High-yield agaricus subrufescens strain cxg001 and cultivation method and application thereof

Abstract

The application belongs to the technical field of edible fungi, and discloses a high-yield thelephora penicillata strain CXG001, a culture and cultivation method and application. The high-yield thelephora penicillata strain is the strain CXG001, which is preserved in the China Center for Type Culture Collection, Wuhan University, Wuhan, China, has a preservation number of CCTCC NO:M 20251055, and a preservation date of May 14, 2025. The application has a yield increase of 71.6% compared with tea 3 and a yield increase of 28.1% compared with tea 5 under the same planting conditions.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of edible fungi, and particularly relates to a pholiota nameko strain CXG001 and a culture cultivation method and application thereof. BACKGROUND

[0002] Pholiota nameko Cyclocybe chaxingu , with a trade name of tea tree mushroom, is one of main cultivated edible fungi, belonging to Basidiomycota, Agaricales, Tubariaceae and Pholiota. Cyclocybe

[0003] The main cultivated varieties of the pholiota nameko are still two varieties of "Tea New Mushroom No. 3" and "Tea New Mushroom No. 5" cultivated more than ten years ago. The two varieties have degenerated in multiple bases, and some pholiota nameko bases even have the phenomenon of not mushrooming at all, so high-quality new varieties of the pholiota nameko are urgently needed. SUMMARY

[0004] In order to overcome the problems in the prior art, the application provides a pholiota nameko strain CXG001 and a culture cultivation method and application thereof.

[0005] The technical solution is as follows: a pholiota nameko strain, which is a strain CXG001, is preserved in the China Center for Type Culture Collection, Wuhan University, Wuhan, China, has a preservation number of CCTCC NO: M 20251055, a preservation date of May 14, 2025, and a classification name of Cyclocybe chaxingu.

[0006] Another purpose of the application is to provide a culture method of the pholiota nameko strain, which is used for the culture of the pholiota nameko strain, and the method comprises the following steps:

[0007] S1, removing the roots of the wild type pholiota nameko CXG001, removing the surface soil and impurities with alcohol cotton, and sterilizing;

[0008] S2, longitudinally cutting the wild type pholiota nameko CXG001 fruiting body to obtain a fruiting body tissue block, placing the fruiting body tissue block into a prepared PDA culture medium, picking the PDA culture medium containing mycelium into a new PDA culture medium dish for culture after the mycelium grows, and repeating the operation multiple times;

[0009] S3, when the mycelium grows on the culture dish, cutting a small piece of the PDA culture medium containing the mycelium, placing multiple pieces of the PDA culture medium cubes into a cryopreservation tube containing sterile water, sealing the cryopreservation tube, and preserving.

[0010] In step S3, a small piece of the PDA culture medium cube containing the mycelium with an edge length of 1-2 mm is cut; and the preservation is performed at 4 DEG C.

[0011] ​Another object of the present application is to provide an application of the Agrocybe aegerita strain as a parent in breeding of Agrocybe aegerita.

[0012] Another object of the present application is to provide an Agrocybe aegerita mycelium, which is obtained by culturing the Agrocybe aegerita strain.

[0013] Another object of the present application is to provide an Agrocybe aegerita fruiting body, which is obtained by culturing the Agrocybe aegerita strain.

[0014] Another object of the present application is to provide a cultivation method of Agrocybe aegerita, which is obtained by culturing the Agrocybe aegerita strain, and the cultivation method comprises the following steps.

[0015] Step 1, preparation of the strain;

[0016] Step 2, preparation and inoculation of the cultivation substrate;

[0017] Step 3, management of the mycelium;

[0018] Step 4, management of the fruiting.

[0019] In step 1, the preparation of the strain comprises: taking out the Agrocybe aegerita strain, transferring to PDA medium in an ultraclean bench, sealing with a sealing film, placing in a constant temperature incubator at 25 DEG C in the dark, and taking out after the mycelium covers the plate for standby;

[0020] In step 2, the preparation and inoculation of the cultivation substrate comprises:

[0021] Preparation of the cultivation substrate: the cultivation substrate is composed of coarse material, fine material and water;

[0022] According to the mass ratio, the coarse material is rape straw 28.5-29.5%, miscellaneous wood chips 14.5-15.5%, wheat bran 21-22%, and chaff 28.5-29.5%, the fine material is sucrose 0.5-1.5%, gypsum 0.5-1.5%, and lime 1.5-2.5%, and the rest is water; wherein, the total of the coarse material and the fine material and the water are in a ratio of 2:3;

[0023] Inoculating the solid mycelium into the sterilized bag, and culturing at 25 DEG C in the dark;

[0024] Placing the cooled bag in an aseptic ultraclean bench, inoculating the prepared strain with an inoculation needle, inoculating 3 pieces of mycelium with a size of (2.5-3.5) cm x (1.4-1.6) cm in each bag, and evenly distributing the mycelium on the top of the culture medium.

[0025] In step 3, the fungus management includes: placing the inoculated fungus bag in a constant-temperature fruiting room for light-avoiding culture, the temperature in the fruiting room is set to 24-26 DEG C, and the relative humidity is set to 65-75%; after the mycelium grows to the top, the sealing film is removed, and the fungus is continuously cultured until the mycelium grows to fill the fungus bag.

[0026] In step 4, the fruiting management includes:

[0027] The temperature is adjusted to 21-23 DEG C, and the original base is stimulated to differentiate under light; after the bag is opened, the layer frame is arranged in a random block to realize the bag-removing fruiting management; during the fruiting, the relative humidity of the air is kept at 90%-95%, and the agaric is harvested after the agaric of the tea fungus grows out and before the film is broken.

[0028] In combination with all the technical solutions described above, the present application has the following beneficial effects:

[0029] Firstly, the strain CXG001 is a variety, and under the same planting conditions, the yield is increased by 71.6% compared with tea 3 (tea new mushroom 3), and the yield is increased by 28.1% compared with tea 5 (tea new mushroom 5). The strain can replace old varieties such as tea 3 and tea 5.

[0030] Secondly, under the same cultivation conditions, the yield of the tea fungus strain of the present application is 28%-71% higher than that of the existing tea fungus strain, and the yield is higher than that of the existing strain under the same cost input. After the technical scheme of the present application is transformed, the expected income is very high, and the producer can increase the income by more than 20% under the same conditions. The yield of the strain of the present application is more than 20% higher than that of the existing strain.

[0031] Thirdly, the strain provided by the present application is a strain domesticated and cultivated by the inventors through a large amount of basic research, a large amount of time and effort, and a large amount of resource investigation and cultivation test, which solves the problem of lack of high-quality varieties and short cycle in the tea fungus industry. BRIEF DESCRIPTION OF DRAWINGS

[0032] The accompanying drawings, which are incorporated into the specification and constitute a part of the specification, illustrate embodiments consistent with the present disclosure and, together with the specification, serve to explain the principles of the present disclosure;

[0033] Figure 1 is a culture method flow chart of the tea fungus strain provided by the embodiment of the present application;

[0034] Figure 2 is a schematic diagram of a molecular phylogenetic tree of the genus Agaricus, in which the LSU and TEF1 alpha sequences of the strain CXG001 are combined together with a reference sequence to construct Cyclocybe is a schematic diagram of a molecular phylogenetic tree of the genus Agaricus, in which the LSU and TEF1 alpha sequences of the strain CXG001 are combined together with a reference sequence to construct

[0035] Figure 3 is an effect diagram of the tea fungus strain provided by the embodiment of the present application. DETAILED DESCRIPTION

[0036] In order to make the above objectives, characteristics and advantages of the present application more apparent, specific embodiments of the present application are described in detail below with reference to the accompanying drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. However, the present application can be practiced in a number of different manners, which are different from those described herein, and it can be apparent that similar modifications can be made by those skilled in the art without departing from the scope of the present application, and therefore the present application is not limited to the specific implementations disclosed below.

[0037] Example 1, the tea fungus strain provided by the embodiment of the present application is strain CXG001, which is preserved in China Center for Type Culture Collection, Wuhan University, Wuhan, China, with a preservation number of CCTCC NO: M 20251055 and a preservation date of May 14, 2025. The detection result is: alive, and the classification name is: Cyclocybe chaxingu CXG001.

[0038] Example 2, as shown in the following table, the culture method of the tea fungus strain provided by the embodiment of the present application includes: Figure 1

[0039] S1, remove the roots of the wild type tea fungus CXG001, remove the surface soil and impurities with alcohol cotton, and sterilize;

[0040] S2, longitudinally cut the wild type tea fungus CXG001 fruiting body to obtain fruiting body tissue blocks, and put them into prepared PDA culture medium. After the mycelium grows, the PDA culture medium containing the mycelium is picked into a new PDA culture medium dish for culture, and the operation is repeated multiple times;

[0041] S3, when the mycelium covers the culture dish, cut the small cubes of the culture medium containing the mycelium, put multiple cubes of the culture medium into a cryopreservation tube containing sterile water, seal the cryopreservation tube, and preserve.

[0042] For example, the culture method of the tea fungus strain specifically includes:

[0043] The wild type tea fungus CXG001 is collected from Nanchang City, Jiangxi Province, and the growth substrate is rotten wood. The fruiting body of the tea fungus CXG001 is collected and brought back to the laboratory after the fruiting body is collected. The tea fungus CXG001 is separated by tissue separation method. The detailed steps are as follows:

[0044] Remove the roots of the tea fungus CXG001 collected in the wild, and remove the surface soil and other impurities with 75% alcohol cotton.

[0045] Put the treated fruiting body into a sampling bag, write the number and take it back to the laboratory.

[0046] ​Operation in the ultraviolet lamp sterilization ultraclean bench: the fruiting body is surface sterilized with 75% alcohol cotton, the fruiting body is longitudinally cut with a knife, a small piece of fruiting body tissue is clamped at the junction of the cap and the stem with sterilized tweezers, and is placed in the prepared PDA culture medium. After 2-5 days of observation, the PDA culture medium containing mycelium is immediately picked with a sterilized needle into a new PDA culture medium dish for culture. After the mycelium grows, the PDA culture medium containing mycelium is again picked with a sterilized needle into a new PDA culture medium dish for culture. When the mycelium grows on the culture dish, 1-2 mm small pieces of culture medium containing mycelium are cut with a sterilized knife, 5-8 small pieces are taken into a cryopreservation tube containing sterile water, the cryopreservation tube is sealed, and is stored at 4°C.

[0047] Example 3, as one possible implementation of the present application, provides a use of a Agaricus bisporus strain as a parent in the breeding of Agaricus bisporus.

[0048] Example 4, as one possible implementation of the present application, provides an Agaricus bisporus mycelium obtained by culturing the Agaricus bisporus strain.

[0049] Example 5, as one possible implementation of the present application, provides an Agaricus bisporus fruiting body obtained by culturing the Agaricus bisporus strain.

[0050] Example 6, identification of strain CXG001.

[0051] 1. Morphological identification of CXG001 strain.

[0052] Colony morphology: on PDA medium, the colony mycelium is villous, round, convex in the middle, white on the front, and brown spots on the back. The aerial mycelium is thin and white, and the growth rate is fast, 0.6-0.8 cm / d.

[0053] Fruiting body morphology: the cap diameter is 20-120.5 mm, initially hemispherical, then convex mirror-shaped to flat, dark brown in youth, gradually changing to brown to light brown, dark in the center, light to nearly white at the edge, not sticky, smooth or wrinkled surface. The flesh is white, thick in the middle and thin at the edge, with a strong and camellia oil aroma. The stem is middle, 44-126 mm long, nearly cylindrical, dirty white, with longitudinal fiber stripes on the middle surface, and sparse at both ends. The annulus is thin, single, white, and spore-bearing on the upper surface, brown, persistent or easily detached. The spore print is brown.

[0054] 2. Molecular identification of strain CXG001.

[0055] 2.1. DNA extraction.

[0056] The specific steps of DNA extraction are as follows:

[0057] 1) About 500 mg of mycelium of the test strain was scraped from the well-grown Petri dish (try not to scrape the culture medium), 1 mL of 60°C preheated 2x CTAB lysis solution was added, and the sample was ground with a grinding rod (add 500 μL of 2x CTAB lysis solution twice; after grinding, add 500 μL of 2x CTAB lysis solution and grind thoroughly). After thorough grinding, transfer to a centrifuge tube, and incubate in a water bath for 30-60 min;

[0058] 2) Add 500 mL of chloroform / isoamyl alcohol (24:1) solution, mix well, and centrifuge at 11400 rpm for 15 min (room temperature);

[0059] 3) Remove the supernatant (about 500 mL, be careful not to suck in the precipitate), add an equal volume of chloroform / isoamyl alcohol solution, mix well, and centrifuge at 11400 rpm for 15 min (room temperature);

[0060] 4) Remove the supernatant (about 400 mL), and repeat step 3);

[0061] 5) Remove the supernatant (about 300 mL), add 10% of the total volume of 5 M KOAc solution, add 2 / 3 of the total volume of isopropanol, mix well, and stand;

[0062] 6) Freeze at -20°C overnight, and centrifuge at 11400 rpm for 5 min;

[0063] 7) Discard the liquid phase (be careful not to pour out the precipitate), add 400 mL of 70% ethanol, shake to wash, and centrifuge at 11400 rpm for 5 min;

[0064] 8) Repeat step 7);

[0065] 9) Discard the liquid phase (be careful not to pour out the precipitate), dry the DNA, add 50-100 μL of 1x TE buffer, dissolve the DNA at 37°C, and store at -20°C for later use.

[0066] 2.2, PCR amplification and sequencing.

[0067] The method of Polymerase Chain Reaction (PCR) is used for amplification, mainly amplifying 28S large subunit ribosomal RNA (LSU) gene and 1-alpha translation elongation factor (TEF1a) gene amplification. The forward / reverse primers used in the application are respectively LSU: LROR / LR5 (forward / reverse), TEF1-a: EF1-983F / EF1-1620R. The primer synthesis and PCR product sequencing of the application are provided by Beijing Qikexing Biotechnology Co., Ltd. (Changsha Branch), and the specific base sequence is as follows:

[0068] LR0R: GTACCCGCTGAACTTAAGC (SEQ ID NO: 3) (Reference: Cai L, Lumyong P, Zhang K, et al. New species of Annulatascus and Saccardoella from the Philippines [J]. Mycotaxon, 2002, 84:255-263.1994);

[0069] LR5: ATCCTGAGGGAAACTTC (SEQ ID NO: 4) (Reference: Cai L, Lumyong P, Zhang K, et al. New species of Annulatascus and Saccardoella from the Philippines [J]. Mycotaxon, 2002, 84:255-263.1994);

[0070] EF1-983F: GCYCCYGGHCAYCGTGAYTTYAT (SEQ ID NO: 5), (wherein, Y represents base C or T, H represents base A or C or T, Reference: TEKPINAR A D, KALMER A. Utility of various molecular markers in fungal identification and phylogeny [J]. Nova Hedwigia, 2019, 109(1-2): 187-224);

[0071] EF1-2218R: ATGACACCRACRGCRACRGTYTG (SEQ ID NO: 6) (wherein, R represents base A or G, reference: TEKPINAR A D, KALMER A. Utility of various molecular markers in fungal identification and phylogeny[J]. Nova Hedwigia, 2019, 109 (1-2): 187-224).

[0072] 2.3, PCR amplification system.

[0073] Amplification reaction system:

[0074] 2x Taq PCR Master Mix 12.5 μL, template DNA 1 μL, upper primer 1 μL, lower primer 1 μL,

[0075] DdH2O 9.5 μL;

[0076] Amplification procedure, including:

[0077] LSU amplification procedure: 95 ℃ pre-denaturation 3 min; 94 ℃ denaturation 30 s, 55 ℃ annealing 50 s, 72 ℃ recombination 1 min, 35 cycles; 72 ℃ extension 1 min, 4 ℃ preservation;

[0078] TEF1-α amplification procedure, 96 ℃ pre-denaturation 2 min; 96 ℃ denaturation 45 s, 52 ℃ annealing 30 s, 72 ℃ recombination 1.5 min, 40 cycles; 72 ℃ extension 5 min, 4 ℃ preservation;

[0079] The PCR product was sent to Beijing Qikong Biological Co., Ltd. for sequencing, and the LSU and TEF1α sequences were obtained, and the nucleotide sequences are shown in SEQ ID NO: 1, SEQ ID NO: 2.

[0080] LSU sequence, SEQ ID NO: 1:

[0081] Actgcgagtgaagcgggaaaagctcaaatttgaaatctggcggtcctttgcggccgtccgagttgtaatctagagaagtgcttacccgcgctggaccgtgtacaagtctcctggaatggagcgtcacagagggtgagaatcccgtctttgacacggactgccagggcttatgtggtgcgctctcgaagagtcgagttgtttgggaatgcagctctaaatgggtggtaaattccatctaaagctaaatacaggcgagagaccgatagcgaacaagtaccgtgagggaaagatgaaaagaactttggaaagagagttaaacagtacgtgaaattgctgaaagggaaacgtttgaagtcagtcgcgttggctggggatcaaccttgctcttgcagggcgcactctccggttcgacgggtcagcatcagtttcggccgctggagaaaggctaggggaatgtggcatcgcttcggtgatgtgttatagcccttggtcgtatgcagcggctgggactgaggaactcagcctgcccttaactgggccgggtcttttgaccacgctaacgggcttaggatgctggcataatggctttaagcgacccgtcttgaaacacggaccaaggagtctaacatgcttgcgagtgtttgggtggaaaacccgagcgcgtaatgaaagtgaaagttgggaaccctgtcgtggggtgcaccgacgcccggaccagaccttcttgtgacggctccgcggtagagcaggtatgttgggacc;

[0082] TEF1 alpha sequence, SEQ ID NO: 2:

[0083] .

[0084] The LSU and TEF1α sequences of strain CXG001 were combined and constructed with a reference sequence. Cyclocybe Molecular phylogenetic tree of the genus ( Figure 2 As can be seen from the phylogenetic tree, strain CXG001 and *Pleurotus ostreatus* (…) Cyclocybe chaxingu The reference sequence of the strain CXG001 is on the same branch and has a very high support rate (99%), therefore, strain CXG001 can be identified as *Pleurotus ostreatus*. The strain CXG001 of this invention has significant genetic uniqueness compared to other *Pleurotus ostreatus* strains.

[0085] Example 7: This invention provides a method for cultivating *Pleurotus ostreatus*, using the *Pleurotus ostreatus* strain described above. The cultivation method includes:

[0086] Step 1, Preparation of inoculum;

[0087] Step 2, preparation of cultivation substrate and inoculation;

[0088] Step 3, Mycelium Management;

[0089] Step 4, mushroom management.

[0090] Example 1, step 1, strain preparation.

[0091] Take out the preserved CXG001 agaric strain, transfer it to PDA medium in a clean bench, seal it with sealing film, and place it in a constant temperature incubator at 25°C in the dark. After the mycelium covers the plate, take it out for standby use.

[0092] Example 1, step 2, preparation and inoculation of cultivation substrate.

[0093] The cultivation formula is as follows: coarse material: rape straw 28.5-29.5%, mixed wood chips 14.5-15.5%, wheat bran 21-22%, and rice husk 28.5-29.5%; fine material: sucrose 0.5-1.5%, gypsum 0.5-1.5%, and lime 1.5-2.5%; and water 2:3. The cultivated solid mycelium is inoculated into sterilized bags, and cultured in the dark at 25°C.

[0094] The size of the bag is 18 cm × 36 cm × 0.005 cm, and the loading capacity is 470-475 kg per bag. After cooling, the bag is placed in a sterile clean bench, and the prepared strain is inoculated with an inoculation needle. Three pieces of mycelium with a size of (2.5-3.5) cm × (1.4-1.6) cm are inoculated into each bag, evenly distributed at the top of the culture material, and inoculated aseptically.

[0095] Example 1, step 3, mycelium management.

[0096] After inoculation, the bag is placed in a constant temperature mushroom house for dark culture. The temperature in the mushroom house is set to 24-26°C, and the relative humidity is set to 65-75%. In the early stage of culture, the strain begins to germinate slowly in the bag. At this time, do not frequently enter and exit the mushroom house to check. After the mycelium germinates for about a week, move the bag to observe whether there is contamination. The contaminated bag is treated in time to prevent contamination of other bags. After ten days, when the mycelium grows to the top, remove the sealing film and continue to cultivate the mycelium. About 45 days after inoculation, the mycelium grows to fill the bag.

[0097] Example 1, step 4, mushroom management.

[0098] Adjust the temperature to 21-23°C, and stimulate the differentiation of primordia with light. After 10 days, open the bag. Randomly arrange the groups to manage the layering and bag removal of mushrooms. During the mushroom period, the relative humidity of the air is maintained at 90%-95%. The humidity is mainly improved by methods such as ground watering, air spraying, and wall spraying. Avoid spraying water on the bag, as it may cause contamination. After the agaric fruiting body grows, it is immediately harvested when the color of the edge of the cap is light and the veil is not broken or slightly broken.

[0099] Example 8, measurement and analysis of agaric yield.

[0100] 1, Agaricus blazei murrill yield and dry weight.

[0101] The first crop of fresh Agaricus blazei murrill and the second crop of fresh Agaricus blazei murrill were weighed and recorded separately.

[0102] First crop yield per bag: the fresh weight of the first crop of Agaricus blazei murrill fruiting bodies;

[0103] Second crop yield per bag: the fresh weight of the second crop of Agaricus blazei murrill fruiting bodies;

[0104] Total yield (total yield of two crops): the sum of the fresh weight of the first crop and the fresh weight of the second crop.

[0105] 2, Biological efficiency.

[0106] The biological efficiency of each bag was calculated according to the following formula:

[0107] Biological efficiency = (total yield of two crops / dry weight of cultivation material) x 100%. Data analysis is shown in Table 1.

[0108] Table 1 Data analysis table

[0109]

[0110] Among them, Figure 3 is the effect picture of the Agaricus blazei murrill strain provided by the embodiments of the present application.

[0111] In the above embodiments, the description of each embodiment has its own emphasis, and the parts not described or recorded in detail in a certain embodiment can be referred to the related description of other embodiments.

[0112] The above is only a preferred specific embodiment of the present application, but the protection scope of the present application is not limited thereto, and any modification, equivalent replacement and improvement made by those skilled in the art within the technical range disclosed by the present application, as long as it is within the spirit and principles of the present application, should be covered within the protection scope of the present application.

Claims

1. A Agaricus subrufescens strain, characterized in that, The tea fungus strain is a strain CXG001, preserved in China Typical Culture Collection Center, with a preservation number of CCTCC NO: M 20251055, a preservation date of May 14, 2025, and a classification name of Cyclocybe chaxingu ; The culture method of the Agrocybe aegerita strain comprises the following steps: S1, removing the roots of the wild Agrocybe aegerita CXG001, removing the surface soil and impurities with alcohol cotton, and sterilizing; S2, longitudinally cutting the wild Agrocybe aegerita CXG001 fruiting body to obtain a fruiting body tissue block, and placing the fruiting body tissue block into a prepared PDA culture medium, picking the PDA culture medium containing mycelium into a new PDA culture medium dish for culture after the mycelium grows, and repeating the operation multiple times; S3, when the mycelium grows on the culture dish, cutting a small piece of the mycelium-containing PDA culture medium cube with a side length of 1-2 mm, and placing the multiple pieces of the mycelium-containing PDA culture medium cube into a cryopreservation tube containing sterile water, sealing the cryopreservation tube, and preserving.

2. The Agaricus sylvinae strain of claim 1, wherein, In step S3, a small piece of the mycelium-containing PDA culture medium cube with a side length of 1-2 mm is cut; and the preservation is carried out at 4℃.

3. The use of Agrocybe aegerita strain as a parent in the breeding of Agrocybe aegerita, characterized in that, The Agrocybe aegerita strain of claim 1 is applied for breeding.

4. A mycelium of Agrocybe aegirit, characterized in that, The Agrocybe aegerita strain of claim 1 is applied for culture.

5. A fruiting body of Agrocybe aegirit, characterized in that, The Agrocybe aegerita strain of claim 1 is applied for culture.

6. A cultivation method of Agrocybe aegerita, characterized by, The Agrocybe aegerita strain of claim 1 is applied for cultivation, and the cultivation method comprises the following steps: Step 1, strain preparation; Step 2, preparation and inoculation of cultivation substrate; Step 3, mycelium management; Step 4, fruiting body management.

7. The cultivation method of Agrocybe aegerita according to claim 6, characterized in that, In step 1, the strain preparation comprises: taking out the Agrocybe aegerita strain, transferring the Agrocybe aegerita strain to a PDA culture medium in a clean bench, sealing the PDA culture medium with a sealing film, placing the PDA culture medium in a 25℃ constant temperature incubator for dark and static culture, and taking out the PDA culture medium for standby use after the mycelium grows on the PDA culture medium; In step 2, the preparation and inoculation of the cultivation substrate comprises: Preparation of the cultivation substrate: the cultivation substrate is composed of coarse material, fine material and water; According to the mass ratio, the coarse material is rapeseed straw 28.5-29.5%, miscellaneous wood chips 14.5-15.5%, wheat bran 21-22% and chaff 28.5-29.5%, the fine material is sucrose 0.5-1.5%, gypsum 0.5-1.5% and lime 1.5-2.5%, and the rest is water; wherein, the total amount of the coarse material and the fine material and the water are in a ratio of 2:3; The prepared solid mycelium block is inoculated into a sterilized fungus bag, and the fungus bag is cultured at 25℃ in the dark; The cooled fungus bag is placed in a sterile clean bench, and the prepared strain is inoculated with an inoculation needle, 3 pieces of mycelium block with a size of (2.5-3.5) cm×(1.4-1.6) cm are inoculated into each fungus bag, and the mycelium block is evenly distributed on the top of the culture material.

8. The cultivation method of Agrocybe aegerita according to claim 6, characterized in that, In step 3, the mycelium management comprises: placing the fungus bag after inoculation in a constant temperature fruiting room for light-proof culture, setting the temperature in the fruiting room to 24-26℃, and setting the relative humidity to 65-75%; after the mycelium grows on the top, the sealing film is removed, the mycelium continues to grow, and the mycelium grows on the fungus bag.

9. The cultivation method of Agrocybe aegerita according to claim 6, characterized in that, In step 4, the fruiting body management comprises: Adjusting the temperature to 21-23℃, stimulating the original base differentiation with light, arranging the fungus bags in a random block group after the fungus bags are opened, and managing the fruiting body to be taken out of the fungus bag in a layer; during the fruiting period, the relative humidity of the air is kept at 90% to 95%, and the Agrocybe aegerita fruiting body is harvested when the Agrocybe aegerita fruiting body grows out and the mycelium film is not broken.

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