JPH0327218A - Production of liquid spawn of basidiomycetes - Google Patents

Abstract

PURPOSE:To advantageously produce liquid spawn and to rationally automatize bed formation by film bag method, etc., by subjecting Basidiomycetes to static culture in a liquid medium, finely cutting and blending the finely cut Basidiomycetes with sterilized water containing a drying suppressing substance. CONSTITUTION:Basidiomycetes are subjected to static culture in a liquid medium and finely cut. Then the finely cut Basidiomycetes are blended with sterilized water containing a drying suppressing substance of the finely cut Basidiomycetes to give the aimed liquid spawn. A substance suppressing water evaporation, a substance swelling by water addition, a water-soluble substance or a substance suspending in water to show a gelatinous state is used as the substance to suppress drying of the finely cut cell and to protect the cell. A substance selected from among starch, saccharide, gelatin, rubber, fats, saponified fats and oils and resins of high degree of polymerization is preferable as the drying suppressing substance of the finely cut cell.

Description

[Detailed Description of the Invention] [Industrial Field of Application] The present invention relates to a method for producing liquid volatilization of castor seedlings [conventional Hi
[Machi] Conventionally, rice fungi 04, which is a basidiomycete, and a scale or saw 31 made from wood were used as the main culture medium (7).

[Problems solved by the invention]

The present inventor also considered a cultivation method using a cultivation bottle as an improvement on the conventional technique, but this method did not yield healthy fruiting bodies (if the bed was small, the fruiting bodies produced would be small, making it impractical for practical use). (Not applicable) Sojin in a regular culture bottle! - A study was conducted on mass production of mushrooms using film bags five times the size of the floor.

Making a floor using the film bag method using solid seeds is as follows:
It took a lot of time and effort to prepare the inoculation port, polyurethane stopper, and rubber cord to attach the ifffi port to the film bag, as well as the collection work.
Kaname is unsuitable for large issue production and 7? :． -tfC, these supplies are also expensive and not suitable for practical use.

In the case of a film bag floor, make a hole in the center of the floor and inoculate the middle part of the floor with the Keyaki bacteria. Therefore, the inoculation can only be carried out on the surface of the bed, and the number of days required for culturing is unavoidable.

In automating the bed setting b using the film bag method, the culture medium is compression molded, wrapped in a film bag, etc., directly sealed, sterilized and cooled, and liquid seed culture is injected into the film using a sieve force method. A book author has analyzed that the best method is to sow rice from the top of the bag to the center of the bed. -'f: We have been conducting research to develop a liquid seed fungus in the human race5.In the past, a considerable amount of research has been conducted on methods of liquid culturing synomycetes and multiplying bacterial bodies. The reason for this is that in order to propagate basidiomycetes in liquid culture, it is difficult to put them into practical use due to many difficulties. [L, The method of propagating by shaking culture and using it as a seed to cultivate in a large tank such as Jar 77 Mentor has been adopted, but this equipment requires a huge amount of cost, and furthermore, There was a problem that personnel costs, maintenance costs, management costs, etc. were too large. In addition, from a technical point of view, compared to common filamentous fungi such as Aspergillus aspergillus, Arachnidium fungi, and hairy molds, the propagation of Spermoid fungi is extremely slow and requires a long period of culture. ejc-
It is easy to become tangled with the pellet and become inconvenient. It had many problems, such as not being able to be glazed, and could not be put into practical use.

Therefore, it is an object of the present invention to provide a system that can rationally automate liquid volatilization.

[Means to solve the problem]

In order to achieve the above objective, in the present invention, after culturing the basidiomycetes in a liquid medium, the resulting liquid is shredded.
The shredded Aiziomycetes were mixed with M@sterilized water containing a substance that inhibits dryness of the fungal cells.

Therefore, as substances that suppress drying of the shredded microbial cells and protect the microbial cells, there are substances that suppress the release of moisture, substances that swell when added with water, substances that dissolve in water, and substances that can be dissolved in water. Use any of the following substances that become suspended and form a gel.

In addition, the dry smoke suppressing substance of the shredded bacterial cells is starch, saccharide (polysaccharide'!A polysaccharide produced by Otapa microorganisms,
sugars including mucopolysaccharides obtained from seaweed etc.), saponified products of gelatinous and rubbery fats and oils, resins with a high degree of polymerization (e.g. resins such as sodium polyacrylate)
Preferably, it is selected from the group consisting of.

[For production]

When the liquid containing the shredded bacterial bodies was directly diluted with sterilized water and placed on the floor, the active M rate of Seeds was poor (/.
Although it is impractical (5' = 2θ excellent or less), it is possible to mix a solution containing shredded basidiomycetes into sterilized water containing a desiccation-inhibiting substance for the shredded fungi. As a result of experiments, it was confirmed that by doing so, the survival rate could be significantly increased (almost to an excellent level of ioθ).

At the time of attachment, the inventor cut it into pieces. Substances that protect the fungal cells by suppressing drying of the fungal cells, such as starches, saccharides (polysaccharides or polysaccharides produced by microorganisms, mucopolysaccharides obtained from S. grass, etc.) (containing sugars), gelatinous and rubbery substances, and fats? 91 fats: saponified products, highly polymerized fats (e.g. sodium polyacrylate, etc.) that swell when added to water, or dissolve or suspend in water. Various tests were conducted using one of the Monoga products, which have a gel-like appearance and suppress water evaporation.

As a result, when diluted with simple sterilized water, most of them wither and do not take root; however, when diluted with Sansui containing a substance that suppresses moisture evaporation, even if diluted to a high magnification, it will not survive. They discovered the surprising fact that even if a very small amount of diluted shredded inoculum solution is injected with one filtrate, lθθ will remain viable and the hyphae will grow vigorously.

The survival rate of Kazuko fungi decreases as the sterilized melon water is shredded. When sterile water is used, the survival rate of basidiomycetes is extremely high, and mycelium forms on the floor. E The joint production that is drawn is also early.

Furthermore, the above-mentioned water evaporation inhibiting substance simply suppresses drying of the shredded bacterial cells and protects the bacterial cells by just 1. It also has the effect of preventing the sedimentation of TJln↑ seedlings and evenly suspending and dispersing K bacteria in the liquid, and this effect is said to be beneficial for production. I'm going to be a child who does k'lJ.

This invention was developed to make a bed for a bag of liquid exfoliated puff pastry obtained by the method of the present invention. Of course, it can be applied to all types of beds, such as those made by spreading the method on the floor, or those made by aseptically mixing liquid seed bacteria with medium L to form blocks.

〔Example〕

Examples of the present invention will be described below.

Example l P. D. A. Platinum loops of Plug jellyfish, Abalone edulis, and T. edulis fungi cultured on a slant medium (medium consisting of potato, dextrose, and agar) were collected by a conventional method.
) lfj, lOθ− in a 300 ml Erlenmeyer flask.
Each strain was inoculated into an onion soy sauce medium, and cultured stationary for 2S in a dark room at 2S0C. p. m. Shred by rotation,
The liquid containing this shredded microbacterial body is approximately lθθme
θθ1d, lθθO mJ, / Kθθ−, 10001, . 2soo- diluted in plain water (see Table 7l) and Sθ0, . i, /θθθ−, lSθ
θ−, λθθθ−, λ. iθθ−’s θ5% BoriakjJ
Diluted with sterile water supplemented with sodium monoxide (see Table 4), remove each using a dispenser and transfer vertically. 20 units x width 20 units x height 10 units, weight II
, k9 bed (bagasse 6, cake J1 rice bran 1 - anhydrous solids weight mixture) was compared for the number of days of infestation on the bed. To prevent this, the inoculation was carried out using a shaker dispenser.

Culture was carried out at 2SOC, humidity of 6θ to 7θ, and CO2 concentration of 7.
The test was carried out under conditions where θθ Heise θO ppm % Illuminance was below SO Lux.

The results of this test are shown in Table 1 by S value. and table Yu Shown here.

In the case of Table 1, when diluted with 5θθmJ of simple sterile water, the survival rate was 7 to 13%, and i7 to 10 at /θθθmJ.
%, l. s；00yrtltehO ~7%teh! )%ko000 1tll, 2! ; At θθd, most of the plants died and did not take root.

As shown in Table-1, is the dilution rate high? The survival rate decreases as the temperature increases. If you carefully observe the floor where the fungal cells have taken root, you will find that the shredded fungal cells wither and die at a rate of 1.b, and it takes time for the mycelium to begin to grow, causing the mycelium to spread on the floor. The days are getting late.

In the case of Table 2, finely shredded diluted body fluid was mixed with sodium polyacrylate for 9. Sterilized water added to 3% K, Yθθ~JSθθdK was mixed, and this was taken out as the inoculum for lOrl, and inoculated on the above-mentioned bed. , Sθθ to 2SOθ
The hyphae were attached to lθθ for all of the ml, and the hyphae were actively elongated, and the number of hyphae spread on the floor was significantly reduced.

4 Let's blaspheme the transformation■ In addition, conventionally 1. Shishi solid bacteria (saw Compare the cases of rotting bacteria). floor surface of Example 1 above to be inoculated with 1C. 32 to 3 days of wood ear fungus, evening o a Wabitake around Sθ day, Manenta The number of culture days is approximately 35 days.

From this point of view, in the case of the liquid inoculum of the Examples of the present invention, it is very early (Example where it is clear that the mycelium spreads). )
Sterilized water added with the desiccation inhibitor shown in Table 11/θθθml
[Mix well and add θ5ml, iyd! , 5m
Similarly to /, /θd and Example /ε, each of them was attached to the floor in 3θ pieces, and the survival rate of k mycelia was observed. Culture conditions id The same conditions as in Example 1 were used. The results are shown in Table 3. The survival rate and the number of days for mycelial spread were each expressed as the average value of 3θ beds.

The shredded solution diluted to a high magnification of b1 shown in Table 3 was added to the as. m
Even when a very small amount of J, zmt・2mttε is taken out and brought into contact with the bed, /θ0 remains active, and the number of bacterial cells is extremely small. Z, I understand this force.

Beds infested with hyphae of liquid fungi such as Makani and Nire were cultivated and tested for mushroom growth, but normal mushrooms were obtained in all cases.

〔Effect of the invention〕

As described above, according to the present invention, it is possible to efficiently produce a liquid polar fungus of Japanese fungi, which is suitable for seedlings as well as for making floors using the film bag method, at a significantly low cost, and to make it possible to use this liquid polar fungus on the floor. When it is grafted, it has many excellent effects such as the number of cultures can be greatly shortened, the grafting of Bacillus can be carried out in a simple and rational manner, it is extremely economical, and it is suitable for mass production.

Inventor Tominaga
Harudo Miura Makoto Okatani Shin Harudo
Takeshi Furukawa patent application 8
Nanei M-gyo Co., Ltd.

Claims (3)

[Claims] (1) After statically culturing the basidiomycetes in a liquid culture, the necessary shredding treatment is performed, and the shredded basidiomycetes are mixed with sterilized water to which a drying inhibitor has been added for the shredded fungi. A method for producing a liquid spawn of basidiomycetes characterized by (2) Substances that protect the microbial cells by suppressing drying of the shredded microbial cells include substances that suppress water evaporation, substances that swell when water is added, substances that dissolve in water, and substances that suspend in water. The method for producing a liquid inoculum of basidiomycetes according to claim 1, characterized in that one of the substances that is converted into a gel-like substance is used. (3) The substance that suppresses drying of the shredded bacterial cells is starch, saccharide (saccharide including polysaccharide or polysaccharide produced by microorganisms, mucopolysaccharide obtained from seaweed, etc.), Maiko fungi characterized by being selected from the group consisting of gelatinous materials, rubbery materials, saponified products of fats and oils, and resins with a high degree of polymerization (for example, resins such as sodium polyacrylate). Method for producing liquid seed culture