body requires a little light of 500-600 lux.

9. Harvest

The white neck harvests mushrooms when it is full.

Hereinafter, the present invention will be described in more detail with reference to Preparation Examples and Experimental Examples, but the following Examples are intended to illustrate the present invention and are not intended to limit the present invention.

Production Example

As a medium for the cultivation, white oak is used as oak sawdust, cottonwood sawdust and sugar watermelon, rice bran, bran, ganban powder of less than 500 mesh, zeolite, calcium carbonate, soy flour, lime, sodium selenite and sugar. After mixing evenly by the weight percent of Table 1, water was added to prepare a media composition having a water content of 65% and pH 6.

division Comparison 1 Comparison 2 Conduct1 Conduct2 Conduct 3 Comparison 3 Comparison 4 Conduct4 Conduct 5 Conduct6 Comparison 5 Conduct7 Default badge Oak sawdust 35 35 33 30 30 28 35 35 35 35 35 30 Cottonwood Sawdust 21.999 21.999 20.999 20.999 19.999 18.999 21.999 21.999 21.999 20.999 20.999 20.999 Sugar watermelon 22 20 20 20 18 18 21.9 21.5 21 21 20 19 additive Rice bran 10 10 10 10 10 10 10 10 10 10 10 10 bran 5 5 5 5 5 5 5 5 5 5 5 5 Other ingredients Calcium carbonate 2 2 2 2 2 2 2 2 2 2 2 2 Soy flour 2 2 2 2 2 2 2 2 2 2 2 2 lime One One One One One One One One One One One One Selenium 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 Sugar One One One One One One One One One One One One Elvan 0 2 5 8 11 14 0 0 0 0 0 8 Zeolite 0 0 0 0 0 0 0.1 0.5 One 2 3 One

Experimental Example  One

After filling the medium composition of Examples 1 to 7 and Comparative Examples 1 to 5 in 900 cc each of the 2000 cc culture bottle, and then aligning for 60 minutes at 100 ℃ by sterilization under high pressure steam and sterilization for 90 minutes at 118 ℃ again 90 minutes After slowly opening the exhaust valve to 100 ° C and taking the medium out of the sterilization pot, it is maintained aseptic state by ultraviolet sterilization and disinfection in the cooling chamber.After sterilization is completed and the steam pressure is removed, the temperature inside the cooling chamber is about 50 degrees. When dropped, the cooling chamber was cooled to rapidly cool the medium to 20 ° C.

When the temperature of the medium was lowered to 20 ° C. or lower, 0.5-1 g of inoculation of white fungus and symbiotic bacteria prepared in advance in the sterile chamber was inoculated, and the lid of the cultivation bottle was closed. After inoculation, the medium is initially cultured in a culture room at 28-25 ° C. After a few days, when the mycelial growth begins to be seen from the inoculation site of the medium, the medium is incubated at 23-25 ° C, which is the optimal temperature for mycelial growth. Fruiting bodies were grown daily.

Table 2 shows the results of measuring the mycelial length and the amount of fruiting bodies grown in the medium compositions of Examples 1 to 7 and Comparative Examples 1 to 5.

division Comparison 1 Comparison 2 Conduct1 Conduct2 Conduct 3 Comparison 3 Comparison 4 Conduct4 Conduct 5 Conduct6 Comparison 5 Conduct7 Elvanite Weight% in Medium Solids 0 2 5 8 11 14 0 0 0 0 0 8 Zeolite Weight% in Medium Solids 0 0 0 0 0 0 0.1 0.5 One 2 3 One Mycelial length (mm / 15 days) 105 105 136 143 131 110 104 128 138 130 125 158 Fruit body weight (g / 900cc medium) 102 102 130 144 142 105 102 140 158 152 120 171

Experimental Example  2

In the same manner as in Experiment 1 using the medium composition of Comparative Example 1, the white-necked mushrooms were grown, but the medium composition was filled up to 1800 cc in a 2000 cc planting bottle, and then the fruiting bodies were grown on the outside of the planting bottle. It was allowed to grow, the inside of the fruiting body growth room maintained an absolute humidity of 90 ~ 95%, the fruiting body picture of the mushroom after 15 days of white wood is shown in FIG.

The photos grown in a bottle using the medium composition of Example 7 of Experimental Example 1 are shown in FIG. 2, and the media composition of Comparative Example 1 of Experimental Example 2 is shown in Table 3 in comparison with the bottleneck cultivation of the conventional method.

Conventional Culture Method (Bottle Cultivation) New culture law (bottle cultivation) Badge Usage (cc) 1800 cc 900 (requires 1/2 only) Humidity Need to maintain humidity of growth room (90-95%) Growth room humidity not required cost Large amount of expenses incurred in temperature and humidity management Reduced costs by not having to maintain humidity in the growth room Pollution rate 5-10% 0-1%

As described above, the medium composition of the present invention is capable of short-term mass production by accelerating mycelial growth of mushrooms and promoting the production of fruiting bodies, and also causing contamination by the external environment due to the bottle growing method of the present invention. White wood without mushrooms will allow mass production of mushrooms.

Claims (3)

delete In the medium composition for mushroom cultivation of white wood comprising at least one selected from oak sawdust, cottonwood sawdust, sugar cane watermelon, as an additive, and at least one selected from rice bran, bran, beer gourd and soybeans as an additive,    From 25 to 35% by weight of oak sawdust, cottonwood sawdust 16-24% by weight and sugar cane watermelon 16-24% by weight as a basic medium component in the solid content of the medium composition; As additives 10% by weight of rice bran and 5% by weight of bran; Other ingredients: 2% calcium carbonate, 2% soy flour, 1% lime, 0.001% selenium and 1% sugar; And 7 to 9% by weight of ganbanite and 0.8 to 1.5% by weight zeolite mushroom hyphae growth and fruiting medium production promotion medium composition comprising. delete

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