A strain of agrocybe aegirit and a breeding method thereof

Abstract

The present application belongs to the field of edible mushroom domestication cultivation, and particularly relates to a new strain of Agrocybe aegerita and a domestication cultivation method. The breeding method of the new strain of Agrocybe aegerita Gan Agrocybe aegerita No.1 comprises the following steps: (1) activation culture of Agrocybe aegerita mother strain, (2) preparation of original strain and cultivation strain, (3) preparation of cultivation bag, (4) cultivation bag culture, (5) bud induction, (6) mushroom cultivation and (7) wild mushroom fruiting body tissue separation. Through fruiting body tissue separation, artificial domestication and system selection, a new excellent variety applicable to production is finally bred, which can be used as an optimal variety for Agrocybe aegerita facility and industrial cultivation. Through domestication cultivation, the suitable growth conditions and cultivation characteristics of Gan Agrocybe aegerita No.1 are determined, and a high-efficiency cultivation technology matched with the variety is established. The Gan Agrocybe aegerita No.1 variety has great application potential.

Description

Technical Field

[0001] This invention belongs to the field of domestication and cultivation of edible fungi, specifically involving a strain of *Agrocybe aegerita* and its breeding method. Background Technology

[0002] All artificially cultivated edible fungi originated from the wild. Under natural conditions, wild edible fungi complete their life cycle through mycelial growth or spore reproduction. With the maturation of pure strain isolation technology, pure culture of edible fungi has become widely applied. People collect fruiting bodies from the wild, and through tissue isolation or spore isolation methods, obtain cultures and wild strains through pure culture. These are then domesticated and repeatedly selected under simulated wild environmental conditions to obtain artificially cultivated strains. As people's understanding of the nutritional modes and growth characteristics of wild edible fungi has deepened, the artificial domestication of wild edible fungi has received increasing attention, and the variety of domesticated and cultivated fungi has become increasingly diverse. Currently, a considerable number of excellent edible fungi varieties in my country originate from the artificial domestication and systematic selection of wild strains. The most prominent advantage of artificially domesticated wild fungi strains lies in the significant differences in geographical conditions at the collection sites of different wild strains. These diverse natural environments can lead to the formation of germplasm materials with significant genetic differences, providing excellent germplasm sources for the domestication, improvement, and breeding of wild edible fungi strains.

[0003] Tea tree mushroom (Agromycetes spp.) belongs to the phylum Basidiomycota, class Agaricales, order Agaricales, family Pseudomonas, and genus Agaricales. It is commercially known as tea tree mushroom because the earliest domesticated and cultivated strains originated from the decaying stump of the tea tree mushroom. Tea tree mushrooms are highly favored by consumers for their beautiful shape, tender cap, crisp stem, and rich, fragrant flavor. According to statistics from the China Edible Fungi Association, in 2022, my country's annual production of tea tree mushrooms exceeded 400,000 tons, with a value exceeding 5 billion yuan, making it one of the most widely cultivated rare edible fungi. Wild tea tree mushrooms typically grow under camellia trees. Changes in the ecological environment have led to an increasing scarcity of wild tea tree mushroom resources. Currently, the tea tree mushroom seed industry in my country faces challenges such as insufficient usable germplasm resources, a lack of superior varieties, and severe strain degradation, which have become bottlenecks restricting the sustainable development of the tea tree mushroom industry. Therefore, the domestication and improvement of collected wild tea tree mushroom resources and the creation of superior cultivated varieties are of great value.

[0004] Edible fungi are abundant in nature, but only a few dozen species are currently cultivated artificially. Therefore, the domestication and cultivation of wild edible fungi has long been a research hotspot for edible fungi researchers. Domestication and cultivation of wild *Agrocybe aegerita* germplasm resources into artificially cultivated strains can not only enrich the varieties of *Agrocybe aegerita* cultivated in my country and provide superior strains for the industry, but also provide breeding materials for the selection of superior varieties, addressing the current situation in my country's breeding field characterized by a lack of usable germplasm resources, a narrow genetic background, and insufficient superior parent lines. Therefore, the discovery and domestication of new *Agrocybe aegerita* strains has immense research and application value. Summary of the Invention

[0005] The purpose of this invention is to provide a method for breeding a new strain of *Agrocybe aegerita*, "Ganchashugu No. 1". The breeding method includes the following steps:

[0006] (1) Activation culture of the mother strain of tea tree mushroom: The purified and preserved wild tea tree mushroom strain mother strain was transferred to sterilized culture dish PDA medium and cultured in a biochemical incubator in the dark for about 10 days to grow into the plate and become the activated mother strain;

[0007] (2) Production of primary and cultivars: The activated mother seed is propagated into primary and cultivars;

[0008] (3) Preparation of cultivation bags: After the cultivation materials are thoroughly mixed according to the formula, the moisture content of the culture medium is controlled at 60% to 65%. Polypropylene plastic bags are used for packaging, with each bag weighing 0.5 to 0.55 kg of dry material. After packaging, the bags are sealed with plastic rings and caps. After packaging, the bags are sterilized by high pressure. After sterilization, the bags are transferred to a pre-sterilized cooling room for cooling. Inoculation is carried out in the inoculation room according to the aseptic operation specifications.

[0009] (4) Cultivation in a cultivation bag: When the mycelium grows to 2 / 3 of the bag, loosen the bag opening and oxygenate the bag. When yellow water droplets begin to appear at the bag opening and brown spots appear on the surface of the bag, the bag is physiologically mature.

[0010] (5) Bud induction: After the mature mushroom bags are moved into the mushroom house, the ambient temperature of the mushroom house is controlled at 20-22℃, the relative humidity of the mushroom house is maintained at 85%-90%, 200-300 lux of diffused light is provided, and ventilation is provided to keep the air in the mushroom house fresh; when rice grain-like primordia appear on the surface of the mushroom bags, straighten the opening of the mushroom bags and then close them slightly. After 1-2 days, the primordia differentiate into mushroom buds.

[0011] (6) Mushroom cultivation: After the mushroom buds are formed, the temperature of the mushroom growing environment is controlled at 22-24℃ and the relative humidity is 90%-95%. The number of ventilations and the time are reduced, the carbon dioxide concentration in the mushroom house is increased, the growth of the tea tree mushroom stipe is promoted, and the opening of the mushroom cap is inhibited.

[0012] (7) Isolation of wild mushroom fruiting body tissues: Before the fruiting bodies of Mushroom Agrocybe aegerita reach 6-7 maturity and the veil is broken, the yield and agronomic traits of the fruiting bodies are observed and statistically analyzed. The fruiting bodies are selected, and their tissues are isolated and purified. After 5 rounds of domestication and repeated selection, the superior Mushroom Agrocybe aegerita strains are finally selected.

[0013] Furthermore, morphological anatomy and molecular phylogenetic analysis techniques were used to identify wild fungi. Morphological characteristics of wild fungal fruiting bodies: Fruiting bodies are solitary, paired, or clustered; the cap is 3-10 cm in diameter, initially hemispherical, dark brown to tan, gradually becoming flattened, turning light brown to pale yellowish-brown, with a lighter edge and a darker center; when moist, it is sticky, smooth or wrinkled, and when young, it is covered with a veil that is rolled inwards; the flesh (except for the surface and base of the stipe) is white or off-white, slightly tough, thicker in the center and thinner at the edge. The stipe is solid, about 3-12 cm long and 0.5-2 cm thick, cylindrical, nearly uniform in thickness, off-white, darker near the base, and fibrous on the surface. At maturity, the stipe hardens, is covered with a dull, sticky substance, and the annulus remains on the stipe or adheres to the edge of the cap and falls off automatically. The gills are adnate to decurrent, dense, and of unequal length, initially white, turning brownish-brown to brown when mature; the annulus is superior, initially white, later becoming dark brownish-brown with basidiospores on the upper surface, membranous to filamentous, and not easily detached. The basidiospores are 7-10 × 4-6 µm in size, elliptical, smooth, and brownish-brown. For detailed morphology, anatomy, and microstructure of the fruiting bodies, please refer to the appendix. Figure 1-10 Using MEGE software and the NJ method to construct a phylogenetic tree based on ITS sequences, the wild strain was identified as *Agrocybe aegerita*.

[0014] Furthermore, in step (2), the original seed and cultivated seed formulas include wood chips with a mass percentage of 39%, cottonseed hulls 33%, wheat bran 18%, corn flour 8%, lime 1.5%, light calcium carbonate 1%, and a moisture content of 58%.

[0015] Further, in step (3), the cultivation material formula includes cottonseed hulls 36%, sawdust 29%, wheat bran 22%, corn flour 10%, soybean meal 1%, lime 1%, and light calcium carbonate 1% by mass; the sterilization conditions for the cultivation material are 0.11-0.14 MPa pressure and steam temperature of 121-126℃ for 2-2.5 hours.

[0016] Furthermore, in step (4), during the first 15 days of cultivation in the cultivation bags, the environment of the mycelium incubation room is adjusted to a temperature of 25-26℃, a relative humidity of 60%-65%, a light intensity of 50-100 Lx, and a CO2 concentration of 0.04-0.1%; during the 16-50 days of cultivation in the mycelium bags, the environment of the mycelium incubation room is adjusted to a temperature of 22-24℃, a relative humidity of 65%-70%, a light intensity of 100-200 Lx, and a CO2 concentration of 0.06-0.08%.

[0017] Furthermore, in step (5), the environment of the mushroom growing room is adjusted to a temperature of 20-22℃, a relative humidity of 85%-90%, a light intensity of 200-300 Lx, and a CO2 concentration of 0.1-0.3%. When rice-grain-shaped primordia appear on the surface of the mushroom bag, the opening of the mushroom bag is straightened, and the primordia differentiate into mushroom buds after 1-2 days.

[0018] Furthermore, in step (6), the mushroom growing environment is adjusted to a temperature of 22-24℃, a relative humidity of 90%-95%, a light intensity of 100-200 Lx, and a CO2 concentration of 0.5-0.7%.

[0019] Furthermore, in step (7), tea tree mushroom fruiting bodies that are early to fruit, have a large number of fruiting bodies with high uniformity, thick caps, thick stems, and slow opening are selected from the mushroom bags, and their tissues are separated and purified for culture.

[0020] Furthermore, in step (8), the new strain of *Agrocybe aegerita* isolated and purified from tissue culture is domesticated and repeatedly selected for 5 generations, and finally a new strain of *Agrocybe aegerita* with excellent traits is selected, clarifying the biological characteristics and agronomic traits of the domesticated new strain of *Agrocybe aegerita*.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0022] (1) Using wild tea tree mushroom resources as materials, a new tea tree mushroom strain, “Gan Tea Tree Mushroom No. 1”, was cultivated. The strain originated from a wild tea tree mushroom fruiting body collected from the stump of a camellia tree. The fruiting body is robust with a thick cap and a thick stem, and has high yield, high quality and stress resistance potential. Through fruiting body tissue isolation, artificial domestication and systematic selection, a superior new variety that can be applied to production was finally cultivated. It can be used as a preferred variety for facility-based and factory-based cultivation of tea tree mushroom.

[0023] (2) Through the analysis of the biological characteristics and agronomic traits of the new strain “Ganchashugu No. 1”, the strain has strong resistance, short cultivation cycle and excellent commercial traits. Through domestication and cultivation, the suitable growth conditions and cultivation characteristics of “Ganchashugu No. 1” have been clarified, and efficient cultivation technology matching the variety has been established. It has the characteristics of tea tree mushroom facility cultivation variety and has great promotion and application value.

[0024] The strain of this invention, "Ganchashugu No. 1" (Ganchashugu 1), was deposited on August 12, 2024, with accession number CGMCC No. 41432. It is classified as *Cyclocybe cylindracea*, and the depository is the China General Microbiological Culture Collection Center, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, 100101, China. Attached Figure Description

[0025] Figure 1 Schematic diagram of the fruiting body and habitat of wild fungi in this invention;

[0026] Figure 2 Schematic diagram of the colony of the wild fungal strain of this invention;

[0027] Figure 3Schematic diagram of the microstructure of the cap epidermis of the wild fungus fruiting body of this invention;

[0028] Figure 4 Schematic diagram of the epidermal structure of the stipe of the wild fungus fruiting body of this invention;

[0029] Figure 5 Schematic diagram of the microstructure of basidiospores of wild fungus fruiting bodies in this invention;

[0030] Figure 6 Schematic diagram of the domestication and cultivation of tea tree mushroom "Gan Tea Tree Mushroom No. 1" according to this invention;

[0031] Figure 7 Schematic diagram of the fruiting body of the domesticated and cultivated "Ganchashugu No. 1" tea tree mushroom of this invention;

[0032] Figure 8 A schematic diagram of the cap of the domesticated and cultivated tea tree mushroom "Gan Tea Tree Mushroom No. 1" according to the present invention;

[0033] Figure 9 Schematic diagram of the gills of the domesticated and cultivated *Tea Tree Mushroom No. 1* in this invention;

[0034] Figure 10 A schematic diagram of the stipe of the domesticated and cultivated tea tree mushroom "Gan Tea Tree Mushroom No. 1" according to the present invention. Detailed Implementation

[0035] The technical solution of the present invention will be clearly and completely described below with reference to embodiments. The described embodiments are only some embodiments of the present invention, not all embodiments. Unless otherwise specified, the methods described are conventional methods, and the raw materials described are all obtainable from publicly available commercial channels unless otherwise specified. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the protection scope of the present invention.

[0036] This invention provides a strain of *Agrocybe aegerita* and a method for its domestication and cultivation, including the following steps: obtaining a wild fungus culture from a wild *Agrocybe aegerita* fruiting body collected from a wild *Camellia oleifera* tree through tissue isolation. The collected wild fungus fruiting body and the isolated wild fungus culture are identified as wild *Agrocybe aegerita* using morphological, anatomical, and molecular phylogenetic analyses. Wild tea tree mushroom cultures were propagated into strains, and a cultivation substrate was prepared using sawdust and cottonseed hulls as the main raw materials. Cultivation was carried out using a pre-grown substrate bag method. Based on the biological characteristics of tea tree mushrooms, the temperature, light, air, and humidity of the mycelium growth and fruiting environment were controlled to cultivate tea tree mushroom fruiting bodies. The growth traits and cultivation characteristics of wild tea tree mushroom strains were analyzed. Fruiting bodies with early fruiting, numerous buds, thick caps, long stipes, good color, and slow cap opening were selected. These were then subjected to tissue isolation and purification culture. Pure cultures with good mycelial growth were screened for propagation into strains and further domestication cultivation. After five rounds of domestication cultivation and systematic selection as described above, a domestication cultivation method for a new strain of wild tea tree mushroom was developed. The domesticated tea tree mushroom strain was named "Gan Cha. Shu Gu No. 1".

[0037] Through this invention, a new strain of *Agrocybe aegerita*, "Ganchashugu No. 1", was obtained. Figure 1-10 As shown, the fruiting body morphology of this strain is as follows: solitary, paired, or clustered; cap diameter 3-10 cm, initially hemispherical, dark brown to tan, gradually becoming flattened, gradually turning light brown to pale yellowish-brown, lighter in color at the edge and darker in the center, sticky when moist, smooth or wrinkled, with a veil that rolls inward when young; flesh (except for the surface and base of the stipe) white or off-white, slightly tough, thicker in the center and thinner at the edge; stipe solid, about 3-12 cm long and 0.5-2 cm thick, round. Columnar, nearly equal in thickness, off-white, darker brownish-black near the base, fibrous surface; stipe hardens at maturity, covered with a dull, sticky substance; annulus remains on the stipe or adheres to the edge of the cap and falls off automatically; gills adnate to decurrent, dense, unequal in length, initially white, turning brownish-brown to brownish-black upon maturity; annulus superior, initially white, later turning dark brownish-brown with basidiospores on the upper surface, membranous to filamentous, not easily detached; basidiospores 7-10 × 4-6 µm in size, elliptical, smooth, spore print brownish-brown. Example

[0038] The domestication and cultivation method for the new strain of *Agrocybe aegerita* includes the following steps: activating and culturing the mother culture of wild *Agrocybe aegerita* strains; propagating the original strain and cultivar using solid-state spawn production methods; preparing cultivation bags using the substrate bag cultivation method; conducting bag cultivation, bud induction, and mushroom management; analyzing the growth traits and cultivation characteristics of wild *Agrocybe aegerita* strains; selecting fruiting bodies that are early-fruiting, have many buds, long stipes, good color, and slow cap opening; performing tissue isolation and purification culture on these fruiting bodies to obtain *Agrocybe aegerita* strains with superior traits; conducting five rounds of domestication and cultivation and systematic selection according to the above cultivation process; and finally selecting a superior *Agrocybe aegerita* strain, which is named "Gan Cha Shu Gu No. 1". The specific methods and steps are as follows:

[0039] (1) Activation culture of the mother strain of tea tree mushroom: The purified and preserved wild tea tree mushroom strain mother strain was transferred to sterilized PDA medium in a culture dish and cultured in a biochemical incubator in the dark for about 10 days to grow mycelium to fill the culture dish and become an activated mother strain.

[0040] (2) Preparation of primary and cultivars: Prepare culture media for primary and cultivars, and propagate the activated mother culture into primary and cultivars. The formula for primary and cultivars includes the following components by weight percentage: 39% sawdust, 33% cottonseed hulls, 18% wheat bran, 8% corn flour, 1.5% lime, 1% light calcium carbonate, and 58% moisture content.

[0041] (3) Preparation of cultivation material bags: The cultivation material is prepared according to the formula, which includes cottonseed hulls 36%, sawdust 29%, wheat bran 22%, corn flour 10%, soybean meal 1%, lime 1%, and light calcium carbonate 1% by weight. After thoroughly mixing the cultivation materials, the moisture content of the cultivation material is controlled at 60%–65%. Polypropylene plastic bags are used for packaging, with each bag containing 0.5–0.55 kg of dry material. The bags are sealed with plastic rings and caps and then sterilized under high pressure. The sterilization conditions are 0.11–0.14 MPa pressure and a steam temperature of 121–126°C, which is maintained for 2–2.5 hours. After sterilization, the bags are transferred to a pre-sterilized cooling room for cooling, and then inoculated in the inoculation room according to the aseptic operation specifications.

[0042] (4) Cultivation in cultivation bags: After inoculation, the bags are transferred to the cultivation room. For the first 15 days of cultivation in the bags, the environment of the mycelium growth room is controlled at a temperature of 25-26℃, relative humidity of 60%-65%, light intensity of 50-100 Lx, and CO2 concentration of 0.04-0.1%. For 16-50 days of cultivation in the bags, the environment of the mycelium growth room is controlled at a temperature of 22-24℃, relative humidity of 65%-70%, light intensity of 100-200 Lx, and CO2 concentration of 0.06-0.08%. When the mycelium grows to 2 / 3 of the bag, the bag opening is loosened and oxygen is added to the bag. When yellow water droplets begin to appear at the bag opening and brown spots appear on the surface, it indicates that the bag is physiologically mature. The mature bags are then transferred to the fruiting room.

[0043] (5) Bud induction management: After the mushroom bags are moved into the fruiting room, the environmental conditions of the fruiting room are adjusted to a temperature of 20-22℃, relative humidity of 85%-90%, light intensity of 200-300 Lx, and CO2 concentration of 0.1-0.3%. When rice-grain-like primordia appear on the surface of the mushroom bags, straighten the opening of the bags. The primordia will differentiate into mushroom buds after 1-2 days.

[0044] (6) Mushroom cultivation management: After the mushroom buds are formed, the mushroom growing environment conditions are adjusted to a temperature of 22-24℃, relative humidity of 90%-95%, light intensity of 100-200 Lx, and CO2 concentration of 0.5-0.7%.

[0045] (7) Fruiting body tissue isolation: Before the fruiting bodies reach 6-7 maturity and the veil is broken, the yield and agronomic traits of the fruiting bodies are observed and statistically analyzed. Fruiting bodies with early fruiting, large number of fruiting bodies with high uniformity, thick stipe and thick cap are selected from the fruiting bags. Tissue isolation and purification culture are carried out on them. After 5 generations of domestication and cultivation experiments and repeated selection, a new strain of tea tree mushroom with excellent traits is finally selected. This strain has strong resistance, short cultivation cycle and excellent commercial traits. It has the characteristics of tea tree mushroom facility cultivation varieties and is named "Gan Tea Tree Mushroom No. 1".

Claims

1. A new strain of *Agrocybe aegerita*, Ganzhou *Agrocybe aegerita* No. 1, with preservation number CGMMCC NO41432, characterized in that... The strain was obtained through a selection method including the following steps: The breeding method includes the following steps: (1) Activation culture of the mother strain of tea tree mushroom: The purified and preserved wild tea tree mushroom strain mother strain was transferred to sterilized PDA medium in culture dishes and cultured in a biochemical incubator in the dark for 10 days to grow to the full size of the plate, thus activating the mother strain; (2) Production of primary and cultivars: The activated mother seed is propagated into primary and cultivars. The formula of primary and cultivars includes the following raw materials by weight percentage: sawdust 39%, cottonseed hulls 33%, wheat bran 18%, corn flour 8%, lime 1.5%, light calcium carbonate 1%, and moisture content 58%. (3) Preparation of cultivation material bags: After thoroughly mixing the cultivation materials according to the formula, control the moisture content of the culture material to 60% to 65%, and pack them into polypropylene plastic bags. Each bag weighs 0.5 to 0.55 kg of dry material. After packing, seal the bags with plastic rings and caps. After packing, sterilize the bags by high pressure. After sterilization, transfer the bags to a pre-sterilized cooling room for cooling. Inoculate the bags in the inoculation room according to the aseptic operation specifications. (4) Cultivation in cultivation bags: For the first 15 days of cultivation in cultivation bags, the temperature in the mycelium incubation room is 25-26℃, the relative humidity is 60%-65%, the light intensity is 50-100Lx, and the CO2 concentration is 0.04-0.1%; for 16-50 days, the temperature in the mycelium incubation room is 22-24℃, the relative humidity is 65%-70%, the light intensity is 100-200Lx, and the CO2 concentration is 0.06-0.08%. When the mycelium grows to 2 / 3 of the bag, loosen the bag opening and oxygenate the bag. When yellow water droplets start to appear at the bag opening and brown spots appear on the surface of the bag, the bag is physiologically mature. (5) Bud induction: After the mature mushroom bags are moved into the mushroom house, the ambient temperature of the mushroom house is controlled at 20-22℃, the relative humidity of the mushroom house is maintained at 85%-90%, 200-300 lux of diffused light is provided, and ventilation is provided to keep the air in the mushroom house fresh; when rice grain-like primordia appear on the surface of the mushroom bags, straighten the opening of the mushroom bags and then close them slightly. After 1-2 days, the primordia differentiate into mushroom buds. (6) Mushroom cultivation: After the mushroom buds are formed, the temperature of the mushroom growing environment is controlled at 22-24℃ and the relative humidity is 90%-95%. The number of ventilations and the time are reduced, and the carbon dioxide concentration in the mushroom house is increased by 0.5-0.7% to promote the growth of the tea tree mushroom stipe and inhibit the opening of the mushroom cap. (7) Tissue isolation of wild mushroom fruiting bodies: Before the fruiting bodies of *Agrocybe aegerita* reach 6-7 minutes of maturity and the veil is broken, the yield and agronomic traits of the fruiting bodies are observed and statistically analyzed. The fruiting bodies are selected, and their tissues are isolated and purified. After 5 rounds of domestication and repeated selection, the superior strain of *Agrocybe aegerita* is finally selected. Furthermore, morphological anatomy and molecular phylogenetic analysis techniques were used to identify wild fungi. The morphological anatomy and microstructure characteristics of the fruiting bodies are as follows: fruiting bodies are solitary, paired, or clustered; the cap is 3-10 cm in diameter, initially hemispherical, dark brown to tan, gradually becoming flattened, turning light brown to pale yellowish-brown, with a lighter edge and a darker center; when moist, it is sticky, smooth or wrinkled, and when young, it is covered with a veil that is rolled inwards; the flesh is white or off-white, slightly tough, thicker in the center and thinner at the edge; the stipe is solid, 3-12 cm long and 0.5-2 cm thick, cylindrical, nearly uniform in thickness, off-white, darker near the base, and fibrous on the surface; at maturity, the stipe hardens, is covered with a dull, sticky substance, and the annulus remains on the stipe or adheres to the edge of the cap and falls off automatically; the gills are adnate to decurrent, dense, and of unequal length, initially white, turning tan to brown when mature. The annulus is superior, initially white, later turning dark brownish-brown with basidiospores on the upper surface. It is membranous to filamentous and not easily detached. The basidiospores are 7-10×4-6µm in size, elliptical, smooth, and brownish-brown.

2. The method for breeding the new strain of *Agrocybe aegerita*, Ganchashugu No. 1, according to claim 1, is characterized in that, In step (3), the cultivation material formula includes the following raw materials by mass percentage: 36% cottonseed hulls, 29% sawdust, 22% wheat bran, 10% corn flour, 1% soybean meal, 1% lime, and 1% light calcium carbonate; the sterilization conditions are 0.11-0.14 MPa pressure, steam temperature of 121-126℃, and maintenance for 2-2.5 hours.

3. The method for breeding the new strain of *Agrocybe aegerita*, Ganchashugu No. 1, according to claim 1, is characterized in that, In step (5), the environment of the mushroom growing room is adjusted to a temperature of 20-22℃, relative humidity of 85%-90%, light intensity of 200-300Lx, and CO2 concentration of 0.1-0.3%. When rice-grain-shaped primordia appear on the surface of the mushroom bag, the opening of the mushroom bag is straightened. After 1-2 days, the primordia differentiate into mushroom buds.

4. The method for breeding the new strain of *Agrocybe aegerita*, Ganchashugu No. 1, according to claim 1, is characterized in that, In step (7), fruiting bodies with early fruiting, many buds, long stipes, good color and slow opening are selected from the fruiting bodies of Mushroom Agrocybe 'Tea Tree', and their tissues are separated and purified for culture.

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