KR100252384B1 - ASI 6046 strain of Hypoxylon Lujibino Island, which is a symbiotic fungus of white-necked mushroom, and artificial cultivation method of white-headed mushroom - Google Patents

Abstract

PURPOSE: Provided is a Hypoxylon rubiginosum ASI 6046 (KFCC-10977) which is symbiont of Tremella fuciformis berk. And a method for cultivating Tremella fuciformis berk in a large quantity using the same is also provided. CONSTITUTION: A preparation method of Tremella fuciformis berk. is composed of the following steps of: (a) preparing a spawn by culturing Hypoxylon rubiginosum ASI 6046 and Tremella fuciformis berk.; and (b) inoculating the spawn into log and then cultivating at 20-25 deg.C for 35-45 days or into a bottle and then cultivating at 25-30 deg.C for 35-45 days.

Description

ASI 6046 strain of Hypoxylon Rubinino island, which is a symbiosis of mushrooms, and artificial cultivation method of mushrooms

The present invention relates to a cultivation method of hypoxylon rubiginosum ASI 6046 strains in which the white neck symbiotic with the mushroom bacteria, and a method for cultivating the white neck mushroom using the same, the mycelial growth of the pure white isolated mushroom is very slow, Since the cultivated sawdust medium hardly grows, the fact that it is not artificially grown in the sawdust medium, which is the most widely used in normal artificial cultivation. After separation, the white-necked black pepper is tested for its interaction with the fungus to isolate the high symbiotic Hypoxylon rubiginosum ASI 6046 strain, It is about.

Tremella fuciformis Berk. Is taxonomically belonging to Heterobasidiomycetes, Tremellales, Tremellaceae, and is commonly known as white jelly fungus or silver. It is also called silver-ear, but it is commonly called white mushroom as distinguished from the woody mushroom belonging to the genus Auricularia sp. White-black mushrooms are widely distributed throughout the world, such as China, Japan, the United States, and the African continent (Sawada, 1942, Ito, 1955; Lowy, 1971). In addition, it has been used as a medicinal and edible mushroom because it has a good taste and contains a large amount of medicinal ingredients, such as linoleic acid, a sulfated substance. Especially, it has been treated as a pole product in China, and has been treated as a lung disease, high blood pressure, cold, beauty and even beauty. It is known as the Buddha's medicine (李, 1596, 三 郞, 1971).

However, because the white-wood mushrooms with these characteristics grow native to small hardwoods that are housed from spring to autumn, it is not enough to be collected outdoors to meet popular demand.

Therefore, as a result of research on white fungi mushrooms in order to supply a large amount of mushrooms, the white fungus mushrooms are gelatinous, so the contamination rate is very high, and mycelial growth is very slow and sterilized. It was found that inoculation into sawdust medium produced little growth. Therefore, in early white mushroom cultivation, the wood to be cultivated is placed next to the alley where fruiting bodies are placed, and the spores scattered from the fruiting bodies are inoculated into the wood and inoculated using a seed called "tissue suspension". Although the cultivation by the method, these methods are too primitive, there was a problem to grow mushrooms in large quantities.

Therefore, the present inventors have conducted a number of studies to artificially cultivate mushrooms in large quantities, the results of the fact that the white mushrooms do not grow on the sawdust medium, but white mushrooms grow in the wild wood, The present invention focuses on the fact that white wood has a symbiotic bacterium that acts as a "biological factor" that whites help mushrooms to degrade the wood and provide surplus nutrients. It was completed.

That is, it is an object of the present invention to provide a Hypoxylon rubiginosum ASI 6046 strain, which is a symbiotic fungus of the white neck.

Still another object of the present invention is to provide a method for artificially cultivating mushrooms (Tremella fuciformis Berk.) Using the aforementioned strains.

Other objects and functions of the present invention will become apparent to those skilled in the art from the following detailed description of the invention.

Figure 1 is a photograph showing the shape of the fruiting body of the mushroom white (A; nut-gall type, B; cock's-comb type).

FIG. 2 is a photograph showing that white-necked estradiol bands of mushrooms and symbiotic bacteria.

[Description of Drawing Symbol]

Lane 1: Albino mushroom ASI 6024, Lane 2: Albino mushroom ASI 6038

Lane 3: Albino mushroom ASI 6039, Lane 4: Albino mushroom ASI 6040

Lane 5: Albino mushroom ASI 6041, Lane 6: Albino mushroom ASI 6042,

Lane 7: Albino Mushroom ASI 6045 (KFCC-10996)

Lane 8: symbiotic ASI 6025, lane 9: symbiotic ASI 6044

Lane 10: symbiotic ASI 6046 (Hypoxylon rubiginosum ASI 6046; KFCC-10997), lane 11: symbiotic ASI 6047, lane 12: symbiotic ASI 6048

3 is a diagram showing the shape (× 650) of the fruiting body of the strain of the present invention, Hypoxylon rubijinosum ASI 6046 (KFCC-10997).

Figure 4 is a photograph showing the follicle spores of the strain of the present invention, Hypoxyllon rubinino island ASI 6046 (KFCC-10997).

FIG. 5 is a band photograph showing DNA polymorphism according to PCR amplification of the ITS portion of the nucleosilon rubinino island ASI 6046 (KFCC-10997), the strain of the present invention.

[Description of Drawing Symbol]

Lane 1: 100 bp DNA ladder, lane 2: hemispherical locus of high-oxylon rubiginous island ASI 6046 (KFCC-10997), lane 3: pulvinate type of high-oxylon rubiginous island ASI 6046 (KFCC-10997) Chair

Figure 6 shows the mycelial growth curve of the strain of the present invention, Hypoxylon Rubinino island ASI 6046 (KFCC-10997).

Figure 7 is a photograph showing the occurrence of the mushroom during the sawdust disease and wood cultivation of the white-necked mushroom using the strain of Hypoxyllon rubinino island ASI 6046 (KFCC-10997), the strain of the present invention.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention mainly grows in the solid wood of solid wood is hardly grown when inoculated on the sawdust medium by incubating the mushroom bacteria purely, because the white wood of the wild alley mushrooms in the area where the mushrooms grow white cellulose in addition to the mushroom bacteria In view of the fact that lignin decomposes white lignin may exist in the mushroom symbiosis, white worms collected from the Jeonnam Haenam County isolates the fungi and their symbiosis from the alley of mushrooms, Of course, through the testing of the suitability of artificial cultivation, the white neck, which can artificially cultivate mushrooms, selected mushroom symbiotic strains, and identified this strain to complete the present invention.

When the white neck of the present invention will be described in detail the process of selecting the symbiotic strain of the mushroom as follows.

(1) Isolation of Tremella fuciformis Berk.

In the mushroom alley, white necks collected from the area of Haenam-gun, Jeollanam-do, are white, similar to chrysanthemum flowers with pure white, translucent, and 3 ~ 10 ears. After washing, the internal tissues were sterilely cut and transplanted into PDA medium and cultured in a 25 ° C thermostat to isolate six white mushrooms (Tremella fuciformis Berk.) With strain Nos. ASI 6424, 6038 ~ 6042, 6045. It was.

(2) Isolation of Albinus Mushroom Symbiosis

White wood collected from Haenam-gun, Jeollanam-do area has mushroom alleys. First, the bark and white wood have been removed from the fruiting body, and the surface of the wood is well wiped with 70% alcohol. Cut the surface of (傳 粉) shallowly, remove the bottom piece, put it in Petri dish containing PDA medium, and incubate in 25 ~ 28 ℃ culture room for 5 days, isolate all grown fruiting bodies and strain No. ASI 6025, 6044, 6046, 6047, 6048 were given.

(3) Homology between isolated strains

In order to confirm the homology between the fungus and the symbiotic fungus isolated from the above, the above-mentioned bacteria were added to MCM medium (peptone 2.0g / L, MgSO ₄ · 7H ₂ O 0.5g / L, KH ₂ except for agar). PO ₄ 0.5g / L, K ₂ HPO ₄ 1.0g / L, malt extract 2.0g / L) for 20 days, and then filtered and washed with distilled water. Then, finely pulverized by adding liquid nitrogen at the induction, and then, the second sample obtained by centrifugation at 4 ° C. for 15 minutes in a centrifuge (12,000 rpm) was placed in an ephendorf tube and centrifuged for 10 minutes ( 12,000 rpm), and the supernatant was electrophoresed. After the completion of the electrophoresis, the gel was stirred three times with 0.2 M phosphate buffer (pH 6.5) to adjust the acidity of the gel, followed by a color developing solution at 35 ° C. (α-naphthyl acetate 20 mg, ethylene glycol monomethyl ether 2 ml, 0.2 120 ml of M phosphate buffer (pH 6.5) and 120 ml of fast blue RR salt (20 mg) were then immersed at 50 rpm in the dark, and developed until the esterase band was clear (about 30 minutes). The result is shown in FIG.

From Fig. 2, six white-necked mushroom strains showed little homology between their respective band patterns, while only ASI 6025 and 6044 had high homology, while ASI 6046, 6047, and 6048 showed different band patterns. Able to know.

(3) Selection of excellent strains

The pure white isolated mushrooms and their symbiotic bacteria were inoculated in PDA medium for each of the strains listed in Table 1, and then cultured for 5 days at 28 ° C. in an incubator to observe the formation of fruit (fructification). Table 1 shows.

TABLE 1

As can be seen from the results in Table 1, the purely isolated 6 species of white-necked cultures were mixed with mushrooms and 5 species of symbiosis, and only when the white-necks were cultured with mushrooms ASI 6045 and symbiotic ASI 6046. It can be seen that a strong fruitage is formed. Therefore, because the white-necked mushroom bacteria ASI 6045 and symbiotic ASI 6045 has a host specificity, for the artificial cultivation of mushrooms and identification of symbiotic bacteria for the white-necked mushroom of the present invention, the white-necked mushroom (Tremella fuciformis) ASI 6045 On October 29, 1997, it was deposited with the Korea Microbiological Preservation Center (KFCC), affiliated with the Korean spawn association, and was given accession number KFCC-10996.

The fruiting body of the fungus ASI 6045 (KFCC-10996) is 5 ~ 15 × 4 ~ 12cm in size and about 0.5 ~ 0.6mm thick. Its shape is similar to chrysanthemum, and the fruiting layer (hymenium) is bidirectional like overlapping leaves. As it occurs, the edges are cordless in a thin torn shape. Mycelia are white to yellow in color, 1.5 to 3.0 μm in diameter, and have a clamp connection where the hyphae are located. The mycelial growth conditions of ASI 6045 (KFCC-10996) of the white-headed mushroom fungus are 25 ~ 30 ℃ and acidity (pH) is 5.5 ~ 6.0.

(4) Symbiosis Effect of Selected Albinus Mushroom ASI 6045 (KFCC-10996) and Symbiosis ASI 6046

In order to confirm the symbiotic effect between the fungus ASI 6045 (KFCC-10996) and the symbiotic ASI 6046 strains, PDA medium (potato 200g / l, dextrose 20g / l, agar 20g / l), oatmeal medium (oatmeal) 30 g / l, agar 20g / l) and malt medium (peptone 5.0g / l, malt extract 20g / l, agar 20g / l) were sterilized at 121 ° C for 20 minutes using autoclave, and then sterile plastic petri dish (diameter). 9 cm) was dispensed in 25 ml, inoculated with the inoculum, and incubated in a thermostat controlled at 25 ± 2 ° C. for 10 days to examine the diameter of the colony. In the above inoculation, the white-necked mushroom ASI 6045 (KFCC-10996), in the case of culturing by scratching (scratching) as in the case of cultivation, and then cut off the slices taken with a cork borer having an inner diameter of 6 mm. In case of commensal bacteria ASI 6046, inoculate hyphae to PDA medium and incubate for 5 days at 28 ℃ incubator.In case of mixed bacteria of fungi ASI 6045 and commensal bacteria ASI 6046, firstly, fungus ASI 6046 6045 (KFCC-10996) was incubated in PDA medium for 10 days, and then inoculated with symbiotic ASI 6046 and cultured for 10 days. The results are shown in Table 2.

TABLE 2

It can be seen from Table 2 that the mycelial growth of white fungus ASI 6045 (KFCC-10996) and symbiotic ASI 6046 was significantly higher than that of fungi. Accordingly, it can be seen that ASI 6046, a commensal bacterium, is a bacterium that promotes the formation of fruiting bodies of mushrooms.

(4) Identification of Symbiotic ASI 6046

White cultivars were cultured with mushroom ASI 6045 (KFCC-10996) and cultivated symbiotic ASI 6046 strains that promoted fruiting of white mushrooms, and observed the morphological characteristics. The mycelium is also shown in FIG. 3.

TABLE 3

In addition, as the culture lengthened, the symbiotic ASI 6046 formed hemispherical stromata lacking a clear asciliar elevation or sparsely sparse pulvinate stromata with a clear asciliar elevation. The vesicles were columnar with a single cyst, and the spermatospores were phaseoliforms, with eight arranged obliquely in a row (see Figure 4).

Genetic characteristics of the symbiotic ASI 6046 were extracted by DNA, and then analyzed using PCR (Polymorase Chain Reaction) -RFLP (Restrition Fragment Length Polymorphisma) for the internal Transcribed Spacer (ITS) portion of the nucleus. That is, the bacteria are transferred to a flask containing 5 g of tap water and yeast extract (Difco) and 20 g / L of glucose, and then the hyphae obtained by shaking culture at 25 ° C. for about 1 week are collected by filtration and wiped with a paper towel. DNA was isolated. PCR amplification of the nuclear ITS1-5.8S-ITS2 rDNA range was performed by a well-known method using an MJ Reasearch DNA thermal cycler (model PTC-150). Primers used for PCR amplification were ITS1 and ITS4, and sequences were prepared using the method described in "Amplification and direct sequencing of fugal ribosomal RNA genes for phylogenetics" (pp 315-322, White et al., 1990). PCR amplification products were subjected to electrophoresis on 3% agarose gel and stained with ethidium bromide to confirm DNA polymorphism (see FIG. 5). From FIG. 5, the size of the amplified DNA fragment of the strain forming a hemispherical locus without asymptomatic angle was about 600 bp, and the size of the pulvinate type amplified DNA fragment with the asymptomatic angle was 580 bp. Can be.

The symbiotic ASI 6046 was identified as Hypoxylon rubiginosum by investigation of the morphological, molecular biological and genetic properties described above. Identified as Hypoxylon truncatum, but as a result of re-identification in connection with the application, identified as Hypoxyllon rubinino island). Therefore, the present inventors named the commensal bacterium ASI 6046 as Hypoxylon rubiginosum ASI 6046, and deposited it on the Korea microbial preservation center (KFCC) affiliated with the Korean spawn association on October 29, 1997, accession number KFCC-10997 was granted.

(5) Optimal Growth Conditions of the High Foxilon Rubinino Island ASI 6046 (KFCC-10997) Strain

The optimum growth conditions of Hypsisilon rubinino island ASI 6046 (KFCC-10997) strains were investigated for the artificial cultivation of fungi.

① Optimum temperature

In order to investigate the optimum temperature for mycelial growth of ASI 6046 (KFCC-10997) strains of Hypoxyllon rubizinom, 25 ml of agar without MCM was constantly dispensed into a Erlenmeyer flask (250ml), Inoculated with the island ASI 6046 (KFCC-10997) strain, and incubated for 15 days in a thermostat controlled to 15, 20, 25, 30, 35 ℃ and then measured the dry cell mass, the results are shown in Table 4.

TABLE 4

As can be seen in Table 4, the optimum incubation temperature for the liquid culture of the strain of Hyposilon rubiginosoma ASI 6046 (KFCC-10997) is 25 ~ 30 ℃.

② optimum pH (pH)

In order to investigate the mycelial growth of Hyposilon Rubinino island ASI 6046 (KFCC-10997) strain, the pH of MCM medium was adjusted to 0.5 to 4.0 ~ 8.0 using 1N hydrochloric acid and 1N sodium hydroxide. After dispensing 25 ml into the flask (250 ml), inoculated with the strain of Hyposilon rubizinos ASI 6046 (KFCC-10997) and incubated for 15 days at 28 ℃ incubator, the dry cell mass was measured and the results are shown in Table 5. .

TABLE 5

As can be seen in Table 5, the optimum acidity of the liquid culture of the strain ASI 6046 (KFCC-10997) of the hypoxylon rubin island is 5.0-5.5.

③ optimal nutrition

In order to investigate the optimal nutrient source for mycelial growth of Hypoxyllon rubinino island ASI 6046 (KFCC-10997) strain, various carbon sources as shown in Table 6 were adjusted to the same amount of carbon as 30 g of sucrose to prepare a medium. In the case of various nitrogen sources as shown in Table 7 below to prepare a medium by adjusting to the same content as 2.0 g of sodium nitrate, 25 ml of this liquid medium is constantly dispensed into a Erlenmeyer flask (250 ml), and Hyposilon rubiginosom ASI 6046 After inoculating (KFCC-10997) strain and incubating for 15 days at 28 ℃ incubator, the dry cell mass was measured, and the results are shown in Table 6 and Table 7. The pH of the medium is 5.0.

TABLE 6

TABLE 7

As can be seen in Tables 6 and 7, the strain of Hyposilon rubinino ASI 6046 (KFCC-10997) of the present invention was well grown in various nitrogen sources and carbon sources, so that various nutrient sources can be widely used. .

④ Growth curve

25 ml of MCM medium was dispensed into a 100 ml Erlenmeyer flask, and then inoculated with strain ASI 6046 (KFCC-10997) of Hypoxilon Rubinus Island. It was. From FIG. 6, the growth curve of the strain ASI 6046 (KFCC-10997) of Hypoxylon Rubinus Island continued to increase until 15 days of cultivation to 14.36 mg / 25ml, but the rate of increase of production was rapidly increased from the third day to the 11th day of cultivation. It increased, and since then showed a gentle increase. Protein content was the highest at 11.2 μg / ml at 11 days of culture, but gradually decreased after that. On the other hand, the optimum density (optical density) was the highest as 1.32 on the 10th day of the culture, it can be seen that gradually decrease thereafter.

The method for artificially cultivating mushrooms using the hypoxylon rubiginus island ASI 6046 (KFCC-10997) strain of the present invention having the culture growth conditions as described above will be described in detail below.

In order for seedlings to grow mushrooms in sawdust and logs, it is necessary to know the seed culture characteristics of A. 6046 (KFCC-10997) strain of Hypoxilon Rubinino island and mixed bacteria of ASI 6045 (KFCC-10996). . Therefore, the spawn culture characteristics of the mixed A. 6046 (KFCC-10997) strains and the white A. mushrooms ASI 6045 (KFCC-10996) strains were examined as follows.

(1) Selection of excellent sawdust badges

In order to select sawdust suitable for mycelial growth of Hyposilon Rubinino island ASI 6046 (KFCC-10997) strains and white-eyed mushroom ASI 6045 (KFCC-10996) mixed bacteria, the sawdust shown in Table 8 in a 250 ml Erlenmeyer flask After adding rice bran to 80: 20 (V / V) and autoclaving by adding 70% water, inoculated mycelia obtained by culturing the mixed bacteria in MCM medium for 12 days, incubating it for 15 days, The diameter of the was investigated and the results are shown in Table 8.

TABLE 8

As can be seen in Table 8, oak sawdust medium is most preferred for growing white mushrooms.

(2) optimum additives, moisture, temperature, acidity

In order to select suitable additives for oak sawdust, freshly dried rice bran, bran, beer foil and soybeans were blended at a rate of 20% (V / V), and the sawdust medium adjusted to a moisture content of 65 ± 2% was tested (Φ3 .0 × 20.0cm) was constantly filled in 50g and autoclaved at 121 ° C. for 30 minutes, and then inoculated with 3 to 5g of mycelia obtained by incubating for 12 days in an MCM medium. After incubating the test tube inoculated with the mixed bacteria for 15 days in a culture room adjusted to 25 ± 2 ℃, the diameter of the colony was examined and the results are shown in Table 9.

TABLE 9

As can be seen in Table 8, beer foil is most preferred as an additive added to oak sawdust in order to grow mushrooms.

In order to investigate the optimum content of beer foil, which is the best additive, the mycelial growth and mycelial growth were carried out in the same way as above, except that a sawdust medium containing 10 to 30% of beer foil was mixed at 0 ~ 50% (V / V). The density was measured and the results are shown in Table 10.

TABLE 10

As can be seen in Table 10, the content of beer gourd added to the oak sawdust medium to grow mushrooms of white wood is most preferably 20-30% in consideration of mycelial growth and density.

In order to investigate the optimum moisture content, after mixing beer foil 20% (V / V) uniformly in oak sawdust medium, adjust the water content to 40 ~ 75% at 5% intervals (Φ3.0 × 20.0cm) Except for filling in, the culture was carried out in the same manner as described above to measure the mycelial growth and mycelial density, and the results are shown in Table 11.

TABLE 11

As can be seen in Table 11, the moisture content of oak sawdust medium for the cultivation of the white wood mushroom is most preferably in consideration of mycelial growth and density 60-65%.

In order to investigate the optimum pH (pH), uniformly mix 20% (V / V) of beer gourd in oak sawdust medium, adjust the water content to 60%, and adjust the pH to 3.0 ~ 11.0 with McILvain buffer. After adjusting the intervals, the test tube (Φ3.0 × 20.0cm) was charged to 50g each, and then autoclaved at 121 ° C. for 30 minutes, incubated for 12 days in MCM medium, and then inoculated with 3 to 5g of the obtained mycelia. Test tubes inoculated with mixed bacteria were incubated at 27 ° C. for 15 days, and the diameters of the colonies were examined and the results are shown in Table 12.

TABLE 12

As can be seen in Table 12, the optimum acidity for growing white mushrooms using oak sawdust medium is preferably 5.0 to 7.0.

To investigate the optimum incubation temperature, uniformly mix 20% (V / V) of beer foil in oak sawdust medium, adjust the water content to 60%, and constantly fill 50 g in a test tube (Φ3.0 × 20.0cm). Then, autoclaved at 121 ° C. for 30 minutes, and inoculated with 3 to 5 g of mycelia obtained by culturing for 12 days in an MCM medium. The test tube inoculated with the mixed bacteria was incubated for 15 days after adjusting the incubation temperature to 5 ~ 10 ℃ to 10 ~ 35 ℃, the diameter of the colony was investigated and the results are shown in Table 13.

TABLE 13

As can be seen in Table 13, the optimum incubation temperature for growing white mushrooms using oak sawdust medium is preferably 25 ~ 30 ℃.

(3) Mass production of white-necked mushroom fruiting body using Hypoxyllon rubinino island ASI 6046 (KFCC-10997) strain

① sawdust cultivation

Quantity according to bottle capacity; To examine the relationship between the container size and the yield during the cultivation of the sawdust jar of mushrooms, the white wood was mixed with oak sawdust and beer foil at a ratio of 80:20 (V / V) and 100 g of sawdust with 65% moisture content. 850ml and 1800ml sized containers were sterilized in a high pressure sterilizer (12 atm) for 40 minutes, inoculated with mixed bacteria and incubated at 28 ° C. for 15 days, and the yield of fruiting bodies was examined, and the results are shown in Table 14.

TABLE 14

As can be seen in Table 14, the yield was best when grown in a 1800ml bottle, but the recovery was reduced compared to the 850ml bottle. In addition, the individual volume was 70.5g in the 1800ml bottle, more than 41.2g in the 850ml bottle, but the recovery rate is 11.5%, lower than 17.1% of the 850ml, so that the size of the container 850ml is more advantageous for the cultivation of mushrooms.

Fruiting body yield according to the blending amount of the strain of Hypoxyllon rubinino ASI 6046 (KFCC-10997); Addition of Mushrooms ASI 6045 (KFCC-10996) and Hyposilon Rubinino Island ASI 6046 (KFCC-10997) Strains in Artificial Cultivation Using Hyposilon Rubinino Island ASI 6046 (KFCC-10997) Strains Mixtures of white fungus ASI 6045 (KFCC-10996) and Hyposilon rubizinomycete ASI 6046 (KFCC-10997) strains to investigate the incubation period, initial development time and yield Mixed bacteria were prepared at a ratio of 100: 100, 100: 50, 100: 25, 100: 12.5, and 100: 6.2. After mixing oak sawdust and beer gourd at 80:20 (V / V) ratio in a 850ml bottle and adding 100g of sawdust adjusted to 65% moisture content, inoculated with mixed bacteria after sterilization for 40 minutes in autoclave (12 atm). , 15 days incubation at 28 ℃, the yield of the fruiting body was investigated and the results are shown in Table 15.

TABLE 15

As can be seen from Table 15, the yield, biological efficiency, etc. of the white agar mushroom were determined by the amount of the A. 6046 (KFCC-10997) strain of Hyposilon Rubinino island, which was mixed with the white agar mushroom ASI 6045 (KFCC-10996). It is not affected. In other words, it can be seen that the effect is high even if a small amount of the combination of the hyalpoxylon rubinino ASI 6046 (KFCC-10997) strain to the mushroom ASI 6045 (KFCC-10996).

② Log cultivation

The wood cultivation of white mushrooms is similar to shiitake mushrooms, and holes are inoculated between January and March, and the size of the holes varies depending on the size of the logs, but the size is 1.0-1.5 cm in diameter and 1.5 cm Inoculate with holes ~ 3.0 cm deep. For the cultivation of logs, as shown in Table 16 below, the number of days of initiation, the number of individuals, and the number of first flowers were examined according to the type of logs, and the number of days of first flowers was 63 days, 83.5 g, and 794 g / m ^{3 of} oak logs. The best yields were found in the order of alder, poplar and acacia, and did not germinate in pine.

TABLE 16

In addition, the optimum temperature for mycelial growth (laying) in the alleys was the best mycelial density at 20-25 ℃ (Table 17), and the incubation period was the best when incubated for 35-45 days (Article 18). Degree). When the relative humidity was raised to 80 ~ 90% to raise the fruiting body, the emergence of the fruiting body was the fastest with 57 days, and the collection period was the longest with 6-7 months (Fig. 19).

TABLE 17

Mycelial density according to mycelial culture temperature (cultivated in oak)

TABLE 18

Mycelial Density According to Incubation Period

TABLE 19

An example of spawn production for artificially cultivating mushrooms using white hypoxylon ribiginosum ASI 6046 (KFCC-10997), whose growth conditions and culture characteristics have been identified in a series of tests. If you look like this:

(1) Spawn production

Discharge material

Oak sawdust is applied, and the additives are suitable to mix beer foil 20 to 30% (V / V).

[Badge Mixing]

While mixing the medium with beer foil added to sawdust, the moisture should be adjusted to 60-65%, and then the acidity should be adjusted to 5.0-7.0.

[Sterilization]

The blended medium material should be filled so that 90% of the volume of glass or plastic bottles of 1,000 ~ 1,500ml size is filled, and then sterilized at 121 ℃ for about 30 minutes.

[Inoculation and Culture]

The sterilized medium was aseptically opened with a stopper, incubated for 10 days with the white fungus ASI 6045 (KFCC-10996) in PDA medium, and then inoculated with the same amount of Hypoxyllon rubizino ASI 6046 (KFCC-10997). After inoculating and transplanting the hyphae generated by incubation for 10 days on the surface of the bottle, the spawn was prepared by culturing for 15 days at 25 ~ 30 ℃.

(2) spawn inoculation and cultivation

After inoculating the seedling prepared in the above (1) with a bottle of 850 ml size or 10 cm diameter oak, 35 to 45 days at a temperature of 25 to 30 ° C. for a bottle and 20 to 25 ° C. for a log Incubated. After the incubation was completed, the mature white necks showed a chrysanthemum-like appearance of the fruiting body of the mushroom, reaching a diameter of 8-12 cm, and the lampshade (see Fig. 7).

When mature white-headed mushrooms grow too early, the yield is low; on the other hand, when too late, the bottom of the fruiting body turns black and starts to rot, which adversely affects the quality of the mushrooms. When it turns from to white, it grows when the outer part of the fruiting body softens and begins to slap downward, so that high-quality white-necked mushrooms can be grown.

As described above, Hypoxylon ribiginosum ASI 6046 (KFCC-10997) grows together with white mushrooms, which promotes mycelial growth and growth of fruiting bodies. Co-cultivation of Foxylon rubijinsum ASI 6046 (KFCC-10997) to prepare a mixed seed, inoculate it into sawdust bottles or logs, and incubate the mushrooms that were not artificially grown. Mass production

Claims (4)

Hypoxylon ribiginosum ASI 6046 (KFCC-10997), which is a symbiotic fungus of white-headed mushrooms. In the way the white tree grows mushrooms Preparing a spawn by mixing the culture medium of Hypoxylon ribiginosum ASI 6046 (KFCC-10997) according to claim 1 with a fungus; And Inoculating the spawn seedlings at a temperature of 20-25 ° C., or inoculating the bottle with 35-45 days of incubation at a temperature of 25-30 ° C .; Whitewood mushroom cultivation method characterized in that it comprises a. According to claim 2, wherein the medium is mixed with oak sawdust medium at 20 to 30% (V / V) of beer foil while adjusting the moisture to 60 to 65%, acidity 5.0 to 7.0 at high pressure at 121 ℃ for about 30 minutes The method of cultivating white mushrooms, characterized in that the sterilized. The method for cultivating a white-headed mushroom according to claim 2, wherein the white-headed mushroom is the white-headed mushroom ASI 6045 (KFCC-10996).

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