Grifola frondosa strain and application thereof

Abstract

The invention belongs to the field of microorganisms, and relates to a grifola frondosa strain and application thereof. The invention provides a novel grifola frondosa strain (Yao ash No.1) which is obtained by brand new separation and successfully preserved, is an excellent strain which is obtained by separating, purifying, qualifying and domesticating sporocarp samples collected from Mengshan County of Guangxi province and has production and application values, and can be normally cultivated and fruiting in Guangxi province. And the sporocarp gradually becomes mature light gray from purple gray which is not too mature. Pilei gradually stretch along with growth and are reversely rolled after maturing, the edges are thin and zigzag, and stipes are short. The strain is suitable for growth, development and cultivation of various raw materials such as weed tree sawdust, ramulus mori scraps and cottonseed hulls, has a good cultivation effect, and is an excellent strain with popularization application value.

Description

Technical Field

[0001] The present application belongs to the field of microorganisms and relates to a Grifola frondosa strain and its application. Background Art

[0002] Grifola frondosa (Dicks. Fr.) SF Gray) belongs to the Basidiomycota, Hymenomycetes, Aphyllomycetes, Polyporaceae, and Genus Grifola. It is also known as Polyporaceae, Thousand Buddha Mushroom, Chestnut Mushroom, Cloud Mushroom, and Lotus Mushroom. It is known as Maitake Mushroom in Japan and Chicken of the Woods in the United States. Grifola frondosa has a crisp, tender texture and a flavor similar to shredded chicken. It can be cooked into a variety of delicious dishes and contains numerous bioactive substances. It is a highly valuable edible and medicinal mushroom, particularly in Japan, Russia, North America, and China's Changbai Mountain area, Sichuan, Yunnan, Zhejiang, and Fujian.

[0003] Currently, reports of maitake mushrooms in Guangxi are rare. Typically, they are more yellowish-brown or dark gray in color. Furthermore, maitake mushrooms are not suitable for cultivation in Guangxi's hot summers and high temperatures. Therefore, researchers are hoping to find a strain of maitake mushrooms that can be cultivated in these hot summers and high temperatures. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this application provides a strain of Grifola frondosa (Yaohui No. 1) that can grow in Guangxi and differs from existing Grifola frondosa strains in color and appearance, making it a Grifola frondosa strain with potential application value. The specific details are as follows:

[0005] A strain of Grifola frondosa (Grifola frondosa) was deposited on March 22, 2024, at the General Microbiology Center of the China Culture Collection Administration (address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing) with the accession number CGMCC No. 41271. This strain, obtained on April 3, 2022, from a fruiting body sample collected from No. 110.43, 24.02, 513 m, Guqian Village, Liuluo Township, Xiayiyao Township, Mengshan County, Guangxi Province, was isolated, purified, characterized, and domesticated to obtain a superior strain with production and application value.

[0006] Furthermore, the described strain of Grifola frondosa has a purple-gray fruiting body, significantly different from the fruiting bodies of existing Grifola frondosa varieties (which are primarily gray, with a small amount of white). This is a brand-new strain of Grifola frondosa, offering excellent ornamental value and potential for development as a high-end food ingredient, better meeting the diverse needs of different consumers and possessing great research and application value.

[0007] Furthermore, the Grifola frondosa strain includes spores, mycelium, and fruiting bodies of the strain.

[0008] The described Grifola frondosa strain is used in food production, pharmaceutical preparation, and agricultural cultivation. The flesh of the Grifola frondosa strain described in this application is tender and delicious, making it a never-ending favorite. Its nutritional benefits are excellent and its medicinal value is high. As a traditional Chinese medicine, it can also be used to invigorate Qi and strengthen the spleen, tonify deficiency, and strengthen the body's health. It is used to treat spleen deficiency, weak Qi, fatigue, lethargy, decreased appetite, and abdominal distension after eating. It is also used to treat cancer patients experiencing these symptoms after radiotherapy or chemotherapy.

[0009] Furthermore, the Grifola frondosa strain includes spores, mycelium, fruiting bodies, and extracts of the Grifola frondosa strain.

[0010] The cultivation method of the Grifola frondosa strain controls the mycelium growth temperature at 25-28 DEG C, the mushroom growing temperature at 18-25 DEG C, the mushroom growing room (shed) temperature at 19-22 DEG C, the air humidity at 80%-90%, the CO2 concentration at below 0.2%, and provides 300lx-500lx of light during the day.

[0011] Furthermore, the culture medium used contains one or more of broad-leaved trees, mulberry branches, cottonseed hulls, and wheat.

[0012] Beneficial effects of this application:

[0013] (1) This application presents a new strain of Grifola frondosa (Yaohui No. 1) that was newly isolated and successfully preserved.

[0014] (2) The new strain of Grifola frondosa (Yaohui No. 1) given in this application is an excellent strain with production application value obtained by separation, purification, characterization and domestication of a fruiting body sample collected from Guqian Village, Liuluo Township, Xiayi Yao Township, Mengshan County, Guangxi on April 3, 2022, and can be normally cultivated and produce mushrooms in Guangxi.

[0015] (3) The new strain of Grifola frondosa (Yaohui No. 1) given in this application was collected from the Dayao Mountains in Guangxi, located in Guqian Village, Liuluo Village, Xiayi Township, Mengshan County. The color of its fruiting body is purple-gray. The cap gradually stretches as it grows, curls up after maturity, has thin, serrated edges, and a short stipe. The fruiting body is coral-like and branched, with fan-shaped, spoon-shaped, and tongue-shaped caps at the end overlapping into clusters, stacked like chrysanthemums. The largest single plant is 20 to 35 cm wide and weighs 1.5 to 2.0 kg ( Figure 1-3 ).

[0016] (4) The new strain of Grifola frondosa (Yaohui No. 1) given in this application is suitable for growth and cultivation on a variety of raw materials such as sawdust, mulberry branch chips, cottonseed hulls, etc., and has good cultivation effects. It is an excellent strain with promotion value. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a photo of the new strain of Grifola frondosa (Yaohui No. 1) growing in the wild.

[0018] Figure 2 This is a photo of the new strain of Grifola frondosa (Yaohui No. 1) growing in the wild.

[0019] Figure 3 This is a photo of the new strain of Grifola frondosa (Yaohui No. 1) growing in the wild.

[0020] Figure 4 The following is a photo of the growth of the new strain of Grifola frondosa (Yaohui No. 1) on PDA slant culture medium.

[0021] Figure 5 The following are photos of the growth and reproduction test of the new strain of Grifola frondosa (Yaohui No. 1) on different raw materials.

[0022] Figure 6 The picture shows the results of antagonism test between the new strain of Grifola frondosa (Yaohui No. 1) and three strains of Grifola frondosa introduced from Hebei and Fujian.

[0023] Figure 7 It is the electrophoresis pattern, in which 8 is Yaohui No. 1.

[0024] Figure 8 This is the identification result of Yaohui No. 1. Yaohui No. 1 (B1) was identified as a strain of Grifola frondosa.

[0025] Figure 9 This is the developmental tree of Yaohui No. 1 (B1). DETAILED DESCRIPTION

[0026] The present application will be further described in detail below with reference to specific embodiments, which are intended to explain rather than limit the present application.

[0027] The invention relates to a Grifola frondosa plant which can utilize starch as a raw material, and belongs to the technical field of edible and medicinal fungi.

[0028] 1. Relevant information of the strain: including function, classification, properties, etc.

[0029] ①Classification status: Grifola frondosa (GF), also known as Polypore, belongs to the Eumycota,

[0030] Basidiomycotina, Hymenomycetes, Homobasidiomycetes, Aphyllophorales, Polyporaceae, Genus Echinops.

[0031] ② Characteristics (including fruiting body morphology, mycelium culture characteristics, biological characteristics, cultivation characteristics, etc.):

[0032] a. Fruiting Body Morphological Characteristics: The fruiting bodies of maitake mushrooms collected from the Dayao Mountains in Guangxi, located in Guqian Village, Liuluo Village, Xiayi Township, Mengshan County, gradually change from a purple-gray color to a light gray color when mature. The cap gradually expands as it grows, curling back at maturity, with thin, serrated edges and a short stipe. The fruiting bodies are generally coral-like and branched, with fan-shaped, spoon-shaped, and tongue-shaped caps at the ends overlapping in clusters, resembling chrysanthemums. The largest individual clusters are 20 to 35 cm wide and weigh 1.5 to 2.0 kg ( Figure 1-3 ).

[0033] b. Mycelial Culture Characteristics: The growth rate of Yaohui No. 1 strain on PDA slant medium is approximately 0.43 cm / day. The mycelium is slender, white, and fuzzy. As the mycelium spreads, dense aerial hyphae gradually fill the entire slant medium and extend toward the tube wall, forming a white cotton-like shape. The matrix hyphae are white, thin, and easy to tear and can be easily separated from the medium. Figure 4 ).

[0034] c. Growth and reproduction test on different raw materials ( Figure 5 )

[0035]

[0036]

[0037] d. Antagonism test

[0038] Antagonistic tests were conducted with three Grifola frondosa strains introduced from Hebei and Fujian. The results showed that all strains produced significant antagonistic reactions ( Figure 6 ).

[0039] e. Identified by BGI as a strain of Grifola frondosa

[0040] Yaohui No. 1 (B1) was identified as a strain of Grifola frondosa ( Figure 7-9 )

[0041] f. Cultivation

[0042] It is mainly used for artificial bag cultivation in spring and autumn or year-round factory cultivation. The optimum temperature for mycelial growth is 25-28°C, and the optimum temperature for fruiting is 18-25°C. The temperature in the fruiting room (shed) should be controlled at 18-25°C, the air humidity at 90%-95%, the CO2 concentration below 700ppm, and 200-500lx of light should be provided during the day.

[0043] ③Functions:

[0044] Grifola frondosa, commonly known as "maitake mushroom," is a edible and medicinal mushroom. Its graceful, layered appearance resembles a chrysanthemum; its refreshing aroma is refreshing and invigorating; and its tender, tender flesh is a delight to savor. It combines nutritional and health benefits with high medicinal value. Containing numerous bioactive substances, it possesses diverse medicinal properties, such as invigorating qi and strengthening the spleen, tonifying deficiency and strengthening the body's health, fighting tumors, lowering cholesterol, providing antioxidant benefits, and treating diabetes. Grifola frondosa is not only a highly nutritious food source, but also boasts a 1709 record in Kaihara Ekiken's "Dahe Bencao." Its medicinal value was first documented in Sakaran's "Jiyupu," which describes it as "sweet, mild, non-toxic, and capable of prolonging life." In recent years, it has become popular in markets such as Japan and Singapore as a health food.

[0045] The Grifola frondosa strain of the present invention is obtained by screening, domesticating and cultivating wild Grifola frondosa from Guangxi. The screening and domestication methods are as follows:

[0046] (1) Five fruiting bodies of Grifola frondosa were collected from the Dayao Mountains of Guangxi (110°43'\24°02'\513m) in Guqian Village, Liuluo Village, Xiayi Township, Mengshan County. One pure strain was obtained by tissue separation and purification. The strain was identified as Grifola frondosa strain by BGI and used as the mother strain HSH-1 for acclimation experiments.

[0047] (2) The mother strain LS-1 was cultivated and domesticated in artificial bags, and 30 strains with fast and dense mycelial growth, strong resistance to weeds, good fruiting performance, and high bioconversion rate were selected based on the mycelial growth trend, resistance to weeds, suitability to feed, fruiting performance, and bioconversion rate. These strains were recorded as the F1 generation.

[0048] (3) The F1 generation strains were acclimated and screened in artificial bag cultivation. Referring to all the indicators in (2), 15 better strains were selected and recorded as F2 generation;

[0049] (4) The F2 generation strains were domesticated and screened by artificial bag cultivation. Referring to all the indicators in (2), 7 excellent strains were screened and recorded as F3 generation;

[0050] (5) The F3 generation strains were subjected to artificial fermentation materials, sunbed covering soil, and domestication and screening in different seasons. Referring to all the indicators in (2), one excellent strain was selected, which had dense, white, cotton-like aerial mycelium, white, thin, and easy-to-tear matrix mycelium, strong anti-weed ability, good mushroom production performance, and a bioconversion rate of 35% to 45% in the first wave of mushrooms. It was named "Yaohui No. 1" after the territorial jurisdiction.

[0051] Wherein, in step (1) to step (5), the tissue separation and purification method of the fruiting body is as follows: select a healthy and disease-free wild maitake mushroom fruiting body, wipe the surface of the maitake mushroom fruiting body lightly with gauze soaked in 75% Vol alcohol solution, put it into a sterilized culture dish and place it in a clean bench, turn on the ultraviolet lamp and the fan for 30 minutes, turn off the ultraviolet lamp, use tweezers, a scalpel and scissors that have been burned and cooled by an alcohol lamp to cut the internal tissue block of about 2mm×2mm at the junction of the handle and cap, insert it into a PAD slant culture medium without water accumulation, and culture it at a constant temperature of 25-28°C until the mycelium grows to 1 / 2-1 / 3 of the slant, pick the frontmost mycelium, and transfer it to a fresh slant culture medium for culture, and continue to pick the frontmost mycelium and transfer it to a fresh slant culture medium for culture in the same way, and repeat this method of selection and transplantation and combine it with microscopic examination until a pure strain is obtained. The test tube slant culture medium comprises: 1000 ml of 20% peeled potato juice, 20 g of glucose, 3 g of potassium dihydrogen phosphate, 1.5 g of magnesium sulfate, 15-17 g of agar powder, and a natural pH value.

[0052] Among them, in step (1) to step (5), the cultivation and domestication method of Grifola frondosa adopts artificial fermentation material and sunbed soil covering cultivation, and the cultivation steps are as follows:

[0053] (1) Preparation of fungus bags: Use 17cm×33cm×0.05mm low-density polyethylene edible fungus bags and place the culture material into the bags according to the conventional method to make fungus bags. Then sterilize: When using normal pressure steam sterilization, the center material temperature should reach above 98℃ within 4-5 hours after the cold air is exhausted and continue to maintain it for 12-15 hours; when using high-pressure steam sterilization, after the cold air is exhausted, the pressure reaches 0.15MPa and the material temperature is maintained at 126℃ for 3-3.5 hours. After pre-cooling and forced cooling to a material temperature of 25-30℃, the sterilized fungus bags are sent to the clean inoculation room and inoculated according to the requirements of the aseptic operation procedures.

[0054] The planting material consists of the following components in parts by weight: 0-78 parts of miscellaneous wood chips, 0-88 parts of mulberry branch chips, 0-90 parts of cottonseed hulls, 7-15 parts of wheat bran, 0-3 parts of corn flour, 0.5-1 part of gypsum and 0.5-1 part of lime.

[0055] (2) Mycelium Culture: After inoculation, the mushroom bags are promptly brought into the culture room and placed on shelves. The indoor environmental parameters are: temperature 22-25°C, humidity 55-65%, dim light, and fresh air. The culture time is 40-45 days. When the surface gradually changes from white to grayish white to purple-gray and secretes yellow water droplets, the fruiting stage can begin.

[0056] (3) Fruiting management: This mainly involves coordinating and controlling various management factors such as temperature, ventilation, humidity, and light. After the mushroom bags are moved into the fruiting room, use a sterilized scalpel to cut vertically and evenly in the plastic below the bag collar to form a circular hole with a diameter of about 2 cm. The temperature in the fruiting room should be maintained at 18-22°C, the humidity at 90%-95%, the illumination at 200-500 lx, and the carbon dioxide concentration at no more than 700 ppm. About 7 days after the cut, a white protrusion will appear at the cut, which is the primordium of the maitake mushroom. As the fruiting body differentiates and grows, spray the fruiting body with small amounts of water multiple times. Do not apply too much water at one time. After 15-20 days of fruiting management, the fruiting body will mature.

[0057] (4) Harvesting: When the outer edge of the fan-shaped cap of the maitake mushroom has no white growth end, the cap is flat, the color is light purple-gray, the whole mushroom looks like a blooming lotus, and emits a strong mushroom fragrance, it can be picked. 1 to 2 days before harvesting, do not directly spray water on the mushroom body. When harvesting, wear gloves, hold the mushroom bag with one hand, and reach the base of the fruiting body with the other hand. Hold the mushroom body and fold it upwards to pick the mushroom body, or use a knife to cut it directly from the base of the maitake mushroom. After harvesting, the maitake mushroom can be stored in a cold storage for sale, or it can be made into a dried product.

[0058] Finally, it should be noted that the above embodiments are merely representative examples of this application. Obviously, the technical solutions of this application are not limited to the above embodiments and are subject to numerous variations. All variations that can be directly derived or conceived by a person of ordinary skill in the art from the disclosure of this application should be considered within the scope of protection of this application.

Claims

1. A Grifola frondosa strain, characterized in that It is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration with the deposit number CGMCC No.41271.

2. The Grifola frondosa strain according to claim 1, characterized in that The fruit body color of the Grifola frondosa strain is purple-grey.

3. The Grifola frondosa strain according to claim 1, characterized in that The Grifola frondosa strain comprises spores, mycelium and fruiting bodies of the strain.

4. Use of the Grifola frondosa strain according to claim 1 in food production.

5. Use of the Grifola frondosa strain according to claim 1 in the preparation of medicines.

6. Use of the Grifola frondosa strain according to claim 1 in agricultural cultivation.

7. The use according to claims 4, 5 and 6, characterized in that: The Grifola frondosa strain comprises spores, mycelium, fruiting bodies and extracts of the Grifola frondosa strain.

8. The method for cultivating the Grifola frondosa strain according to claim 1, characterized in that: The mycelium growth temperature is 25-28℃, the ear temperature is 18-25℃, the temperature of the mushroom room (shed) is controlled at 19℃-22℃, the air humidity is 80%-90%, the CO2 concentration is controlled below 0.2%, and 300lx-500lx of light is provided during the day.

9. The cultivation method of Grifola frondosa strain according to claim 8, characterized in that: The culture medium used contains one or more of broad-leaved trees, mulberry branches, cottonseed hulls and wheat.

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