JPH0747A - Method for cultivating mushroom - Google Patents

Abstract

PURPOSE:To provide a method for cultivating a mushroom in which a spawn prepared by the solution culture can be inoculated into a sterilized culture medium by using a slender injection needle without requiring the use of a thickener and shredding of sawdust and prevent a malformed mushroom which has hitherto been in the fear from occurring at all. CONSTITUTION:A mushroom fungus is proliferated in a culture medium obtained by blending a nutrient source derived from cereals and beans with sawdust and the resultant primarily cultured spawn is then added to a secondary culture medium for culturing a spawn containing preferably saccharides and water-soluble proteins added to an extract solution of the cereals, beans and seed coats thereof to carry out the solution culture. The obtained secondarily cultured spawn is further injected and inoculated into a culture medium for cultivating the mushroom fungus prepared by filling the culture medium in a cultivation bag, closing the bag mouth and allowing the culture medium to cool with an injection syringe. Holes left in the injection port for the secondarily cultured spawn of the cultivation bag are closed by jetting a molten plastic or a hot-melt adhesive to carry out the cultivation of the mushroom fungus.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to inoculum cultivation of mushrooms such as shiitake mushrooms, maitake mushrooms, etc., in the inoculum injection method using a syringe for the purpose of improving the efficiency, preventing the formation of malformations, and The present invention relates to a method for cultivating mushrooms, which facilitates the injection work by injecting.

[0002]

2. Description of the Related Art Conventionally, in mushroom bed cultivation of mushrooms, a plastic medium provided with a filter member having an air permeability and a bacteria-blocking property is prepared by mixing a culture medium containing nutrients such as sawdust, rice bran, and corn bran. After being stored in a bag and sterilized, inoculum was sprayed on the medium. Recently, efficiency improvement by automating the cultivation process has been studied, and a method of closing the bag mouth and sterilizing after filling the medium has been attempted. In this case, the bag mouth is closed so that steam generated during sterilization is released without breaking the bag and air is not passed during culture.
In this case, inoculation of the inoculum is carried out by sterilizing and then directly injecting the medium from the outside of the bag using a syringe.

As a method of injecting seeds into a medium using a syringe, seeds cultured in a medium mainly containing sawdust are generally used. Therefore, the inoculum itself contains a large amount of sawdust, and upon injection, the sawdust is crushed into small pieces, or a thickener derived from cellulose such as carboxymethylcellulose and methylcellulose is blended to obtain a uniform suspension. It must be injected as a liquid. Moreover, it is necessary to use an injection needle having an inner diameter of 5 to 6 mm, and the perforation left in the culture bag after injection is large. It has also been found that shreds of sawdust as an inoculum tend to form mononuclear hyphae and induce the development of malformed mushrooms.

[0004]

A method of inoculating a seed culture by solution culture was also investigated, but the seed culture obtained by solution culture could not be adopted because the incidence of malformed mushrooms was abnormally high. . Therefore, even though it is a solution-cultured inoculum that gives a uniform solution without using a thickener,
There was a need for an inoculum that does not cause malformed mushrooms.

[0005]

The present invention is intended to solve the above problems, and its constitution is a secondary culture inoculum medium in which an extract of cereals, beans or seed coats thereof is preferably added with a saccharide and a water-soluble protein. In addition, the secondary culture inoculum obtained by adding the primary culture inoculum in which the mushroom is propagated in a medium in which the nutrient source derived from cereals and beans is mixed with sawdust is added to the culture bag, and the medium is placed in a cultivation bag. Using a syringe, inject and inoculate the fungus culture medium that has been filled, closed the bag mouth and sterilized, and then inject the remaining holes in the secondary culture inoculum injection port of the cultivation bag with molten plastic or hot melt adhesive. It is then closed to cultivate fungi.

The primary culture inoculum in the present invention is an inoculum used in conventional bed culture, and is mainly composed of sawdust, preferably a pulverized product having a diameter of about 2 to 5 mm obtained from the wood of broadleaf trees. It is a dense body of fungi which is propagated in a medium in which seed coat of grain such as bran, rice bran, corn bran or seed coat of legume such as soybean skin is added as a nutrient source. The use of seed coat is a problem in terms of commercial value, and in some cases, grains or various beans themselves may be crushed before use. Since cereals and beans are used as nutrient sources, any seeds can be propagated smoothly, and there is no particular limitation.

As the solution medium, at least one hot water extract selected from the above-mentioned grains, beans, and seed coats thereof is used. The concentration is 1 to 20 g by weight when left in a natural state.
Is boiled with about 100 g of water for 30 minutes to 2 hours and filtered if necessary before use. Generally, about 1 hour is sufficient. A carbohydrate and / or a water-soluble protein is preferably mixed therein as a nutrient source. Glucose is added as the carbohydrate. Other monosaccharides, disaccharides, trisaccharides, oligosaccharides and the like can be used, but glucose is common. As the protein source, a substance containing the essential amino acids evenly and having a low fat content is preferable. For example, meat extract powder and yeast powder are preferably used. Crude fat content 2% by weight or less, crude protein content 40% by weight or more, preferably crude fat content 1% by weight or less, crude protein content 60% by weight or more, more preferably crude fat content 0.5.
It is a water-soluble protein having a weight percentage of not more than 70% and a crude protein content of not less than 70% by weight.

When glucose is used, the saccharide added to the solution medium is 2% by weight more than 1% by weight added.
To 10% by weight, preferably 3% to 5% by weight
The added one quickly reaches the concentration of the secondary culture inoculum that can be inoculated. Similarly, the crude protein also reaches the required secondary culture inoculum concentration more quickly in a medium containing 1% by weight of the crude protein in terms of protein than that of 0.1% by weight. Therefore, the amount of the water-soluble carbohydrate or protein added, including the type of mushroom, can be appropriately increased or decreased according to the working process of the entire factory.

The culture medium varies depending on the type of mushroom, but in the case of shiitake or maitake mushrooms, sawdust, particularly preferably hardwood sawdust, bran, rice bran, corn bran, etc. A mixture containing 15%, preferably 6 to 10% is used.

For inoculation of the secondary culture inoculum, a solution in which the mushroom is sufficiently propagated is used. A culture medium is filled into a plastic bag provided with a filter member that is capable of blocking various bacteria but allows air to pass through, and the bag is closed by closing the bag mouth. When closing the bag mouth, do not completely seal it, leave a vent hole to release it without breaking even if a large amount of water vapor is generated by heating in the sterilization process, and sterilization is completed, and the internal pressure of the bag is If it decreases, the bag mouth should be in close contact with the bag so that the air permeability disappears.

For example, there is a method in which the bag mouth is bent, both ends are further bent into triangles, and a part is heat-sealed. In this case, the bag mouth is opened by the internal pressure during sterilization, but when the internal pressure is reduced, the bag mouth is naturally closed. In addition, two or more linear fusion bonding lines are provided at the bag mouth with a constant interval. Each fusion line is provided with a portion that is not fused, that is, a missing portion is provided, and this missing portion is provided at a position different from the adjacent fusion line. During heat sterilization, steam is discharged without breaking the bag through the missing portions of the inner fusion line, the gaps between the fusion lines, and the missing portions of the outer fusion line. After the sterilization is completed, the internal pressure is lowered and the two films are kept in close contact with each other so that air cannot pass through.

A culture bag is filled with a medium, steam can be discharged during sterilization, and the bag mouth is closed by a method in which air does not pass after sterilization, followed by steam sterilization. After the sterilization is finished, the mixture is allowed to cool and the secondary culture inoculum of the present invention is automatically injected from the side of the medium filled with a syringe to inoculate. The inoculation site may be an arbitrary one site, but the injection may be performed from two sites or four sites or more evenly arranged on the side surface.

After inserting the injection needle deep into the medium, the injection is started while removing the injection needle, and the injection is completed before the injection needle is completely removed. By injecting in this way, it is possible to inject the secondary culture inoculum into the entire medium as uniformly as possible.
The injection needle has an inner diameter of 1.5 because the secondary culture inoculum is in solution.
A needle of ~ 4 mm, preferably 2.0-3.5 mm can be used.

After inoculation with the inoculum, perforations remain in the cultivation bag. If this hole is left as it is, it will be contaminated by various bacteria, so there is a method of attaching an adhesive tape. In this case, the adhesive tape needs to be further sterilized. A more preferable method is to apply a heat-melted plastic or a hot-melt adhesive to the perforations in a molten state. These molten plastics,
The hot melt adhesive may be injected in an amount that covers the perforations. Even if miscellaneous bacteria are attached to the perforations, they are sterilized by the heat of the molten plastic or hot melt adhesive, and there is no risk of contamination from this site. The molten plastic material is preferably a plastic material of the same quality as the bag material or a plastic material having an affinity with the bag material.

[0015]

The present invention uses solution-cultured inoculum when inoculating the inoculum with mushrooms from the side of the cultivation bag in mushroom bed cultivation. However, this inoculum is not an inoculum that has been repeatedly subcultured in solution culture, but is an inoculum of a secondary culture inoculum that has been propagated in a solution medium as an inoculum that has propagated in a sawdust medium as a primary culture inoculum. According to the experiments by the present inventors, the secondary culture inoculum in which the primary culture inoculum cultivated in the sawdust medium was propagated only once in the solution has no teratogenicity, and only normal-shaped mushrooms are collected. did it. It is considered that this is because, as a result of repeated solution culture, mutation occurs and malformation occurs, but mutation occurs only once in solution culture.

[0016]

EXAMPLES Examples 1 to 3 98 g of water was added to 2 g of bran and the mixture was boiled for 1 hour and the bran was removed using gauze to obtain a bran extract. Separately, glucose and meat extract powder were prepared. This meat extract powder contains 85.9% crude protein, 0.2% crude fat and 3.4 ash.
%, The water content was 7.5%, and the amino acid composition of the protein contained the essential amino acids almost uniformly. 90 ml of a solution medium having the composition shown in Table 1 was added to sawdust: bran = 8: 1, water content 6
0.3 g of primary cultivated inoculum of shiitake mushrooms that had been sufficiently propagated in a medium adjusted to 4% was added, and shake culture was performed at 28 ° C. and 120 rpm for 7 days.

[0017]

[Table 1]

The cell concentration gradually increased, and after 7 days, the dry weight was 0.8 g / 100 ml in the case of Example 1, 0.55 g / 100 ml in the case of Example 2, and 0 in the case of Example 3. Reached 0.35 g / 100 ml.

Example 4 A test was conducted in the same manner as in Example 1 except that Maitake fungus was used as the inoculum. The cell concentration gradually increased, and the cell concentration after 7 days was 0.7 g / 100 ml in dry weight. Example 5 A test was conducted in the same manner as in Example 2 except that Maitake fungus was used as the inoculum. The cell concentration gradually increased, and the cell concentration after 7 days was 0.4 g / 100 ml in dry weight. Example 6 A test was conducted in the same manner as in Example 3 except that Maitake fungus was used as the inoculum. The cell concentration gradually increased, and the cell concentration after 7 days was 0.35 g / 100 ml in dry weight.

Example 7 7.5 ml of the secondary culture inoculum obtained in Example 1 after 4 days of culture was inoculated into 2.5 kg of a medium of the following formulation at 4 sites. Sawdust 44g Bran 5g Corn skin 1.3g Soybean skin 2.7g Water 47g The cultivation bag has a gusset fold of 32cm in circumference, 45cm in depth, and 6cm on both sides, and is in a position where it does not come in contact with the medium after filling with medium. A high-density polyethylene bag provided with a porous film that blocks out bacteria and allows air to pass was used. After the medium was filled, the gusset fold was opened to form a rectangular parallelepiped, and one side of the cuboid was inoculated with the secondary cultivated inoculum of shiitake mushroom of the present invention from four locations at equal intervals.

For perforation after inoculation, a hot melt applicator system (manufactured by Nordson Co., Ltd.) was immediately used to inject the hot melt adhesive to close the fusion. This culture bag was allowed to stand in a dark place at 24 ° C. for a certain number of days, and then moved to a room at 18 ° C. and a humidity of 94% to generate shiitake mushrooms. 992 g of shiitake mushroom was collected from the culture bag left for 90 days, and 737 g of shiitake mushroom was collected from the culture bag left for 101 days. No malformation appeared in this example.

Comparative Example 1 Shiitake mushrooms were cultivated in the same manner as in Example 7 except that 8 g of the primary culture inoculum was sprayed from above the culture medium as the inoculum. 9
467 g of shiitake mushrooms were collected from the culture bag left for 0 days, and 487 g of shiitake mushrooms were collected from the culture bag left for 101 days. However,
The size of each shiitake mushroom was larger in Comparative Example 1 on average.

Example 8 Example 2 except that 7.5 ml of the secondary culture inoculum after 6 days of culture obtained in Example 1 was used in a medium of the following formulation and 2.5 kg of polybropyrene film was used as a bag material. Inoculation was carried out at 4 sites in the same manner as in 7. Sorghum powder 37 g Bran 5 g Corn skin 1.5 g Soybean skin 1.5 g Water 55 g Culture conditions were left in the dark at 24 ° C for about 48 days, then 18
Maitake mushrooms were generated by moving to a bright place at ℃ and humidity of 83%. The number of samples was 12, and an average of 421 g of maitake mushrooms was obtained. No malformation appeared in this example.

Comparative Examples 2 and 3 As Comparative Example 2, Maitake mushrooms were cultivated in the same manner as in Example 8 except that 8 g of the primary culture inoculum as the inoculum was sprayed from the upper part of the culture medium. The Maitake mushrooms were collected. In Comparative Example 3, Maitake mushrooms were cultivated in the same manner as in Comparative Example 2 except that 16 g of the primary culture inoculum was sprayed from the upper part of the culture medium as an inoculum, and an average of 400 g of Maitake mushrooms was collected with 15 samples.

Example 9 2% glucose was added to the 2% bran extract as Experiment 1 and 0.33% of the meat extract powder used in Example 1 was added as Experiment 2. Glucose 3% and the meat used in Example 1 were added as Experiment 2. Addition of 0.5% of extract powder In Experiment 3, 4% of glucose was added, and 0.66% of meat extract powder used in Example 1 was added. In Experiment 4, 5% of glucose was added and 0.83% of meat extract powder used in Example 1. The primary inoculum of Maitake was cultured for 10 days in the same manner as in Example 4 except that the addition was used.

The cell concentration increased proportionally up to the 4th day of culture, and in Experiment 1, the concentration was 0.45 g / 100 ml, and in Experiment 2
In the case of Experiment 6, it was 0.65 g / 100 ml, in the case of Experiment 3, it was 0.80 g / 100 ml, and in the case of Experiment 4, it was 0.95 g.
/ 100 ml, there was not much change thereafter. The secondary culture inoculum of any culture days could be used as a solution inoculum inoculum, but around the 4th day, particularly good results were obtained.

[0027]

EFFECTS OF THE INVENTION According to the present invention, since the solution-cultured inoculum can be used, it is not necessary to use a thickening agent, it is not necessary to shred sawdust, and a thin injection needle is used. The sterile medium can be inoculated with the inoculum. Moreover, the malformed mushroom that was feared in the past does not occur at all.

Claims (4)

[Claims] 1. A primary culture inoculum in which fungi are propagated in a medium in which at least one extract selected from cereals, beans and seed coats thereof is mixed with sawdust and a nutrient source derived from cereals and beans. The secondary culture inoculum obtained by adding and culturing, the medium was filled in a cultivation bag, the bag was closed, and the bag was closed and sterilized. A method for cultivating mushrooms, which comprises cultivating mushrooms by closing the perforations left in the secondary culture inoculum injection port of the bag. 2. The method for cultivating mushrooms according to claim 1, wherein saccharides are added to the solution medium to which the primary culture inoculum is added. 3. The method for cultivating mushrooms according to claim 1 or 2, wherein a water-soluble protein is added to the solution medium to which the primary culture inoculum is added. 4. The molten plastic or hot-melt adhesive is sprayed and sealed at the injection port when closing the holes left in the secondary culture inoculum injection port of the cultivation bag. A method for cultivating a mushroom according to any one of items.