Armillaria fungus agent and application thereof

Abstract

The application provides an armillaria mellea agent and application thereof, and belongs to the technical field of microbial agents; the armillaria mellea agent is a solid symbiotic agent, raw materials include armillaria mellea mycelium, an organic carrier and a composite functional additive; according to mass fractions, the armillaria mellea mycelium dry powder accounts for 12-28 parts, the organic carrier accounts for 50-75 parts, the composite functional additive accounts for 6-18 parts, and the balance is water; the armillaria mellea is an artificial domestication symbiotic armillaria mellea, and is not limited to a specific strain, and a conventional commercial armillaria mellea strain can be used. The application optimizes the raw material ratio, improves the fermentation, pretreatment and mixing process, does not need a specific preserved strain, and can be produced by using a conventional artificial domestication armillaria mellea, so that the process is simple, the cost is low, and the application is suitable for industrialized batch production.

Description

Technical Field

[0001] This invention provides an inoculant for Armillaria mellea and its application, belonging to the field of microbial inoculant technology. Background Technology

[0002] Armillaria mellea is a typical facultative parasitic fungus that can form a symbiotic relationship with Gastrodia elata and the roots of forest trees. It is widely used in the cultivation of Gastrodia elata in artificial forests, the rejuvenation of forestry seedlings, and soil improvement. Armillaria mellea relies on its hyphae to entwine the host's roots, forming mycelial cords to transport mineral elements and organic matter to the host, while simultaneously relying on the host's organic matter to sustain its own growth. It is a core functional fungal species for the cultivation of medicinal herbs under forest cover and for ecological forestry restoration.

[0003] Currently, the preparation process of conventional Armillaria mellea inoculants on the market is simple, but it generally suffers from the following technical defects: First, the traditional strain cultivation method is constant temperature culture, resulting in uneven mycelial growth, low mycelial cord formation, and poor strain activity; Second, the high-temperature drying process is mostly used after mycelial collection, which severely damages mycelial cells and results in a low viability rate; Third, ordinary inoculants only use simple physical mixing, resulting in poor carrier adsorption, short storage period, and susceptibility to moisture and contamination by other microorganisms; Fourth, the existing inoculants do not have a pre-activation process, resulting in slow colonization of mycelium after entering the soil, weak environmental resistance, and seriously affecting the yield of Gastrodia elata and the effect of improving the root system of forest trees.

[0004] Most existing Armillaria mellea inoculants have simple formulations and crude preparation processes. Insufficient dissolved oxygen during fermentation and low mycelial enrichment rates prevent large-scale, standardized production. Furthermore, these inoculants have poor resistance to low temperatures and drought, and are poorly adaptable to complex forest environments. Therefore, it is necessary to optimize the formulation system, improve the inoculum activation, liquid fermentation, mycelial treatment, and mixing processes, and develop an Armillaria mellea inoculant with high viability, strong stability, and suitability for forest understory cultivation. Utility Model Content

[0005] This invention aims to solve the technical problems of existing Armillaria mellea inoculants, such as low viability, low mycelial content, poor storage stability, weak adaptability to forest environments, and crude preparation processes. It provides an Armillaria mellea inoculant and its application. This invention optimizes the raw material ratio, improves the fermentation, pretreatment, and mixing processes, and does not require specific preservation strains. It can be produced using conventional commercially available artificially domesticated Armillaria mellea. The process is simple, low-cost, and suitable for industrial mass production.

[0006] To address the aforementioned problems, the present invention proposes the following technical solution: a *Armillaria mellea* inoculant and its application, wherein the *Armillaria mellea* inoculant is a solid symbiotic inoculant, and the raw materials include *Armillaria mellea* mycelium, an organic carrier, and a composite functional adjuvant; by mass, the proportions of each raw material are: 12-28 parts of dry *Armillaria mellea* mycelium powder, 50-75 parts of organic carrier, 6-18 parts of composite functional adjuvant, and the remainder is water; the *Armillaria mellea* is an artificially domesticated symbiotic type, without specific strain limitations, and conventional commercial *Armillaria mellea* strains can be used.

[0007] Furthermore, the organic carrier is a mixture of two or more of oak sawdust, corn cob powder, wheat bran, and humus, and the carrier is pulverized into particles of 40-90 mesh; the composite functional additive includes nutritional additives, stress-resistant protective additives, and slow-release binding additives.

[0008] Furthermore, by mass percentage, the composite functional additive comprises 40%–60% nutritional additives, 20%–35% stress-resistant protective additives, and 15%–25% slow-release binding additives; the nutritional additives are a mixture of glucose, potassium humate, and soluble phosphates; the stress-resistant protective additives are a mixture of seaweed polysaccharides, mannitol, and B vitamins; and the slow-release binding additives are xanthan gum and lignin sulfonate.

[0009] Furthermore, the preparation steps include the following: S1. Stepwise activation treatment of strains: Select healthy Armillaria mellea mother strains free from contamination, inoculate them into modified solid culture medium, and use variable temperature activation culture at 21-26℃. Incubate in the dark for 6-10 days to obtain activated mycelia. Add trace amounts of magnesium salts and vitamin nutrient solution to the modified solid culture medium. S2. Liquid deep layer expansion culture: The activated mycelium is chopped and inoculated into the liquid fermentation medium. The fermentation process is carried out by segmented variable speed stirring + gradient temperature control. The fermentation cycle is 5 to 9 days to obtain a uniform mycelium fermentation broth containing mycelium cords. S3. Low-temperature enrichment pretreatment of mycelia: The fermentation broth is filtered to remove impurities, and mycelia and mycelial cords are collected. They are then subjected to low-temperature gentle cell wall disruption treatment at a temperature of 0-5℃ for 10-30 min to preserve the biological activity of the mycelia. S4. Low-temperature vacuum drying and powdering: The pretreated mycelium is spread out and dried under low-temperature vacuum. The drying temperature is not higher than 38℃ and the vacuum degree is controlled between -0.06 and -0.095MPa. The material is dried until the moisture content is ≤5%. After pulverizing, it is passed through an 80-120 mesh sieve to obtain dry powder of Armillaria mellea mycelium. S5. Closed-loop slow-release mixing: The mycelial dry powder, organic carrier, and compound functional additives are sequentially added to a closed mixing equipment. The mixing method is adopted in stages: first, mix at low speed, then spray and humidify at low speed. The mixing speed is 30-70 r / min, the mixing time is 20-40 min, and the moisture content of the material is adjusted to 5%-12%. After packaging, the Armillaria mellea inoculant is obtained.

[0010] Furthermore, in step S2, the liquid fermentation culture medium comprises, by mass percentage: 15%–25% potato extract, 2%–4% glucose, 0.5%–1.5% peptone, 0.2%–0.8% yeast extract, with the remainder being sterile water, and the initial pH of the culture medium is adjusted to 5.0–6.0.

[0011] Furthermore, the segmented variable-speed stirring + gradient temperature-controlled fermentation process in step S2 is specifically as follows: During the initial fermentation stage (1-2 days): maintain a constant temperature of 24-26℃ and a stirring speed of 60-90 r / min to promote mycelial germination; During the middle stage of fermentation (3-5 days): cool down to 22-24℃, stir at 100-140 r / min, and enhance dissolved oxygen propagation. During the later stage of fermentation (6-9 days): cool down to 19-22℃, stir at 40-70 r / min, and let stand to enrich mycelial cords.

[0012] Furthermore, the segmented mixing method in step S5 is as follows: first, dry mix for 15-25 minutes, then slowly humidify with atomized pure water spray, continuously humidify and mix for 5-15 minutes, in a closed and sterile environment throughout the process to avoid contamination by other bacteria.

[0013] Furthermore, the modified solid culture medium raw materials mentioned in step S1 include potatoes, agar, glucose, magnesium sulfate, and vitamin B1. The pH of the culture medium is adjusted to 5.2-5.7, and it is sterilized by high-pressure steam before use.

[0014] Furthermore, the Armillaria mellea inoculant is used for the companion cultivation of Gastrodia elata under forest cover and for promoting and improving the root system of forest trees. The application methods include hole application mixed with soil, wrapping the inoculant material, and covering with soil and spreading. When used for Gastrodia elata cultivation, the Armillaria mellea inoculant is laid in layers with broad-leaved tree segments at intervals, with a thickness of 2-5 cm, covered with soil and compacted, and the humidity of the cultivation environment is controlled at 60%-85%. When used for forest tree improvement, a circular trench is dug around the root system of the forest trees, and the inoculant is mixed with garden soil at a mass ratio of 1:5 to 1:10 and backfilled.

[0015] Furthermore, before applying the Armillaria mellea inoculant, it is pre-activated by sterile water mist at room temperature for 12-24 hours at an activation temperature of 20-25°C. After the Gastrodia elata cultivation is completed, it is treated with shading and moisture retention, with the shading rate controlled at 70%-90%.

[0016] Due to the adoption of the above technical solution, the beneficial effects of the Armillaria mellea inoculant and its application of the present invention are as follows: 1. Wide adaptability of strains, no strain restrictions: This invention does not require specific preservation strains. It can be produced using conventional artificial domestication of commercial Armillaria mellea, which lowers the production threshold and is suitable for large-scale production by small and medium-sized processing enterprises. It has extremely strong versatility.

[0017] 2. Optimized variable temperature fermentation process: This invention adopts a segmented temperature control + variable speed stirring fermentation process. The initial high temperature promotes germination, the middle medium temperature promotes propagation, and the later low temperature enriches mycelial cords, which solves the problems of uneven mycelial growth and low mycelial cord yield in traditional constant temperature fermentation, and increases mycelial biomass by more than 25%.

[0018] 3. Low-temperature treatment preserves activity: This invention uses low-temperature gentle cell disruption and low-temperature vacuum drying technology to avoid denaturation of mycelial proteins caused by high temperature, thus preserving mycelial activity to the maximum extent. The viable bacteria rate inside the agent can reach more than 92%, which is far higher than that of conventional high-temperature dried agents.

[0019] 4. Scientifically formulated and highly resistant to adverse conditions: This invention adds seaweed polysaccharides, mannitol and other stress-resistant additives to enhance the mycelium's resistance to low temperatures, drought and poor soil conditions; combined with slow-release binding agents, it extends the shelf life of the inoculant, with a shelf life of 6 to 8 months at room temperature, and it is not easily affected by moisture and mold.

[0020] 5. Closed-loop segmented mixing, low rate of contaminants: The closed-loop mixing method of first dry mixing and then atomized humidification avoids dust pollution and contaminant invasion, resulting in high purity of the inoculant and suitability for complex understory cultivation environments.

[0021] 6. Improved application process and fast planting speed: This invention adds a pre-activation process for the inoculant, which restores the activity of the mycelium before it enters the soil, shortening the root planting time by 3 to 5 days; when used for Gastrodia elata cultivation, the symbiotic combination rate is high, and the yield of Gastrodia elata is increased by 15% to 22%; when used for forest tree improvement, it can promote the germination of fibrous roots and improve the survival rate of seedlings. Attached Figure Description

[0022] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used in conjunction with embodiments of the invention to explain the invention and do not constitute a limitation thereof. In the drawings: Figure 1 This is a flowchart illustrating the preparation process of an Armillaria mellea inoculant and its applications, as described in this invention.

[0023] Figure 2 This is a fermentation process diagram of an Armillaria mellea inoculant and its application according to the present invention.

[0024] Figure 3 This diagram illustrates the application of an Armillaria mellea inoculant and its application methods according to the present invention. Detailed Implementation

[0025] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0026] Example 1 1. Raw material ratio (parts by mass) The mixture consists of 20 parts of dry powdered Armillaria mellea mycelium, 65 parts of organic carrier, 10 parts of compound functional additives, and the remainder is water. The organic carrier is composed of oak sawdust, corn cob powder, and wheat bran in a ratio of 5:3:2, with a particle size of 60 mesh. The compound additives consist of 50% nutrient additives, 30% stress-resistant protective additives, and 20% slow-release binding additives.

[0027] 2. Preparation process S1. Strain activation: Using conventional commercially domesticated Armillaria mellea mother culture, inoculated with modified solid culture medium containing potato, agar, glucose, magnesium sulfate, vitamin B1, pH 5.5, autoclaved; cultured at variable temperature, 22-25℃, in the dark for 8 days.

[0028] S2, Liquid fermentation: Liquid culture medium consists of 20% potato extract, 3% glucose, 1% peptone, 0.5% yeast extract, and the remainder is sterile water, pH 5.5; Segmented fermentation: 1-2 days, 25℃, 80r / min; 3-5 days, 23℃, 120r / min; 6-8 days, 20℃, 50r / min.

[0029] S3. Low-temperature pretreatment: Filter and collect mycelium, then gently heat and break the cell wall at 3℃ for 20 minutes.

[0030] S4. Drying and powdering: Drying temperature 35℃, vacuum degree -0.08MPa, dry to moisture content 4%, and pass through a 100-mesh sieve.

[0031] S5. Closed mixing: Dry mix for 20 minutes, atomize and humidify for 10 minutes, stir at 50 r / min, adjust the moisture content to 8%, and dispense to obtain the finished bacterial agent.

[0032] 3. Application Method Pre-activate at 20℃ for 18 hours before application; when cultivating Gastrodia elata, lay the fungicide and broad-leaved tree segments in layers with a thickness of 3cm, cover with soil and compact, maintain an ambient humidity of 75% and a shading rate of 80%; when improving forest trees, mix the fungicide with garden soil at a ratio of 1:8 and backfill.

[0033] Example 2 1. Raw material ratio (parts by mass) The mixture consists of 12 parts of dry powder of Armillaria mellea mycelium, 75 parts of organic carrier, 6 parts of compound functional additives, and the remainder is water; the organic carrier is corn cob powder: humus = 4:1, with a particle size of 40 mesh; the compound additives include 40% nutrient additives, 35% stress-resistant protective additives, and 25% slow-release binding additives.

[0034] 2. Preparation process S1. Activation of strains: Using conventional commercially domesticated Armillaria mellea mother strain, inoculated with modified solid culture medium, pH 5.2, autoclaved, and cultured at variable temperature (21-23℃) for 6 days in the dark.

[0035] S2, Liquid fermentation: Liquid culture medium consists of 15% potato extract, 2% glucose, 0.5% peptone, 0.2% yeast extract, and the remainder is sterile water, pH 5.0; Segmented fermentation: 1-2 days, 24℃, 60r / min; 3-5 days, 22℃, 100r / min; Fermentation ends after 5 days, and the mycelial cords are enriched by static incubation at low temperature.

[0036] S3. Low-temperature pretreatment: Filter and collect mycelia, then gently warm and break the cell walls at 0℃ for 10 minutes to preserve the complete mycelial structure.

[0037] S4. Drying and powdering: Drying temperature 32℃, vacuum degree -0.06MPa, dry to moisture content 3.2%, and pass through an 80-mesh sieve.

[0038] S5. Sealed mixing: Dry mix for 15 minutes, atomize and humidify for 5 minutes, stir at 30 r / min, adjust the moisture content to 5%, and seal and package to obtain the finished bacterial agent.

[0039] 3. Application Method Pre-activate at 22℃ for 12 hours before application; when cultivating Gastrodia elata in mountainous areas, lay the inoculant and broad-leaved tree segments in layers with a thickness of 2cm, cover with soil and compact, maintain an ambient humidity of 60% and a shading rate of 70%; when improving barren mountain forests, mix the inoculant with garden soil at a ratio of 1:5 and backfill, which is suitable for improving barren mountain soil.

[0040] Example 3 1. Raw material ratio (parts by mass) The mixture consists of 28 parts of dry powder of Armillaria mellea mycelium, 50 parts of organic carrier, 18 parts of compound functional additives, and the remainder is water; the organic carrier is oak wood chips: wheat bran = 3:2, with a particle size of 90 mesh; the compound additives consist of 60% nutrient additives, 20% stress-resistant protective additives, and 20% slow-release binding additives.

[0041] 2. Preparation process S1. Activation of strains: Using conventional commercially domesticated Armillaria mellea mother strain, inoculated with modified solid culture medium, pH 5.7, autoclaved, and cultured at variable temperature (24-26℃) for 10 days in the dark.

[0042] S2, Liquid fermentation: Liquid culture medium consists of 25% potato extract, 4% glucose, 1.5% peptone, 0.8% yeast extract, and the remainder is sterile water, pH 6.0; Segmented fermentation: 1-2 days, 26℃, 90r / min; 3-5 days, 24℃, 140r / min; 6-9 days, 22℃, 70r / min, extending the fermentation time to enrich a large number of mycelial cords.

[0043] S3. Low-temperature pretreatment: Filter and collect mycelia, gently warm and break the cell walls at 5℃ for 30 minutes to break the mycelial cell walls and release endogenous nutrients.

[0044] S4. Drying and powdering: Drying temperature 38℃, vacuum degree -0.095MPa, dry to moisture content 4.5%, and pass through a 120-mesh sieve.

[0045] S5. Sealed mixing: Dry mix for 25 minutes, atomize and humidify for 15 minutes, stir at 70 r / min, adjust the moisture content to 12%, and seal and package to obtain the finished bacterial agent.

[0046] 3. Application Method Pre-activate at 25℃ for 24 hours before application; when cultivating high-quality Gastrodia elata, lay the inoculant and broad-leaved tree segments in layers with a thickness of 5cm, cover with soil and compact, maintain an ambient humidity of 85% and a shading rate of 90%; when cultivating precious seedlings, mix the inoculant with garden soil at a ratio of 1:10 and backfill to enhance the root symbiotic effect.

[0047] Example 4 (General-purpose forest land improvement microbial agent with medium ratio) 1. Raw material ratio (parts by mass) The mixture consists of 20 parts of dry powdered Armillaria mellea mycelium, 62 parts of organic carrier, 13 parts of compound functional additives, and the remainder is water. The organic carrier is composed of oak sawdust, corn cob powder, wheat bran, and humus in a mass ratio of 3:2:2:3, with a particle size of 65 mesh. The compound additives consist of 52% nutrient additives, 28% stress-resistant protective additives, and 20% slow-release binding additives.

[0048] 2. Preparation process S1. Activation of strains: Using conventional commercially domesticated Armillaria mellea mother strain, inoculated with modified solid culture medium, pH 5.4, autoclaved, and cultured at variable temperature (22-24℃) for 7 days in the dark.

[0049] S2, Liquid fermentation: Liquid culture medium consists of 20% potato extract, 3% glucose, 1.0% peptone, 0.6% yeast extract, and the remainder is sterile water, pH 5.5; Segmented fermentation: 1-2 days, 25℃, 75r / min; 3-5 days, 23℃, 110r / min; 6-8 days, 21℃, 60r / min.

[0050] S3. Low-temperature pretreatment: Filter to collect mycelium, then gently heat and break the cell wall at 2℃ for 22 min.

[0051] S4. Drying and powdering: Drying temperature 34℃, vacuum degree -0.08MPa, dry to moisture content 3.8%, and pass through a 100-mesh sieve.

[0052] S5. Sealed mixing: Dry mix for 22 minutes, atomize and humidify for 12 minutes, stir at 45 r / min, adjust the moisture content to 9%, and seal and package to obtain the finished bacterial agent.

[0053] 3. Application Method Pre-activate at 22℃ for 15 hours before application; suitable for ordinary Gastrodia elata field cultivation and conventional economic forest improvement. For Gastrodia elata cultivation, lay the mixture 3.5cm thick, with an ambient humidity of 72% and a shading rate of 82%; mix the forest improvement microbial agent with garden soil at a ratio of 1:7 for backfilling. It is highly versatile and adaptable to most forest soil environments.

[0054] The mixture consisted of 28 parts of dried Armillaria mellea mycelium powder, 50 parts of organic carrier, and 18 parts of compound functional additives. The organic carrier was made of oak sawdust and wheat bran with a particle size of 90 mesh. The fermentation cycle was 9 days, and dissolved oxygen stirring was enhanced. The remaining process parameters were the same as in Example 1.

[0055] Effect comparison test A comparative experiment was conducted between the fungal agents prepared in Examples 1, 2, and 3 of this invention and commercially available common Armillaria mellea fungal agents: 1. Viability test: The viability rate of commercially available bacterial agents is 65.2%, while the viability rate of Example 1 of this invention is 93.5%; 2. Storage stability: After 6 months of storage at room temperature, the mold rate of commercially available microbial agents is 18.6%, while the mold rate of this invention is less than 2.1%. 3. Gastrodia elata cultivation experiment: Under the same cultivation conditions, the yield of commercially available inoculant for Gastrodia elata is 128 kg per mu, while the yield of Example 1 of this invention is 152 kg per mu; 4. Tree survival rate: When transplanting seedlings from barren hills, the survival rate of commercially available inoculants is 78.3%, while the survival rate of this invention is 94.7%.

[0056] The present invention and its embodiments have been described above. This description is not restrictive. In short, if a person skilled in the art is inspired by this description and designs a similar structure and embodiment without departing from the spirit of the present invention, such design should fall within the protection scope of the present invention.

Claims

1. A fungal agent for Armillaria mellea and its application, characterized in that, The Armillaria mellea inoculant is a solid symbiotic inoculant, and the raw materials include Armillaria mellea mycelium, organic carrier, and compound functional adjuvants. By mass, the proportions of each raw material are: 12-28 parts of dry powdered Armillaria mellea mycelium, 50-75 parts of organic carrier, 6-18 parts of compound functional adjuvants, and the remainder is water. The Armillaria mellea is an artificially domesticated symbiotic type, and there is no specific strain limitation; conventional commercial Armillaria mellea strains can be used.

2. The inoculum agent of Armillaria mellea according to claim 1 and its application, characterized in that: The organic carrier is a mixture of two or more of oak sawdust, corn cob powder, wheat bran, and humus, and the carrier is pulverized to a particle size of 40-90 mesh; the composite functional additive includes a nutritional additive, a stress-resistant protective additive, and a slow-release binding additive.

3. The inoculum agent of Armillaria mellea according to claim 1 and its application, characterized in that: By mass percentage, the composite functional additive comprises 40%–60% nutritional additives, 20%–35% stress-resistant protective additives, and 15%–25% slow-release binding additives; the nutritional additives are a mixture of glucose, potassium humate, and soluble phosphates; the stress-resistant protective additives are a mixture of seaweed polysaccharides, mannitol, and B vitamins; and the slow-release binding additives are xanthan gum and lignin sulfonate.

4. The inoculum agent of Armillaria mellea according to claim 1 and its application, comprising the following preparation steps: S1. Stepwise activation treatment of strains: Select healthy Armillaria mellea mother strains free from contamination, inoculate them into modified solid culture medium, and use variable temperature activation culture at 21-26℃. Incubate in the dark for 6-10 days to obtain activated mycelia. Add trace amounts of magnesium salts and vitamin nutrient solution to the modified solid culture medium. S2. Liquid deep layer expansion culture: The activated mycelium is chopped and inoculated into the liquid fermentation medium. The fermentation process is carried out by segmented variable speed stirring + gradient temperature control. The fermentation cycle is 5 to 9 days to obtain a uniform mycelium fermentation broth containing mycelium cords. S3. Low-temperature enrichment pretreatment of mycelia: The fermentation broth is filtered to remove impurities, and mycelia and mycelial cords are collected. They are then subjected to low-temperature gentle cell wall disruption treatment at a temperature of 0-5℃ for 10-30 min to preserve the biological activity of the mycelia. S4. Low-temperature vacuum drying and powdering: The pretreated mycelium is spread out and dried under low-temperature vacuum. The drying temperature is not higher than 38℃ and the vacuum degree is controlled between -0.06 and -0.095MPa. The material is dried until the moisture content is ≤5%. After pulverizing, it is passed through an 80-120 mesh sieve to obtain dry powder of Armillaria mellea mycelium. S5. Closed-loop slow-release mixing: The mycelial dry powder, organic carrier, and compound functional additives are sequentially added to a closed mixing equipment. The mixing method is adopted in stages: first, mix at low speed, then spray and humidify at low speed. The mixing speed is 30-70 r / min, the mixing time is 20-40 min, and the moisture content of the material is adjusted to 5%-12%. After packaging, the Armillaria mellea inoculant is obtained.

5. The inoculum agent of Armillaria mellea according to claim 4 and its application, characterized in that: In step S2, the liquid fermentation culture medium comprises, by mass percentage: 15%–25% potato extract, 2%–4% glucose, 0.5%–1.5% peptone, 0.2%–0.8% yeast extract, with the remainder being sterile water, and the initial pH of the culture medium is adjusted to 5.0–6.

0.

6. The inoculum agent of Armillaria mellea according to claim 4 and its application, characterized in that: The segmented variable-speed stirring + gradient temperature-controlled fermentation process in step S2 is as follows: During the initial fermentation stage (1-2 days): maintain a constant temperature of 24-26℃ and a stirring speed of 60-90 r / min to promote mycelial germination; During the middle stage of fermentation (3-5 days): cool down to 22-24℃, stir at 100-140 r / min, and enhance dissolved oxygen propagation. During the later stage of fermentation (6-9 days): cool down to 19-22℃, stir at 40-70 r / min, and let stand to enrich mycelial cords.

7. The inoculum agent of Armillaria mellea according to claim 4 and its application, characterized in that: The segmented mixing method in step S5 is as follows: first, dry mix for 15-25 minutes, then slowly humidify with atomized pure water spray, and continue to humidify and mix for 5-15 minutes. The whole process is carried out in a closed and sterile environment to avoid contamination by other bacteria.

8. The inoculum agent of Armillaria mellea according to claim 4 and its application, characterized in that: The modified solid culture medium raw materials mentioned in step S1 include potato, agar, glucose, magnesium sulfate, and vitamin B1. The pH of the culture medium is adjusted to 5.2-5.7, and it is sterilized by high-pressure steam before use.

9. The inoculum agent of Armillaria mellea according to claim 1 and its application, characterized in that: The Armillaria mellea inoculant is used for the companion cultivation of Gastrodia elata under forests and for promoting and improving the root system of forest trees. The application methods include hole application mixed with soil, wrapping the inoculant material, and covering with soil and spreading. When used for Gastrodia elata cultivation, the Armillaria mellea inoculant is laid in layers with broad-leaved tree segments at intervals, with a thickness of 2-5 cm. The soil is then covered and compacted, and the humidity of the cultivation environment is controlled at 60%-85%. When used for forest tree improvement, a circular trench is dug around the root system of the forest trees, and the inoculant is mixed with garden soil at a mass ratio of 1:5 to 1:10 and backfilled.

10. The inoculum agent of Armillaria mellea according to claim 1 and its application, characterized in that: Before applying the Armillaria mellea inoculant, it is pre-activated by sterile water mist at room temperature for 12-24 hours at an activation temperature of 20-25°C. After the Gastrodia elata cultivation is completed, it is treated with shading and moisture retention, with the shading rate controlled at 70%-90%.

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