JPH0662661A - Medium for culturing basidiomycete belonging to genus armillariella - Google Patents

Abstract

PURPOSE:To promote formation of a rhizomorph from an inoculated spawn in a mushroom bed culture of a basidiomycete belonging to the genus Armillariella using sawdust by adding carotene in a specific concentration to a medium. CONSTITUTION:A medium is mixed with carotene or a carotene-containing substance in an amount corresponding to 0.0002-0.5wt.% calculated as carotene based on total weight of the medium. Carotene is much contained in pericarp of carrot, pumpkin, parsley and citrus fruits. In preparing the medium, the medium is blended with carotene powder or the ground material of the carotene- containing substance or an extract of the ground material with a solvent. In a method for blending, the whole medium is uniformly mixed with the ground material or extract during the preparation of the medium. Or before or after sterilization, the ground material or extract is collectively added to part to which spawn are to be inoculated. By using the medium thus obtained, the number of days required to form a rhizomorph from spawn can be reduced.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a culture medium for Basidiomycetes of the genus Armillaria.

[0002]

2. Description of the Related Art The fruiting body of Basidiomycetes of the genus Armillaria (hereinafter abbreviated as Armillaria) is known as an excellent edible fungus with excellent texture and good soup stock in Hokkaido. It is prized as one of the tastes of autumn. For this reason, although various attempts have been made to cultivate a large amount of this armillaria by artificial bed cultivation, stable cultivation conditions cannot be found and it has not been successful yet.

That is, a culture medium is prepared by mixing sawdust, nutrients such as rice bran or bran, and water as a medium.
This is heat-sterilized and then cooled, inoculated with inoculum of the genus Armillaria,
When the culture is performed at a temperature of 20 to 25 ° C., the initial stage of the culture,
That is, when the inoculum is regenerated, the inoculum is frequently contaminated with various bacteria such as penicillium, and the subsequent cultivation becomes impossible.

Fortunately, if the above problems do not occur, after culturing for 30 to 60 days, the temperature should be 16 to
When the bacterial bed is developed in a generating chamber at 18 ° C., relative humidity of about 85% and an illuminance of about 300 | x, aratake mushrooms germinate after about 20 days, and aratake mushrooms can be collected after about 12 days.

[0005]

The feature of the basidiomycetes of the genus Armillaria is that during the growth of the fungus, in addition to the normal mycelium (mycelium), a bundle of root hyphae in which the hyphae are gathered in a string form, Although the growth rate is slow, the growth rate of the root hyphae is as high as 13 times the growth rate of mycelia. However, it takes a long time to form the root hyphae bundles, and in the above-mentioned fungal bed cultivation, it takes 7 hours for the root mycelium bundles to be formed from the inoculated inoculum.
It takes ~ 11 days.

[0006] In artificial cultivation of the genus Armillaria, the cause of the above-mentioned troubles in the early stage of culture is that the hyphae growth rate from the inoculated inoculum is slow and it takes a long time to form root-shaped hyphae bundles. This is because the seed bacteria become dry and weak, and eventually contaminated with various bacteria such as penicillium. As for the root hyphae, although it takes time to form,
Growth is rapid after formation. Therefore, in order to enable stable artificial cultivation of basidiomycetes of the genus Armillaria, it is only necessary to find out a method for speeding up the formation of root hyphae bundles from the inoculated inoculum.

The problem to be solved by the present invention is to find this strategy.

[0008]

[Means for Solving the Problems] In order to solve the above problems, first, a substance that promotes the formation of a root hyphae bundle of Bacillus subtilis genus Armillaria is found, and then the substance is mixed with a medium to inoculate. The method of promoting the formation of root hyphae bundles from the selected inoculum was examined.

In selecting the substance that promotes the formation of root-shaped hyphae, it is an essential condition that the performance is not impaired even when heated to about 120 ° C., considering the heat sterilization operation of the medium. Furthermore, considering the production cost of mushrooms, it is desirable that the substance be inexpensive such as agricultural waste or forest waste.

From a variety of vitamins, hormones, and inorganic substances that are thought to promote the growth of microorganisms, a substance that promotes the formation of root hyphae was carefully searched. It has been found that it has the property of promoting formation.

[0011] Carotene promotes the formation of root hyphae of Bacillus subtilis genus and further increases the growth rate and production amount of the formed root hyphae. It has no effect on the growth rate or production amount. The effect is that the concentration of carotene in the medium is 0.0002%.
It is efficiently expressed in the range of 0.5%.

Carotene is abundant in vegetables and fruits such as carrots, pumpkins, parsley and citrus peels. Therefore, as a result of conducting a cultivation test by mixing such a carotene-containing material in a medium, when the concentration converted to carotene is within the above range, the effect of promoting the formation of root hyphae of Naratake basidiomycetes Was recognized.

In actual use, when the medium is prepared, carotene powder or a ground product of a substance containing carotene or a solvent extract of the ground product is mixed with the medium. As a method of mixing, either mix uniformly throughout the medium preparation,
Alternatively, before the sterilization of the medium or after the sterilization, concentrate it only on the part to be inoculated with the inoculum.

When the carotene-containing material is added and cultivated as described above, the number of days from the seed fungus to the formation of root-shaped hyphae bundles is shortened by 18 to 38% as compared with the case where the material is not added. In addition, the incidence of miscellaneous bacteria at the initial stage of culture is about 30% when not added, but when added, it is significantly reduced to about 4%.

Among those containing carotene, carrots and pumpkins are cultivated in large quantities as agricultural products, and the carotene content based on fresh weight is 0.007 each.
3% and 0.00085% (Western pumpkin). In the production process of both of them, a large amount of nonstandard products are produced at the time of harvest due to malformation and poor color, and this utilization is a big issue. The carotene-containing material used in the present invention is
Since it is ground or extracted and used, its shape and color are irrelevant. Even nonstandard products can be fully used.

[0016]

[Function] Although the details of how carotene is involved in the formation of root hyphae bundles of Bacillus subtilis belonging to the genus Armillaria is unknown, it is thought that some mechanism may promote the metabolism of the fungus.

[0017]

EXAMPLES Examples are shown below together with the control examples. Examples 1 to 3 show the case where the whole medium was mixed with the carotene powder and the ground carotene-containing material, and Examples 4 to 5 show the case where the whole medium was mixed with the solvent extract of the carotene-containing material. 6-8 shows the case where the carotene-containing material was added only to the inoculated portion of the inoculum.

[0018]

[Example 1] 35 g of air-dried birch, 16 rice bran
g, 106 ml of water, and 0.75 g of β-carotene powder (manufactured by Wako Pure Chemical Industries, Ltd.) were mixed to prepare a medium having a water content of 69%. This was filled in a 200 ml wide-mouth glass culture bottle with a screw cap, and the mixture was kept at 120 ° C. for 20 minutes.
It was sterilized under high pressure for a minute. After cooling, it was inoculated with 2 g of the genus Armillaria spp. And cultured in a culture room at a temperature of 22 ° C and a relative humidity of 70% for 45 days.
Sprouting and growth were carried out in an OOlx generation chamber, and after 32 days, an average of 22 g of Japanese armillaria was harvested per culture bottle. The carotene concentration relative to the total weight of the medium at this time was 0.
48%.

It took 6 days to form root-shaped hyphae bundles from the inoculum and 18 days around the fungus in the medium, including these 6 days. The number of test bottles in the culture bottle was 12, and the number of culture bottles in which trouble of germs occurred in the initial stage of culture was 1.

[0020]

[Comparative Example 1] Cultivation was carried out in the same manner as in Example 1 except that the β-carotene powder was not mixed when the medium was prepared. And on average 1 per culture bottle
5 g of armillaria were harvested.

It took 8 days to form the root hyphae bundles from the inoculum and 22 days including the 8 days around the fungus around the medium. The number of test bottles in the culture bottle was 12, and the number of culture bottles in which germ trouble occurred in the initial stage of culture was 4.

[0022]

[Example 2] 25 g of air-dried birch, 15 rice bran
Cultivation was carried out in the same manner as in Example 1 except that a medium having a water content of 72% was prepared by mixing g, ground ginseng 35 g, and water 73 ml. Then, an average of 24 g of armillaria was harvested per culture bottle. The carotene conversion concentration based on the total weight of the medium at this time was 0.0017%.

It took 6 days for the formation of root hyphae bundles from the inoculum, and 18 days for the culture of the medium, including these 6 days. The number of tested culture bottles was 12, and none of the culture bottles had miscellaneous bacteria troubles in the initial stage of culture.

[0024]

[Example 3] Air-dried birch 23 g, rice bran 14
Cultivation was carried out in the same manner as in Example 1 except that a medium having a water content of 69% was prepared by mixing g, 53 g of ground fresh pumpkin, and 57 ml of water. Then, an average of 23 g of armillaria per one culture bottle was harvested. The carotene conversion concentration based on the total weight of the medium at this time was 0.0003%.

It took 6 days for the formation of root hyphae bundles from the inoculum and 18 days around the fungus in the medium including these 6 days. The number of culture bottles tested was 12, and the number of culture bottles with miscellaneous bacteria trouble in the initial stage of culture was 1.

[0026]

[Example 4] In Comparative Example 1, 56 m of water was added when the medium was prepared.
1, the same cultivation as in Control Example 1 except that 50 ml of a hot water extract of carrot (400 g of crushed raw carrots was boiled and extracted in water for 2 hours, filtered, and the filtrate was diluted to 1000 ml) was mixed. went. And culture bottle 1
An average of 24 g of armillaria was harvested per book. At this time, the converted concentration of carotene with respect to the total weight of the medium was 0.
It is 0009%.

It took 6 days for the formation of root-shaped hyphae bundles from the inoculum, and 19 days for the culture of the medium, including these 6 days. The number of tested culture bottles was 12, and none of the culture bottles had miscellaneous bacteria troubles in the initial stage of culture.

[0028]

[Example 5] In Comparative Example 1, 6 m of water was added when the medium was prepared.
l, pumpkin hot water extract (400 g of crushed raw pumpkin boiled and extracted with water for 2 hours, filtered, and filtered 1000 times
Cultivation was carried out in the same manner as in Control Example 1 except that 100 ml (made up to ml) was mixed. Then, an average of 23 g of armillaria per one culture bottle was harvested. The carotene conversion concentration based on the total weight of the medium at this time was 0.0002%.

It took 6 days for the formation of root-shaped hyphae bundles from the inoculum and 19 days for the culture of the medium including these 6 days. The number of test bottles in the culture bottle was 12, and the number of culture bottles in which trouble of germs occurred in the initial stage of culture was 1.

[0030]

[Example 6] In Comparative Example 1, the cultivation was performed in the same manner as in Comparative Example 1 except that after the medium was filled in the culture bottle, 4.5 g of the ground carrot ground product was placed on the seed inoculation site and sterilized. . Then, an average of 25 g of armillaria was harvested per culture bottle.

It took 5 days to form root-shaped hyphae bundles from the inoculum and 20 days including the 5 days around the fungus around the medium. The number of tested culture bottles was 12, and none of the culture bottles had miscellaneous bacteria troubles in the initial stage of culture.

[0032]

[Example 7] Same as Control Example 1 except that in Control Example 1, after sterilizing and cooling the medium, 4.5 g of sterilized raw pumpkin grind was placed on the medium and inoculated with the inoculum. Was cultivated. Then, an average of 23 g of armillaria per one culture bottle was harvested.

It took 5 days to form root-shaped hyphae bundles from the inoculum, and 20 days including the 5 days around the bacteria in the medium. The number of test bottles in the culture bottle was 12, and the number of culture bottles in which trouble of germs occurred in the initial stage of culture was 1.

[0034]

[Comparative Example 2] 140 g of air-dried birch bark, rice bran 65
g and 425 ml of water were mixed to fill a medium having a water content of 69% into an 800 ml polypropylene culture bottle, which was capped with polypropylene and sterilized under high pressure at 120 ° C. for 1 hour. After cooling, 8 g of the mushroom species of the genus Armillaria
Inoculate with 60 ° C in a culture room at a temperature of 22 ° C and a relative humidity of 70%.
After culturing for a day, temperature 16 ° C, relative humidity 85%, illuminance 300
Sprouting and growth were performed in the lx generation chamber, and after 32 days, an average of 60 g of armillaria was harvested per culture bottle.

It took 11 days to form the root hyphae bundles from the inoculum and 35 days including the 11 days around the fungus around the medium. The number of test bottles in the culture bottle was 12, and the number of culture bottles in which germ trouble occurred in the initial stage of culture was 4.

[0036]

[Example 8] In Control Example 2, raw carrot 18 ground in an inoculum inoculation site after filling the medium in an 800 ml culture bottle
Cultivation was carried out in the same manner as in Control Example 2 except that the cells were sterilized after adding g and the culture period was 30 days. And
After 32 days, an average of 98 g of armillaria was harvested per culture bottle.

It took 9 days to form root hyphae bundles from the inoculum and 30 days including the 9 days around the culture medium. The number of tested culture bottles was 12, and none of the culture bottles had miscellaneous bacteria troubles in the initial stage of culture.

[0038]

INDUSTRIAL APPLICABILITY In the bed cultivation of basidiomycetes of the genus Armillaria using sawdust, by using a culture medium to which an appropriate amount of carotene or a carotene-containing substance is used, the roots from the inoculated inoculum are The number of days until the formation of mycelial bundles was shortened by 18 to 38%. Furthermore, the incidence of miscellaneous bacteria at the beginning of culture is approximately 30% when not added.
However, the addition significantly reduced it to about 4%. As a result, stable artificial cultivation of armillaria has become possible. In addition, the utilization of carotene-containing materials also leads to effective utilization of agricultural waste of low utility value. Therefore, the present invention not only helps to develop the demand for new mushrooms, but also promotes effective utilization of waste resources, and greatly contributes to the development of the mushroom industry and the improvement of profits of agricultural products.

Claims (1)

[Claims] 1. In bed cultivation of basidiomycetes belonging to the genus Armillaria using sawdust, a total weight of the medium of 0.
A cultivating medium for Bacillus subtilis of the genus Armillaria, which comprises mixing an amount of carotene or a substance containing carotene corresponding to a concentration of 0002% to 0.5%.