JP3522202B2 - Culture method of Matsutake fungi for transplantation - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for culturing matsutake fungi for transplantation, which is a pure culture of matsutake fungi for transplantation using an agar medium. The present invention also relates to a culture tube and a culture device suitable for use in a method for culturing Matsutake mushrooms for transplantation. Further, the present invention relates to a method for transplanting purely cultured Matsutake fungus for transplantation.

[0002]

2. Description of the Related Art Various methods have been tried for a long time as a method for artificially culturing and growing Matsutake mushrooms. As one of these methods, pure culture of matsutake fungi for transplantation is performed using an agar medium, and the cultured matsutake fungus is transplanted to a natural medium such as Mt. Matsutake to artificially grow and culture Matsutake. The method is known.

In the pure culture method, a solid medium or a liquid medium is formed on the bottom of a test tube, a flask, a bottle, etc., and after sterilization,
Inoculate Matsutake inoculum such as spores into the medium, and after that,
The matsutake mushroom is allowed to stand in an atmosphere suitable for growth and culture.

It has been confirmed that fruit body primordia can be produced from the matsutake hyphae thus purely cultured.
No further achievements have been confirmed or reported. Therefore, in general, attempts have been made to artificially cultivate or multiply Matsutake mushrooms by transplanting purely cultured Matsutake hyphae into a natural medium (white) to form mycorrhizae in the roots of Japanese red pine. .

[0005]

Here, it is necessary to take out the matsutake hyphae purely cultivated in this way from the test tube or the flask, transfer them to the transplant destination, and implant them therein. However, such porting work is tedious,
In addition, there is a high possibility that miscellaneous bacteria will be mixed in at the time of transplantation and the matsutake hyphae will not grow after transplantation.

In view of the above points, an object of the present invention is to
Another object of the present invention is to propose a method for culturing Matsutake fungi for transplantation, which allows pure culture of Matsutake fungi for transplantation to be carried out easily.

Another object of the present invention is to propose a culture tube and a culture device suitable for use in such a culture method.

A further object of the present invention is to propose a method for transplanting matsutake bacterium for transplantation, which has been cultivated using the culture cylinder and the culture device according to the present invention.

[0009]

In order to solve the above problems, the present invention relates to a culture tube used in a method for culturing matsutake fungi for transplantation, in which matsutake inoculum is purely cultivated using a solid medium. The culture tube is: a tube body with a bottom;
It is characterized in that it has a lid detachably attached to the upper end opening of the cylinder body; and a slit formed on the outer peripheral wall of the cylinder body.

[0010] Here, the present invention, in order to be able to carry out the solid medium were placed in the cylinder body easily planting matsutake inoculum, as the lid, the rod can be inserted into the cylinder body that features a seed planting lid with.

Further, the present invention, at the time of pure culture, in order to prevent bacteria from entering while ensuring air permeability to a solid medium Matsutake inoculum is planted, as the lid, that features a vented lid .

Furthermore, in order to protect the part of the matsutake mycelium when the culture container after the growth of the matsutake mycelium is transported to the transplant destination, etc., in order to protect the part of the matsutake mycelium, a protection with a bottom into which the slit forming part of the cylinder body can be inserted It is desirable to have a cylinder.

Next, the present invention relates to a culture device used in a method for culturing matsutake fungus for transplantation, which is a pure culture of a matsutake inoculum using a solid medium. The culture device comprises a culture container and at least one culture tube. have.

The culture vessel includes: a culture tube; a culture tube mounting recess into which the culture tube can be inserted from the upper side; a container main body in which the culture tube mounting recess is formed; It is characterized in that it has a container lid detachably covered on the opening side of the recess for use; a vent hole formed in the container body; and a vent hole formed in the container lid. The culture tube includes: a tube body having a bottom; a lid detachably attached to an upper end opening of the tube body; a slit formed on an outer peripheral wall of the tube body; and a slit forming portion of the tube body. It is characterized by having a protective cylinder with a bottom that can be inserted.

On the other hand, the present invention relates to a method for culturing matsutake fungus for transplantation using the culturing apparatus having the above-mentioned structure, wherein the culturing method is: A step of inserting a solid medium into the body of the tube after inserting it into the recess for mounting the culture tube; and a lid for inoculum seeding equipped with a rod that can be coaxially inserted into the body of the tube as the lid to cover the body of the tube And covering the tube body with the lid, and performing sterilization treatment in that state; removing the lid for seed germ implantation,
Attaching matsutake inoculum to the rod, and inoculating the matsutake inoculum of the rod tip into a solid medium; and sterilizing the tube body with the vent hole portion sealed with a cotton plug. And a step of covering the cylinder body with the lid; and a step of covering the container body with the container lid that has been sterilized in a state where the ventilation hole portion is closed with a cotton plug. In this state, by leaving the culture container under a predetermined atmosphere temperature and humidity for a predetermined period of time to grow Matsutake hyphae on a solid medium in each culture cylinder. There is.

Next, the method for transplanting Matsutake fungus of the present invention is as follows:
The culture tube in the state of growing Matsutake hyphae by the method of culturing the above-mentioned matsutake fungus for transplantation is taken out from the culture tube mounting recess of the culture container, and the culture tube is put in the protection tube, in this state, The feature is that the culture tube is transported to the transplant destination.

In this case, at the transplant destination, the culture tube may be extracted from the protective tube and embedded in the transplant destination medium.

[0018]

BEST MODE FOR CARRYING OUT THE INVENTION An example of a method for culturing matsutake bacterium for transplantation to which the present invention is applied and a method for transplanting matsutake bacterium obtained by the method will be described below with reference to the drawings.

(Cultivation device for matsutake fungus for transplantation) An example of a culture device suitable for use in the method for culturing matsutake fungus for transplantation of this example will be described with reference to FIGS. 1 to 4.

FIG. 1 is a perspective view showing an example of a culture tube which is a component of the culture apparatus of this example, FIG. 2 is an exploded perspective view thereof, and FIG. 3 is a sectional view.

The culture tube 1 of this example is made of plastic,
A cylinder body 2, a lid 3, and a protective cylinder 4 are provided. Tube body 2
Has a cylindrical upper portion 21 and a lower truncated cone-shaped lower portion 22, and a large-diameter collar 23 is formed therebetween. Further, the lower portion 22 includes a circular bottom plate portion 24 and an outer peripheral wall portion 25, and the outer peripheral wall portion 25 is formed with slits 26 extending in the vertical direction at a plurality of positions at equal angular intervals.

The lid 3 is provided with a circular lid body portion 31, a cylindrical portion 32, and a knob plate portion 33 formed on the upper surface of the lid body portion 31. This lid 3 is attached to the upper end opening 2 of the cylinder body 2.
The size of each part is set so that the upper part 21 of the cylinder body is just fitted into the inside of the cylindrical part 32 and the lower end edge of the cylindrical part 32 is in contact with the collar 23 in the state of being covered with 7. .

On the other hand, the protective cylinder 4 is a truncated cone-shaped cylinder with an open upper end, into which the lower part 22 of the cylinder body can be inserted, and the lower part 22 can be inserted. In this state, the lower surface of the collar 23 rests on the upper opening edge of the protective cylinder 4.

Here, the culture tube 1 of the present example is also provided with a lid 3A for inoculation of seeds which can be put on the tube body 2 similarly to the lid 3, and a lid 3B with a vent hole. In addition to the structure of the lid 3 described above, the seed-inoculation implanting lid 3A is provided with a rod 34 for inoculating inoculum at a right angle from the center of the back surface of the circular lid body 31. This rod 34 is a cylinder body 2
The length is set such that the tip of the lid 3A is inserted to a position close to the bottom plate portion 24 of the cylinder body when the lid 3A is covered.

The vented lid 3B has a cylindrical portion 35.
And a partition plate portion 36 partitioning the middle portion of the cylindrical portion 35 into upper and lower parts, and a plurality of ventilation holes 37 formed in the partition plate portion 36. The upper portion of the cylinder body 2 is inserted into the lower portion of the cylindrical portion 35, and the upper portion of the cylindrical portion 35 is filled with a cotton plug as described later.

FIG. 4 is an exploded perspective view and a sectional view showing an example of a culture container which is a component of the culture apparatus of this example. The culture container 5 of this example is also made of plastic and is composed of a container body 6 and a container lid 7 which covers the container body 6 on the upper side. A possible culture tube mounting recess 8 is formed.

The container body 6 is provided with a hexagonal support plate portion 61, and a through cylinder portion 62 extends vertically upward at the center thereof, and the through cylinder portion 62 extends through the support plate portion 61. The ventilation holes 63 are defined. Further, in the support plate portion 61, concavity-shaped recesses 8 for mounting the culture tube are formed at six locations.

The culture tube mounting recess 8 of this embodiment has a cylindrical portion 81 extending vertically upward from the support plate portion 61 and a cylindrical portion with a bottom extending downward from the support plate portion 61 in a tapered state. It is formed from 82. When the tube main body portion 2 of the culture tube 1 is inserted from above, the collar 23 rides on the upper edge of the cylindrical portion 81, and the periphery of the lower portion 22 with the slit in the tube main body portion 2 is completely covered. Is formed.

The container lid 7 of the container body 6 is composed of a hexagonal lid plate portion 71 and a tubular portion 72 having a hexagonal cross section which is continuous with the outer peripheral edge thereof. A through cylinder portion 73 that extends vertically upward is formed at the center of the lid plate portion 71.
The through-cylinder portion 73 defines a ventilation hole 74 that extends through the cover plate portion 71. In the state where the container lid 7 is covered with the container body 6, as shown in FIG. 4B, the outer peripheral edge of the lower end of the container lid 7 is in close contact with the support plate portion 61, whereby the culture tube mounting recess 8 is formed. The space at the upper end opening side of is covered.

Next, the culture tube 1 thus constructed
An example of a method for culturing matsutake fungus for transplantation using a culturing device including the culturing container 5 will be described.

First, as shown in FIG. 5, the tube body 2 of each culture tube 1 is inserted into the culture tube mounting recess 8 of the culture container 5, the agar medium 11 is put therein, and the lid 3A is covered.
In this state, the whole is sterilized or sterilized. At this time, the cotton plug 14 filling the ventilation hole 63 of the culture container 5 is also sterilized or sterilized at the same time. In addition to this, cotton plug 1
The lid 13B filled with 3 and the protective cylinder 4 are also sterilized or sterilized. Similarly, the container lid 7 of the culture container 5,
The ventilation hole 74 is sterilized or sterilized with the cotton plug 15 being filled therein. As a sterilization method, for example, an autoclave can be adopted.

Next, the lid 3A for inoculating the inoculum is pulled out,
At the tip of the rod 34, matsutake spores, matsutake inoculum tissue such as matsutake fruiting body tissue, etc. are attached, and inoculated on the agar medium 11 in each tube body 2.

After this, as shown in FIG. 6, each cylinder main body 2
Then, the lid 3B with a vent hole, which has been sterilized with the cotton plug 13 attached, is covered.

Thereafter, the container body 6 is covered with the container lid 7, and the culture container 5 is left for a predetermined period under a predetermined atmosphere condition and culture condition. Generally, it is left for a predetermined period in an atmosphere of 23 degrees Celsius and 70% humidity. As a result,
The matsutake inoculum 12 planted in the agar medium 11 of the tube body 2 of each culture tube 1 grows to grow the matsutake hyphae, and a state is formed in which the slit 26 of the tube body 2 extends outward.
Here, although sufficient oxygen is required for the growth of Matsutake fungi, in this example, the lid 3B with a vent is used as the lid that closes each agar medium 11, and the culture vessel 5 has a vent 6
3, 74 are formed, which are provided with breathable cotton plugs 13, 14, 15. Therefore, sufficient oxygen can be supplied to the matsutake bacterium during the culture, and invasion of foreign bacteria and the like from the outside can be reliably prevented.

Next, a procedure for transplanting the cultured Matsutake fungus to an artificial medium or a natural medium will be described. First, after the matsutake fungus has grown sufficiently, remove the lid of the culture container 5,
The culture tube 1 is taken out from each culture tube mounting recess 8. Next, the lid 3B of each culture tube 1 is removed, and instead, the lid 3 is covered as shown in FIG. Further, the protective cylinder 4 is attached to the lower portion of the cylinder body 2. As a result, the grown Matsutake fungus is covered with the protective cylinder 4.

The culture tube 1 is carried to, for example, a cultivation place where a large number of cultivation containers 17 in which an artificial medium 16 as shown in FIG. 9 is formed are arranged, and the tube body 2 is taken out,
Embed in artificial medium 16. This completes the porting work.

Alternatively, the transplant destination may be a portion (natural medium) where the fine roots near the surface of the root of the red pine 18 in the red pine forest as shown in FIG. In this case, for example, as shown in FIG. 8, if the culture tube 1 is put in a bag 19 with a seal and is carried to the transplant destination in this state, invasion of various bacteria can be surely prevented. preferable.

[0038]

As described above, according to the present invention,
The matsutake inoculum can be easily planted in a solid medium, and the main body of the culture cylinder containing the cultured matsutake bacterium for transplantation can be directly transferred to the transplant destination and transplanted. Further, there is very little risk of being infected by various bacteria in the culture and transplantation work.

[Brief description of drawings]

FIG. 1 is a perspective view showing an example of a culture tube to which the present invention is applied.

FIG. 2 is an exploded perspective view of the culture tube of FIG. 1, which also shows three types of lids used for the culture tube.

FIG. 3 (a) is a cross-sectional view showing a culture tube covered with a seed-implanting lid, and FIG. 3 (b) is a cross-sectional view showing a state in which a tube body of the culture tube is covered with a vented lid.

FIG. 4 is a perspective view and a sectional view showing a culture container to which the present invention is applied.

FIG. 5 is an explanatory view for explaining a method of culturing Matsutake fungus for transplantation to which the present invention is applied.

FIG. 6 is a cross-sectional view showing a state of a matsutake bacterium for transplantation, to which the present invention is applied, during culturing.

FIG. 7 is an explanatory view showing a state in which a lid and a protective tube are put on a tube body of a culture tube after culturing.

FIG. 8 is an explanatory view showing a state in which a culture tube after culture is housed in a bag.

FIG. 9 is an explanatory view showing a state in which matsutake bacterium for transplantation is transplanted to an artificial medium.

FIG. 10 is an explanatory view showing a state in which matsutake bacterium for transplantation is transplanted to a natural medium.

[Explanation of symbols]

1 culture tube 2 tube body 21 Upper part 22 Lower part 23 Tsuba 24 Bottom plate part 25 Peripheral wall 26 slits 27 Top opening 3, 3A, 3B lid 31 Lid body part 32 Cylindrical part 33 Plate part 34 Rod 37 Vent 4 Protective tube 5 culture vessels 6 container body 61 Support plate part 63 Vent 7 Container lid 74 Vents 8 Recess for mounting culture tube 11 agar 12 Matsutake inoculum 13, 14, 15 cotton plugs 16 artificial medium 17 cultivation container 19 bags

Claims (8)

(57) [Claims] 1. A culture tube for use in a method for culturing matsutake fungus for transplantation, in which matsutake inoculum is purely cultivated using a solid medium, wherein a tube body having a bottom and an upper end opening of the tube body is detachably covered. has a lid, and a slit formed on the outer peripheral wall of the tubular body, as the lid has a lid for seed planting, the seed planting lid insertable into the tubular body
A culture tube for matsutake fungi for transplantation, which is equipped with various rods . 2. A pure culture of Matsutake inoculum using a solid medium.
A culture tube used for a method of cultivating transplanted Matsutake fungus.
Can be attached to and detached from the bottom cylinder body and the top opening of this cylinder body.
A lid and a slit formed on the outer peripheral wall of the cylinder body
And a lid with a vent hole as the lid, which is a culture tube for matsutake fungus for transplantation. 3. The culture tube for matsutake fungus for transplantation according to claim 1, wherein the lid includes a lid with a vent hole. 4. The matsutake fungus for transplantation according to any one of claims 1 to 3, further comprising a protective cylinder with a bottom into which a slit forming portion of the cylinder body can be inserted. Culture tube. 5. A culture device for use in a method for culturing matsutake fungus for transplantation, in which matsutake inoculum is purely cultivated using a solid medium, comprising a culture vessel and at least one culture tube, wherein the culture vessel comprises: A culture tube mounting recess into which the culture tube can be inserted from above, a container body in which the culture tube mounting recess is formed, and an opening side of the culture tube mounting recess in the container body are detachably covered. A container lid, a vent hole formed in the container body, and a vent hole formed in the container lid, the culture tube is a tube body with a bottom, and the tube body is attached to and detached from an upper end opening of the tube body. A matsutake fungus for transplantation characterized by comprising a lid that can be covered, a slit formed on the outer peripheral wall of the cylinder body, and a protective cylinder with a bottom into which the slit forming portion of the cylinder body can be inserted. Culture device. 6. A method for cultivating Matsutake fungus for transplantation using the culture apparatus according to claim 5, wherein the tube body of the culture tube is inserted into the culture tube mounting recess of the container body of the culture vessel. After that, a solid culture medium is put into the cylinder body, and as the lid for covering the cylinder body, a seed-implanting lid having a rod that can be coaxially inserted into the cylinder body is prepared, and the lid is aforesaid. Cover the cylinder body, sterilize in that state, remove the lid for seeding inoculum, attach Matsutake inoculum to the rod, plant the Matsutake inoculum of the rod tip in a solid medium, and cover the tube body As the lid, prepare a lid with ventilation holes that has been sterilized with the ventilation holes closed with a cotton plug, cover the cylinder body with the lid, and then close the ventilation holes with a cotton plug. Sterilized By covering the container body with the treated container lid, and in this state, by leaving the culture container under a predetermined atmospheric temperature and humidity for a predetermined period, a solid medium in each culture cylinder is obtained. A method for cultivating matsutake fungus for transplantation, which comprises growing matsutake hyphae on. 7. The culture tube in which the matsutake hyphae are proliferated by the method according to claim 6, is taken out from the culture tube mounting recess of the culture vessel, and the culture tube is put into the protection tube, Then, a method for transplanting matsutake fungus, which comprises transporting the culture tube to a transplant destination. 8. The method for transplanting matsutake bacterium according to claim 7, wherein the culture tube is extracted from the protective tube at the transplantation destination and embedded in the culture medium at the transplantation destination.