A culture medium for culturing Cortinaria sinensis and its preparation method and application
By adding specific ingredients to potato glucose agar medium and adjusting pH, the problem that Chinese Silkworm cannot grow under laboratory conditions was successfully solved, and the effective cultivation and growth of this bacteria was achieved.
Patent Information
- Application Number
- CN202410509644.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-04-25
- Publication Date
- 2025-05-09
- Estimated Expiration
- 2044-04-25
AI Technical Summary
Chinese Silkworm cannot grow on commonly used potato glucose agar medium and lacks effective culture methods.
By adding malt leach powder, vermiase, potassium dihydrogen phosphate, magnesium sulfate and Qinghai spruce pine needle extraction solution to the potato glucose agar medium, the pH of the medium is adjusted to weak acidity, thereby mimicking the natural growth environment of Chinese Silkworm.
This method successfully promoted the growth of Chinese Silkworm and realized the possibility of large-scale domestication and cultivation under laboratory conditions.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of Cortinaria culture medium, and specifically relates to a culture medium for culturing Cortinaria sinensis and a preparation method and application thereof. Background Art
[0002] Cortinarius sinensis was first reported to grow in northwest China in 2020. It is mainly consumed in large quantities in Ningxia and the Helan Mountain area of Inner Mongolia, and is also distributed in small quantities in parts of Sichuan. It has high edible and economic value. Large fungi are usually cultured in the laboratory on potato dextrose agar (PDA) medium at 25°C. However, Cortinarius sinensis cannot be cultured under these conditions, let alone large-scale artificial domestication and cultivation. Therefore, how to change the culture conditions to make Cortinarius sinensis grow on the culture medium is the primary problem we face.
[0003] The collection site of Cortinaria sinensis is at a high altitude, ranging from 2000m to 4000m; the collection time is concentrated in August and September each year. The above growth conditions may result in a lower temperature for the growth of Cortinaria sinensis; it usually lives on the ground of coniferous forests, and the nutrients it requires are also different from those of ordinary large fungi. At present, there are few publications on effective cultivation methods for Cortinaria sinensis. Summary of the invention
[0004] In view of this, the object of the present invention is to provide a culture medium for culturing Cortinaria sinensis and a preparation method and application thereof, wherein the culture medium can promote the growth of Cortinaria sinensis.
[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:
[0006] The invention provides a culture medium for culturing Chinese Cortinaria sinensis, comprising the following components in parts by weight: 180-220 g of potato, 8-15 g of glucose, 15-25 g of agar, 5-10 g of malt extract, 0.003-0.008 g of earthworm enzyme, 0.3-0.8 g of potassium dihydrogen phosphate, 0.3-0.8 g of magnesium sulfate, 1-2 g of beef extract, 150-250 mL of Qinghai spruce pine needle extract, and 600-1000 mL of distilled water.
[0007] Preferably, the preparation method of the Qinghai spruce pine needle extract comprises the following steps: washing the Qinghai spruce needles, drying and grinding at 60-70°C; putting the ground pine needles and water in a water bath at a weight ratio of 1:8-15, extracting at 85-95°C for 0.5-3h, extracting 2-3 times, combining the extracts and filtering.
[0008] Preferably, the pH of the culture medium is 6.5-7.
[0009] The present invention also provides a method for preparing the culture medium, comprising the following steps:
[0010] Peel and cut the potatoes, boil them in water for 20 to 30 minutes, filter them with gauze, add glucose, agar, malt extract, potassium dihydrogen phosphate, magnesium sulfate and beef extract, heat and stir evenly, add Qinghai spruce pine needle extract after cooling slightly, and finally add distilled water to make up the volume, dispense them into conical bottles, add stoppers, bandage, and sterilize them by high pressure to obtain sterilized culture medium;
[0011] Add sterile water to earthworm enzyme to obtain earthworm enzyme dilution solution. Filter the earthworm enzyme dilution solution to make it sterile, then add it into the cooled sterilized culture medium, shake it evenly and pour it into the culture dish.
[0012] Preferably, the culture medium is cooled to 35-45°C after sterilization.
[0013] Preferably, the high pressure sterilization condition is 101 kPa, 120-121° C., and sterilization for 15-25 min.
[0014] Preferably, the potatoes are cut into 0.5-2 cm 3 of small pieces.
[0015] The present invention also provides a method for culturing Cortinaria sinensis, comprising the following steps: inoculating the fruiting bodies of Cortinaria sinensis in the culture medium or the culture medium prepared by the preparation method and culturing for 20 to 35 days until colonies grow; picking mycelium blocks from the above colonies and transferring them to the above culture medium and culturing for 45 to 60 days.
[0016] Preferably, the culture temperature is 15-25°C.
[0017] Preferably, the mycelium block has an area of 3 mm 2 ~15mm 2 .
[0018] Compared with the prior art, the present invention has the following beneficial effects:
[0019] According to the characteristics that Chinese Cortinaria often grows on the ground of coniferous forests and the growth environment is acidic, a certain amount of malt extract powder, earthworm enzyme, potassium dihydrogen phosphate (KH2PO4) and magnesium sulfate (MgSO4) and Qinghai spruce pine needle extract are added to the commonly used potato dextrose agar (PDA) medium to form a PDA+ medium, the pH of the medium is weakly acidic, and a certain amount of mineral elements can be provided for mycelial growth to promote the growth of Chinese Cortinaria. The present invention inoculates sterilized Chinese Cortinaria fruiting bodies on a cooled medium, cultures at 15 to 20°C, and mycelia grows after 10 to 20 days; when the mycelia grows into a colony with a diameter of about 1 cm, pick the mycelial blocks of Chinese Cortinaria in the culture medium, and grow into a circular colony with a diameter of 4 to 6 cm after 45 to 60 days, the color gradually becomes lighter in a ring shape from the inside to the outside, the center is white, the inner circle close to the center is purple-brown, and the outermost circle is white. The culture medium and the culture method of the present invention can effectively solve the problem that Cortinaria sinensis cannot grow under laboratory conditions. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] Figure 1 For the present invention to transfer 4mm 2 The growth of Chinese Cortinaria sinensis after the mycelium block; in the figure, A is the transfer of 4mm 2 Mycelium block day 0, B is transferred 4mm 2 Mycelium block on the 10th day, C is transferred to 4mm 2 Mycelial block day 45;
[0021] Figure 2 For the present invention, 15mm 2 The growth of Chinese Cortinaria after mycelial block; in the figure, A is the transfer of 15mm 2 Mycelium block day 0, B is transferred 15mm 2 Mycelium block on the 10th day, C is transferred to 15mm 2 Mycelium block on the 15th day, D is transferred to 15mm 2 Mycelium block on the 30th day, E is transferred to 15mm 2 Mycelial block day 60;
[0022] Figure 3 This is the growth of Cortinarius sinensis inoculated in SRG medium for 60 days;
[0023] Figure 4 This is the growth of Cortinarius sinensis inoculated in MEA medium for 60 days;
[0024] Figure 5 This is the growth of Cortinarius sinensis inoculated in FIM medium for 60 days;
[0025] Figure 6 This is the growth of Cortinarius sinensis inoculated in CMA medium for 60 days;
[0026] Figure 7 This is the growth of Cortinarius sinensis inoculated in CAM medium for 60 days. DETAILED DESCRIPTION
[0027] The present invention provides a culture medium for culturing Chinese Cortinaria, preferably comprising the following components by weight: 180-220g of potato, 8-15g of glucose, 15-25g of agar, 5-10g of malt extract, 0.003-0.008g of earthworm enzyme, 0.3-0.8g of potassium dihydrogen phosphate, 0.3-0.8g of magnesium sulfate, 1-2g of beef extract, 150-250mL of Qinghai spruce pine needle extract, and 600-1000mL of distilled water. More preferably, the culture medium comprises the following components: 200g of potato, 10g of glucose, 20g of agar, 6g of malt extract, 0.005g of earthworm enzyme, 0.5g of potassium dihydrogen phosphate, 0.5g of magnesium sulfate, 1.5g of beef extract, 200mL of Qinghai spruce pine needle extract, and 800mL of distilled water. The growth area of Chinese Cortinaria sinensis is at a high altitude, ranging from 2000m to 4000m; the collection time is concentrated in August-September every year; it usually lives on the ground of coniferous forests, and the nutrition required is different from that of ordinary large fungi. According to the characteristics that it often grows on the ground of coniferous forests and the growth environment is acidic, the present invention adds a certain amount of malt extract powder, earthworm enzyme, potassium dihydrogen phosphate (KH2PO4) and magnesium sulfate (MgSO4) and Qinghai spruce pine needle extract to the commonly used potato dextrose agar (PDA) culture medium to prepare PDA+ culture medium. The pH of the culture medium is weakly acidic, and it can also provide a certain amount of mineral elements for mycelial growth.
[0028] In the present invention, the preparation method of the Qinghai spruce pine needle extract preferably includes the following steps: the Qinghai spruce needles are washed, dried and ground at 60-70°C; the ground pine needles and water are placed in a water bath at a weight ratio of 1:8-15, extracted at 85-95°C for 0.5-3h, extracted 2-3 times, and the extracts are combined and filtered. More preferably, the following steps are included: the Qinghai spruce needles are washed, placed in an oven, dried at 65°C and ground. The ground pine needles and water are placed in a water bath at a weight ratio of 1:10, extracted at 90°C for 1h, extracted 2 times, the extracts are combined, and filtered with four layers of gauze to prepare the Qinghai spruce pine needle extract. Pine needles are natural litter of pine trees, and studies have shown that pine needles are a substance with strong allelopathic effects. Chinese Cortinaria usually lives on the ground of coniferous forests, and there are a large number of pine needles in the growth environment. Therefore, when preparing the culture medium of Chinese Cortinaria, the pine needle extract of Qinghai spruce is added to restore the growth environment of Chinese Cortinaria. The pine needle extract will provide growth hormone substances for Chinese Cortinaria and promote the growth of mycelium. Among them, allelopathic effect is also called allelopathic effect or allelopathic effect, which refers to a plant affecting the growth of other plants or microorganisms by secreting chemical substances in the metabolic process outside the body.
[0029] In the present invention, the earthworm enzyme contains active ingredients such as fibrinolytic enzyme, cholinesterase, and peroxidase, which are beneficial to the growth of mycelium. The present invention does not specifically limit the source of the earthworm enzyme, and conventional commercial products can be used. The earthworm enzyme of the present invention is purchased from Inner Mongolia Zhengshi Biotechnology Co., Ltd.
[0030] In the present invention, the pH of the culture medium is preferably 6.5-7, more preferably 6.8.
[0031] The present invention also provides a method for preparing the culture medium, comprising the following steps:
[0032] Cut the peeled potatoes into small pieces, boil in boiling water for 20 to 30 minutes, filter with gauze, add glucose, agar, malt extract powder, KH2PO4, MgSO4 and beef extract, heat and stir evenly, add Qinghai spruce pine needle extract after cooling slightly, finally add distilled water to make up the volume, divide into conical bottles, stopper and bandage; sterilize under high pressure.
[0033] Add 0.6-1.6 mL of sterile water to the earthworm enzyme, and use a pre-sterilized filter to filter the earthworm enzyme dilution to make it sterile. After the sterilized culture medium has cooled, add it to the culture medium and shake it evenly; pour the culture medium into a culture dish and cool it overnight.
[0034] In the present invention, the sterilized culture medium is cooled to 35-45°C, more preferably 40°C.
[0035] In the present invention, the high pressure sterilization condition is 101 kPa, 120-121° C. sterilization for 15-25 min, more preferably 121° C. sterilization for 20 min.
[0036] In the present invention, the potatoes are cut into 0.5-2 cm 3 Small pieces, more preferably 1 cm 3 of small pieces.
[0037] The present invention also provides a method for culturing Cortinaria sinensis, comprising the following steps: inoculating the fruiting bodies of Cortinaria sinensis in the culture medium or the culture medium prepared by the preparation method and culturing for 20 to 35 days until colonies grow; picking mycelium blocks from the above colonies and transferring them to the above culture medium and culturing for 45 to 60 days. The present invention does not specifically limit the source of the fruiting bodies of Cortinaria sinensis used for cultivation, and conventional commercial products can be used. The fruiting bodies of Cortinaria sinensis described in the present invention were collected from Helan Mountain.
[0038] In the present invention, the culture temperature is preferably 15 to 25°C.
[0039] In the present invention, the area of the mycelium block is preferably 3 mm 2 ~15mm 2 , preferably 4 mm 2 ~12mm 2 Different transfer areas directly affect the growth of the fungus species. Within the transfer area range of the present invention, the larger the mycelium block area, the faster the growth rate.
[0040] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.
[0041] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.
[0042] Example 1
[0043] A culture medium for culturing Chinese Cortinaria, comprising the following components: 200g potato, 10g glucose, 20g agar, 6g malt extract, 0.005g earthworm enzyme, 0.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 1.5g beef extract, 200mL Qinghai spruce pine needle extract, and 800mL distilled water. The pH is adjusted to 6.5. The preparation method of the Qinghai spruce pine needle extract comprises the following steps: the Qinghai spruce needles are washed, placed in an oven, dried at 65°C and ground. The ground pine needles and water are placed in a water bath at a weight ratio of 1:10, extracted at 90°C for 1h, extracted twice, the extracts are combined, and filtered with four layers of gauze to prepare the Qinghai spruce pine needle extract.
[0044] Example 2
[0045] A culture medium for culturing Chinese Cortinaria, comprising the following components: 180g potato, 8g glucose, 15g agar, 5g malt extract powder, 0.003g earthworm enzyme, 0.3g potassium dihydrogen phosphate, 0.3g magnesium sulfate, 1g beef extract, 150mL Qinghai spruce pine needle extract, and 600mL distilled water. The pH is adjusted to 6.8. The preparation method of the Qinghai spruce pine needle extract comprises the following steps: the Qinghai spruce needles are washed, placed in an oven, dried at 60°C and ground. The ground pine needles and water are placed in a water bath at a weight ratio of 1:8, extracted at 85°C for 0.5h, extracted twice, the extracts are combined, and filtered with four layers of gauze to prepare the Qinghai spruce pine needle extract.
[0046] Example 3
[0047] A culture medium for culturing Chinese Cortinaria, comprising the following components: 220g potato, 15g glucose, 25g agar, 10g malt extract powder, 0.008g earthworm enzyme, 0.8g potassium dihydrogen phosphate, 0.8g magnesium sulfate, 2g beef extract, 250mL Qinghai spruce pine needle extract, and 1000mL distilled water. The pH is adjusted to 7. The preparation method of the Qinghai spruce pine needle extract comprises the following steps: the Qinghai spruce needles are washed, placed in an oven, dried at 70°C and ground. The ground pine needles and water are placed in a water bath at a weight ratio of 1:15, extracted at 95°C for 3h, extracted 3 times, the extracts are combined, and filtered with four layers of gauze to prepare the Qinghai spruce pine needle extract.
[0048] Example 4
[0049] A method for preparing a culture medium of Cortinaria sinensis comprises the following steps: firstly, cutting 200 g of peeled potatoes into pieces with a volume of about 1 cm 3, boil the water for 20-30 minutes, filter with gauze, add 10g glucose, 20g agar, 6g malt extract, 0.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate and 1.5g beef extract, heat and stir evenly, add 200mL Qinghai spruce pine needle extract after cooling slightly, and finally add distilled water to 1000mL, divide into conical bottles, add stoppers and bandage; sterilize in an autoclave at 101kPa and 121℃ for 20min. 0.005g earthworm enzyme is added to 1mL sterile water, and the earthworm enzyme dilution is filtered to sterility with a pre-sterilized filter head, and the sterilized culture medium is added to the culture medium when it cools to about 40℃, and shakes evenly; the culture medium is poured into a culture dish and cooled overnight. Among them, the preparation method of the Qinghai spruce pine needle extract comprises the following steps: wash the Qinghai spruce needles, put them in an oven, dry them at 65℃ and grind them. Put the ground pine needles and water in a water bath at a weight ratio of 1:10, extract at 90°C for 1 hour, extract twice, combine the extracts, filter with four layers of gauze to make the Qinghai spruce pine needle extract.
[0050] Example 5
[0051] A method for culturing Cortinaria sinensis comprises the following steps:
[0052] (1) Cultivation of Cortinarius sinensis
[0053] A culture medium was obtained according to the preparation method of Example 4, and the sterilized fruiting bodies of Cortinaria sinensis were inoculated on the cooled culture medium and cultured at 20° C. Mycelium began to grow after 20 days, and mycelium grew into colonies with a diameter of about 1 cm after 30 days.
[0054] (2) Spreading of Cortinarius sinensis
[0055] The culture medium was prepared according to the method of Example 4, and the transfer area on the cooled culture medium was 4 mm 2 Mycelial blocks of Cortinarius sinensis ( Figure 1 A), cultured at 20℃, and after 10 days, it grew into a circular colony with a diameter of about 1 cm ( Figure 1 B); after 45 days, it grows into a circular colony with a diameter of 4 to 6 cm. The color gradually becomes lighter from the inside to the outside in a ring shape. The center is white, the inner circle near the center is purple-brown, and the outermost circle is white ( Figure 1 C).
[0056] Example 6
[0057] The difference from Example 5 is that the transfer area on the cooled culture medium is 15 mm 2 Mycelial blocks of Cortinarius sinensis ( Figure 2 A), cultured at 20°C, grew to an area of about 1.5 cm after 10 days 2 The colony ( Figure 2B); after 15 days, the colony area is about 2cm 2 , and the surface of the colony appears obvious purple ( Figure 2 C); After 30 days, the colony area is about 4 cm 2 , the surface of the colony is still purple, and the bottom begins to turn brown ( Figure 2 D); After 60 days, the colony has grown all over the culture dish, and the color gradually becomes lighter from the inside to the outside in a ring shape, with white hyphae in the center, purple-brown in the inner circle near the center, and white in the outer circle ( Figure 2 E).
[0058] Comparative Example 1
[0059] Using SRG medium to culture Cortinarius sinensis:
[0060] The formula of SRG medium is: 100g potato, 15g glucose, 18g agar, 20mL malt extract, 1g potassium dihydrogen phosphate (KH2PO4), 0.5g magnesium sulfate (MgSO4), 1g peptone, 1g ammonium chloride (NH4Cl), 0.15g vitamin B1, and 100mL distilled water.
[0061] The SRG medium was prepared as follows: First, 100 g of peeled potatoes were cut into pieces with a volume of about 1 cm 3 Take small pieces, boil for 20-30 minutes after the water boils, filter with gauze, add 15g glucose, 18g agar, 1g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 1g peptone, 1g ammonium chloride and 0.15g vitamin B1, heat and stir evenly, adjust the pH to 6.0 after cooling slightly, add distilled water to 1000mL, divide into conical flasks, stopper and bandage; sterilize in an autoclave at 101kPa and 121℃ for 20min. Pour the sterilized culture medium into a culture dish and cool overnight. Inoculate the sterilized Chinese Cortinaria fruiting body on the cooled culture medium and culture at 20℃. No mycelium grows after 60d ( Figure 3 ).
[0062] Comparative Example 2
[0063] Cultivation of Cortinarius sinensis using MEA medium:
[0064] The formula of MEA culture medium is: 30 g malt powder, 3 g soy peptone, 15 g agar, and 1000 mL distilled water.
[0065] The preparation method of MEA medium is as follows: first, add 30g malt powder, 3g soy peptone and 15g agar in a beaker, heat and stir evenly, add distilled water to 1000mL after cooling slightly, divide into conical bottles, add stoppers and bandage; sterilize in an autoclave at 101kPa and 121℃ for 20min. Pour the sterilized medium into a culture dish and cool overnight. Inoculate the sterilized Chinese Cortinaria fruiting body on the cooled medium and culture at 20℃. No mycelium grows after 60d ( Figure 4 ).
[0066] Comparative Example 3
[0067] Cultivation of Cortinarius sinensis using FIM medium:
[0068] The formula of FIM medium is: 0.5 g of calcium nitrate tetrahydrate [Ca(NO3)2·4H2O], 0.2 g of potassium dihydrogen phosphate (KH2PO4), 0.1 g of potassium chloride (KCl), 0.1 g of magnesium sulfate (MgSO4·7H2O), 0.1 g of yeast extract, 5.0 g of sucrose, 12 g of agar, and 1000 mL of distilled water.
[0069] First, add 0.5g of calcium nitrate tetrahydrate, 0.2g of potassium dihydrogen phosphate, 0.1g of potassium chloride, 0.1g of magnesium sulfate, 0.1g of yeast extract, 5g of sucrose and 12g of agar to a beaker, heat and stir evenly, add distilled water to 1000mL after cooling slightly, divide into conical flasks, add stoppers and bandage; sterilize in an autoclave at 101kPa and 121℃ for 20min. Pour the sterilized culture medium into a culture dish and cool overnight. Inoculate the sterilized Chinese Cortinaria fruiting body on the cooled culture medium and culture at 20℃. No mycelium grows after 60d ( Figure 5 ).
[0070] Comparative Example 4
[0071] Cultivation of Cortinarius sinensis using CMA medium:
[0072] The formula of CMA medium is: 40g corn flour, 12g agar, 1000mL distilled water
[0073] First, add 40g corn flour and 12g agar to a beaker, heat and stir evenly, add distilled water to 1000mL after cooling slightly, dispense into conical flasks, add stoppers and bandage; sterilize in an autoclave at 101kPa and 121℃ for 20min. Pour the sterilized culture medium into a culture dish and cool overnight. Inoculate the sterilized Chinese Cortinaria fruiting bodies on the cooled culture medium and culture at 20℃. No mycelium grows after 60d ( Figure 6 ).
[0074] Comparative Example 5
[0075] Cultivation of Cortinarius sinensis using CAM medium:
[0076] The formula of CAM medium is: 200 g carrot, 20 g glucose, 12 g agar, and 1000 mL distilled water.
[0077] The preparation method of CAM culture medium is as follows: first, cut 200g peeled carrots into small pieces with a volume of about 1cm3, boil them in water for 20-30min, filter them with gauze, add 20g glucose and 12g agar, heat and stir evenly, add distilled water to 1000mL after cooling slightly, divide them into conical bottles, add stoppers and bandages; sterilize them in an autoclave at 101kPa and 121℃ for 20min. Pour the sterilized culture medium into a culture dish and cool it overnight. Inoculate the sterilized Chinese Cortinaria fruiting bodies on the cooled culture medium and culture them at 20℃. No mycelium grows after 60d ( Figure 7 ).
[0078] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.
Claims
1. A culture medium for culturing Cortinarius sinensis, characterized in that: The invention is composed of the following components: 180-220 g of potato, 8-15 g of glucose, 15-25 g of agar, 5-10 g of malt extract powder, 0.003-0.008 g of earthworm enzyme, 0.3-0.8 g of potassium dihydrogen phosphate, 0.3-0.8 g of magnesium sulfate, 1-2 g of beef extract, 150-250 mL of Qinghai spruce needle extract, and 600-1000 mL of distilled water; The preparation method of the Qinghai spruce needle extract comprises the following steps: washing the Qinghai spruce needles, drying and grinding at 60-70° C.; putting the ground needles and water in a water bath at a weight ratio of 1:8-15, extracting at 85-95° C. for 0.5-3 hours, extracting 2-3 times, combining the extracts, filtering, and then finishing; The pH of the culture medium is 6.5-7.
2. The method for preparing the culture medium according to claim 1, characterized in that: The method comprises the following steps: peeling and cutting potatoes, boiling for 20 to 30 minutes after the water is boiled, filtering with gauze, adding glucose, agar, malt extract powder, potassium dihydrogen phosphate, magnesium sulfate and beef extract, heating and stirring evenly, adding Qinghai spruce needle extract after cooling slightly, finally adding distilled water to make up the volume, dispensing into conical bottles, plugging, bandaging, and high-pressure sterilization to obtain sterilized culture medium; adding earthworm enzyme to sterile water to obtain earthworm enzyme dilution solution, filtering the earthworm enzyme dilution solution to sterile, adding the cooled sterilized culture medium, shaking evenly and pouring into a culture dish.
3. The preparation method according to claim 2, characterized in that: The sterilized culture medium is cooled to 35-45°C.
4. The preparation method according to claim 2, characterized in that: The high pressure sterilization conditions are 101 kPa, 120-121° C., and 15-25 min.
5. The preparation method according to claim 2, characterized in that: The potatoes are cut into 0.5-2 cm 3 of small pieces.
6. A method for culturing Cortinarius sinensis, characterized in that: The method comprises the following steps: inoculating the fruiting bodies of the Chinese Cortinaria tenuifolia into the culture medium described in claim 1 or the culture medium prepared by any one of the preparation methods described in claims 2 to 5 and culturing them for 20 to 35 days until colonies grow; picking mycelium blocks from the above colonies and transferring them to the above culture medium and culturing them for 45 to 60 days.
7. The culture method according to claim 6, characterized in that: The culture temperature is 15-25°C.
8. The culture method according to claim 6, characterized in that: The mycelium block area is 3mm 2 ~15mm 2 .