JPH0576242A - Method for culturing basidiomycete and spawn and mushroom bed useful in the same method - Google Patents

Abstract

PURPOSE:To facilitate inoculating operation, to promote and to increase harvesting of fruit body and to efficiently culture a basidiomycete such as Lentinus edodes by culturing the basidiomycete in a base having given length and hardness, grinding the base to give spawn, packing the spawn into a container and culturing. CONSTITUTION:An approximately cylindrical woody bar having 7mm diameter and 15cm length is used as a base having given length and given hardness, sterilized, placed on a potato dextrose agar medium in which a basidiomycete such as lentinus edodes is cultured and the basidiomycete is cultured at 25 deg.C under a dark condition for 7-10 days and multiplied until the whole surface of the woody bar is covered white by observation with the naked eye to give cylindrical spawn. Then, the cylindrical spawn 1 is thrust, stood and inoculated into a wood flour medium 2 and cultured until the whole medium looks while by observation with the naked eye. The medium is ground, mixed with water, packed into a container such as polypropylene bag and cultured. At a stage when the medium is browned and hardened and is made into a mushroom bed of a cylindrical state, the medium is taken out from the container, fruit bodies are induced by a conventional method and the basidiomycete is efficiently cultured.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a fungal species suitable for cultivating basidiomycetes, a microbial bed, and a basidiomycete cultivation method using the same.

[0002]

2. Description of the Related Art Conventionally, shiitake, enoki mushroom, nameko,
In fungal bed cultivation such as Bunnamjiji, the secondary hyphae fully infested on agar or wood flour medium is used as an inoculum, and this inoculum is inoculated into a medium such as sawdust and cultivated to form a bacterial bed,
Fruit bodies were induced from this bed.

[0003]

However, since it is necessary to make a hole in the medium for inoculum such as conventional wood flour and inoculate the inoculum into the hole, the inoculation work is complicated and the inoculum in the medium is When the inoculum was formed, the exposed surface area of the medium at the time of inoculation became larger, so that various bacteria were easily mixed in the medium at the time of inoculation. Further, even when the hole was dug deep to the bottom of the medium, the hole was likely to collapse before the inoculation of the inoculum, and it was difficult to inoculate the inoculum to the bottom of the medium. The conventional fungal bed required a long period of time for hyphae to spread, and the fruiting body yield was not sufficient. Therefore, an object of the present invention is to provide a basidiomycete cultivation method that is easier to inoculate than before, and promotes mycelial spread than before, and can also improve fruiting body yield. Is to provide an inoculum and a fungal bed used in the basidiomycete cultivation method.

[0004]

[Means for Solving the Problems] In order to solve the above problems,
The inoculum according to claim 1 is characterized in that a basidiomycete is cultivated on a substrate having a predetermined length and a predetermined hardness.
The microbial bed is prepared by crushing a medium in which basidiomycete is cultivated in a container, and the method for cultivating basidiomycetes uses the inoculum of claim 1 and the bed of claim 2. It is characterized by

In the claim 1, the "predetermined length" means a length which is substantially the same as or longer than the depth of the medium inoculated with the inoculum.

Further, the "predetermined hardness" means a hardness that is more than sufficient to allow the medium to be inoculated with the inoculum.

The substrate according to claim 1 may be made of a material that can be inoculated with basidiomycete and cultured, and its shape and size can be arbitrarily determined according to the shape and size of the medium inoculated with the inoculum. Can be set. For example, examples of the substrate include a rod made of wood, plastics, or paper.

In the claims, the basidiomycete means fungi such as shiitake, maitake, nameko, oyster mushroom and enoki mushroom. A basidiomycete in the claims also includes a bacterium that is a new species and is taxonomically classified as a basidiomycete.

Any method may be used to inoculate the substrate with basidiomycetes. Further, the degree of culturing of basidiomycetes can be made to be a better inoculum as the hyphae occupy more substrates. Specifically, when the entire surface of the substrate is visually covered with white, it is considered that the mycelium has spread almost to the substrate. If inoculum in this state is used for inoculation, the mycelium spread rate in the medium is faster. Therefore, it is more preferable. The whiteness of the surface of the substrate is 50 points or more when the entire surface of the substrate is whitely covered with the naked eye. In the present specification, the whiteness means a color tristimulus value (X.Y., using a colorimetric color difference meter (Minolta CR-200 manufactured by Minolta Co., Ltd.).
It means a value of Z / 1.18 when Z) is measured.

When inoculating the medium with the inoculum according to claim 1, the inoculum can be easily set because the inoculum can be set up by simply inserting the inoculum into the medium, and the inoculation can be automated by using a piston type inoculator or the like. Is also easy. It is more preferable to insert the inoculum of claim 1 deeper into the medium because the hyphae are evenly inoculated into the entire bed, and most preferably, the miscellaneous bacteria are introduced to the bottom of the medium. It is also possible to inoculate a plurality of seeds per medium.

The medium in which the basidiomycete is cultivated in claim 2 may be a medium inoculated with the basidiomycete and cultured by any method, or may be any kind of medium.

When basidiomycetes are cultivated in a medium and the mycelium is almost infested in the medium, the entire medium looks white to the naked eye. The whiteness of the medium surface in this state is 50 points or more. When the mycelium further spreads, the medium turns brown and hardens, but it is considered that the mycelium almost completely spread in the medium in this state.

In the second aspect, the degree of basidiomycete culturing in the medium in which the basidiomycete is cultivated is such that the more the hyphae occupy the medium until the whole medium looks white, It is more preferable because the amount of bacterial cells is large. Also,
The medium according to claim 2, wherein the medium in which the hyphae are more monopolized than the medium in which the whole medium looks white or the hyphae are completely or almost completely spread and browned, and the hardened medium is crushed You may get However, it is better to crush the medium at the stage where the mycelium has been cultivated to a state where the whole medium looks white to the naked eye and obtain the fungal bed of claim 2 after inoculating the fungus bed with the inoculum and then inducing fruiting bodies. It is more preferable because the period required to obtain a suitable bacterial bed can be shortened.

The crushed material is intended to include any material of various sizes ranging from a large one in which the shape of the medium is simply broken to a fine powder. The preferred size of the crushed product is a particle size that can be fractionated using a standard sieve of 8 mesh or more and 40 mesh or less, more preferably 10 mesh or more and 20 mesh or less. This is because if it is too fine, the flow of air to the fungal bed will deteriorate, while
This is because if it is too coarse, water will easily evaporate from the fungal bed.

In the fungus bed of claim 2, it is preferable that an appropriate number of crushed products of the medium in which the basidiomycete is cultivated is packed in a container by applying an appropriate pressure. The mushroom bed weight per unit volume of the preferred mushroom bed is ^{0.5g / cm 3 ~2.0g / cm 3} , more preferably from ^{0.8g / cm 3 ~1.6g / cm 3} . This is because if the above value is too large, the ventilation of the air to the fungus bed will be poor, and if the above value is too small, the evaporation of water from the fungal bed will be too much.

Then, the number of culture media used for preparing the bacterial bed of claim 2 is arbitrarily determined according to the size of the culture media in consideration of the size of the bacterial bed to be obtained in the work. be able to. However, by crushing 1 part of the medium in which the basidiomycete was cultivated in the container in which the medium was originally contained, A bacterial bed obtained by crushing the medium before or after removing it from the original container and repacking the crushed product into the original container is also included in the bacterial bed of claim 2. These fungi beds can be a fungus bed that is effective in promoting hyphae infestation and increasing fruiting body yield because the hyphae are stimulated and oxygen is supplied at the stage of crushing the medium. In addition, it is advantageous to administer an appropriate amount of a nucleic acid-related substance, a hypha activator of saccharides or water, and nutrients to a crushed product of a basidiomycete culture, because it promotes hyphal spread and increases fruit body yield.

In claim 2, the shape of the container for refilling the medium is arbitrary, but if it is refilled into a cylindrical shape having a diameter of about 10 cm and a height of about 1 m, and the fungus bed is browned and hardened, normal shiitake log cultivation is performed. It is almost the same size and shape as the log used and is lighter than the log, so it is easy to handle, and since the length is a cylindrical shape, the fruiting body is less likely to be deformed and the surface area of the fungal bed is large. The fruiting body yield is also large and preferable.

In claim 3, the basidiomycete cultivation method using the inoculum of claim 1 and the inoculum of claim 2 means that the crushed product of the medium obtained by inoculating and culturing the inoculum of claim 1 in a container It means a method of cultivating a basidiomycete using a fungus bed packed in.

The time for inducing fruiting bodies from the fungus bed obtained in claim 2 is that when the hyphae are almost completely infested and the fungus bed is browned and hardened, it is taken out of the container or the container is removed and the surface thereof is removed. It is more preferable to expose the fruit to the atmosphere to induce the fruit body formation. However, even if the hyphae are not completely infested and not hardened, the fruiting bodies may be induced by punching the container (for example, punching holes having a diameter of 7.8 mm at intervals of about 5 cm). Moreover, more fruiting bodies can be obtained than before.

[0020]

According to the inoculum of the first aspect, since the substrate has a predetermined length and a predetermined hardness, it can be inoculated simply by standing it in a medium. According to the fungus bed of claim 2, the mycelium is stimulated by crushing the medium in which the basidiomycete is cultivated, and oxygen is also supplied. According to the basidiomycete cultivation method of claim 3, both the above-mentioned actions occur because the inoculum of claim 1 and the bed of claim 2 are used.

Next, examples of the present invention will be described. The shiitake mycelia used in this example were selected from those with stable growth and were strains preserved in the Kyushu University Faculty of Agriculture experimental forest produced by fusion with mononuclear spores, and subculture was repeated on potato-dextrose agar medium. It is still active.

【Example】

Example 1 Production of rod-shaped inoculum (a) 120 substantially cylindrical wooden rods having a diameter of 7 mm and a length of 15 cm
After sterilizing at 1.2 ° C / cm ² for 15 minutes and allowing to cool, this wooden stick was placed on a potato-dextrose agar medium in which shiitake hyphae were cultured, and cultured at 25 ° C in the dark.
It was observed with the naked eye that the whole surface of the wooden stick was covered with white shiitake mycelium from 1 week to 10 days after the culture, and it was found that the mycelium spread to the wooden stick. The wooden rod in which the mycelium spread was used as a rod-shaped inoculum in Examples 2 to 4 below. (B) Bamboo materials for skewers (diameter: 2 mm, length: 20 cm) or squares of beech (4 mm square, length: 20 cm) are used as the substrate in claim 1, and these are soaked in water to moisten the whole corn. Sprinkle with corn flour to get the powder. Next, pack this in a heat-resistant polypropylene bag or polypropylene bottle, and use a high-pressure kettle at 120 ° C, 1.2 kg / cm ² ,
Sterilize for 15 minutes. Next, in a polypropylene bag or bottle, put the well-crushed wood flour inoculum prepared by the conventional method using shiitake hyphae, mix with the substrate in the bag or bottle, and culture at 25 ° C in the dark. did. Approximately one week after the culture, it was visually observed that hyphae spread to the substrate and the entire surface of the substrate was covered with white shiitake hyphae. The substrate cultivated to this state is also one of the specific examples of rod-shaped inoculum. In the present example, it is not necessary to uniformly apply the conventional wood flour seed bacteria to the entire substrate, and the substrate and the wood flour seed bacteria were simply mixed and cultured for about one week, and the bacteria were substantially spread over the entire substrate. Example 2 Production of bacterial bed using rod-shaped inoculum Beech wood flour, rice bran, bran, and corn powder were mixed in a weight ratio of 7: 1: 1: 1, and after stirring, distilled water (pH 6.8) was added. The water content of the medium was adjusted to 60%. When 1.2 kg of this was packed in a polypropylene bag, it became a substantially cylindrical shape with a diameter of 12 cm and a height of 12 cm. This was autoclaved at 120 ° C. and 1.2 kg / cm ^{2 for} 20 minutes and allowed to cool to obtain a medium. Next, the rod-shaped inoculum 1 produced in (i) of Example 1 was inoculated with the shiitake bacterium by standing up against the bottom of the medium 2 as shown in FIG. After inoculation, dark culture was carried out at 25 ° C. for 20 days, and the rate of mycelial spread and the whiteness of the surface of the obtained fungus bed (hereinafter referred to as the rod-shaped inoculum bed) were measured. As a control, as shown in FIG. 2, the wood flour inoculum 3 (10 g) conventionally produced using Shiitake hyphae was inoculated into the above-mentioned medium 2 by a usual method and cultured under the same conditions. With respect to the bed (hereinafter referred to as the conventional fungal bed), the mycelial spread rate and the whiteness of the surface were measured in the same manner. The results are shown in Table 1.

[Table 1]

In Table 1, the numerical units of the mycelial spread rate and whiteness are mm / day and points, respectively. As shown in FIG. 3, the whiteness of the surface of the bacterial bed is proportional to the density of basidiomycetes in the bacterial bed (Miyata, Kyushu University Master's thesis 1985). In FIG. 3, the vertical axis represents whiteness (points), and the horizontal axis represents bacterial cell amount (mg / cm ² ). White circles are shiitake mushrooms and black circles are willow matsutake mushrooms.

As shown in Table 1, the fungal bed of rod-shaped inoculum was larger than the fungal bed of the conventional example due to the mycelial spread rate and the bacterial cell density. Also, by inserting rod-shaped seeds to the bottom of the medium, the whole medium could be inoculated with the seeds evenly. Furthermore, according to the rod-shaped inoculum of this example, the mycelia are in a state of spreading on the substrate, so that the amount of bacterial cells is approximately proportional to the volume of the wooden rod that is the substrate. Therefore, if a wooden rod of the same volume is used as a base, the amount of cells to be inoculated can be easily equalized, and it is convenient to manage multiple cells at one time, and there is less variation in the fruiting body production between the beds. became.

When the culture was further continued under the same conditions, when inoculated with a conventional wood flour fungus species, it took about one month from the culture until the entire bed became white and the mycelia were almost infested. Further, it took about 3 months from the culture until the mycelium almost completely spread and the medium became brown and hardened due to the activity of redox oxygen. On the other hand, when rod-shaped inoculum is inoculated, it takes only 2-3 weeks from the culture until the above-mentioned whole bed becomes white, and about 1.5-2 months from the culture until the medium turns brown. There wasn't. That is, by using the rod-shaped inoculum, the time required for the hyphae to spread was remarkably shortened. Therefore, by using the rod-shaped inoculum, it is considered that the mycelium quickly occupies the medium, and therefore it is difficult for other miscellaneous bacteria to be mixed due to the biological defense mechanism.

Example 3 Production of a log-shaped fungal bed The rod-shaped inoculum was inoculated and cultivated by the method described in Example 2, and the medium turned brown and hardened, and it was considered that shiitake hyphae were almost completely infested. Remove the medium from the polypropylene bag, crush it, and add about 500 ml of water to the 5 bacterial beds (total of about 6.0 kg) and pack it in another larger polypropylene bag to obtain a diameter of 12 cm and a height of 50 cm.
A cylindrical fungus bed was obtained. When culturing was continued under the same conditions as in Example 2, the mycelium was browned and hardened due to almost complete spread of mycelium in 1 to 2 weeks, and a raw woody bacterial bed was obtained. As a control, the conventional fungal bed in Example 2 was also cultivated under the same conditions until the fungal bed became brown and hardened. In order to compare the bacterial cell amount of the conventional example cultivated up to this state and the raw woody bacterial bed, the content of the components chitin, ergosterol, phospholipids and glycogen in the mycelium (per weight of the bacterial bed) ) Was measured. Chitin is measured by Braid, GH and Line,
HA (1981): Holzforschung 35: 10-15., Ergosterol is described in Seitz, LM (1977): Cereal Chem. 54: 1207-1.
217. Phospholipids are described by Derilex, PJL et al. (1990): Appl. E.
nviron.Microbiol. 56: 3029-3034, glycogen is Ro
e, JH and Dailey, RE (1966): Anal. Biochem. 15: 2.
It was measured by the method described in 45-250. The results are shown in Table 2.
Shown in.

[0026]

[Table 2]

In Table 2, A shows the result of the conventional example of the bacterial bed in Example 2, and B shows the result of the raw woody bacterial bed. The numerical units are mg / g for the content of chitin, ergosterol and glycogen, and μmol / g for the content of phospholipids.

As shown in Table 2, it was found that the log-shaped fungal bed had a remarkably large cell content per the bacterial bed amount (g) as compared with the conventional bacterial bed. In the original fungal bed of this example, shiitake hyphae were almost completely infested, browned, and prepared by crushing the hardened medium, but it was considered that the entire medium became white with the naked eye and shiitake hyphae were almost infested. It is better to crush the resulting medium to shorten the period from inoculation of the inoculum to obtaining the raw woody bacterial bed. In addition, it is better to handle about 10 of the fungus beds (total of about 12 kg) and to use a cylindrical raw wood-like fungus bed with a diameter of 12 cm and a height of 100 cm, because the size is closer to that of a normal log. More convenient. Example 4 Induction of fruiting body generation Next, the polypropylene bag of the raw woody fungus bed produced in Example 3 was completely peeled off, and 17 ° C., 80% RH, 100-15.
Fruit body development was induced in an environment of 0 lux, and the fruit body yield (per kg of bacterial bed) was measured 2 weeks after the induction. The results are shown in Table 3, column B. The raw wood produced as described in Example 3 using 5 of the medium obtained by culturing the bacterial bed of the conventional example in Example 2 under the culture conditions of Example 2 until browning and hardening. Table 3 shows the results for the fungus bed.
Shown in column C. For comparison, with respect to the conventional fungal bed in Example 2, the fruit bodies were similarly induced from the state where the mycelium was almost completely infested and browned and hardened, and the yield was measured. The results are shown as A in Table 3.

[0029]

[Table 3]

As shown in Table 3, the raw woody fungus beds B and C have a fruit body generation amount of about 1.5 times that of the conventional fungus bed A. In the fruit bodies obtained in B and C, the thickness of the fungal umbrella was increased and the quality was improved.
Further, as to the original fungal bed B prepared by using the rod-shaped inoculum, the mycelial spread rate is faster than that in the case of using the conventional wood flour inoculum as the inoculum (AC), as described in Examples.
After inoculation, the period was further shortened until fruiting bodies could be obtained.

The size and weight of the raw woody fungus beds B and C of this embodiment can be made uniform, so that the fungi beds can be easily handled. In addition, since it is in the form of raw wood, the fungus bed can be set up diagonally or assembled as in normal raw wood cultivation, and the cultivation place is not so wide. Furthermore, since the fungal bed has a large surface area because it is in the form of a log, it is easy to harvest upper fruiting bodies that can induce many fruiting bodies.

[0032]

According to the inoculum of claim 1, the inoculation work is facilitated. According to the fungus bed of claim 2, the harvest of fruiting bodies is promoted more than before and the yield thereof is also increased. According to the method for cultivating basidiomycetes according to claim 2, the inoculation work is facilitated, the harvest of fruiting bodies is promoted more than before, and the yield thereof is also increased.

[Brief description of drawings]

FIG. 1 is a cross-sectional view showing a state in which a rod-shaped inoculum is inoculated into a wood flour medium.

FIG. 2 is a cross-sectional view showing a state in which a normal wood flour inoculum is inoculated into a wood powder medium.

FIG. 3 is a graph showing the relationship between the whiteness of the surface of a bacterial bed and the density of bacterial cells in the bacterial bed.

[Explanation of symbols]

 1 Rod-shaped inoculum 2 Medium 3 Wood flour inoculum

Claims (3)

[Claims] 1. An inoculum comprising a basidiomycete cultivated on a substrate having a predetermined length and a predetermined hardness. 2. A microbial bed, characterized in that a crushed product of a medium in which basidiomycete is cultured is packed in a container. 3. A basidiomycete cultivation method using the inoculum according to claim 1 and the fungus bed according to claim 2.