WO2013161939A1 - Method for culturing fungus - Google Patents

Description

Fungal culture method

The present invention relates to a fungal culture method for growing basidiomycetes from mycelium to fruit bodies in layers on a sheet using a culture sheet supporting a solid medium or a liquid medium.

Various fungi such as venix nokitake (gyudonoshiba), garlic mushroom (Ganoderma lucidum), eucommia serrata (Cycium edulis), and agaricus are used as medicinal ingredients for herbal medicines and as active ingredients for cosmetics and health foods.

Natural basidiomycetes are grown on the back of a base material such as a raw tree of a plant that is a nutrient source. Among these natural basidiomycetes, Taiwan's endemic species, Benix nokitake, grows exclusively on the natural wood of raw gyudon, and is extremely slow growing, coupled with the rough cutting of the base gyudon. It is a rare natural resource that is difficult to collect.

Benix nokitake contains almost no medicinal ingredients in its mycelium, and as disclosed in Non-Patent Document 1, triterpenoids exhibiting pharmacological actions such as liver function improving function and antitumor action in the fruit body. Such a medicinal component is contained in many kinds and at a high concentration. Benix nokitake is considered to be artificially produced in liquid culture or solid culture because it has a small collection amount from nature. However, in liquid culture, the culture rate is somewhat faster than natural growth, and a large amount of culture solution must be used. Even if a large-capacity tank is used, the production volume is small for the scale and the production efficiency is poor, and it is not suitable for industrial production. On the other hand, conventional solid culture requires a lot of agar medium containers to be cultivated in an aseptic room and is troublesome. In addition, there is a large difference in the growth rate and medicinal component content among the agar medium containers, and the volume is limited. It is not suitable for industrial production because the agar containers cannot be arranged so much in the room.

Edible mushrooms, such as shiitake mushrooms, are much easier to cultivate than Benicus mushrooms, and are cultivated by culturing fungi in a medium and transplanting them into raw wood. Patent Document 1 is used for solid culture of fungi such as shiitake mushrooms, and has a plurality of stitch-like pores formed of synthetic fibers as a whole, and has a plurality of thin top portions by press molding and a thin top portion. A molded sheet for fungal culture having a recess is disclosed. Such a molded sheet for culturing fungi is cultivated for transplanting fungi to raw wood, so that solid fungi such as Benicus mushrooms that become membranous fruit bodies from mycelium are not transplanted to the raw wood. It cannot be used for culturing.

Evidence-Based Complementary and Alternative Medicine (Evidence-Based Complementary and Alternative Medicine), Volume 2011, ID 212641

Japanese Patent No. 3139837

The present invention has been made in order to solve the above-mentioned problems, and a basidiomycete can be grown from a mycelium to a fruit body in a solid medium or a liquid medium, and can be cultured efficiently and easily with uniform quality. It is an object to provide a method and a culture sheet used therefor.

The fungal culture method according to claim 1, which has been made to achieve the above object, is made of metal, synthetic resin, natural resin, and / or paper, from mesh, woven fabric, non-woven fabric, and porous sheet. After forming the selected reinforcing body and attaching a medium for basidiomycete mycelium, the mycelium of basidiomycetes is inoculated on the front side by coating or spraying the culture solution, and the mycelium is grown After forming the culture sheet, the mycelium growth step for growing the mycelium, and the mycelium grown on the front surface side is attached with a medium for basidiomycetous fruit bodies, And having a fruiting body growth step for growing the mycelium from the mycelium to the fruiting body.

The fungal culture method according to claim 2 is the method according to claim 1, wherein the basidiomycete mycelium medium and the basidiomycete fruit body medium are respectively a liquid medium or a solid medium. And

The fungal culture method according to claim 3 is the method according to claim 2, wherein the basidiomycete mycelium medium comprising the liquid medium is supported on the reinforcing body during the mycelium growth step, During the fruiting body growth step, the medium for basidiomycetous fruiting bodies comprising the liquid medium is supported on the front surface side on the reinforcing body, or the medium for basidiomycetous fruiting bodies comprising the solid medium is the reinforcing body. Supported on the back side, or
During the mycelium growth step, the basidiomycetous mycelium medium comprising the solid medium is supported on the front side on the reinforcing body, and during the fruiting body growth step, the basidiomycetes comprising the liquid medium A medium for fruiting bodies is carried on the medium for basidiomycetous mycelium or the medium for basidiomycetous fruit bodies consisting of the solid medium is carried on the back side of the reinforcing body, or
During the mycelium growth step, the basidiomycete mycelium medium comprising the solid medium is supported on the front side on the reinforcing body, and during the fruit body growth step, from the basidiomycete mycelium medium The basidiomycete fruit body culture medium comprising the solid medium is supported on the back side of the reinforcing body from which the reinforcing body has been peeled off together with the mycelium.

The fungal culture method according to claim 4 is the method according to claim 3, wherein during the mycelium growth step, the basidiomycete mycelium medium comprising the layered solid medium on the mycelium film, After the reinforcing body is mounted and crimped and integrated, the mycelium culture solution of the basidiomycetous fungus is sprayed to form the mycelium growth culture sheet, and during the fruit body growth step, the front surface side The reinforcing body together with the mycelium grown in step 1 is peeled off from the basidiomycete mycelium medium and the mycelium film, and then the basidiomycetous fruit body medium comprising the layered solid medium on the fruit body film is used. The fruit body growing culture sheet is formed by pressure-bonding the reinforcing body on the back surface side.

The method for culturing a fungus according to claim 5 is the method according to any one of claims 1 to 4, wherein the mycelium-growing culture sheet is rolled up to grow the mycelium, and / or The fruit body growing culture sheet is wound up in a roll shape to grow the fruit body.

The method for culturing a fungus according to claim 6 is the method according to any one of claims 1 to 5, wherein the basidiomycete is a fungus of the order Hydrangea.

The method for culturing fungi according to claim 7 is the method according to claim 6, wherein the fungus of the class Agaricaceae is a chanterelle family, a white mushroom family, a broom bamboo family, a red-branch family, a coral family, a sorghum family, a tobacco scale. It is characterized by being a fungus of the family Bambooceae, Idotake, Shirotake, Nikuharitake, Ezoharitake, Ibotake, Ningyotake, Pleurotus, Amanita, or Miyamatonbiidae.

The method for culturing fungi according to claim 8 is the method according to any one of claims 1 to 7, wherein the mycelium and / or the fruiting body is subjected to physical stimulation, electrical stimulation, optical stimulation, chemical It is characterized in that at least one treatment selected from a physical stimulus and a biological stimulus is performed.

The culture sheet according to claim 9 is selected from a mesh, a woven fabric, a non-woven fabric, and a porous sheet formed of metal, synthetic resin, natural resin, and / or paper, and has a basidiomycetous fungus on its front surface. The reinforcing body for growing the mycelium carries the basidiomycete fruit body culture medium grown from the mycelium to the fruit body.

The culture sheet according to claim 10 is the one described in claim 9, wherein the reinforcing body is made of a liquid medium or a solid medium, and the medium for basidiomycetous mycelium in which the mycelium is grown is an unsupported reinforcing body. The basidiomycete fruiting body medium comprising a liquid medium or a solid medium is supported on or supported by the reinforcing body.

The culture sheet according to claim 11 is the culture sheet according to any one of claims 9 to 10, and has air permeability due to a mesh-like, lattice-like, gathered-like, irregular wave-like shape, and / or a saw-like gap. A spacer is attached to the reinforcing body.

According to the fungal culture method using the culture sheet of the present invention, basidiomycetes can be grown from mycelium to fruiting bodies in a short period of time on a solid culture sheet. By winding the solid culture sheet and allowing it to stand at a constant temperature, it can be cultured in a large amount in a narrow space. Since a solid culture sheet is used, it can be cultured efficiently and easily with uniform quality.

It is a schematic diagram which shows the fungi culture | cultivation method to which this invention is applied. It is a schematic diagram which shows another fungi culture | cultivation method to which this invention is applied.

Hereinafter, modes for carrying out the present invention will be described in detail, but the scope of the present invention is not limited to these modes.

The fungal culture method of the present invention will be described with reference to FIG. This fungal culture method is performed using the mycelium growing solid culture sheet 1 and the fruit body growing solid culture sheet 2 according to the following procedure.

First, the mycelium growth step [I] is performed. A reinforcing body 15 capable of supporting a culture medium is formed and wound up with metal, synthetic resin, natural resin, paper, mesh, woven fabric, nonwoven fabric, and irregularities or holes of a porous sheet. The mycelium film 11 is pulled out from the non-permeable resinous mycelium film roll. The basidiomycete mycelium medium coating solution 13 is applied or sprayed to the mycelium film 11 on the front surface side with a basidiomycete mycelium medium application nozzle 12 to form a mycelium growth agar medium layer which is a solid medium layer. A layered basidiomycete mycelium medium 14 is formed. When the roll-shaped reinforcing body 15 is pulled out and pressed onto the basidiomycete mycelium medium 14, the reinforcing body 15 and the basidiomycete mycelium medium 14 are integrated. When a mycelium culture spray solution 17 of basidiomycetous liquid cultured on the basidiomycete mycelium medium 14 is sprayed onto the basidiomycete mycelium medium 14 from the mycelium culture solution spray nozzle 16, the basidiomycetes 20 become basidiomytes. The medium 14 for fungal mycelium is inoculated. If necessary, a net-like / lattice-like or saw-like spacer (not shown) for preventing adhesion during winding is further provided thereon. Thereby, the mycelium growing solid culture sheet 1 is obtained. The mycelium growing solid culture sheet 1 is loosely wound up in a roll shape so that the sheets do not adhere to each other, and is left to stand. Meanwhile, the basidiomycetes 20 are grown on the basidiomycete mycelium medium 14 and are planarly grown in all directions to form a mycelium layer of the basidiomycetes 20.

Next, the fruiting body growth step [II] is performed. The mycelium film 11 is peeled off from the mycelium-growing solid culture sheet 1 together with the basidiomycete mycelium medium 14. Then, the mycelium layer of the basidiomycetes 20 remains on the front side on the reinforcing body 15. The fruiting body film 21 is pulled out from the resinous fruiting body film roll which is impermeable. A basidiomycetous fruit body medium coating liquid 23 is applied or sprayed onto the fruit body film 21 with the basidiomycetous fruit body medium nozzle 22 on the front surface to form a fruit body growing agar medium layer. An entity medium 24 is formed. When the reinforcing body 15 is pressure-bonded to the basidiomycetous fruit body medium 24 on the fruit body film 21 on the back side, the reinforcing body 15 and the basidiomycete fruit body medium 24 are integrated. The mycelium layer of the basidiomycetes 20 comes into contact with the basidiomycetous fruiting medium 24 and can be grown and grows. Thereby, the fruit body growing solid culture sheet 2 is obtained. The fruit body growing solid culture sheet 2 is loosely wound up in a roll shape so that the sheets do not adhere to each other, and is left to stand. Meanwhile, the basidiomycetes 20 grow and grow from mycelium to fruiting bodies in the basidiomycetous fruiting medium 24, and grow in layers on the sheet in all directions to form a fruiting body layer of the basidiomycetes 20.

The mycelium-growing solid culture sheet 1 and the fruit body-growing solid culture sheet 2 can maintain a humidity and aeration state because a gap is generated by a spacer during winding and the adhesion is inhibited. The mycelium layer and fruiting body layer of the basidiomycetes 20 grow while avoiding the mesh-like, lattice-like, or saw-like gaps of the spacers.

In the mycelium growth step [I] and the fruiting body growth step [II], both showed examples in which a solid medium was used as a basidiomycete mycelium medium and basidiomycete fruiting body medium. The medium may be a solid medium, and the other may be a liquid medium. For example, another fungal culture method will be described with reference to FIG. 2 according to the following procedure.

First, by performing the mycelium growth step [I] in the same manner as described above, the mycelium-growing solid culture sheet 1 is prepared, wound and allowed to stand, and the basidiomycetes 20 are grown and propagated. 20 mycelium layers are formed (see FIG. 1). Next, as shown in FIG. 2, a fruit body growth step [II] is performed. The basidiomycetous fungus body medium coating solution 23 is sprayed onto the basidiomycete mycelium medium 14 on the front side of the mycelium-growing solid culture sheet 1 with the basidiomycetous fruit body medium nozzle 22 to spray the fruiting body growing liquid medium. The basidiomycetous fruiting body medium 24 to be layered is formed. Thereby, the fruit body growing solid culture sheet 2 is obtained. The fruit body growing solid culture sheet 2 is loosely wound up in a roll shape so that the sheets do not adhere to each other, and is left to stand. Meanwhile, the basidiomycetes 20 grow from the mycelium to the fruiting bodies and grow in the basidiomycetous fruiting medium 24 and multiply in a planar manner to form a fruiting body layer of the basidiomycetes 20.

The fungal culture method is suitable for culturing basidiomycetes. Basidiomycetes can be expressed according to the classification of Mushrooms, Newly-developed Sankei Field Books 7 (June 10, 2006, first edition, first edition, published by Yamato Valley Co., Ltd.). It is preferable that the order is Lepidoptera-H. Subtilis-Hymenaceae. For example, chanterelles, sphagnum mushrooms, blossoms mushrooms, blossoms mushrooms, coral mushrooms, sorghum mushrooms, cigarette mushrooms, iridaceae, mushroom mushrooms, mushroom mushrooms, mushroom mushrooms, mushroom mushrooms, Family fungi of the family, Koyakutake department, Manentake department, Miyamatonbimai family. More specifically, chanterelles (Cantharellus cibarius), white-spotted bamboo (Clavaria vermicularis), broom bamboo (Ramaria ハ botrytis), red-bellied bamboo (Sparassis crispa), coral harimatake (Hericium ramosun), fusahari cyno (bat) Beech haritake (Mycoleptodonoides aitcbisonii), Maitake mushroom (Grifola 、 frondosa), Giant mushroom (Trametes gibbosa), Agaric mushroom (Phanerochaete chrysorbiza), Olive mushroom (Lopharia cinerascens), Pepper mushroom (patite) ), Ningyotake (Albatrellus confluens), Aurouji (Albatrellus caeruleoporus), Numeraitake (Albatrellus yasudai), Kotake (Sarcodon aspratus), Kurokawa (Boletopsis leucomelas), Sennintake (Albatrellus pas) caprae), choreimaitake (Polyporus umbellatus), mustard (Laetiporus sulphureus), oyster mushroom (Pycnoporus coccineus), Japanese oyster mushroom (Antrodia albida), yamashiroamitake (Antrodia beteromorpha) No Japanese name), Melanoporia castanea, Anatake (Schizopora flavipora), Tsunonomanentake (Ganoderma mastporum), Bamboo shoot (Inonotuslicoblicuus), Kawatake (Peniophora quercina), and the like.

The medium for basidiomycete mycelium and the medium for basidiomycete fruit bodies may be a solid medium such as an agar medium or a liquid medium.

The solid medium is, for example, a hydrogel-forming polymer such as polyalginate and agar in 1 L of medium alone or mixed, 3 to 20 g thereof, 3 to 5 g of potato extract, glucose, dextrose, fructose, sucrose , Cellobiose, or trehalose alone, or 10 to 30 g of a saccharide such as a mixture of any of them. As necessary, pH regulators, inorganic salts that serve as nitrogen and mineral sources, sawdust, pigeons, oats and wheat, barley, straw, and other animals and plants, their extracts, secondary metabolites, etc. You may add suitably.

The liquid medium is, for example, 10 to 30 g each of yeast extract and malt extract in 1 L of medium, dextrose, dextrose, fructose, sucrose, cellobiose, trehalose alone, or a mixture of any of them. Contains 10-30 g of sugar. Adjust pH as necessary, inorganic salts as nitrogen and mineral sources, animal and plant extracts and sawdust, animal and plant bodies such as wheat, oats, wheat, barley and straw, their extracts, secondary metabolites Etc. may be added as appropriate.

When the fruit body is grown, it is formed on the solid body-cultivating solid culture sheet 2, or during the formation of the fruit body, or after the formation of one month, physical stimulation, electrical stimulation, optical stimulation, biological stimulation, chemistry You may perform the process of at least any one of an irritation | stimulation. As physical stimulation, electrical stimulation, and optical stimulation, irradiation with pulse laser at 20 to 100 mW at intervals of 10 nanoseconds to 100 milliseconds, preferably several tens of nanoseconds to several tens of milliseconds, 20 to Examples include plasma treatment with a pulsed plasma generator at 90 kV, preferably 30 to 70 kV and 100 μA to 1 mA at intervals of 100 to 500 milliseconds for 30 to 60 seconds, and light irradiation treatment with a wavelength of 500 to 800 nm. Among physical stimulation, electrical stimulation, and optical stimulation, physical stimulation and electrical stimulation may not be applied directly to the fruit body of the fruit body growing solid culture sheet 2. For example, as shown in FIG. 2, a pulse voltage 26a is generated by the pulse voltage electrode 25a toward the fruit body growing solid culture sheet 2, and / or the pulse voltage 26b is generated by the pulse voltage electrode 25b of the fruit body growing solid culture sheet 2. It may be generated for air sterilization without directing it upwards. As an example, plasma treatment with a pulsed plasma generator at 50 to 90 kV, preferably 70 to 90 kV, 100 μA to 1 mA and 100 to 500 milliseconds for 1 to 10 seconds is continued once per minute. As biological and chemical stimuli, coexisting heat-treated products of heterogeneous fungi such as lactic acid bacteria, yeast, and koji molds that do not inhibit fruit body growth, or live bacteria and heat-treated products of different lots of the same type of fungi to grow fruit bodies And treatment of mixing terpenes and sawdust from beef cake producing terpenes and terpenes. Processes such as formation physical stimulation, electrical stimulation, optical stimulation, biological stimulation, and chemical stimulation significantly improve the production amount, generation rate, and generation density of fruiting bodies. With the pulsed laser irradiation treatment, the production amount is increased by about 20 to 50%.

The reinforcing body 15 is a cell culture mesh, for example, having an opening of 50 to 1000 microns and a thickness of 30 to 300 microns. The material is a metal (for example, stainless steel), a synthetic resin (for example, nylon, Polyester, polyethylene, polypropylene, fluorine fiber, polyvinylidene chloride, polyvinyl alcohol, ethylene-vinyl acetate copolymer, polyurethane, polycellulose, etc.) mesh cloth and porous sheet, woven fabric of natural fibers (eg silk, cotton) , Paper, and non-woven fabric. The width is, for example, 200 to 1100 mm.

The mycelium film 11 and the fruit body film 21 are support films, and are formed of a heat-resistant resin exemplified by polyester, polypropylene, and polyvinylidene chloride. The thickness is, for example, 10 to 200 microns, and the width is, for example, 200 to 1100 mm.

In the fungus culture method, pretreatment such as pulse voltage treatment, pulse laser irradiation treatment, plasma treatment, and light irradiation treatment may be performed as necessary for sterilization and germination promotion of basidiomycetes.

The spacer is a mesh or lattice, for example, a gathered shape that is creased at intervals of 1 to 3 cm, and a concavo-convex wave having continuous irregularities of sine wave shape, cone shape, bell shape, or truncated cone shape at intervals of 0.1 to 3 cm. It may be air permeable due to its shape and / or slat-like gap. This unevenness is to allow the mycelium-growing solid culture sheets 1 or the fruiting body-growing solid culture sheets 2 to be in line contact / point contact with each other to give air permeability.

The mycelium-growing solid culture sheet 1 and the fruit body-growing solid culture sheet 2 may be used while being stretched, or may be used by being folded or wound, or may be used after being cut to an appropriate size. When a continuous long sheet is rolled up and used, it contributes to an improvement in productivity in a narrow space.

An example of cultivating Benicaria mushroom by a fungal culture method using a culture sheet to which the present invention is applied is shown. In addition, a reagent special grade which is commercially available was used except for the reagent in which the trade name and manufacturer were written.

(Preparation example)
(1) Preparation of medium Peel 200 g of potato, cut into 1 cm square, boiled in 1 L boiling water for 20 minutes, filtered with gauze, added ion exchange water to 20 g of glucose to a total volume of 1 L, 121 ° C., 20 A potato dextrose medium (hereinafter referred to as PDB medium) was prepared by autoclaving for minutes. At this time, the pH was adjusted to 5 to 7 with a phosphate buffer.
Also, peel off 200 g of potato, cut into 1 cm square, boiled in 1 L boiling water for 20 minutes, filtered with gauze, added 15 to 20 g of agar (Nissui Pharmaceutical Co., Ltd.) to 20 g of glucose, and stirred. Ion exchanged water was added to make the total volume 1 L, transferred to a flask, and autoclaved at 121 ° C. for 15 minutes to prepare an agar medium (hereinafter referred to as PDA medium). At this time, the pH was adjusted to 5 to 7 with a phosphate buffer.
The obtained medium was cooled in a sterile draft by adding 15 to 20 mL of PDA medium to a petri dish having a diameter of 9 cm in the case of (i) solid culture. (Ii) In the case of liquid culture, the flask containing the PDB medium was inoculated and cultured after cooling to room temperature. All the cultures were performed at a constant temperature of 25 ° C. in an incubator.

(2) Solid Passage Culture of Wild Species of Benicus mushroom A 2 mm x 3 mm piece of the tissue behind the wild Species of Benicus mushroom was peeled off with tweezers sterilized by a flame of a gas burner, and inoculated into PDA medium. Ten days later, the mycelium that became a colony having a diameter of about 2.5 cm was cut out as a 5 mm square piece and inoculated into a new PDA medium. Thereafter, the passage culture was carried out approximately every 21 days and used for the fungal culture method of the present invention.
In addition, during the subculture, no change was observed in the growth rate from the first generation to the tenth generation. As a result of DNA analysis of the second generation mycelium using a DNA analyzer, the base sequence showed a homology of 96% (266/276), and this mycelium was identified as Benicus nokitake.

(3) Optimization of solid medium (3-1) Selection of medium In PDA medium, glucose, which is a saccharide, and dextrose, fructose, or sucrose instead of 0.5% and 1.0%, respectively, as a carbon source , 1.5%, 2.0%, and 3.0% test samples were prepared separately, and after passage culture, the diameter of the mycelium colony was measured. did. Each colony diameter was about 50 mmΦ in the case of 1.0% or more in a medium containing glucose and dextrose or fructose, and about 25 mmΦ in a concentration-independent medium in a medium containing sucrose. For this reason, glucose, dextrose, fructose, and particularly inexpensive glucose were suitable for cultivation of Benix noctum mycelium. The mixture of camphor with 500 ppm both had a diameter of 60 mm, which was suitable for promoting growth.

(3-2) Selection of fruiting body growth medium In 1 L of medium, Benix nokitake strain that has been subcultured in a medium consisting of 10 g of yeast extract, malt extract, 20 g of dextrose, 15 g of agar, and 100 g of crushed pigeons. After culturing for 3 months after inoculation, a part of the cells were observed with a microscope, and the presence of clamps and spores indicating fruiting body formation could be confirmed. Moreover, when the obtained microbial cells were dried and extracted with methanol and subjected to high performance liquid chromatography (HPLC) analysis, ergosterols as fruit body-containing components were also detected. From these results, it was found that PDA medium is suitable for mycelium growth, and that the medium is suitable for fruiting body growth.

(4) Optimization of liquid medium (4-1) Examination of liquid culture feasibility Add 200 mL of PDB medium to a 300 mL Erlenmeyer flask, and after sterilization, add five 5 mmΦ PDA medium carrying Benix nokitake. Then, liquid culture was performed in a thermostatic chamber maintained at 25 ° C. with shaking at 150 rpm for 7 days to obtain mycelia.

(Example 1: Culture on a planar solid culture sheet)
A polyethylene terephthalate (PET) film, which is a mycelium film 11 having a thickness of 100 microns and a thickness of 20 × 25 cm, was laid on a bat having a size of 25 × 30 cm. After flowing about 150 g of PDA medium, which is a basidiomycete mycelium medium 14, which was autoclaved at 121 ° C. for 30 minutes in advance and cooled to 60 ° C. or less so as to be approximately 15 × 20 cm, 15 × 20 cm A medium was prepared by laying a PET mesh, which is a reinforcing member 15 of the above. The medium was cooled to room temperature. A liquid culture of Benix nokitake, which had been previously cultured in a PDB medium and homogenized with a homogenizer, was evenly applied to this medium to obtain a mycelium-growing solid culture sheet 1. The vat was cultured at 25 ° C. under aseptic conditions for 30 days.
The resulting mycelium-attached mesh was transferred together with the basidiomycetes 20 onto the medium described in Example 3 (3-2) to obtain a fruit body growing solid culture sheet 2. This was cultured for 60 days. At this time, plasma treatment with a pulsed plasma generator at 30 kV and 400 μA at intervals of 100 milliseconds for 30 seconds was performed once every day for 24 days from the start of the culture for 2 days. Below, spores and clamps were observed, and the obtained microbial cells were dried, extracted with methanol, and subjected to HPLC analysis. As a result, ergosterols, which are fruit body-containing components, were also detected. This confirmed that a child entity was generated.

(Example 2: Culture in a roll-shaped solid culture sheet)
In Example 1, a PET film, a PET film and a rolled PET film having a width of 300 mm are used, and a 1 cm square mesh spacer made of porous vinyl chloride resin having a diameter of 1 to 2 mm is wound between layers in each step. In the same manner as in Example 1, the mycelium growing solid culture sheet 1 and the fruiting body growing solid culture sheet 2 were prepared and subjected to plasma treatment with a pulse plasma generator for 30 seconds at 400 μA at 30 kV and at intervals of 100 milliseconds. After 24 days from the start of the culture, it was applied once a day for 2 days. After 60 days, the culture on the mesh was observed for spores and clamps under a microscope. When extracted and analyzed by HPLC, ergosterols, which are fruit body-containing components, were also detected. This confirmed that a child entity was generated.

The method for culturing fungi using the solid culture sheet of the present invention is a method of artificially cultivating basidiomycetes that grow and produce a film-like fruit body on a smooth or rough surface and used as a crude drug in medicines or health foods It is used to be used as an active ingredient.

1 is a solid culture sheet for growing mycelium, 2 is a solid culture sheet for growing fruit bodies, 11 is a film for mycelium, 12 is a medium application nozzle for basidiomycete mycelium, 13 is a medium application solution for basidiomycetous mycelium, and 14 is a basidiomycete Medium for fungal mycelium, 15 reinforcement, 16 mycelium culture spray nozzle, 17 mycelium culture spray nozzle, 20 basidiomycetes, 21 fruit body film, 22 basidiomycete culture medium nozzle , 23 is a basidiomycete fruit body medium coating solution, 24 is a basidiomycete fruit body medium, 25a and 25b are pulse voltage electrodes, and 26a and 26b are pulse voltages.

Claims (11)

Form a reinforcement selected from mesh, woven fabric, non-woven fabric, and porous sheet with metal, synthetic resin, natural resin, and / or paper, attach a medium for basidiomycete mycelium, After inoculating the mycelium of basidiomycetes on the surface side by coating or spraying the culture solution thereof to form a mycelium-growing solid culture sheet, a mycelium growth step for growing the mycelium,
The mycelium grown on the front side is attached with a basidiomycetous fruit body medium to form a fruit body growth culture sheet, and then a fruit body growth step for growing from the mycelium to the fruit body ,
A method for culturing fungi, comprising: 2. The fungal culture method according to claim 1, wherein the basidiomycete mycelium medium and the basidiomycete fruit body medium are respectively a liquid medium or a solid medium. During the mycelium growth step, the basidiomycetous mycelium medium composed of the liquid medium is supported on the reinforcing body, and during the fruiting body propagation step, the basidiomycetous fruit body culture medium composed of the liquid medium is reinforced. The body is carried on the front side, or the basidiomycete fruiting body medium comprising the solid medium is carried on the back side on the reinforcing body, or
During the mycelium growth step, the basidiomycetous mycelium medium comprising the solid medium is supported on the front side on the reinforcing body, and during the fruiting body growth step, the basidiomycetes comprising the liquid medium A medium for fruiting bodies is carried on the medium for basidiomycetous mycelium or the medium for basidiomycetous fruit bodies consisting of the solid medium is carried on the back side of the reinforcing body, or
During the mycelium growth step, the basidiomycete mycelium medium comprising the solid medium is supported on the front side on the reinforcing body, and during the fruit body growth step, from the basidiomycete mycelium medium 3. The fungal culture method according to claim 2, wherein the basidiomycete fruiting body medium comprising a solid medium is supported on the back side of the reinforcing body from which the reinforcing body has been peeled off together with the mycelium. During the mycelium growth step, the basidiomycetous mycelium medium comprising the layered solid medium on the mycelium film is mounted by pressing and integrating the reinforcement, and then the mycelium of the basidiomycetes is integrated. Spray the culture solution to form the mycelium growth culture sheet,
During the fruiting body growth step, after peeling the reinforcing body together with the mycelium grown on the front surface side from the basidiomycete mycelium medium and mycelium film, The fungus cultivation method according to claim 3, wherein the basidiomycete fruit body culture medium comprising the solid medium is pressure-bonded to the reinforcing body on the back surface side to form the fruit body growth culture sheet. The mycelium growing culture sheet is wound up in a roll shape to grow the mycelium, and / or the fruit body growing culture sheet is wound up in a roll shape to grow the fruit body. Item 5. The fungal culture method according to any one of Items 1 to 4. The method for culturing fungi according to any one of claims 1 to 5, wherein the basidiomycetous fungi are fungi of the order Cleoptera. The fungus of the Agaricaceae is chanterelles, white mushrooms, brooms, coral, family coral, cigarettes, cigarettes, iridaceae, agaricaceae, agaricaceae, agaricaceae, agaricaceae, 7. The method for culturing fungi according to claim 6, wherein the fungus is a member of the family Ningyotake, Amanita, Amanentake, or Miyamatonbiidae. The mycelium and / or the fruiting body is subjected to at least one treatment selected from physical stimulation, electrical stimulation, optical stimulation, chemical stimulation, and biological stimulation. The fungal culture method according to any one of 7. A reinforcement body made of metal, synthetic resin, natural resin, and / or paper, selected from mesh, woven fabric, non-woven fabric, and porous sheet, and growing mycelium of basidiomycetes on its front surface A culture sheet, which carries a medium for basidiomycete fruit bodies grown from the mycelium to the fruit bodies. The reinforcing body is composed of a liquid medium or a solid medium, and the basidiomycetous mycelium medium in which the mycelium is grown is supported on or unloaded from the unsupported reinforcing body, and is composed of a liquid medium or a solid medium. The culture sheet according to claim 9, wherein the medium for basidiomycete fruit bodies is supported on the reinforcing body. 11. The spacer having air permeability by a mesh-like, lattice-like, gather-like, uneven wave-like shape, and / or slat-like gap is attached to the reinforcing body. The culture sheet as described.

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