JPH04190714A - Production of mycorrhiza of mycorrhiza fungus - Google Patents

Abstract

PURPOSE:To efficiently produce the subject mycorrhiza by artificially rooting cultured myceliums of a mycorrhiza fungus buried in a sterilized humectant- containing gel in the living root of a host plant and protecting various germs pollution for a long period. CONSTITUTION:Cultured myceliums of a mycorrhiza fungus such as T. matsutake or L. aggregatum, are buried in a sterilized gel such as agar-agar or devil's tongue containing 0.1-10wt.% (based on whole gel) humectant, e.g. glycerin preferably in a state where the myceliums are thoroughly wrapped up in the gel. The myceliums are rooted in a state where the myceliums cover the living root of a host plant, e.g. preferably pine or Quercus serrata Thunb, thus producing the objective mycorrhiza.

Description

Detailed Description of the Invention (Industrial Application Field) The present invention is directed to artificially injecting mycorrhizal fungi of living host plants such as Matsutake and Honshimeji into the living roots of host plants (host plants) such as pine, Quercus serrata, and oak oak. The present invention relates to a method for forming mycorrhizae by colonizing the mycorrhizae.

(Prior art) The main thing to keep in mind when artificially forming mycorrhizae of Matsutake, Honshimeji, etc. is to prevent the propagation of various bacteria. When burying host plants underground in forest areas, the roots of the host plants are sterilized or replaced with sterilized soil.

(Problem to be solved by the invention) Even if the roots and soil of the host plant are sterilized, there are still countless bacteria in the soil, and the soil contains a considerable amount of organic matter that serves as nutrients. It is not possible to sufficiently prevent the proliferation of mycorrhizas, and the formation of mycorrhizae becomes inefficient.

In this regard, the present inventors have investigated the use of cultured mycelia of mycorrhizal fungi embedded in sterilized gel, and have succeeded in increasing the efficiency of mycorrhizal formation.

However, if a gel-embedded object is buried underground and left for a long period of time, the water in the gel will be absorbed by the soil and the gel will become thinner.
It was found that there is a high possibility of bacterial contamination.

Therefore, the present invention aims to provide a means by which the advantages of gel embedding can be maintained for a long period of time.

(Means for Solving the Problems) The present invention is a method for forming mycorrhizae by artificially attaching mycorrhizal fungi to the living roots of a host plant, in which the mycorrhizae are embedded in a sterilized humectant-containing gel. The gist is a method for forming mycorrhizae of mycorrhizal fungi by inoculating the cultured mycelium of the fungus onto the living roots of a host plant.

The details will be explained below.

Various mycorrhizal fungi that can be used include Matsutake fungi, Honshimeji fungi, Kōtake fungi, Hatutake fungi, Amitake fungi, and these mycorrhizal fungi that have been cultured under appropriate conditions are used. Here, culture can be carried out by either liquid culture or solid culture, but for example, cultured mycelium of mycorrhizal fungi is dispersed using a homogenizer, etc., and solid culture is carried out using a culture carrier such as vermiculite. and mycelium can propagate well.

In addition, the gel that embeds the cultured mycelium of mycorrhizal fungi may contain natural polymers such as agar, alginic acid, konjac, carrageenan, gellan, pullulan, guar gum, locust bean gum, xanthan gum, pectin, starch, etc. Synthetic polymers such as acrylamide and polyvinyl alcohol can be used. They may be used alone or in combination. These polymers can be dispersed in water, buffer solutions, salt solutions, etc., and then gelled by heat, light, etc. These gels are used for the purpose of culturing mycorrhizal fungi to survive, attach themselves to the fine roots of host plants, and form mycorrhizae, and are not necessarily limited to the following conditions:
It is preferable to use one that satisfies the following conditions.

(1) It is difficult for bacteria in the soil to decompose it for six months to a year.

(2) It must have sufficient strength for ease of handling during inoculation and stability against soil formation in the soil.

(3) When embedding the cultured mycelium of mycorrhizal fungi, there should be no adverse effects such as excessive heat.

(4) It must be soft enough to allow the fine roots of the host plant to root and grow.

(5) The polymer to be gelled must be stable against sterilization by heating, radiation, etc.

In addition, the concentration of the gel when embedding the cultured mycelium of mycorrhizal fungi is appropriate depending on the type of liquid, but it is generally 0.2 to 5.0% by weight, preferably 0.2 to 5.0% by weight relative to water. is 1.0-3.
It is preferable to set it to about 0% by weight.

In addition, as a moisturizing agent to be included in the gel to be embedded, glycerin, ethylene glycol, polyethylene glycol (for example, #200. #300, #400. #6)
00. #1000. #1540, #2000, 8400
0,86000° #10000. #20000), polyacrylamide, super absorbent resins (e.g., grafted starch type (starch/acrylate graft copolymer crosslinked product), polyacrylic acid type (crosslinked polyacrylate)
, cellulose type, synthetic polymer type, poval type, polyoxyethylene type), etc. These humectants can be used alone or in combination. The concentration used should be such that gel solidification is possible and does not inhibit the growth of cultured mycelium.
Although it cannot be determined specifically due to the type of gel, etc.
Generally, it is preferable that the amount is 0.1 to 10% by weight of the entire gel. In addition, nutrients for the growth of the roots of the host plant and nutrients for the cultured mycelium of mycorrhizal fungi may be mixed in the embedding gel. In addition, to embed the cultured mycelium of mycorrhizal fungi in gel, for example, a test tube or beaker, etc. that is thicker than the cultured mycelium can be used to form a gel layer of an appropriate thickness. .

At this time, the cultured mycelium may be suspended with a wire or the like and placed at a desired position. In addition, operations are performed under sterile conditions to avoid contamination with bacteria. Furthermore, it is preferable to embed the gel so that all of the cultured mycelium is encapsulated, and if necessary, multiple embeddings of different types of mycelium can be performed, but it is recommended that it be used in combination with other germ isolation methods. It may be partial if possible. When multiple embedding is performed, a moisturizing agent can be contained only in the gel of an appropriate layer such as the outermost layer.

In this way, the cultured mycelium of mycorrhizal fungi embedded in the gel is used to inoculate the living roots of the host plant.

Although it may be buried in the ground near the tip of the living root, it is preferable to cover the living root. Just insert the raw root directly. The tip can be placed on the edge of the embedding gel, or it can be placed so that it touches the cultured mycelium of mycorrhizal fungi. Here, as a host plant,
For example, for the Matsutake fungus, plants of the Pinaceae family such as Japanese red pine are used, and for the Honshimeji fungus, Quercus Quercus, Sawtooth oak, Japanese red pine, and the like can be used. These host plants are preferably about 10 to 30 years old and have well-developed fine roots, and the preferred living roots for inoculation are about 2 to 15 mo in diameter, more preferably 5 to 15 mo in diameter.
It is about 10W1, and it is preferable to sterilize the fresh roots themselves using alcohol, flame, etc. during inoculation. Furthermore, the roots can be left as they are, but it is better to cut them. More fine roots will grow than the cut surface.

After covering with cultured mycelium of mycorrhizal fungi embedded in gel, the roots of the host plant are returned to their original location and covered with clean soil such as masa top.

The best time to inoculate the host plant with mycorrhizal fungi is to avoid summer, but when the host plant's roots are actively growing, for example, in the case of pine trees,
It is preferable to set the period between ~April and June.

(Function) The gel protects from bacterial contamination, and the humectant contained in this gel suppresses water movement from the gel to the soil and increases the stability of the gel.

(Example) Hereinafter, parts simply refer to parts by weight.

N loss yeast extract 0.15 parts Soiton (manufactured by Difco) 0.15 parts glucose
2.0 parts distilled water 100
Matsutake fungus was cultured at 23° C. for 4 weeks in a medium consisting of the above components adjusted to pH 5, dispersed using a homogenizer, and 5 mQ of the culture was placed in a polypropylene beaker with a capacity of 50 mQ. This beaker is made of horticultural vermiculite 8
g, sealed with aluminum foil and heated to 120'C.
It was heat sterilized for 15 minutes. Further, 5 mQ of a medium containing 0.2 parts of agar was added to the above ingredients, and cultured at 23°C for 4 weeks.

After confirming that the mycelium is well-propagated, the obtained Matsutake cultured mycelium was taken out from the beaker, heated and sterilized at 120℃ for 15 minutes, and mixed with a 2% agar solution (using distilled water) and 1% Biogel P (Biogel P). Once the agar solution has cooled below 40°C, quickly pour it into a polypropylene beaker with a 100mA capacity containing 50m12 of polyacrylamide manufactured by Rudd (containing a humectant), cool it with ice water, and make an agar gel approximately 10 cm thick. Matsutake cultured mycelium embedded with

In mid-March, we dug 150 holes 5 meters away from the base of a 25-year-old pine tree and dug out roots about 8 squares in diameter.
Cut with a knife and sterilize with flame. This was inserted into the agar gel-embedded Matsutake cultured mycelium obtained above to about half the depth, returned to its original place, and then covered with masa soil from above.

After 2 months, it was confirmed that many fine roots had sprouted from the cross section of the cut pine roots in the agar gel-embedded Matsutake cultured mycelium, and furthermore, after 1 month, the formation of mycorrhizae was confirmed. Admitted. Also, at this time, the thickness of the agar gel is still approximately 611I1.
1 remained, confirming that sufficient protection from bacterial contamination was achieved.

In Example 1, instead of Biogel P (moisturizer) used in the agar solution, Sunwet IM-2300 (Sanyo Chemical Co., Ltd. chewed starch/acrylate graft copolymer crosslinked product), Sumikagel 5P-520 (vinyl alcohol/acrylic acid copolymer manufactured by Sumikagaku Kogyo ■), Arasorb S
-100 (polyacrylate-based super water-absorbent resin manufactured by Arayo Kagaku Kogyo ■) was used in each case, but everything was the same as in Example 1.

The results regarding mycorrhiza formation and protection from bacterial contamination by agar gel were both as good as in Example 1 (3
Thickness of remaining agar gel after drying: approx. 6-7 m+).

In Example 1, everything was the same as in Example 1 except that the agar gel to be embedded was doubled. Here, the inner layer does not contain biogel P, and the outer layer contains Arasorb S-
100 (mentioned above), each containing approximately 1
0m++, thickness of approximately 15cm (approx. 25cm for the whole agar gel)
).

The results regarding mycorrhiza formation and protection from bacterial contamination by agar gel were both as good as in Example 1 (3
Total thickness of the remaining agar gel after the moon: approx. 15cm).

In Example 5, Arasorb S-1 was added to the agar gel in the outer layer.
Everything was the same as in Example 5 except that 5% glycerin and 2% polyethylene glycol (#6000) were contained instead of 00.

The results regarding mycorrhiza formation and protection from bacterial contamination by agar gel were both as good as in Example 5 (3
Overall thickness of remaining agar gel after heating: about Low in Example 6, about 1211II+ in Example 7).

1. In Example 1, instead of using an agar solution as the embedding gel, a solution consisting of the following components was used.
Everything was the same as in Example 1 except that mQ was used.

Carrageenan 0.7 parts Locust bean gum 0.3 parts Potassium chloride
0.24 parts Biogel P 1.0
Partly distilled water 100 parts The results of mycorrhizal formation and protection from bacterial contamination by the agar gel were both as good as in Example 1 (thickness of gel after 3 months: about 5 rm).

The same procedure as in Example 1 was carried out except that the Matsutake fungus was replaced by the Honshimeji fungus and the pH adjustment of the culture medium was changed from 5 to 6.

The results regarding mycorrhiza formation and protection from bacterial contamination by agar gel were both as good as in Example 1 (3
Thickness of agar gel after drying: Approximately 4 kan).

When the mycelium of the Matsutake fungus cultured in Example 1 was inoculated near the roots of a pine tree similar to those in Examples 1 and 2, without embedding it in gel, 2. rear,
The matsutake fungus was almost extinct.

(Effects of the Invention) As described above, according to the present invention, it is possible to artificially attach mycorrhizal fungi to the living roots of host plants to efficiently form mycorrhizae, and it is also possible to protect them from bacterial contamination. Can be secured over a long period of time.

To patent appearance Pendel Co., Ltd.

Claims (1)

[Claims] This is a method of artificially attaching mycorrhizal fungi to the living roots of a host plant to form mycorrhizas, and the method involves inoculating the cultured mycelium of mycorrhizal fungi embedded in a sterilized gel containing a humectant into the living roots of the host plant. Mycorrhizal formation method of mycorrhizal fungi.