JP2808227B2 - Inoculation method of liquorice mushroom - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for inoculating a culture medium for liquorice mushrooms with a liquorice mushroom species capable of spreading the hyphae of liquorice mushrooms on the culture medium in a short time.

[0002]

2. Description of the Related Art Licorice ( Fistulinah)
Epatica ) is also called beef mushroom and occurs in the wild in the natural world from May to June or around October. The shape of the fruiting body (umbrella) is liver-like or bovine tongue-like, and the surface exhibits a reddish red to a dark reddish brown. The size of the fruiting body is about 10 to 20 cm in diameter. This liquorice mushroom is known to be very tasteless because it can be cut into slices of the fruit body and eaten raw or fried with butter or the like. Therefore, if this liquorice mushroom is cultivated and can be produced in large quantities throughout the year, it is possible to enrich the diet and provide excellent processed foods. However, in recent years, the production amount of mushrooms has increased due to artificial cultivation, but the types thereof are limited, and no artificial cultivation technology has been established for liquorice mushrooms. Conventionally, the culture of liquorice mushrooms is disclosed in JP-B-52-44603 and JP-B-54-27912,
These techniques are intended to be from the vegetative mycelium obtained by medium and the culture was used either for the cultivation of beefsteak fungus obtaining a substance having an anti-neoplastic, a large amount of vegetative mycelium of beefsteak fungus It is a technique for culturing, not for obtaining fruiting bodies,
Therefore, a liquid medium was employed as a culture medium. On the other hand, when using a solid medium, it is common to inoculate a mushroom inoculum on the surface of the medium, but in the case of liquorice mushroom, inoculation on the surface of the medium in this way slows the spread of mycelia and produces fruiting bodies. To take time.

[0003]

SUMMARY OF THE INVENTION An object of the present invention is to provide a method for inoculating liquorice mushrooms, which can spread hyphae into a culture medium in a short time in artificial cultivation of liquorice mushrooms.

[0004]

The present invention is different from the usual inoculation method in mushrooms in that the inoculation is carried out by dispersing a seed bacterium throughout a solid medium, and then culturing the medium, the mycelia of Licorice mushrooms are cultured in a short time. It was based on the finding that it could spread. That is, the present invention provides a method for inoculating liquorice mushrooms, which comprises dispersing the inoculum throughout the whole culture medium for liquorice mushrooms by mixing the inoculum of liquorice mushroom with a sterilized solid culture medium for liquorice mushrooms. In the present invention, any solid medium can be used, but it contains (a) a woody raw material, (b) a nutrient containing saccharides and organic nitrogen, and has a pH of 3.5.
The use of a solid medium in the range of 5.5 to 5.5 is preferable in that artificial hypha mushroom cultivation can be practically used because the mycelia of liquorice mushroom can be spread in the culture medium in a shorter period of time. Here, as the woody raw material (a), for example,
It is preferable to use wood powder such as sawdust, but it is also possible to use a material obtained by crushing or breaking a tree with a crusher, or flaked with a plane, a slicer, or the like.

[0005] The tree used as the woody raw material may be any of broad-leaved trees such as beech, sardine, oak, oak, zelkova, oak, alder, birch, etc., and conifers such as cedar, fir, larch, red pine, and hinoki. However, beech, shrimp, oak and oak are particularly preferred. The wood raw material can pass through a sieve with a mesh of 11.10 mm and
Those that cannot pass through a 850 μm sieve, preferably 6.
What can pass through a sieve of 00 mm and cannot pass through a sieve with an opening of 1.00 mm is 50% by weight or more, preferably 70% by weight of the whole
The content is more preferably 80% by weight or more. This is because the hypha of the liquorice mushroom spreads in the culture medium in a shorter time.

[0006] The nutrient (b) contains sugars and organic nitrogen. Glucose,
A saccharide such as sucrose, starch, pregelatinized starch, and malt extract, and an organic nitrogen such as amino acids, peptone, and yeast extract, each of which may be used alone or in combination of two or more, may be used. Also, as a nutrient containing sugars and organic nitrogen from the beginning, for example, corn cob or corn obtained by grinding corn core into granules or powder
Maize-based nutritional additives such as bran , okara, defatted soybeans, soybean-based nutritional additives such as soybean flour, wheat germ, rye whole wheat, oats whole wheat and other wheat-based nutritional additives, rice bran, Bran, koji and the like can be mentioned, and among these, koji is particularly preferred. Further, a saccharide and / or organic nitrogen may be added to a nutrient substance containing saccharide and organic nitrogen from the beginning. The content of the saccharide and the organic nitrogen contained in the nutrient is not particularly limited, but the saccharide is 0.1 to 90% by weight of the whole nutrient and the organic nitrogen is 0.02 to 50% by weight. Is preferred.
Further, the usage ratio of the woody raw material to the nutrient substance is not particularly limited, but is set to 1: 1 to 9: 1, preferably 3: 1 to 9: 1 as a dry weight. In addition, minerals and metal salts such as phosphates, magnesium salts, and calcium salts may be added to the solid medium as needed.

[0007] The water content of the solid medium containing the woody material and nutrients is 50 to 70% by weight, preferably 50 to 60% by weight, particularly preferably 5 to 60% by weight, based on the weight of the whole medium.
6 to 58% by weight. In a solid medium, the pH after heat sterilization is 3.5 to 5.5, preferably 4 to 5.5.
2, more preferably 4.2 to 5.0. Medium p
H changes depending on the type and amount of the nutrient added. Therefore, the pH of this medium is adjusted, for example, by adding an appropriate inorganic acid and organic acid, for example, a hydrochloric acid solution, a lactic acid solution, an acetic acid solution, a succinate buffer, etc. to the culture medium before or after heat sterilization. It is done by doing. In addition, pH measurement of the culture medium,
Suspending the media 5g of distilled water 50 ml, it performed more on measuring the pH of the suspension after standing for 10 minutes.
In the present invention, the solid medium is sterilized by a conventional method, and then a solid or liquid inoculum is aseptically mixed with the solid medium so as to be substantially uniform, and the bacteria are dispersed in the solid medium and cultured. After the sterilized solid medium is aseptically mixed with the inoculum under an aseptic environment such as a clean bench, a solid medium in which the inoculum is mixed so as to be substantially uniform is prepared and culture is started. Alternatively, in the case of using a seed in a form that can be poured into the space of the medium such as a liquid seed, by injecting the seed into the sterilized solid medium, even if a solid medium in which the seed is present is almost uniformly prepared and the culture is started. Good. In addition, any amount of inoculum can be dispersed in the solid medium,
Preferably, the amount is 0.4 parts by weight or more, preferably 0.4 to 20 parts by weight, more preferably 1 to 10 parts by weight, per 100 parts by weight of the solid medium.

[0008] Specifically, the solid medium is filled in a predetermined container, preferably a container with a filter through which gas such as oxygen or carbon dioxide can pass but which can prevent invasion of spores and the like . Subsequently, the medium is heated at, for example, 100 to 120 ° C. and 30 to
Heat sterilization is performed under the condition of 300 minutes. Thereafter, the seed medium is aseptically mixed with the sterilized solid medium, and the seed medium is inoculated so as to be dispersed evenly throughout the solid medium.
This solid medium is passed through a gas such as oxygen or carbon dioxide as described above, but is filled in a container with a filter capable of preventing bacteria from entering, and the temperature is 10 to 30 ° C, preferably 20 to 25 ° C,
Culture is performed under a dark condition at a humidity of 50 to 90%, preferably 60 to 90%. After inoculation of the inoculum, the hypha spreads throughout the solid medium in a shorter time than with conventional methods.

[0009]

According to the present invention, it is possible to spread mycelia of liquorice in a short time to obtain a fruiting body generating group.
In addition, since hyphae spread in a short period of time, the resistance of the fruiting body generating group to harmful bacteria is increased, and the growth of harmful bacteria that inhibit cultivation of liquorice mushrooms can be reduced. Therefore, the inoculation method of the present invention is a method extremely suitable for carrying out artificial cultivation of liquorice mushrooms. Next, the contents of the present invention will be described with reference to examples.

[0010]

Example 1 Beech chips (100% by weight of the whole) 534 which do not pass through a sieve having an opening of 2 mm and pass through a sieve having an opening of 6 mm
g and 178 g of dried koji, water was added so that the water content was 60% by weight to obtain a solid medium. Subsequently, the obtained solid medium was filled into a mushroom culture pouch (trade name; Kinopack, manufacturer; Nissho Corporation) with a 2.5 kg capacity filter so as to have a density of 0.5 g / cm ^3. And this has 12
Heat sterilization was performed at 1 ° C. for 60 minutes. The solid medium after heat sterilization had a pH of 4.9. Thereafter, 17 g of a liquorice mushroom inoculum (an inoculum obtained by culturing on a medium having the same composition as the solid medium) was added to the heat-sterilized solid medium under aseptic conditions and mixed, and the inoculum was homogenized in the solid medium. Was dispersed. This solid medium was heated at a temperature of 25 ° C. and a humidity of 85.
% Of the liquorice mushroom was spread on a culture medium for 25 days under dark conditions.

Comparative Example 1 775 g of the beech chips described in Example 1 and dried koji 17
Using 8 g, a solid medium was prepared in the same manner as in Example 1 (water content of the medium: 60%).
A mushroom culture pouch (trade name; Kinopack, manufacturer; Nissho Corporation) with a filter capacity of g capacity is 0.
5 g / cm ³ was charged at 121 ° C. and 60 ° C.
A heat sterilization treatment was performed under the conditions of minutes. The solid medium after heat sterilization had a pH of 4.9. Thereafter, 17 g of liquorice mushroom inoculum (inoculation obtained by culturing on a medium having the same composition as the solid medium) was inoculated on the surface of the heat-sterilized solid medium under aseptic conditions. This solid medium was cultured under conditions of a temperature of 25 ° C. and a humidity of 85% in a dark place until the hyphae of liquorice mushroom spread, and it took 55 days. As described above, according to this method, it takes a total of 55 days for the hyphae of the liquorice mushroom to spread on the culture medium.
Within 5 days, the mycelium of Licorice could not be spread on the culture medium.

──────────────────────────────────────────────────続 き Continuation of front page (56) References JP-A-48-61240 (JP, A) JP-A-63-17629 (JP, A) JP-A-63-3730 (JP, A) JP-A-3-3 292816 (JP, A) JP-B-47-38992 (JP, B1) JP-B-54-27912 (JP, B1) JP-B-52-44603 (JP, B1) (58) Fields investigated (Int. ⁶ , DB name) A01G 1/04

Claims (1)

(57) [Claims] 1. A method for inoculating a liquorice mushroom, comprising mixing a seed of a liquorice mushroom with a solid culture medium for a killed liquorice mushroom containing a koji and dispersing the seed bacterium throughout the culture medium for a liquorice mushroom.