JPH0671391B2 - Cultivation method of Netsuke Numerita - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention is directed to oudemansiella canari.
i) Related to the artificial cultivation method. More specifically, the present invention relates to a method for cultivating a nutmeg mushroom having excellent taste in a short period of time with high yield.

[Conventional technology]

As the name suggests, N. edulis is a mushroom that lives on fallen or dead trees of broad-leaved trees in the subtropical to tropical regions, and is known for its delicious texture and good taste.
However, there is no example of successful artificial cultivation of Aedes aegypti, and the only natural growth of Aedes aegypti is now collected and used as food. Shiitake mushrooms, oyster mushrooms, enoki mushrooms, nameko mushrooms, and shimeji mushrooms have been artificially cultivated on an industrial scale since ancient times, and it is expected that the demand for mushrooms will increase in the future, judging from the recent tendency of the Japanese people toward health foods. It

[Problems to be solved by the invention]

It is an object of the present invention to provide a method for industrially inexpensively producing an Aedes aegypti mushroom having an excellent taste by artificially cultivating it using a solid culture medium.

[Means for solving problems]

Briefly describing the present invention, the present invention relates to an industrially inexpensive method for artificially cultivating Nettu numeritake, and in the artificial cultivation of Netu Numeritake, (A) an inoculum of Netu numeritake is used as sawdust and rice bran. After inoculating into a solid culture medium containing as a main component, culturing at 20 to 35 ° C. for 16 to 32 days to obtain a fruiting body generating group (B) Humidity of the fruiting body generating group of 80% or more, temperature of 15 to Medium culture step of forming fruiting body primordium from fruiting body generating group by culturing at 25 ° C for 4 to 10 days (C) Humidity of 80% or more, temperature of 15 to 25 ° C, illuminance of 100 lux As described above, the method is characterized by including the following steps of the post-culturing step of forming a mature fruiting body from the fruiting body primordium by culturing at 500 lux or less for 5 to 8 days.

In view of the present situation, the present inventors have conducted intensive studies on the artificial cultivation method of Netsuke numeritake for the purpose of developing a new edible mushroom, and as a result, a cultivation method using a solid culture medium containing sawdust and rice bran as main components. In addition, the inventors have found that artificial cultivation can be performed with high yield in a very short period by a cultivation method in which cellulose or hemicellulose is added to the solid culture medium, and the present invention has been completed.

The strain of K. mellifera K-35 used in the present invention is purely separated from the fruiting body growing on the fallen tree of the beech forest on Ogasawara Island, and is usually cultivated on an Ebios agar slant medium and then at 10 ° C. Store under moderate temperature. It is desirable to transfer the strain to a new medium every 6 to 12 months.

The morphological characteristics of fruiting bodies and spores of this strain are as follows.

Fruiting bodies are colonies or solitaries, umbrellas are 1.5 to 5 cm in diameter, initially hemispherical and later wide round mountain-shaped, surface viscous, white, pale yellowish white or grayish brown with fragments. The edges show striations when smooth and moist. The meat is white and about 7mm thick. The folds are slightly gelatinous, white, and later cream or light brown in color. Sparse and powdery. The stem is 1.5 to 5 cm x 1.5 to 8 mm, the base is slightly bulbous, white, slightly viscous and hairless, and rarely hollow. Spores are spherical, smooth, colorless and 12.5 to 22.5
μ, spore pattern is white, cystisia is spindle-shaped and 150 μm in length,
Maximum width 40μ, colorless and thin film. Comparing the above characteristics with the description in Seiya Ito, "Magazine of Japan", Vol. 2, No. 5, 1955, Yokendo Publishing, it is clear that this bacterium is Netuinumeritake. This bacterium has been deposited in the Institute of Microbial Science and Technology of the Agency of Industrial Science and Technology as Micromachine Research Institute No. 8557 (FERM P-8557).

The present invention will be described in more detail below.

First, the method for preparing the inoculum to inoculate the solid culture medium is shown below. A liquid culture may be used as the inoculum, but a solid culture inoculum may be used. Liquid inoculum is glucose 2.0%,
0.2% peptone, 0.2% yeast extract, 28 ° C. in a medium of the composition of 0.05% KH ₂ PO ₄ and 0.05% of _{MgSO 4 · 7H 2 O (pH5.5} )
It can be easily adjusted by culturing for about 7 days. The solid inoculum is a mixture of hardwood sawdust or coniferous sawdust or an appropriate mixture thereof and rice bran in a dry weight ratio of 1: 1 and water is further added to adjust the water content to about 63%. Compress into a culture bottle (850 ml). Then, after making a hole with a diameter of about 1 cm at one central location, it is stoppered with a urethane stopper and sterilized at 120 ° C. for 90 minutes to prepare a solid culture medium for inoculum. Inoculate this with about 20 ml of the above liquid inoculum, and then
After culturing in the dark for 15 to 25 days, an inoculum in which the mycelium of Aedes aegypti spread throughout the culture medium is obtained.

Next, the preculture step in the present invention is an adjusting step for obtaining a fruiting body-generating group from a solid culture medium. In the culturing step, the solid culture medium is inoculated with an inoculum, and the culturing is carried out at a temperature of 20 to 35 ° C., preferably at a humidity of 40 to 70% and in a dark place for 15 to 25
Apparently, mycelium spreads throughout the culture medium by day. When culturing is continued for another 1 to 7 days and the culture medium is aged, a good fruiting body-generating group can be obtained. The components of the solid culture medium used in the present invention include sawdust and rice bran commonly used for artificial cultivation of mushrooms,
A mixture of soybean meal or bran is suitable, but it is preferable to use a mixture of sawdust and rice bran. The mixing ratio of sawdust and rice bran is 1 to 3: 1 by weight ratio, preferably 1: 1 is the best, and the water content is 60 to 70% of the total weight of the medium, preferably
Around 63% is appropriate. Regarding sawdust, it is appropriate to use hardwood sawdust (eg beech, oak, oak, zelkova, birch, alder, etc.) alone, but coniferous sawdust (eg cedar, cypress, larch, red pine, etc.)
May be used alone. Further, even if hardwood sawdust and coniferous sawdust are mixed and used at an appropriate ratio, a fruit body having a good shape can be cultivated with high yield.

Incidentally, when adjusting the solid culture medium, cellulose, hemicellulose, starch, dextrin, glucose, carbon sources such as maltose, yeast extract, peptone, defatted soybean,
A nitrogen source such as soybean flour and inorganic or metal salts such as phosphate, potassium salt, magnesium salt and the like may be added. In particular, the addition of cellulose or hemicellulose has the effect of shortening the period of the medium culture step from the formation of the fruiting body primordium to the fruiting body primordium, and particularly increasing the yield. The amount of cellulose or hemicellulose added is 0.5 to 8%, preferably 1 to 3% of the mixture (dry weight) of sawdust and rice bran.
Is desirable.

Powdered cellulose, microcrystalline cellulose and the like are suitable as the cellulose used in the present invention, and xylan and the like are preferably used as the hemicellulose. A mixture of cellulose and hemicellulose may be added to the solid culture medium. Table 1 shows the effect of addition of cellulose and hemicellulose on fruit body formation. The addition test was conducted as follows. 50 g of hardwood sawdust (beech wood), 50 g of softwood sawdust (cedar wood), 100 g of rice bran, 350 ml of tap water, powder cellulose or xylan (both manufactured by Hanai Chemical Co., Ltd.) 0,
Solid culture medium and powdered cellulose with 2, 4, 6 or 8g added
A solid culture medium containing 2 g and 2 g of xylan and 4 g of powdered cellulose and 1 g of xylan was prepared, and a hole having a diameter of about 1 cm was opened in the center of each, and after stoppering, it was sterilized at 120 ° C. for 90 minutes. After cooling
Inoculate 20 ml each of liquid inoculum, dark place, temperature 28 ℃, temperature 55%
Under the conditions described in (1), the culture medium was cultivated until the mycelium was apparently sprinkled, and the culture was continued for further 4 days for aging. Next, after removing the stopper and scraping the fungus about 1 cm from the upper part of the fruiting body-generating group to remove the mycelium layer, 20 ml of tap water was added to sufficiently absorb water. After leaving it for 3 hours, the water remaining on the top was discarded, and the culture was continued under the conditions of an illuminance of 10 lux, a temperature of 23 ° C and a humidity of 90% until the fruit body primordia were formed, and then an illuminance of 500 lux and a temperature of 2
Cultivation was continued under the condition of 3 ℃ and 90% humidity until mature fruiting bodies were obtained, and the effects of powdered cellulose and xylan on the fruiting body yield and the total number of culture days were examined.

As is clear from Table 1, the addition of powdered cellulose or xylan to the solid culture medium increases the yield of Aedes aegypti and further shortens the total cultivation days.

The medium culture step is a step of obtaining a fruiting body primordium from the fruiting body generating group, and the fruiting body generating group obtained in the pre-culturing step has a humidity of 80%.
Or more, preferably 85-95%, temperature 15-25 ℃, preferably
By culturing for about 4 to 10 days under the conditions of about 23 ° C. and an illuminance of 1 to 200 lux, preferably 5 to 50 lux, the primordium of Nostoc commune is formed. In addition, in the cultivation using a polypropylene wide-mouth culture bottle, after removing the mycelium layer of about 1 cm from the upper part of the fruiting body-generating group obtained in the pre-culture (bacteria scraping step), add water for several hours. It is desirable to leave the solution to stand and allow it to absorb water sufficiently, then remove the remaining upper aqueous layer and transfer to the above-mentioned medium culture. It is known that the oyster oyster promotes the enzymatic activity of the glycogen-degrading system in the hypha layer around it. Therefore, glycogen accumulated inside the mycelium is rapidly decomposed and the decomposed product is utilized for the formation of the fruiting body primordia, so that the primordia formation is greatly promoted.
In addition, the soil may be covered with soil and then the above conditions may be applied. In other words, covering the soil improves the water retention effect,
It prevents the hypha layer from drying out, and the activity of the hypha becomes active. Suitable soil covering materials are sand, natural soil such as Kanuma soil, and soil conditioners such as vermiculite.

The post-culture step is a process of forming a mature fruiting body from a fruiting body primordium. Humidity of fruit body primordia obtained in medium culture process is 80%
Or more, preferably 85-95%, temperature 15-25 ℃, preferably
23 ° C, illuminance 100 lux or more, preferably 300-500
If the culture is continued for 5 to 8 days under the condition of luctus, a mature fruiting body of Aedes aegypti can be obtained. The culturing step is the same as the culturing conditions in the medium culturing step except that the illuminance is increased, and a fruit body having a height of 6 to 10 cm in which umbrella differentiation has occurred in 4 to 6 days from a fruit body primordia having a diameter of about 1 mm. Is obtained. When culturing is continued for further 1 to 2 days, a fully matured fruiting body with a height of 8 to 15 cm is obtained. If all steps from pre-culture to post-culture are required for 27 to 40 days, a mature fruit body with good texture and good taste can be obtained.

〔Example〕

Hereinafter, the method for artificially cultivating N. chinensis according to the present invention will be shown with examples, but the present invention is not limited to the scope of the following examples.

Example 1 Glucose 2.0%, peptone 0.2%, yeast extract 0.2%,
0.05% of KH ₂ PO ₄ and 0.05% of MgSO ₄ .7H ₂ O (pH 5.
5) 100 ml of medium having the composition of
iella canarii) K-35 (Microtechnology Research Institute No. 8557) was inoculated and cultured at 28 ° C. for 6 days to give an inoculum. On the other hand, 50 g of softwood sawdust (cedar wood), 50 g of hardwood sawdust (beech wood), 100 g of rice bran
After adding 350 ml of tap water to the mixture and mixing well to make it wet, this is a polypropylene wide-mouth culture bottle (850 ml volume)
Then, a hole having a diameter of 1 cm was opened at one central position, and a urethane stopper was plugged to prepare a solid culture medium. The solid culture medium
After sterilizing at 120 ° C. for 90 minutes and cooling, about 20 ml of the above-mentioned inoculum was inoculated to artificially cultivate Netume Nutritake. In the pre-culturing step, after culturing for 18 days in the dark at a temperature of 28 ° C. and a humidity of 55%, apparent mycelia were spread over the entire culture medium, and further culturing was continued under the same conditions for 4 days to ripen. Next, remove the stopper and scrape the fungus to remove the mycelium layer of about 1 cm from the top of the fruiting body-generating base, add 20 ml of tap water and allow it to absorb water for 4 hours.
The water remaining on the top was discarded and the medium was transferred to the medium culture step. The medium culture step was carried out under the conditions of an illuminance of 10 lux, a temperature of 23 ° C. and a humidity of 90% for 7 days to form a fruit body primordia of N. mellifera. When this was further cultured for 7 days under the conditions of the post-cultivation process at an illuminance of 500 lux, a temperature of 23 ° C. and a humidity of 90%, a fruiting body of Netsuke Nomuratake mushroom with good texture was obtained. The total weight of this fruiting body was 62 g, and the total number of cultivation days was 36 days.

Example 2 100 g of hardwood sawdust (beech wood), 100 g of rice bran and 350 ml of tap water
Was mixed well, then pressed into a polypropylene wide-mouth culture bottle, a hole having a diameter of 1 cm was opened in the center, and then stoppered with a urethane stopper and sterilized at 120 ° C. for 90 minutes, and then obtained in Example 1. After inoculating 20 ml of the liquid inoculum and culturing for 18 days in the dark at a temperature of 28 ° C and a humidity of 55%, a solid culture medium for an inoculum was obtained. On the other hand, 100 g of hardwood sawdust (beech wood), 100 g of rice bran, 350 m of tap water
Example 1 was a solid culture medium in which 6 g of powdered cellulose or xylan (both manufactured by Hanai Chemical Co., Ltd.) was added to 1 l.
It was adjusted like. This was inoculated with the inoculum from the above-mentioned solid culture medium for inoculum, and artificial cultivation of Aedes aegypti was carried out as in Example 1. With the powdered cellulose-containing culture medium, 80 g of xylan-containing culture medium in a total cultivation period of 28 days was used. In the case of total cultivation days of 27 days, 78 g of Netsuke Numetake mushrooms with good texture and delicious fruiting bodies were obtained.

Example 3 100 g of softwood sawdust (cedar wood), 100 g of rice bran, 4 g of powdered cellulose and 1 g of xylan in 350 m of tap water [both are Hanai Chemical Co., Ltd.]
Was added to prepare a solid culture medium as in Example 1. When the solid inoculum prepared in Example 2 was inoculated into this and artificial cultivation of Netsuke Numetaritake was carried out, 79 g of crunchy and delicious fruiting bodies were obtained in 27 days of total cultivation.

〔The invention's effect〕

As described in detail above, according to the cultivation method of the present invention, it is possible to cultivate mushrooms with good texture and good taste in a very short period of time and with high yield.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Tsutomu Taniguchi 3-4-1 Seta, Otsu City, Shiga Prefecture Central Brewery Co., Ltd. (72) Inventor Akira Obayashi 3-4-1 Seta, Otsu City, Shiga Prefecture Central Research Institute Co., Ltd.

Claims (3)

[Claims] 1. In the artificial cultivation of Netsuke numeritake, (A) after inoculating a solid culture medium mainly composed of sawdust and rice bran with an inoculum of Netume numeritake, culturing at 20 to 35 ° C. for 16 to 32 days, Pre-culturing step for obtaining fruiting body generating groups (B) The fruiting body generating groups are formed by culturing the fruiting body generating groups at a humidity of 80% or more and a temperature of 15 to 25 ° C. for 4 to 10 days. Medium culturing step (C) By culturing the fruiting body primordia at a humidity of 80% or more, a temperature of 15 to 25 ° C., an illuminance of 100 lux or more and 500 luxes or less for 5 to 8 days, mature fruiting bodies are cultivated from the fruiting body primordia. Post-cultivation step for forming A method for cultivating Aedes aegypti, characterized by including the above steps. 2. The cultivation method according to claim 1, wherein the outer solid culture medium contains cellulose and / or hemicellulose. 3. The method according to claim 1, wherein the nutmeg mushroom is a nutmeg K-35 (FERM P-8557).
Item or the cultivation method according to Item 2.