High-temperature-resistant lepista sordida strain and application thereof

Abstract

The invention belongs to the field of microorganisms, and relates to a high-temperature-resistant lepista sordida strain and application thereof. The invention provides a brand-new lepista sordida new strain (Yao xiang mill No.1) which is obtained through separation and successfully preserved, and the lepista sordida can grow and fruit under a high-temperature condition, so that the lepista sordida can still be normally cultivated and fruit at a high-temperature period in Guangxi regions, especially regions around Nanning. The fungus is rich in nutrition, delicious in taste and rich in protein, 18 kinds of amino acid and other nutrient substances, not only is a good dining table item, but also has the effects of oxidation resistance, bacteria resistance, tumor resistance and the like, is rich in selenium, germanium, iron, zinc, calcium and other microelements, polysaccharide and trehalose (higher than shiitake mushrooms, lucid ganoderma and the like), and is an edible and medicinal fungus with high commercial value and medicinal research and development potential.

Description

Technical Field

[0001] The present application belongs to the field of microorganisms and relates to a high temperature resistant strain of Oyster mushroom and its application. Background Art

[0002] Lepista sordida (Fr.) Singer is a fungus of the family Tricholomataceae and the genus Lepista. The cap is hemispherical when young, then flattened, translucent or water-soaked when wet. The margins are involute, with inconspicuous stripes, often wavy or petal-shaped. The flesh is pale violet and thin. The gills are straight, sometimes slightly curved or slightly extended. Pale purple. The stipe is violet, solid, and mostly curved at the base. The basidiospores are broadly elliptical to oval, rough to pitted, and colorless.

[0003] It is usually distributed in Inner Mongolia, Heilongjiang, Henan, Gansu, Qinghai, Sichuan, Xinjiang, Tibet, Shanxi, Jilin, etc. In summer and autumn, it grows in groups or near clusters on hillside grasslands, grasslands, vegetable gardens, village roadsides, burned land, compost, etc.

[0004] At present, there are few reports of flower-faced shiitake mushrooms in Guangxi. This is because flower-faced shiitake mushrooms belong to mesophilic edible fungi. The suitable temperature for mycelium growth is 5-26℃, and the best is 18-22℃; the mycelium growth slows down significantly when the temperature is lower than 15℃ or higher than 24℃, and the mycelium growth stops when the temperature is higher than 30℃. If the temperature is above 30℃ for a long time, the mycelium will autolyze. The suitable temperature for the growth and development of fruiting bodies is 5-22℃, and the best is 12-17℃; it cannot produce mushrooms when the temperature is lower than 3℃ or higher than 25℃. Although some flower-faced shiitake mushrooms can grow and produce mushrooms above 26℃, the temperature they can accept does not exceed 30℃, and they cannot grow and produce mushrooms above 30℃. Therefore, it is rarely reported in Guangxi, and it is not suitable for cultivation in the hot summer and high temperature areas of Guangxi. Therefore, researchers hope to find a flower-faced shiitake mushroom strain that can be cultivated in the hot summer and high temperature areas of Guangxi. Summary of the invention

[0005] In view of the shortcomings of the prior art, the present application provides a high temperature resistant strain of the lily mushroom (Yaoxiangmo No. 1), which can grow at temperatures above 30°C and is a strain of the lily mushroom with potential value for promotion and application. The specific contents are as follows:

[0006] A thermostable strain of Lepista sordida (Yaoxiangmo No. 1), which was deposited in the General Microbiological Center of China Microbiological Culture Collection Administration (address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing) on ​​March 22, 2024, with a deposit number of CGMCC NO.41272. It was isolated from 110 Changtan Village, Changer Village, Jinxiu Town, Jinxiu County, Guangxi on June 10, 2022. 13 ， \24.9 ，The fruiting body samples collected from N\1153m were separated, purified, characterized and domesticated to obtain excellent strains with production application value.

[0007] Furthermore, the heat-resistant Oleander strain grows well in an environment of 28-32°C.

[0008] Furthermore, the heat-resistant Oleander strain includes spores, mycelium and fruiting bodies of the strain.

[0009] The high temperature resistant shimeji mushroom strain is applied in food production, drug preparation, agricultural planting and the like. Shimeji mushroom is rich in nutrition, and contains 2.9 grams of protein, 0.2 grams of fat, 3 grams of carbohydrates, 0.6 grams of crude fiber, 8 milligrams of calcium, 6.6 milligrams of phosphorus, 1.3 milligrams of iron, 4 milligrams of vitamin C, 3.3 milligrams of niacin, sodium, potassium, manganese, copper, zinc, fluorine, iodine, tyrosine, vitamin A, vitamin B, vitamin D, vitamin E, vitamin K, 5-adenosine phosphate and various amino acids, such as threonine, aspartic acid, leucine, alanine, hydroxylysine, etc., and contains non-specific plant lectins, etc., and can be fully applied in food production, drug preparation, agricultural planting and the like.

[0010] Furthermore, the strain includes its spores, mycelium, fruiting bodies, and extracts.

[0011] The method for cultivating the high-temperature-resistant shimeji mushroom strain controls the temperature in the range of 28-32°C.

[0012] Furthermore, the culture medium used is: 200 g of potato (peeled), 20 g of glucose, 3.0 g of potassium dihydrogen phosphate, 1.5 g of magnesium sulfate, 10 mg of vitamin B1, 1000 ml of water, and natural pH.

[0013] Beneficial effects of this application:

[0014] (1) This application provides a new strain of Oleander (Yaoxiangmo No. 1) that was newly isolated and successfully preserved.

[0015] (2) The new strain of the flower-faced shimeji (Yaoxiangmo No. 1) given in this application is an excellent strain with production and application value obtained by separation, purification, characterization and domestication of the fruiting body sample collected from 110.13, \24.9, N\1153m, Changtan Village, Changer Village, Jinxiu Town, Jinxiu County, Guangxi on June 10, 2022. It can grow and produce mushrooms under high temperature conditions, so that Guangxi, especially the surrounding areas of Nanning, can still be cultivated and produce mushrooms normally during high temperature periods. However, the current flower-faced shimeji stops growing mycelium above 30°C, and if it is above 30°C for a long time, the mycelium will autolyze.

[0016] (3) The new strain of the flower-faced shiitake mushroom (Yaoxiangmo No. 1) given in this application was collected from the hinterland of Dayao Mountain in Jinxiu, Guangxi in 2022. When young, it is hemispherical, purple or light red diamond-shaped ( Figure 1-4 ), as the cap gradually unfolds, there is an obvious bulge in the middle, and the color gradually fades from the inside to the outside, and becomes water-soaked ( Figure 2 , 3 4). After being flattened, the flesh is thin, fragile, light purple, with the same color as the stem and cap, and has a fragrant smell. After losing water, the color gradually fades to grayish white. Figure 5 , 6 ), with involute margins, indistinct striations, often wavy or petal-shaped ( Figure 5 ). The cap diameter is 2cm to 9cm, the cap thickness is 0.2cm to 0.3cm, the stipe is 2.5cm to 5.5cm long, the diameter is 0.3cm to 0.9cm, solid, and the base is mostly curved. The gills are straight ( Figure 7 ) or bend into a corrugated shape ( Figure 8 ), sparse, lavender. Grown in groups or nearly in clusters or singly in fields from early summer to summer.

[0017] (4) The new strain of Shiitake mushroom (Yaoxiangmo No. 1) provided in the present application is rich in nutrition, delicious, rich in protein and 18 kinds of amino acids and other nutrients. It is not only a good table delicacy but also has antioxidant, antibacterial, anti-tumor and other effects. It is rich in trace elements such as selenium, germanium, iron, zinc, calcium, polysaccharides, and trehalose (higher than shiitake mushrooms, ganoderma lucidum, etc.). It is an edible and medicinal fungus with high commercial value and medicinal research and development potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a photo of wild shiitake mushrooms growing in a relatively high temperature environment (25-32℃).

[0019] Figure 2 This is a photo of wild shiitake mushrooms growing in a relatively high temperature environment (25-32℃).

[0020] Figure 3 This is a photo of wild shiitake mushrooms growing in a relatively high temperature environment (25-32℃).

[0021] Figure 4 This is a photo of wild shiitake mushrooms growing in a relatively high temperature environment (25-32℃).

[0022] Figure 5 This is a photo of wild shiitake mushrooms growing in a relatively high temperature environment (25-32℃).

[0023] Figure 6 This is a photo of wild shiitake mushrooms growing in a relatively high temperature environment (25-32℃).

[0024] Figure 7 This is a photo of wild shiitake mushrooms growing in a relatively high temperature environment (25-32℃).

[0025] Figure 8 This is a photo of wild shiitake mushrooms growing in a relatively high temperature environment (25-32℃).

[0026] Fig. 9 These are photos of the mycelium growth of Yaoxiangmo No. 1 mycelium in a conventional comprehensive potato culture medium in a relatively high temperature environment (28-32℃).

[0027] Fig.10 These are photos of the mycelium growth of Yaoxiangmo No. 1 mycelium in a conventional comprehensive potato culture medium in a relatively high temperature environment (28-32℃).

[0028] Fig.11 This is the antagonistic reaction between Yaoxiangmo No. 1 and the flower-faced mushroom introduced from Northeast China.

[0029] Fig.12 This is the antagonistic reaction between Yaoxiangmo No. 1 and the flower-faced mushroom introduced from Northeast China.

[0030] Fig.13 It is an electrophoresis spectrum, among which 12 is Yaoxiangmo No. 1, and 13 is the flower-faced fragrant mushroom introduced from Northeast China.

[0031] Fig.14 This is the identification result of Yaoxiangmo No. 1. Yaoxiangmo No. 1 (C1) was identified as a strain of the Cephalexin class.

[0032] Fig.15 It is the development tree of Yaoxiangmo No. 1 (C1). Note: C2 is the flower-faced mushroom introduced from Northeast China. DETAILED DESCRIPTION

[0033] The present application is further described in detail below in conjunction with specific embodiments, which are intended to explain the present application rather than to limit it.

[0034] The invention relates to a pleurotus eryngii which can utilize starch as a raw material, and belongs to the technical field of edible and medicinal fungi.

[0035] 1. Relevant information of the strain: including function, classification, properties, etc.

[0036] ①Classification status: Lepista sordida (Fr.) Singer belongs to the kingdom Fungi, the phylum Basidiomycota, the class Agaricomycetes, the order Agaricales, the family Tricholomatacete, the genus Lepista, and the genus Lepista.

[0037] ② Characteristics (including fruiting body morphology, mycelium culture characteristics, biological characteristics, cultivation characteristics, etc.):

[0038] a. Morphological characteristics of fruiting bodies Flower-faced mushrooms are also known as clove mushrooms, flower-faced mushrooms, purple crystal mushrooms, purple flower-faced mushrooms, etc. They are widely distributed in Northeast China, Northwest China, Inner Mongolia, Central China and other regions, but are rarely reported in Guangxi. The flower-faced mushrooms collected from the hinterland of Dayao Mountain in Jinxiu, Guangxi in 2023 are hemispherical, purple or light red diamond-shaped when young ( Figure 1-4 ), as the cap gradually unfolds, there is an obvious bulge in the middle, and the color gradually fades from the inside to the outside, and becomes water-soaked ( Figure 2 , 3 4). After being flattened, the flesh is thin, fragile, light purple, with the same color as the stem and cap, and has a fragrant smell. After losing water, the color gradually fades to grayish white. Figure 5 , 6 ), with involute margins, indistinct striations, often wavy or petal-shaped ( Figure 5 ). The cap diameter is 2cm to 9cm, the cap thickness is 0.2cm to 0.3cm, the stipe is 2.5cm to 5.5cm long, the diameter is 0.3cm to 0.9cm, solid, and the base is mostly curved. The gills are straight ( Figure 7 ) or bend into a corrugated shape ( Figure 8 ), sparse, lavender. Grown in groups or nearly in clusters or singly in fields from early summer to summer.

[0039] b. Mycelium culture characteristics Yaoxiangmo No. 1 mycelium is cultured in conventional comprehensive potato medium [200g potatoes (peeled), 20g glucose, 3.0g potassium dihydrogen phosphate, 1.5g magnesium sulfate, 10mg vitamin B1, 1000ml water, natural pH.] Culture characteristics: Mycelium is initially white and gradually changes to light purple, dense, round, strong and powerful, with short aerial mycelium ( Fig. 9 , 10 ); its mycelium can still grow above 30℃, and the suitable growth temperature range is 25~28℃.

[0040] Compared with the flower-faced mushroom introduced from Northeast China, it has a significant antagonistic reaction [Note: Individuals with significantly different genetic backgrounds will form a clear dividing line in the junction area, which is an antagonistic reaction. Antagonistic reaction is a specific manifestation of somatic cell incompatibility. It has been used in the preliminary study of fungal kinship because of its rapid, convenient and intuitive advantages. It was once the main means of fungal species identification. ]( Fig.11 , 12 ).

[0041] The new strain of the flower-faced mushroom (Yaoxiangmo No. 1) of the present application is identified by BGI as a species of the flower-faced mushroom ( Figure 13-15 ).

[0042] ③Function:

[0043] The flower-faced shiitake mushroom is rich in nutrients and tastes delicious. It is rich in protein and 18 kinds of amino acids and other nutrients. It is not only a great delicacy on the table, but also has antioxidant, antibacterial and anti-tumor effects. It is rich in trace elements such as selenium, germanium, iron, zinc, calcium, polysaccharides, and trehalose (higher than shiitake mushrooms, Ganoderma lucidum, etc.). It is an edible and medicinal fungus with high commercial value and medicinal research and development potential.

[0044] The cultivar of the present invention is obtained by screening, domesticating and cultivating wild cultivar of the present invention in Guangxi, and the screening and domestication methods are as follows:

[0045] (1) From 110.13, Changtan Village, Changer Village, Jinxiu Town, Jinxiu County, Guangxi ， \24.9 ， Seven wild tricholoma fruiting bodies were collected at N\1153m, and five pure strains were obtained through tissue separation and purification, which were identified as tricholoma strains by BGI.

[0046] (2) Five strains of wild shiitake mushrooms were subjected to strain antagonism tests. After excluding the same strains (no antagonism lines), three different strains with obvious antagonism lines were obtained. Then, mycelium growth rate, strength, feed suitability and other tests as well as mushroom production evaluation tests were carried out. Based on the mycelium growth trend, resistance to impurities, feed suitability, mushroom production performance, and bioconversion rate as indicators, the strains with faster mycelium growth rate, dense and strong, purple color, strong resistance to impurities, wide feed suitability, good mushroom production performance, and higher bioconversion rate were selected as the mother strain LS-1;

[0047] (3) The mother strain LS-1 was subjected to artificial fermentation, soil-covered cultivation in sunbeds, and domestication and screening in different seasons. Referring to the various indicators in (2), 15 strains with excellent indicators, wide temperature adaptability, and good high temperature resistance were selected and recorded as F1 generation;

[0048] (4) The F1 generation strains were subjected to artificial fermentation, soil covering in the sunbeds, and acclimatization and screening in different seasons. Referring to all the indicators in (2), 8 better strains were screened out and recorded as the F2 generation;

[0049] (5) The F2 generation strains were subjected to artificial fermentation, soil covering in the sunbeds, and acclimatization and screening in different seasons. With reference to all the indicators in (2), 5 excellent strains were selected and recorded as the F3 generation;

[0050] (6) The F3 generation strains were subjected to artificial fermentation, soil covering in sunny beds, and domestication and screening in different seasons. Referring to all the indicators in (2), an excellent strain with purple mycelium, dense and strong, thick and large mycelial cords, strong resistance to weeds, and a fresh mushroom yield of more than 6 kg per square meter (equivalent to a bioconversion rate of more than 35%) was selected and named "Yaohui No. 1" after the territorial jurisdiction.

[0051] Among them, in steps (1) to (6), the tissue separation and purification methods of the fruiting bodies are all as follows: select healthy and disease-free wild violet mushroom fruiting bodies, lightly wipe the surface of the black fungus fruiting bodies with gauze soaked in 75% Vol alcohol solution, then put them into a sterilized culture dish and place them in a clean bench, turn on the ultraviolet lamp and the fan for 30 minutes, turn off the ultraviolet lamp, use tweezers, scalpels 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, and 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 front mycelium, and transfer it to a fresh slant culture medium for culture, and continue to pick the front 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 formula of the test tube slant culture medium is: 1000ml of 20% peeled potato juice, 20g of glucose, 3g of potassium dihydrogen phosphate, 1.5g of magnesium sulfate, 15-17g of agar powder, and natural pH.

[0052] Among them, in step (1) to step (6), the cultivation and domestication method of the flower-faced mushroom adopts artificial fermentation materials and sunbed soil-covered cultivation, and the cultivation steps are as follows:

[0053] (1) Raw materials and composting: Flower-faced mushrooms are suitable for cultivation with agricultural and sideline products such as cottonseed hulls, rice straw, wheat straw, sugarcane leaves (dregs), corn cobs (stalks), wheat straw (bean straw), mushroom chaff, cow dung and other raw materials. However, the raw materials must be processed by composting. Method: Mix the raw materials according to the formula components, build a pile (length is not limited, width and height are 1.0-1.2m), make holes, cover with straw mats or films for fermentation. After 3-5 days, when the temperature in the middle of the pile rises to 60℃, keep it for about 24 hours and turn it over. When turning the pile, turn it inside out, up and down, inside out, turn it thoroughly and evenly, build a pile again, make holes, cover with straw mats or films, and keep it for 24 hours when the material temperature rises to 60℃ again. Repeat this 3-5 times. When a large number of actinomycete hyphae are formed in the pile, the fermentation is over. Lime is added in two times, 1 / 2 at first and 1 / 2 at the end of the pile turning. Adjust the moisture content to 60%-65% and the pH value to 6.5-7.

[0054] The cultivation material consists of the following components in parts by weight: 0-70 parts of rice straw, 0-50 parts of mulberry branch chips, 0-45 parts of cottonseed hulls, 0-30 parts of corn cobs (stalks), 0-50 parts of sugarcane bagasse (leaves), 0-45 parts of mushroom chaff, 0-25 parts of wheat straw (bean straw), 15-25 parts of cow dung, 7-15 parts of wheat bran, 2-5 parts of corn flour, 0.5-1.5 parts of gypsum and 1.0-1.5 parts of lime.

[0055] (2) Spreading materials and sowing: Prepare the mushroom bed as required (length is unlimited, width is 1.0-1.1m). Before spreading materials, spray water on the bed to keep the soil on the bed surface moist to prevent the culture medium from losing water, which is not conducive to the fungus. Then, spread the fermented culture medium on the bed. When the material temperature drops below 32℃, sowing can be carried out. Generally, the layer sowing method is used for sowing. The total thickness of the fermentation material is 20-25cm. The seed amount per square meter is about 1-2 bags (15 fungus bags). The sowing method adopts layer sowing + broadcast sowing, that is, sowing in two layers. The first layer is sown in the middle layer of the material, and the second layer is spread on the surface of the material. Cover with black film after sowing.

[0056] (3) Management of spawning: Keep the spawning temperature at 22-28℃, not exceeding 30℃. Open the film on both sides of the greenhouse for ventilation in the morning and evening. 2-3 days after sowing, open the film on the bed for ventilation in the morning and afternoon every day.

[0057] (4) Covering with soil: After 25 to 28 days of mycelium cultivation, when mycelium grows to 15 cm above the material bed and the surface mycelium begins to spit out yellow water, the soil can be covered. Loose, fertile sandy loam is preferred, with a particle size of 2 to 3 cm and a moisture content of 65 to 75%. Appropriate disinfection should be carried out and the thickness of the covering soil should be 3 to 5 cm.

[0058] (5) Management after covering the soil: After covering the soil, keep the temperature at 20-25℃, and remove the film once in the morning and afternoon for ventilation, each time for 15-30 minutes.

[0059] (6) Promoting budding: When the mycelium grows to 2-3 cm above the soil layer, remove the black film covering it, control the relative humidity of the air in the greenhouse to 85%-95%, the temperature to 20-25°C, and the scattered light intensity to 100-500lx, and spray water on the bed 3-5 times a day to maintain the appropriate moisture content of the soil particles. After about 30 days of covering the soil, the mycelium in the soil layer begins to form mycelial bundles, twist into light purple primordia, and swell into small mushroom buds. Stop spraying water on the bed, and then focus on controlling temperature and humidity and enhancing ventilation.

[0060] (7) Management of mushroom production: As the mushroom buds differentiate and grow, the management of mushroom production begins. The water management during the mushroom production period should be determined according to the climate conditions, the density of mushrooms on the bed, and the size of the fruiting bodies. The amount of water sprayed should be enough to keep the covering soil moist. The relative humidity of the air should be maintained at 90% to 98%, and the light intensity should be adjusted appropriately. The droplets of water should be fine and uniform when spraying.

[0061] (8) Harvesting: The fruiting tide of Hualianxiang is not obvious, so it needs to be harvested according to the growth situation. Timely harvesting is one of the key links to ensure quality. The mushroom body is round, the cap diameter is 5-6cm, the edge is slightly curled, the stem is 3-4cm long, and the mushroom body is purple-brown, then it can be harvested. When harvesting, pick the big ones and leave the small ones. Do not spray water on the mushroom bed 3 days before harvesting. When picking, gently twist the stems. Put the picked mushrooms in the transfer box and stack them in 3-4 layers. Do not press them heavily. Fill the holes after picking with soil. The fruiting body of Hualianxiang has low toughness. The more over-mature the fruiting body is, the easier it is to break. If harvested too late, the cap will be flat and curled, the color will become lighter, and over-maturity will greatly reduce its commercial value. At the same time, it is necessary to handle with care during the harvesting process to ensure its quality.

[0062] Finally, it should be pointed out that the above embodiments are only representative examples of the present application. Obviously, the technical solution of the present application is not limited to the above embodiments, and there may be many variations. All variations that can be directly derived or associated with the contents disclosed in the present application by a person of ordinary skill in the art should be considered as the protection scope of the present application.

Claims

1. A thermostable strain of Mushroom, characterized in that: It is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration with the deposit number CGMCC NO.41272.

2. The heat-resistant mushroom strain according to claim 1, characterized in that: The heat-resistant shimeji mushroom strain grows well in an environment of 28-32°C.

3. The heat-resistant mushroom strain according to claim 1, characterized in that: The high temperature resistant Lepidium scaly-leaf strain comprises spores, mycelium and fruiting bodies of the strain.

4. Use of the heat-resistant Mushroom strain according to claim 1 in food production.

5. Use of the heat-resistant Lepidium scaly-leafed mushroom strain according to claim 1 in the preparation of medicines.

6. Application of the heat-resistant Lepidium edodes strain according to claim 1 in agricultural planting.

7. The use according to claims 4, 5 and 6, characterized in that: The heat-resistant leucopus scaly strain comprises spores, mycelium, fruiting bodies and extracts of the heat-resistant leucopus scaly strain.

8. The method for cultivating the heat-resistant shimeji mushroom strain according to claim 1, characterized in that: The controlled temperature range is 28-32℃.

9. The cultivation method of the heat-resistant shimeji mushroom strain according to claim 8, characterized in that: The culture medium used is: 200 g potatoes (peeled), 20 g glucose, 3.0 g potassium dihydrogen phosphate, 1.5 g magnesium sulfate, 10 mg vitamin B1, 1000 ml water, pH natural.

Images