JPH0458826A - Method for forming mycorrhiza by mycorrhizal fungi - Google Patents

Abstract

PURPOSE:To artificially and efficiently form mycorrhiza of matsutake mushroom, Lyophyllum aggregation, etc., in prevented proliferation of various germs by inoculating a living root of a host plant with cultured mycelium of mycorrhizal fungi packed and embedded with sterilized gel so as to directly cover the live root and rooting mycorrhizal fungi on the living root of host plant. CONSTITUTION:A cultured mycelium of mycorrhizic fungi (e.g. matsutake mushroom) is packed and embedded with a sterilized gel (e.g. agar) having 0.2-5.0% concentration and the treated mycelium is directly applied to living root, preferably having 5-10mm diameter of a host plant (e.g. pine). The coated living root is returned to original state and covered with Masa soil. Inoculation period of the mycorrhizal fungi is effective in March to April and June.

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 the host plant in the ground in the host plant's woodland, the roots of the host plant 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.

(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, and the method includes culturing mycorrhizal fungi embedded in a sterilized gel. The gist is a method for mycorrhizal formation using mycorrhizal fungi, which involves inoculating the living roots of a host plant with mycelium so as to directly cover them.

The details will be explained below.

Various mycorrhizal fungi that can be used include Matsutake fungi, Honshimeji fungi, Kōtake fungi, Haratake 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.1 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, examples of gels for embedding cultured mycelium of mycorrhizal fungi include agar, alginic acid, konjac, carrageenan, gellan, pullulan, and guar gum.

Natural polymers such as locust bean gum, xanthan gum, pectin, and starch, and synthetic polymers such as polyacrylamide 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 meet the following criteria.

(1) It is difficult for bacteria in the soil to decompose it for six months to one 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 for water etc., it is generally 0.2 to 5.0%, preferably 1%. It is preferable to set it to about .0 to 3.0%.

In addition, nutrients for the growth of the roots of the host plant and nutrients for the cultured mycelium of mycorrhizal fungi can be mixed in the gel to be embedded. 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 and placed in a desired position, and the operation is performed under sterile conditions to avoid contamination with bacteria. Furthermore,
It is preferable that the gel embedding covers all of the cultured mycelium, but it may be partially embedded as long as it can be used in combination with other bacteria isolation means.

In this way, the cultured mycelium of mycorrhizal fungi embedded in gel is covered with the living roots of the host plant. Just insert the raw root directly. The tip can be positioned in a layer of embedding gel;
Moreover, it can also be positioned so that the cultured mycelium of mycorrhizal fungi comes into contact with it. Here, examples of the host plants include plants of the Pinaceae family such as Japanese red pine for the Matsutake fungus, and Quercus serrata, Sawtooth oak, Japanese red pine, and the like for the Honshimeji fungus. These host plants are preferably about 10 to 30 years old and have well-developed fine roots.

The living root of the host plant to be inoculated should have a diameter of 2~
15■, preferably 5 to Loan, and
During inoculation, it is preferable to sterilize the roots themselves using alcohol, flame, etc.

Furthermore, the roots can be left as they are, but it is better to cut them. This is because more fine roots 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 soil.

The timing of inoculating the host plant with mycorrhizal fungi is preferably around March to April and June, when the roots of the host plant are actively growing, avoiding summer.

(Function) The gel protects the cultured mycelium and the fine roots of the host plant from bacterial contamination.

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

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, seal with aluminum foil and heat at 120℃ for 1
It was heat sterilized for 5 minutes. Further, 5 mA of a medium containing the above ingredients and 0.2 part of agar was added thereto, 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 and placed in a 100 m 12 volume polypropylene tube containing 50 mQ of a 2% agar solution (using distilled water) that had been heat sterilized at 120°C for 15 minutes. When the agar solution reached 40° C. or lower, it was quickly poured into a beaker manufactured by Matsutake Corporation and cooled with ice water to obtain agar-gel-embedded Matsutake cultured mycelium.

In mid-March, a 15-acre hole was dug 5 meters away from the base of a 25-year-old pine tree, and the roots, which were about 8 squares in diameter, were dug out, cut with a knife, and sterilized 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 observed. Admitted.

In mid-March, a 20-an trench was dug at a distance of 10 degrees Celsius from the base of a 30-year-old pine tree, and the roots of about 5 ridges in diameter were dug out. After sterilizing the tip of this root with flame without cutting it, insert it into the agar gel-embedded Matsutake cultured mycelium obtained in Example 1 to about half the depth, return it to its original place, and insert the root into the agar gel-embedded Matsutake cultured mycelium obtained in Example 1. I covered it with soil.

After 2 months, it was confirmed that pine fine roots had taken root in the agar gel-embedded Matsutake cultured mycelium, and after 1 month, although it was not the case in Example 1, mycorrhizal growth was observed. Formation was enshrined.

At the beginning of the white month, a hole of 25 aa was dug at a depth of 3 m from the base of a pine tree of about 20 years old, and roots with a diameter of about ICan were dug out, cut with a knife, and sterilized with flame. Next, two pieces of the agar gel-embedded Matsutake cultured mycelium obtained in Example 1 were placed so that the first one was completely skewered, and the second one was placed at about half the depth. Insert the cut side of a pine root.

After 2 months, fine roots were observed in both agar gel-embedded Matsutake cultured mycelia.

In particular, many fine roots sprouted from the cut surface at the second agar gel-embedded Matsutake cultured mycelium that was inserted halfway. After another month, the development of mycorrhizae was observed in the agar gel-embedded Matsutake cultured mycelium into which the cut surface was inserted.

In Example 1, everything was the same as in Example 1, except that instead of using an agar solution as the embedding gel, 50 mQ of a solution consisting of the following components was used.

Carrageenan 0.7 parts Locust bean gum 0.3 parts Potassium chloride
0.24 parts distilled water 100
The results of mycorrhizal formation were as good as in Example 1. 11" Possible In Example 1, a solution consisting of the following components was used at 50 mA instead of using an agar solution as the embedding gel. Everything was the same as in Example 1.

rhomethoxy pectin 2 parts thiabendazole 0.1 part streptomycin
0.2 parts distilled water 100
Potassium chloride (Note) (Note) Potassium chloride is prepared by adjusting a solution consisting of other substances to pH 5,
Heat sterilize at 120°C for 15 minutes, and when the temperature drops below 40°C, mix equal amounts of 0.5% solution and use.

The mycorrhiza formation results were as good as in Example 1.

In Example 1, Honshimeji fungus was used in place of Matsutake fungus, and the pH of the culture medium was also changed. Everything was the same as in Example 1 except that the number 11 was changed from 5 to 6.

Mycorrhiza formation results were still good.

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

(Effects of the Invention) As described above, according to the present invention, mycorrhizal fungi can be artificially attached to the living roots of host plants to efficiently form mycorrhizae.

Claims (1)

[Claims] This is a method of artificially attaching mycorrhizal fungi to the living roots of a host plant to form mycorrhizae, and involves inoculating the living roots of the host plant so that they are directly covered with cultured mycelium of mycorrhizal fungi embedded in sterilized gel. Mycorrhizal formation method of mycorrhizal fungi.