JP6265083B2 - Method of symbiosis of choreimaitake and naritatake, and cultivation method of choreimaitake using the method - Google Patents

Description

  The present invention relates to a symbiotic method of Choreimaitake and naritatake, and a method of cultivating Choreimaitake using the method.

  Choreimaitake (Polyporus umbellatus) is a fungus belonging to the family Sarnocosiaceae, and its mycorrhiza (猪苓: chorei) has been used as a herbal medicine since ancient times, and has been incorporated into Kampo prescriptions such as Gojo-to and Tsukuyu. It was.

  It has been reported that moths collected from the natural world coexist with aratake in most cases (Non-patent Document 1). It is thought that. Therefore, the symbiosis of Choreimaitake and Nararatake is considered to be the key to the cultivation of strawberries. In the field cultivation of persimmon performed in China etc., the persimmon cultivation using this symbiotic relationship has already been performed (Non-patent Document 2). In China's field cultivation method, we lay down the Japanese oak tree inoculated with aratake, inoculate it with the fungal nuclei of seedlings, and symbiotic with it. Harvest. This method is about half a year to one year before inoculating oyster mushrooms and spreading them, half a year to one year after the inoculation of choreimaitake mushroom nuclei and larvae of choreimaitake, and the fungus of choreimaitake after symbiosis It takes two to three years for the enlargement of the nucleus, a total of nearly four years.

  Patent Document 1 discloses a method for cultivating choreimaitake in the field, in which choreimaitake is symbiotic with aratake, which has low phytopathogenicity, in order to efficiently proliferate sclerotia of choreimaitake in a short period of time.

  However, the symbiosis rate when larvae and choreimaitake are symbiotic in the soil is 40% to 50%, which is one factor that decreases the yield per unit area of chorei.

JP 2007-319104 A

Kikuchi G. and Yamaji H. (2010) Identification of Armillaria species associated with Polyporus umbellatus using ITS sequences of nuclear ribosomal DNA.Mycoscience 51 (5): 366-372 Kyung-Da Choi, Kyung-Tae Lee, Jae Ouk Shim, Youn-Su Lee, Tae-Soo Lee, Sang Sun Lee, Shun-Xing Guo and Min Woong Lee (2003) A New Method for Cultivation of Sclerotium of Grifola umbellata Mycobiology 31 (2): 105-112

  An object of the present invention is to increase the symbiosis rate of oyster mushrooms and choreimaitake, to shorten the cultivation period of choreimaitake and to increase the yield per unit area.

  As a result of earnest examination focusing on the symbiotic process of choreimaitake and narake to solve the above-mentioned problems, the inventor inoculated choreimaitake on the upper part after the formation of arbuscular mycelial bundle, In order to complete the present invention, it was found that by cultivating crushed hardwood derived from hardwood, the symbiosis rate can be significantly increased compared to the conventional method, and the cultivation period can be shortened by symbiosis in winter. It came.

That is, the gist of the present invention is as follows.
(1) After inoculating oyster mushrooms on a fungus bed containing hardwood-derived pulverized wood, and after forming arbuscular mycelial bundles, larvae are inoculated with choreimaitake on the upper part of the root mycelium bundle forming part. A method for symbiosis of choreimaitake and narake, which includes cultivating a crushed hardwood-derived timber and cultivating it.
(2) Bacterial beds containing hardwood-derived pulverized wood were inoculated with aratake, and after the larvae formed root mycelium bundles, mixed with hardwood-derived wood pulverized material at the top of the root mycelium bundle forming part. A method of coexistence of Choreimaitake and Narratake, including inoculating and cultivating Choreimaitake.
(3) After inoculating oyster mushrooms on a fungus bed containing hardwood-derived pulverized wood, and after larvae forming root mycelium bundles, inoculating choreimaitake on the upper part of the root mycelium bundle forming part, The method according to (1) or (2) above, wherein the cultivated product is cultivated at 7 to 17 ° C. with a hardwood-derived crushed wood.
(4) Choreimaitake, comprising adhering choreimaitake mushrooms and larvae symbiotically by the method according to any one of (1) to (3) to a hardwood log and burying them in soil and cultivating them Cultivation method.

  According to the present invention, the symbiotic period of Choreimaitake and Narratake is shortened, and the cultivation period of Choreimaitake can also be shortened. Moreover, the yield per unit area can also be raised by raising the symbiosis rate of Choreimaitake and Nararatake.

FIG. 1 shows a schematic diagram of the relationship between the large sawdust occupancy rate of larva mycorrhizal mycelium bundles and the symbiosis of choreimaitake and larvae. FIG. 2 is an optical micrograph showing the state of the symbiotic part of Choreimaitake and Narratake. FIG. 3 is a photograph showing the root mycelium bundle forming part of narushrooms in the upper part of the fungus bed.

  Hereinafter, the present invention will be described in detail.

  Choreimaitake mushrooms form black-brown irregular nuclei in the soil and produce fruit bodies from the sclerotia. In the present invention, these sclerotia are used for culturing. The sclerotia of Choreimaitake used in the present invention is not particularly limited in terms of size, weight, production area, etc., but preferably has a diameter of about 2 to 10 cm, and larger sclerotia is divided into several. May be used.

  There are no particular limitations on the narcissus used in the present invention, as long as it belongs to the genus Agaricaceae, but there is a syrup that has low phytopathogenicity in terms of the symbiosis rate with choreimaitake and the proliferation rate of mycorrhiza. preferable. In the present specification, “phytopathogenicity” means that the mycelium of larvae is parasitic on a healthy growing tree and is killed or weakened. It is known that there are many species with different properties in the genus Fungus, and in this sense, there are those having high and low phytopathogenicity. Specific examples of such low phytopathogenic oysters include Armillaria sinapina, Armillaria gallica, Armillaria cepistipes, Armillaria calvescens, etc. It is done.

  In the present invention, larvae are inoculated into “a fungus bed containing a crushed wood derived from hardwood”.

  Although there is no restriction | limiting in particular as said hard-leaved tree, For example, a beech, a Quercus, a maple, a cucumber, a cherry tree, a shrimp, a willow, elm etc. are mentioned. In addition, crushed wood derived from conifers such as red pine may be added. It is preferable that the crushed wood derived from conifers is less than 30%, preferably less than 10% of the entire crushed wood.

  The pulverized wood is not particularly limited, and examples thereof include large sawdust, chips, chip dust, shavings, bark shreds, bark shavings, and the like.

  As for the particle size of the pulverized wood, the major axis is usually about 0.1 mm to 100 mm, preferably about 1 mm to 5 mm.

  As the narcissus used in the present invention, the narcissus collected from nature may be used as it is, or the larva inoculum marketed by Kagawa Shiitake Co., Ltd. or the like for the production of mushrooms may be used. Furthermore, they may be pre-cultured with a PDA medium or the like. The method of inoculating oyster mushrooms on the pulverized wood can be performed by a conventional method. For example, the fungus bed made of the pulverized wood is inoculated with nasal bamboo cultivated in advance in a medium and cultured for a certain period of time (for example, , Room temperature for 3 to 4 months under dark conditions), narrator forms a brown needle-shaped mycelial mycelium bundle on the fungus bed (FIG. 3).

  When inoculating larvae on the fungus bed containing the above-mentioned pulverized wood, the sawdust and chips are preferable as the pulverized wood used for the fungus bed. Nutrient sources such as wheat bran, sucrose, rice bran and the like may be included as a carbon source or nitrogen source. The water content of the fungus bed is usually adjusted to 50 to 80% by weight, preferably 55 to 65% by weight.

  For the fungus bed, various nutrient sources are added to the pulverized wood, water is added and mixed, and for example, a fungus bed having a diameter of about 15 cm and a height of about 12 cm may be prepared and used (special feature). No. 8-13225). It is preferable to make an inoculation hole in the fungus bed mass.

  Since aratake can be aseptically cultured, the bacterial bed used here is preferably sterilized from the viewpoint of preventing the propagation of various bacteria.

  After inoculating aratake on a fungus bed containing hardwood-derived pulverized wood, it is usually cultured at 5-32 ° C., preferably 20-25 ° C., usually for 70-110 days, preferably about 3-4 months. Form a mycorrhizal bundle.

  In the present invention, after larvae form a root mycelium bundle as described above, (i) choreimaitake is inoculated on the upper part of the root mycelium bundle forming part, and hardwood-derived wood crushing on the upper part (Ii) Inoculate and cultivate choreimaitake mixed with crushed wood derived from hardwood.

  Here, the state in which larvae formed a root mycelium bundle means a state in which a white, brown or dark brown wire-like root mycelium bundle has risen above the fungus bed in a sword mountain shape. Inoculate Choreimaitake on the top of the bundle.

  Although the same thing as the above can be used as the hardwood pulverized material used here, the water content is usually adjusted to 50 to 80% by weight, preferably 55 to 65% by weight. The pulverized wood used here may be used as a fungus bed mixed with the above-mentioned various nutrient sources, if necessary, but does not contain various nutrient sources in order to increase the symbiotic ratio of choreimaitake and narita mushrooms. A pulverized wood is preferable. If the crushed wood used here is sterilized and sterilized, certain miscellaneous bacteria adhering to the sclerotia of Choreimaitake may proliferate and adversely affect the growth of Choreimaitake. It is preferable not to sterilize in terms of suppressing the growth of miscellaneous bacteria that inhibit the growth of maitake.

  The choreimaitake mixed with the hardwood-derived wood pulverized product in (ii) is cultivated after inoculating the hardwood-derived wood pulverized product with choreimaitake, and the choreimaitake mushroom nuclei are formed in the wood pulverized product. It may be.

  The temperature at the time of symbiosis of choreimaitake and aratake varies depending on the cultivation method, and is preferably 7 to 17 ° C., more preferably 10 to 10 in indoor cultivation in which aratake is inoculated into a fungus bed containing crushed wood derived from hardwood. 16 ° C.

  In the present invention, larvae and larvae are symbiotic, when the mycelium of larvae enters into the fungal nuclei of the larvae, and when one end of the root mycelium bundle of larvae is lifted, It shall be said that it is in the state which was firmly tied so that the mycorrhiza of Choreimaitake was not separated from the mycelial bundle.

  The enlargement of chorei due to the coexistence of larvae and choreimaitake proceeds through the following steps 1 to 3.

  In addition, the symbiotic state of Choreimaitake and Nararatake described in the examples means the following 1-3.

1 Adhesion A stage in which larvae mycelial bundles adhere to the outside of the fungal nucleus because larva penetrates into the fungal nucleus of Choreimaitake.

2 Start of symbiosis / hypertrophy The stage in which Nararatake enters the inside while dissolving the fungal nuclei of Choreimaitake.

3. Progression of hypertrophy The stage where chorei and aratake take symbiosis, and nutrients are obtained from narake, and choreimaitake enlarges.

  By treating as described above, choreimaitake and aratake can be symbiotic normally in half a year to one year.

  By burying and cultivating cholay maitake and aratake, which are symbiotic in this way, in the soil together with hardwood logs, it is possible to cultivate chorei maitake in a short period of time.

  In general, larva and choreimaitake cease to grow in a low temperature soil environment such as in winter. Therefore, in the present invention, since it is possible to grow in summer soil after symbiosis indoors in winter, the fungal nuclei of choreimaitake can be efficiently enlarged.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated further more concretely, the scope of the present invention is not limited to these Examples.

[Example 1] Confirmation of indoor symbiosis conditions (materials)
Narratake: Armillaria gallica (from Shizuoka Prefecture)
Mycorrhiza of Choreimaitake (猪苓): Purchased inoculum nuclei from Shanxi Province, China and inoculum nuclei collected from various parts of Japan (from Aomori Prefecture, Hokkaido, and Ibaraki Prefecture) were used.
Grinded wood: large sawdust mixed with broad-leaved trees (beech, Japanese oak, maple) and coniferous trees (red pine)

(Method)
Culture of oyster mushrooms 1) Bacteria bed Sawdust: Wheat bran = 5: 1, mixed with 2% sucrose, and finally adjusted the water content of the sawdust to 60-65%, 1.7 kg of fungus Created the floor.
2) Sterilization was performed at 105 ° C. for 120 minutes.
3) Agaricaria galica previously cultured in a PDA medium was used as the inoculum. Aluminaria galica is known to have low pathogenicity among narcissus, so it has the advantage of having a low environmental impact during large-scale cultivation in the field.
4) After inoculation, the cells were cultured for about 3 months in an air bath incubator at 20 to 25 ° C.
5) When the larva mycelium grows in the majority of the fungus bed, and when the growth of the root mycelium bundle is confirmed in the upper part of the fungus bed, the mycorrhiza of Choraymaitake mushroom forms the root fungus of the larvae of the upper fungus bed. Inoculated to the department. Then, adjust the saw blade used in preparing the fungus bed in 1) so that the water content is 60 to 65%, and place it on the inoculum of the inoculated choraimaitake mushroom. Covered.
6) 4 ° C, 10 ° C, 14 ° C, 18 ° C, 20 ° C, 22 ° C, and 24 ° C incubator were prepared in the state of 5), and the symbiotic state of each temperature was confirmed (see Fig. 1).

Survey item 1) Observation of symbiosis (survey on enlargement rate of choreimaitake)
2) Presence or absence of symbiosis (microscopic observation)

(result)
Symbiosis test Investigation was carried out on the 197-218th day (about six months later) and the 338-362rd day (about one year later) from the beginning of planting. The results are shown in Table 1.

  At 4 ° C., the growth of oyster mushrooms was slow, and adhesion of choreimaitake mushrooms and larvae was not confirmed in one year.

It was confirmed that symbiosis occurred at 10-14 ° C in half a year to one year.
At 10 ° C., the growth of larvae was slow, but the growth of root mycelium bundles spread to about 90% of the fungus bed in one year. As a result of investigating the fungal bed, some symbiotic hypertrophy and symbiotic hypertrophy were observed in all 5 fungal beds (Table 1, symbiotic hypertrophy ◎, partial symbiotic hypertrophy ○).

  The maximum enlargement rate of choreimaitake at 10 ° C. was 121% in about one year of culture and 110% in about half a year of culture.

  At 14 ° C., larvae mycorrhizal bundles spread to about 90% to 90% of the fungus bed in one year. As a result of investigating the fungal bed, some symbiotic hypertrophy and symbiotic hypertrophy were observed in all 8 fungal beds (Table 1, Symbiotic hypertrophy ◎, Partial symbiotic hypertrophy ○).

  The maximum enlargement rate at 14 ° C. was 166% in about one year of culture and about 103% in about half a year of culture.

  At 18 ° C., the mycelial growth of larvae was vigorous, and the adhesion between the fungus mushroom nuclei and the larvae was confirmed in one microbial bed, but none of the symbiotic symbiosis was found (Table 1, adhesion confirmation Δ).

  Even at 20 ° C. or higher, the mycelial growth of larvae was vigorous, and the fungal nuclei of the inner chrysanthemum rotted and did not coexist (Table 1, adhesion confirmation Δ, not symbiotic ×).

  From the above results, it was found that the temperature range of 10 to 14 ° C. is suitable for symbiotic choreimaitake mushrooms and oyster mushrooms in indoor cultivation. As a consideration, I think that it is necessary for cholaymaitake symbiotic system construction to moderately suppress the growth of arbuscular mycorrhizal bundles.

  Furthermore, the state of the symbiotic part was observed (FIG. 2). Slices of glued oyster mushrooms that were confirmed to be enlarged were sliced and observed with an optical microscope. Naratake was found to be present in the black-browned partition in Choreimaitake.

[Example 2] Indoor symbiosis of Choreimaitake and Nararatake with expanded scale (experimental method)
1. Ingredients: Araraaria gallica: A mycorrhiza of choraimaitake from Shizuoka Prefecture (猪苓): from Shanxi, China

2. Creation of the large sawdust fungus bed The large sawdust is a mixture of maple, beech, konara and kunugi in an equal amount, and an appropriate amount of water is added to adjust the water content to about 55 to 75%. A fungus bed was prepared, and natura mushrooms were grown in the fungus bed in the same manner as in Example 1 and cultured until a root mycelium bundle formed at the upper part of the fungus bed.

3. Inoculation of Choreimaitake Opening a fungus bed bag infested with oyster mushrooms, mixing maple, beech, Japanese oak and cucumber in equal amounts in the root mycelium bundle forming part and adding an appropriate amount of water to a water content of about 65-75% A mixture of unsterilized large sawdust and chorimaitake mushroom nuclei prepared in the above was inoculated to a thickness of about 20 cm. The inoculation amount of the fungal nucleus of Choreimaitake was three types of 200 g, 100 g, and 25 g. Sealed except for the vents, placed in cardboard and stored in the warehouse. The temperature of the medium was approximately 10 ° C to 14 ° C.

  As a result of irrigation so that the fungus bed did not become too dry and symbiotic for a total of 4 months from December to March, the growth of larvae was good, and the symbiosis of choreimaitake and larvae was inoculated with the fungus of inoculated choreimaitake It was confirmed in more than 90% of the nuclei.

[Example 3] Soil cultivation after symbiosis indoors (contents)
An inoculum obtained by symbiosis of larvae and choreimaitake obtained by symbiosis indoors in the same manner as in Example 1 (however, the symbiotic temperature is 14 ° C. to 15 ° C.) was buried in soil and cultivated with logs. .

(experimental method)
(1) A hole with a depth of about 15 cm was dug in a field in Aomori Prefecture, and (Konara) logs (diameter 3 cm to 5 cm, length 60 cm) were arranged so as to form four sides of a rectangle. A fungus bed that coexisted with oyster mushrooms and choreimaitake was placed as it was between the raw wood, and the raw wood (diameter 3 cm to 5 cm, length 30 cm) was placed on top to cover the fungus bed and laid with soil.

  After 10 months, when the buried log was dug up, the fungal nuclei of Choreimaitake, which was enlarged from the time of inoculation, were obtained.

(2) A hole with a depth of about 15 cm was dug in a field in Aomori Prefecture, and (Konara) logs (diameter 3 cm to 5 cm, length 60 cm) were arranged in parallel. Naratake and choreimaitake symbiotically symbiotically obtained by the method described in Example 1 (symbiotic temperature is 14 ° C. to 15 ° C.) were taken out from the fungus bed, divided into 2 to 4 pieces, and inoculated between raw trees. The soil was placed on top and buried.

  After 10 months, when the buried log was dug up, the fungal nuclei of Choreimaitake, which was enlarged from the time of inoculation, were obtained.

Claims (4)

  After inoculating oyster mushrooms on a fungus bed containing hardwood-derived pulverized wood, and after forming arbuscular mycelium bundles, inoculated choreimaitake on the upper part of the root mycelium bundle forming part A method of symbiosis of choreimaitake and aratake, which includes cultivating a crushed crushed wood.   After inoculating oyster mushrooms on the fungus bed containing hardwood-derived pulverized wood, and after forming the root mycelium bundle, larvae mixed with hardwood-derived wood crushed at the upper part of the root mycelium bundle formation part A method of coexistence of Choreimaitake and Nararatake, including inoculating and culturing   After inoculating oyster mushrooms on a fungus bed containing hardwood-derived pulverized wood, and after forming arbuscular mycelium bundles, inoculated choreimaitake on the upper part of the root mycelium bundle forming part The method according to claim 1 or 2, wherein the cultivated product is cultivated at 7 to 17 ° C after being covered with a pulverized product of wood.   A method for cultivating Choreimaitake, comprising adhering Choreimaitake and narratake symbiotically by the method according to any one of Claims 1 to 3 to a hardwood log and burying them in soil and cultivating them together.

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