KR20100136895A - Methods for producing hyphae of mycorrhizal mushrooms and solid mediums for the methods, and methods for producing mycorrhiza-infected seedlings of woody plants using the hyphae formed on the solid medium as inoculum sources - Google Patents

Abstract

PURPOSE: A producing method of a mycorrhizal mushroom, a solid medium for the method, and a producing method of a mycorrhiza infected young tree using thereof are provided to use a hypha of the mycorrhizal mushroom for infecting a young tree. CONSTITUTION: A producing method of a mycorrhizal mushroom comprises a step of injecting a hypha of the mycorrhizal mushroom to a solid medium including a pumice with the diameter of 0.1~10centimeters, and an elvan stone with the diameter of 0.1~10centimeters. The solid medium additionally contains a surfactant, and an element selected from rice, wheat, barley, starch, and corn.

Description

Method for producing mycorrhizal mycelium and solid medium for use in the method, and method for producing mycorrhizal seedlings of trees using mycelium formed on the solid medium as inoculum. , and methods for producing mycorrhiza-infected seedlings of woody plants using the hyphae formed on the solid medium as inoculum sources}

The present invention relates to the production of mycorrhizal mushroom mycelium and the production of mycorrhizal seedlings using the produced mycelium, and more particularly, to a method for producing mycorrhizal mushroom mycelium and a solid medium for use in the method, and the solid medium. The present invention relates to a method for producing mycorrhizal infectious seedlings of trees in containers, facilities, seedlings packaging, etc., which are not sterile using the mycelium formed on the inoculator.

Fungal mushrooms represented by pine mushrooms, oyster mushrooms, tuber mushrooms, al mushrooms, etc., unlike by-product mushrooms such as enoki mushrooms, oyster mushrooms, shiitake mushrooms, which can be stably produced by artificial cultures, are difficult to artificially cultivate. For the fungal mushrooms, artificial cultivation experiments using spores, cultured mycelia and mycorrhizal fungi have been conducted. However, no effective means of production has been established for mushrooms other than Lyophyllum shimeji (Kawam.) Hongo. to be. In general, the artificial cultivation of mushrooms is usually aimed to produce fruiting bodies by inoculating and cultivating the spawn produced from purely cultured mycelia. However, unlike fungal mushrooms, most mycorrhizal mushrooms, which are representative of pine mushrooms, have very low physiological characteristics such as cellulose and starch, which makes them difficult to apply artificial culture technology for byproduct mushrooms. In addition, mycorrhizal mushrooms such as Matsutake mushrooms have a slow growth rate due to their physiological characteristics, which makes it difficult to produce only a mycelium in large quantities. This is because non-sterile outdoor soils are home to many microorganisms that inhibit mycelial growth and soil animals that feed on mushroom hyphae. Therefore, the ability to produce inoculated mycelium that is not only resistant to soil microorganisms but also harmed by soil microbes is an important factor for the success of artificial cultivation.

In Korea, the only mushroom fungus mushroom has been studied relatively much, and a culture method using a liquid medium of the pine mushroom mycelium has been developed. Korean Laid-Open Patent Publication No. 2007-0099362 (published Oct. 9, 2007) includes mixing at least one of masato, pearlite, vermiculite, sawdust, and leafy soil, and then barley or barley flour, corn flour and rice bran alone or mixed. Further disclosed is a method for mass production of cluster mycelium by culturing the cluster mycelium on a solid medium further comprising. However, it is doubtful whether their cultured mycelium will have viability in unstained outdoor soil. In addition, although perlite and vermiculite are porous, they are made artificially at high temperatures, so there are no nutrients such as trace elements necessary for the growth of bacteria. In particular, the pearlite does not fully penetrate the water even if soaked in enough water, and since the air always remains inside, there is a feature that floats up in the water. Therefore, since the function of adsorption surface of the medium, which is a nutrient source necessary for cultivation of bacteria, is reduced, the efficiency of long-term culture of mycelia is inevitably deteriorated. In addition, vermiculite may be brittle, and even if the mycelium grows, the form of the medium may be brittle, and the mycelium may be broken, thereby adversely affecting the growth of the bacteria. Therefore, there has been a great demand for the development of a method for supplementing the disadvantages of existing culture media and methods and producing mycelia of mycorrhizal mushroom more efficiently.

Pumice, on the other hand, is a kind of volcanic debris, which refers to a pale color among porous ones. When the rhyolite-andesite magma erupts, it rises from the deep underground and decreases the pressure, so that volatiles such as water dissolved in the magma foam and become porous. The color of pumice is generally pale color of white, gray, yellow, etc., and contains some colorless-black mineral crystal. As a general use, finely crushed and processed into small sections are used to scrub the dead skin of the heel. Moreover, since it is porous, water retention is good and it is used as a gardening soil. However, there have been no examples of using pumice as a medium for culturing mycelium of mycorrhizal mushrooms.

As a method of cultivating mycorrhizal mushrooms, it is known that fruiting bodies sometimes occur during the growth of mycorrhizal seedlings based on mycorrhizal synthesis experiments using the ecological function of mycelium symbiosis with host plants. Therefore, the artificial cultivation of mycorrhizal mushrooms aims to produce fruiting bodies by seedlings of mycorrhizal seedlings and outdoor transplants. Tuber spp., Which is a common mycorrhizal fungus, along with shoulders, has been used in many countries in Europe for many years. In addition, the research on the culture of mycelia for inoculation to create mycorrhizal seedlings has been performed for a long time, but most of them have been performed in aseptic condition, and the study on the outdoor experiment in the natural state is insufficient. Korean Patent No. 390,048 (registered on June 21, 2003) relates to a method of co-cultivating mycelia of pine seedlings and pine mushroom strains to form pine seedlings infected with pine mushroom alone. Inoculating the mycelial pulverized product obtained by pulverization at the bottom of the sterilized culture vessel; Pouring mixed clay prepared by mixing pearlite and spegnum pitmos on the pulverized mycelium mycelium pulverized product; Dispensing a K-medium onto the mixed clay; Densifying pine sterile germinating seedlings on the infected medium including the mixed clay and K-medium and closing the lid of the culture vessel; Disclosed is a method comprising co-culturing pine seedlings and pine mushroom culture prepared by the above step. However, there have been no reports on the mycelial synthesis of seedlings that can be planted in natural, non-sterile, and non-germinated seedlings.

The present inventors carried out continuous research in order to develop a technology that can produce a large amount of mycelia more quickly for the artificial cultivation of mycorrhizal mushrooms. As a result, it was confirmed that the above object can be successfully achieved by culturing mycelium of mycorrhizal mushrooms in a solid medium containing pumice which is a porous natural stone. Furthermore, the present inventors have succeeded in creating and mass producing mycorrhizal infectious seedlings of trees using natural mycelium formed on the solid medium as the inoculum by the above method, thus completing the present invention.

Accordingly, a first object of the present invention is to provide a method for producing mycelium of mycorrhizal mushrooms by culturing mycelium of mycorrhizal mushrooms.

A second object of the present invention is to provide a solid medium for use in the method for producing mycelium of mycorrhizal mushrooms.

A third object of the present invention is to provide a method for producing mycorrhizal infectious seedlings of trees using natural mycelium of mycorrhizal mushrooms formed on the solid medium as an inoculum.

The first aspect of the present invention relates to a method for producing mycelia of mycorrhizal mushroom, comprising inoculating and incubating the mycelia of mycorrhiza on a solid medium comprising pumice having a diameter of 0.1 to 10 cm. The solid medium may further comprise elvan stones with a diameter of 0.1 to 10 cm. In addition, the polymer carbon source may further include rice, wheat, barley, starch, corn alone or mixed. In addition, it may further comprise an cooking oil and a surfactant. The solid medium according to the invention may be to comprise pumice in a breathable pouch.

The second aspect of the present invention relates to a solid medium for use in the method for producing mycelium of mycorrhizal mushrooms, which comprises pumice having a diameter of 0.1 to 10 cm.

The third aspect of the present invention

1) inoculating the mycelia of mycorrhizal mushrooms to the roots of the seedlings using the mycelia of mycorrhizal mushrooms formed on the solid medium according to the above method;

2) Planting seedlings inoculated with mycelium:

It relates to a method for producing mycorrhizal seedlings of trees, comprising the step. The method may be carried out in a non-aseptic condition, for example in a container, facility, seedling packaging and the like. Preferably, the method may further include irrigating the planted seedlings with pesticides, fungicides and plant growth promoters at intervals of 1-4 weeks at 1000-200 times the respective amounts.

According to the present invention, it is possible to rapidly produce a large amount of mycelium for use as an inoculum for mycorrhizal synthesis and fungal (shiro, shiro) formation in the roots of trees that are symbiotic mushrooms. In addition, even if the mycelia of mushroom bacteria penetrate into the porous gaps, they are not damaged by soil microorganisms and soil animals even when used as an inoculum in the field, and natural minerals contain a large number of minerals. Long term survival In addition, it is expected to be able to produce mycorrhizal infectious seedlings in a natural state, that is, not in a sterile state, so that they can contribute greatly to the artificial cultivation of mycorrhizal mushrooms represented by pine mushrooms, eryngii mushrooms, etc. which were considered impossible until now.

In addition, mycorrhizal seedlings produced according to the present invention can not only be used for greening of forests and forest roads and road slopes disturbed by forest fires or floods, but also used for greening of deserts where the survival of plants is almost impossible. It is expected to contribute greatly to greening. In addition, it can be used as a physiological and ecological research data of trees and mycorrhizal fungi as it can produce mycorrhizal infection freely.

Hereinafter, the present invention will be described in detail.

One. Production method and medium of mycorrhizal mycelium

The inoculum production method of mycorrhizal mushroom mycelium according to the present invention is characterized in that the mycelia of mycorrhizal mushroom are cultured in a solid medium containing pumice which is a porous natural stone.

In the present invention, the "mycorrhizal mushroom" encompasses the mycorrhizal mushroom forming a symbiotic relationship with the host tree, and there are no particular limitations on the kind as long as the medium and method of the present invention can be cultured and produced. , For example, Tricholoma matsutake (S. Ito et Imai) Sing., T. bakamatsutake Hongo, Sarcodon aspratus (Berk.) S. Ito, Tuberculosis [ Tuber] spp.], Rhizopogen rubescens (Tul.) Tul., Sullus grevillei (Klotzsch) Sing., S. bovines (L .: Fr.) O. Kuntze, Mushrooms Boletus edulis Bull. Ex Fr.], Lyophyllum shimeji (Kawam.) Hongo, Hydrangea mushroom [ Hygrophorus spp.], Sage mushroom [ Lyophyllum spp.], Matsutake mushroom [ Tricholoma spp.], Cucumber mushroom [ Leucopaxillus spp. ] vast mushroom [Amanita spp.], the net mushroom [Boletus spp.], silk net mushrooms [Sullus spp.], rattling reluctant net mushrooms [Leccinum spp.], the shaman mushrooms [Russula spp.], milk mushrooms [Lactarius spp ., Cantharellus spp., Ramaria spp., Morel mushroom Morchella esculenta (L .: Fr.) Pers. ver. Esculenta ], Pisolithus tinctorius Cokr & Couch, Scleroderma spp. And Morchella spp., And may be matsutake or thigh mushroom.

The solid medium for mycelial mushroom mycelium culture of the present invention is characterized in that it comprises a pumice of 0.1 to 10 cm in diameter. Pumice can be used as it is, such as natural stone (for example, ovary), or absorbed nutrients such as inorganic salts in natural stone (for example, gold arsenite), or may be mixed and used. In case of using pumice less than 0.1 cm in diameter, there may be a problem that it is easy to be exposed to feeding damage by soil animals such as earthworms. Sterilization may be somewhat inconvenient.

In addition to the pumice, the medium according to the present invention may further include elvan rock, thereby achieving an even better effect. Elvanite belongs to granite calcite, which is a mixture of quartz and feldspar, and its main components are silicic anhydride and aluminum oxide. It is used as a filter, anti-inflammatory, and so on, and it is called weak stone. Elvan also contains a large amount of minerals, which not only promote the growth of mycorrhizal mushrooms, but also have the ability to purify microorganisms. Can be represented. In particular, mycorrhizal mushrooms, such as pine mushrooms, dissolve rocks by secreting organic acids from mycelia, so that they effectively and continuously utilize large amounts of minerals contained in elvan to promote mycelial growth and increase the lifespan of the inoculum. You can increase it considerably.

In a specific example of the present invention, an excellent effect can be obtained by mixing and using an arsenic having a diameter of 0.1 to 10 cm, a gold ore having a diameter of 0.1 to 10 cm, and a hepatite having a diameter of 0.1 to 10 cm. At this time, it is preferable to mix the ovum, gold ovary and elvan in a volume ratio of 2-5: 2-5: 1, especially 3: 3: 1. Further, in the present invention, the solid medium, i.e., pumice, or pumice and elvan, is sterilized (e.g., 121) in a liquid medium containing a carbon source and a nitrogen source (e.g., a liquid medium containing dry yeast and glucose in distilled water) before use. 60 minutes at < RTI ID = 0.0 >

The solid medium of the present invention may further comprise rice, wheat, barley, starch, corn alone or mixed as a polymer carbon source. Even when barley, corn, etc. are added, a good effect can be obtained, but when rice is added, an excellent effect can be obtained. The polymer carbon source may be used in 10 to 25 parts by volume with respect to 100 parts by volume of pumice. Specifically, ovarian stone, gold ovary, elvan and rice may be used in a volume ratio of 3: 3: 1: 1 .

The solid medium of the present invention may further comprise cooking oil and / or surfactant. Edible oil may be used vegetable oils such as olive oil, sesame oil, perilla oil, soybean oil, peanut oil and animal oils such as diesel oil and fish oil, and in particular, vegetable oils such as olive oil, sesame oil and perilla oil may be used. Surfactants include polyethyleneglycolated sorbitan fatty acid esters such as polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan monooleate In particular, you can use Tween 20 or Tween 80. Cooking oil and surfactant may be used in 1 to 5 parts by volume based on 100 parts by weight of pumice, respectively. By adding the cooking oil and the surfactant, the mycelium can be greatly increased.

In the present invention, mycelium can be cultured more efficiently and conveniently by including pumice in a breathable bag. There is no particular limitation on the breathable pouches that can be used, but for example pouches made of burlap, ramie, nonwovens, in particular burlap pouches, can be used. When using a breathable bag as described above, the inoculum can be produced in large quantities quantitatively, there is an advantage that the inoculum can be quantified during the mycorrhizal inoculation experiment. In addition, since the inoculator can be directly attached to the pine roots of pine seedlings or forests, it is possible to inoculate the desired root portion, for example, it can be inoculated by fixing directly to the desired roots with a stapler or the like. In addition, mycelia are not destroyed during outdoor inoculation, and pumice medium is present as one large mycelial mass, and there is no breakage of mycelia and inoculation during inoculation, so the initial growth of mycelia after inoculation is fast and the feeding of soil animals is avoided. . In addition, the mycelia are grown in a bundle of mycelia, the antibacterial components are concentrated to increase the resistance to soil microorganisms during inoculation.

Mycelium used as an inoculum of the solid medium in the present invention is purely isolated from wild matsutake mushrooms. To cultivate the pure isolated strain, 5 g of dry yeast or 2 g of yeast extract, 20 g of glucose, and 20 g of agar were prepared per 1 liter of distilled water, and a solid medium was prepared. Incubate at 20-25 ° C., especially 24-25 ° C. for 4-5 weeks. The cultured mycelium is put into a beaker containing sterile water by chopped solid medium pieces using a scalpel, and then pulverized with a homogenizer and used as an inoculum of the medium according to the present invention. In addition, the mycelium firstly cultured in a solid medium prepared by mixing 5 g of dry yeast or 2 g of yeast extract, 20 g of glucose, and 20 g of agar per liter of distilled water was pre-cultured in a liquid medium obtained by removing agar from the medium for primary culture. It is also possible.

The incubation rate varies according to the inoculation amount of the ground mycelia, and the inoculation amount is effective for rapid culture.

The culture temperature in the medium according to the present invention is preferably 20-25 ° C., in particular 24-25 ° C., and the incubation period depends on the medium and the inoculated mycelia, but is approximately 1 to 3 months, for example, 2 months. Mycelia are rampant throughout the medium.

2. Production of Mycorrhizal Seedlings Using Mycelium Formed on Solid Media

Furthermore, in the present invention, the mycelia of mycorrhizal mushrooms formed on the solid medium are inoculated according to the method as described above, inoculating the mycelia of mycorrhizal mushrooms on the roots of the seedlings, and planting seedlings inoculated with the mycelia. Produce mycorrhizal seedlings. In the present invention, the tree is a host plant that forms a symbiotic relationship with the mycorrhizal fungi, and pine pine is typically the case for the pine mushroom. In this invention, the seedling of 10 years old or less, for example, 5 years old or less, for example, 6 months or more and 3 years old or less, for example, 1 year old can be used. In addition, it is preferable to irrigate pesticides, fungicides and plant growth promoters at intervals of 1-4 weeks at 1000-200 times to planted seedlings, respectively.

According to the present invention, it is possible to produce mycorrhizal infectious seedlings in a container, a facility or a seedling packaging, which is not aseptic. About 3 months after planting by inoculating mycelial inoculum, mycorrhizal formation is confirmed in the root of seedlings. In order to identify mycorrhiza infected with seedlings, DNA was extracted from mycorrhizal fungus and then amplified by polymerase chain reaction (PCR) using specific primers of mycorrhizal fungi. It can be confirmed that the DNA is consistent with that of the inoculated fungal mushroom.

Hereinafter, the present invention will be described in detail with reference to examples, but these are merely to aid the understanding of the present invention and are not intended to limit the scope of the present invention in any way.

Example 1 Cultivation of Matsutake Mycelium for Inoculation in Solid Medium

Mycelia were purely isolated from wild pine mushrooms. A solid medium was prepared by combining 5 g of dry yeast or 2 g of yeast extract, 20 g of glucose, and 20 g of agar per liter of distilled water. Pure matsutake bacteria were inoculated into a curry containing a solid medium. The mycelium cultured at 24-25 ° C. for 4-5 weeks was placed in a beaker containing sterile water by using a scalpel, and then pulverized with a homogenizer and used as an inoculum of a porous natural stone medium.

Example 2 Cultivation of Mycelia in a Porous Natural Stone Medium of Matsutake Mushroom

Pumice 3 cm in diameter was placed in a vessel containing a liquid medium containing 5 g of dry yeast and 20 g of glucose per liter of distilled water and sterilized at 121 ° C. for 60 minutes. Thereafter, the liquid medium in the container was decanted, and the pumice was transferred to a sterile shale, followed by inoculation by inoculation of the mycelium prepared in Example 1. Culture conditions were the same as in Example 1.

Figure 1a is a photograph showing the mushroom mycelium grown in pumice. FIG. 1B is a photograph of the cluster mycelium (lower left) and its enlarged (lower right) infiltrating the inner gap of the pumice (top) in which the cluster mycelium is widespread and the transverse section of the pumice. As shown in these figures, the pine mushroom mycelium was widespread not only on the surface of the pumice but also in the inner gap, and it was confirmed that the growth of the pine mushroom mycelium was very excellent.

Example 3 Cultivation of Matsutake Mycelium in Pouches Containing Pumice

After mixing the ovite, gold ovary, elvan and rice in a volume ratio of 3: 3: 1: 1, and filling them in a bag made of natural fiber burlap (5 cm x 5 cm), sealing the liquid medium of Example 2, Sterilized by the same method in a container. After sterilization, the fungus mycelium was inoculated using the inoculating mycelium prepared in Example 1 in each bag and incubated at 24-25 ° C. for 2 months. The results are shown in FIG. As shown in Figure 2, it was confirmed that the pine mushroom mycelium is rampant not only on the surface of the bag but also inside the medium.

Example 4 Cultivation of Matsutake Mycelium in Pumice Medium Containing Edible Oil or Surfactant

In Example 2, pine mushroom mycelium was cultured by adding 0.01 to 0.05 volume of olive oil and Tween 20 to pumice, respectively. The results are shown in FIG. 3 (left: when adding Tween 20, right: when adding olive oil). As shown in FIG. 3, when cooking oil or a surfactant was added, it was confirmed that the mycelium and the mycelium grew more abundantly and the culture mycelium was greatly increased.

Example 5 Creation of Pine Pine Infection Seedlings in Natural State Using Pumice Marrow Mycelia Inoculator

The roots were cut in half using annual pine. The inoculum of the pine mushroom mycelium formed on the solid medium obtained according to Example 3 was placed in a container, and then pine roots were placed thereon and covered with soil to plant pine. Insecticides (Smithion, Hapa Chemical, Israel), fungicides (Benomil, SMBT) and plant growth promoters (Multifeed, Dongbang Agro Co., Ltd.) at 1000-200 times intervals after 1-2 weeks of planting. Was sufficiently dispensed. Fig. 4 shows the seedling state after 3 months after planting with Matsutake mycelium inoculum, and Fig. 5 shows the mycorrhizal fungus formed on the pine root, and FIGS. 6 and 7 are micrographs of the mycorrhizal fungus.

Example 6 Detection of Matsutake Bacteria from Mycorrhizal Formed in Matsutake Infected Seedlings

In Example 4, the roots of the pine seedlings 3 months after planting were identified, the roots of the roots were collected, DNA was extracted, and the polymerase chain reaction (PCR) was performed using a pine mushroom specific primer. Was detected. The results are shown in FIG.

Figure 1a is a photograph showing the mushroom mycelium infested in pumice, a porous natural stone medium;

FIG. 1B is a photograph of cluster mycelium (bottom left) and enlarged (bottom right) photographs of the pumice mycelium infested with pumice (top) and the inner gap of the transverse section of the pumice;

Figure 2 is a photograph showing the mushroom mycelium infested in the burlap pouch containing the pumice which is a porous natural stone medium;

Figure 3 is a photograph showing the mycelial bundles and mycelia of the Matsutake mushroom grown in pumice media containing cooking oil and surfactant (left: surfactant, right: cooking oil);

Figure 4 is a photograph showing the pine seedlings 3 months after planting with the pine mushroom mycelium inoculum infested in pumice which is a porous natural stone medium;

5 is a photograph showing the pine roots formed in pine roots;

6 is a micrograph showing the morphology of pine mushroom;

7 is a micrograph by cross-section of M. aeruginosa;

Figure 8 is an electrophoresis picture showing the results of detecting the pine mushroom from the mycorrhizal formed by PCR using the pine mushroom specific primers.

Claims (10)

Inoculating a mycelium of mycorrhizal mushrooms in a solid medium containing pumice having a diameter of 0.1 to 10 cm, culturing the mycelium of mycorrhizal mushrooms. The method of claim 1, wherein the solid medium further comprises gantherite of 0.1 to 10 cm in diameter. The method of claim 1, wherein the solid medium further comprises rice, wheat, barley, starch, corn alone or in combination. The method of claim 1, wherein the solid medium further comprises cooking oil or a surfactant. The method of claim 1, wherein the solid medium comprises pumice in a breathable pouch. According to claim 1, the mycorrhizal mushrooms are Tricholoma matsutake (S. Ito et Imai) Sing., T. bakamatsutake Hongo, Sarcodon aspratus (Berk.) S. Ito , Tuber spp., Rhizopogen rubescens (Tul.) Tul., Sullus grevillei (Klotzsch) Sing., S. bovines (L .: Fr.) O. Kuntze, Boletus edulis Bull. Ex Fr.], Lyophyllum shimeji (Kawam.) Hongo, Hydrangea mushroom [ Hygrophorus spp.], Sage mushroom [ Lyophyllum spp.], Matsutake mushroom [ Tricholoma spp.], Cucumber mushroom [ Leucopaxillus spp. ] vast mushroom [Amanita spp.], the net mushroom [Boletus spp.], silk net mushrooms [Sullus spp.], rattling reluctant net mushrooms [Leccinum spp.], the shaman mushrooms [Russula spp.], milk mushrooms [Lactarius spp ., Cantharellus spp., Ramaria spp., Morel mushroom Morchella esculenta (L .: Fr.) Pers. ver. A method for producing a mycelium of mycorrhizal fungi, which is at least one selected from the group consisting of Esculenta , Pisolithus tinctorius Cokr & Couch, Scleroderma spp., And Morchella spp. A solid medium for use in the method for producing a mycelium of mycorrhizal mushrooms according to any one of claims 1 to 6, characterized in that it comprises pumice having a diameter of 0.1 to 10 cm. 1) inoculating mycelia of mycorrhizal mushrooms into the roots of seedlings using mycelia of mycorrhizal mushrooms formed on the solid medium obtained according to the method according to any one of claims 1 to 6; 2) Planting seedlings inoculated with mycelium: A method of producing fungal infection seedlings of trees, comprising the step. The method of producing a mycorrhizal infectious seedling of tree, according to claim 8, which is carried out in a non-sterile state. The method according to claim 8, further comprising irrigation of the planted seedlings with pesticides, fungicides and plant growth promoters at intervals of 1-4 weeks at 1000-200 fold, respectively.

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