JP2002369635A - New lyophyllum ulmarium strain - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new Lyophyllum ulmarium strain in which period from inoculation of strain to harvest of fruit body is shortened. SOLUTION: About 1,000 wild Lyophyllum ulmarium strains are collected and mycelium of each strain is cultured in a liquid medium and activity of cellulase produced in the culture medium is sequentially measured and 4 strains each having a high cellulase activity from initial stage of culture is primarily selected. Culture test of the primarily selected strains is repeatedly practiced to observe occurrence conditions and characteristic of fruit body and one strain having good conditions and characteristics is secondarily selected. A fruit body produced without aging step is obtained from the secondarily selected strains. Selection of the fruit body is repeated to provide a finally selected strain having good shape of fruit body and high yield. The finally selected strain is repeatedly cultured to obtain the new strain of Lyophyllum ulmarium in which period from inoculation of strain to harvest of fruit body is 50-75 days, and the occurrence, the character and the yield are stabilized.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a novel strain of Bunashimeji (Agaricaceae: Shirotamagitake). More specifically, the present invention, in artificial cultivation,
The present invention relates to a new strain of Bunashimeji having a period from inoculation to fruiting body harvesting of 50 to 75 days.

[0002]

2. Description of the Related Art Conventionally, artificial cultivation of Bunashimeji has been carried out in a medium in which sawdust and nutrients such as rice bran are mixed, and the cultivation technique has been established. However, conventionally used strains require at least 85 days from inoculation to harvest, and usually require around 100 days. The current situation is that when the culture period is shortened, the yield is extremely reduced.

[0003]

DISCLOSURE OF THE INVENTION An object of the present invention is to provide a new strain of Bunashimeji, which makes it possible to shorten the period from inoculation to harvest of Bunashimeji in view of the above situation. It is a further object of the present invention to provide a method for culturing a mycelium and a method for cultivating a fruit body of the new Bunashimeji strain.

[0004]

Means for Solving the Problems The artificial cultivation of Bunashimeji consists of three steps: fungal turning, ripening and development. Conventional strains require 30 to 40 days for fungus turnover, 30 to 60 days for ripening, 20 to 25 days for development, and the total cultivation period is about 80 to 125 days. Here, it is very difficult to reduce the period of the bacterial turnover and generation process. Therefore, the present inventors have thought of shortening the cultivation period by shortening the ripening process period, and have found and created a new strain by the following procedure using a method of selecting a natural mutant strain. (1) Approximately 1,000 strains from all over the country
Strains were collected and tissue separated. (2) The mycelium of each strain is cultured in a liquid medium, and the cellulase activity produced in the culture solution is measured over time. A strain having a high cellulase activity from the initial stage of culture is a strain capable of shortening the maturation period. It was presumed that there was, and four strains presumed to have a high possibility of shortening the ripening period were selected primarily. (3) Repeat the cultivation test on the 4 primary selected strains,
Observations were made on the occurrence status, the characteristics of the fruiting bodies, and the like, and these favorable ones were selected. As a result, one strain that hardly required the ripening step and had a good occurrence was selected secondarily. (4) As a result of repeating the cultivation of the secondary selected bacterial strain, fruiting bodies which are generated even without a maturing step were obtained. The tissue separation of this fruiting body was repeated several times and the selection was repeated.As a result of intensive studies, a strain with a good fruiting body shape and high yield appeared.
This was the final selection. (5) The last selected strain (line) was repeatedly cultivated, and it was confirmed that generation, traits, and yield were stable without a ripening step, and a new strain was found. The characteristics of the fruiting bodies and spores of the new strain thus obtained are as follows.

[0005] The fruiting body is crowded, the umbrella is 3.0 to 10 cm in diameter, circular or irregular and bun-shaped, the surface is smooth, wet, dark gray-brown to gray-brown, and the center of the umbrella is a fine marbled mottled pattern. There is. The meat is white and powdery. The folds are white and dense, the stem is eccentric or central 3-8 × 1-2 cm in the umbrella, white, and a little soft hair is found at the root. The spores are spherical, smooth, colorless, 4-5.5 × 3.5-4.5 μm, and the spores are white.

[0006] The above characteristics are described in "Primary Color Japanese New Fungi Illustration Book I", edited by Rikuya Imaseki and Tsugio Hongo, 1988, Hoikusha (Tokyo)
It is clear that this bacterium is Bunashimeji as compared with the description in the above. This new strain is Bunashimeji Nagano Agricultural Research Institute B-1
And deposited at the National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary under the accession number FERM P-18338 on May 22, 2001.

[0007] Next, as a judgment of the difference between the new strain of Bunashimeji and another strain of Bunashimeji, based on the taxonomic fact that the hyphae are different hyphae if the genetic factors possessed by both hyphae are different from each other. Factor differences were investigated by confronting culture on agar medium. The Bunashimeji strains tested were cultivated commercial varieties, “Hokuto Industrial Production” and “Treasure Flower M”
8171 "," Treasure Flower K0259 ", and" Hakata Bunashimeji ". The results are shown in Table 1 (+ when banding occurs,
If not-).

[0008]

[Table 1]

[0009] As shown in Table 1, each of the above-mentioned strains produced all bands in the confrontation culture with the new strain, which clearly shows that the new strain is a new strain.

The culture and mycological properties of the new strain are as follows. (1) Malt extract agar medium (25 ° C.) Vigorous growth is exhibited, and mycelial elongation for 15 days is 44 mm, and a large number of white, dense mycelial and aerial mycelia are produced. (2) Potato-glucose agar medium (25 ° C.) Vigorous growth is exhibited, mycelial elongation for 15 days is 41 mm, and a large number of white, dense mycelia and aerial mycelia are generated. (3) Optimal growth temperature A seed bacterium having a diameter of 5 mm was inoculated on a potato-glucose agar medium and cultured at 5 ° C intervals between 5 ° C and 35 ° C. When the hyphal elongation was measured 15 days later, the culture temperature at which the elongation was the fastest was 25 ° C. At 35 ° C., it hardly grew. (4) Optimum growth pH Potato-glucose agar medium was adjusted to a pH of 4.0 to 8.0 at intervals of 0.2, and a 5 mm diameter inoculum was inoculated.
Incubated at ℃. When the hyphal elongation was measured after 15 days,
The optimal growth pH was between 5.6 and 6.6. Medium with a pH higher than 7.0 grew extremely slow.

Next, the feature of the present strain that has a short ripening period will be described. The new strain has a high yield and is about the same as Takara No. 2 (Takara Hana M8171) and has a similar yield.
59) higher. Further, if the ripening days are short in the conventional strains, the generation of fruiting bodies is delayed, especially in a high nutrient medium (a medium in which several kinds of materials such as bran, beech, corn cob meal, etc. are added in addition to rice bran), or at all. Failures such as no occurrence, different growth speeds for different bottles, different shapes, etc. occur. However,
The new strain stably reproduces good fruiting body development even in a high nutrient medium even if the culture days are short.

Table 2 shows the yields by aging days. The new strains tend to have a good outbreak regardless of the length of maturation and have a constant yield. On the other hand, treasure flower M8171 appears to have started from the aging days of 21 days, but the commercial value is recognized after the 35th day. In addition, treasure flower K0259 requires more aging days, and its generation stabilizes after the 49th day.

[0013]

[Table 2]

The test conditions are as follows. Cedar sawfish 1
00g, rice bran 60g, bran 20g, mamekawa 20
g, corn cob meal 30 g, TAKARA CLEAN 2.5 g
Was mixed well, and tap water was added thereto to adjust the water content to 63%. The medium was pressed into 32 850 ml polypropylene cultivation bottles, facing downward from the center of the mouth of the bottle and having a diameter of 2 cm.
After drilling holes, they were stoppered with cultivation caps. The culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 30 minutes, and then inoculated with a new strain of Bunashimeji and a conventional inoculum. When the culture medium was cultured for 30 to 35 days in a dark place at 23 ° C. and a humidity of 60%, the fungal turnover was completed. Next, the culture medium is aged for the number of days indicated in the test plot, and after completion of the maturation, the bacteria are scraped (generation treatment).
The fruiting body primordium is formed in an environment at 15 ° C., a humidity of 95% or more, and an illuminance of 100 lux, and then the illuminance is further increased to 1000
Luxed to form fruiting bodies. The fruiting bodies were harvested at the stage when the umbrella was opened for 8 minutes after the germ scraping 20 days later.

[0015]

EXAMPLES Examples of artificial cultivation of the new strain of Bunashimeji according to the present invention will be described below, but the present invention is not limited only to the scope of the following examples. Example 1 100 g of cedar sawfish and 95 g of rice bran were mixed well, and a medium whose water content was adjusted to 63% by adding tap water was pressed into 32 850 ml polypropylene cultivation bottles, and the pressure was lowered below the center of the bottle mouth. After drilling a hole with a diameter of 2 cm opposite, it was stoppered with a cap exclusively for cultivation. The culture medium was autoclaved at 120 ° C. for 30 minutes, and then inoculated with a seed of a new strain of Bunashimeji, and as a control, Takara No. 3 was inoculated. When the culture medium was cultured for 30 days in a dark place at 23 ° C. and a humidity of 60%, the turnover was completed. Next, the culture medium is scraped (generation treatment),
The fruiting body primordium is formed in an environment at 15 ° C., a humidity of 95% or more, and an illuminance of 100 lux, and then the illuminance is further increased to 1000.
Luxed to form fruiting bodies. After 20 days after scraping, fruiting bodies were harvested when the umbrella was opened for 8 minutes. The fruiting bodies of the new strain were all harvested on the 20th and 21st days after the scraping, and the average yield was 117 g / bin. On the other hand, in the case of Takara No. 3 bacteria, those that did not form a primordia were half or more of the bottles, and even those that did form a primordia showed poor growth.

Example 2 (Test in high nutrient medium) 100 g of Japanese cedar, 60 g of rice bran, 20 g of bran, 20 g of mameka, 20 g of corn cob meal, and 2.5 g of Takara clean are mixed well, and tap water is added to determine the water content. The medium adjusted to 63% was pressed into 32 850 ml polypropylene cultivation bottles, a hole having a diameter of 2 cm was made downward from the center of the mouth of the bottle, and stoppered with a cap exclusively for cultivation. The culture medium was autoclaved at 120 ° C. for 30 minutes and inoculated with a seed of a new strain of Bunashimeji.
No. Bacteria was inoculated. When the culture medium was cultured for 35 days in a dark place at 23 ° C. and a humidity of 60%, the bacterial turnover was completed. Next, the culture medium is scraped (generation treatment), and the temperature is 15 ° C.
As described above, the fruit body primordium was formed in an environment having an illuminance of 100 lux or less, and then the illuminance was further increased to 1000 lux to form a fruit body. After 20 days after scraping, fruiting bodies were harvested when the umbrella was opened for 8 minutes. The fruiting bodies of the new strain were all harvested on the 20th and 21st days after the scraping, and the average yield was 2
It was 17 g / 1 bottle. On the other hand, in the case of Takara No. 3 bacteria, those that did not form primordia were 3/4 or more bottles, and even those that did form primordia showed poor growth.

[0017]

According to the present invention, a new strain of Bunashimeji can be obtained in which the period from inoculation of fungi to harvest of fruiting bodies is 50 to 75 days in artificial cultivation. The new strain of Bunashimeji in the present invention has good yield and does not require a ripening period,
Since the cultivation period is significantly shorter than the conventional 85 to 100 days, it is extremely economically advantageous.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Rikiya Tokoo 787-1 Osaka Suzaka, Suzaka City, Nagano Prefecture F-term in the Nagano Prefectural Rural Industry Research Institute (reference) 2B030 AA05 AD06 CB02 CD03 CD07 CD10 CD28 4B065 AA71X AC12 AC13 BA22 BB26 BC41 CA60

Claims (3)

[Claims] 1. A new Bunashimeji strain having a period of 50 to 75 days from inoculation of a bacterial species to harvesting of fruiting bodies in artificial cultivation. 2. The method for culturing a mycelium of a new fungus of Bunashimeji according to claim 1, wherein the mycelium is produced by inoculating a species of the new fungus of Bunashimeji according to claim 1 into a culture medium. 3. The method for cultivating the fruiting body of a new strain of Bunashimejiji according to claim 1, wherein the seed of the new strain of Bunashimejiji according to claim 1 is inoculated into a medium to form fruiting bodies.