Method for obtaining cordyceps militaris sporocarp through liquid culture
By controlling parameters such as temperature, humidity, and light through liquid culture and optimizing the culture medium formula, the problem that liquid fermentation cannot directly cultivate Cordyceps militaris fruiting bodies has been solved, achieving efficient and low-cost production of Cordyceps militaris fruiting bodies and improving product quality and large-scale production capacity.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SHANXI FUNCTIONAL FOOD RES INST OF SHANXI AGRI UNIV
- Filing Date
- 2026-01-23
- Publication Date
- 2026-04-28
AI Technical Summary
In existing technologies, liquid fermentation of Cordyceps militaris can only obtain mycelium, and cannot directly cultivate complete Cordyceps militaris fruiting bodies in liquid culture medium, resulting in a complex production process, high cost, and difficulty in achieving large-scale production.
The liquid culture method was used to cultivate Cordyceps militaris fruiting bodies in a liquid culture medium by controlling environmental parameters such as temperature, humidity and light in multiple steps. This included stages such as strain activation, mycelial growth, aerial mycelial formation and fruiting body maturation. The culture medium formula was optimized to promote the growth of Cordyceps militaris fruiting bodies.
This method enables the direct cultivation of complete Cordyceps militaris fruiting bodies in liquid culture medium, improving production efficiency and biomass harvest, reducing pollution risks, and enhancing the medicinal and economic value of the product, which aligns with the concept of green and environmentally friendly production.
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Figure CN121926085A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of artificial fungal cultivation technology, specifically a method for obtaining Cordyceps militaris fruiting bodies through liquid culture. Background Technology
[0002] Cordyceps militaris belongs to the Ascomycota, Clavicipitaceae, and Cordyceps genus. As a precious medicinal and edible fungus, it boasts extremely high nutritional value, containing active ingredients such as ergosterol, minerals, vitamins, cordycepic acid, cordycepin, and cordyceps polysaccharides. Modern research indicates that Cordyceps militaris possesses various pharmacological effects, including relieving fatigue, regulating immunity, antioxidation, lowering blood sugar, anti-tumor activity, anti-neurological disease activity, and antiviral activity. Cordyceps militaris is a unique resource in my country and has obtained a new food ingredient approval number due to its rich nutritional value. Besides medicinal uses, Cordyceps militaris extract is also used as an ingredient in various functional foods, nutritional supplements, and cosmetics. Currently, Cordyceps militaris mainly appears on the market in the form of health products and foods, such as tea, wine, oral liquids, and vinegar. Wild Cordyceps militaris is rare and expensive, thus failing to meet market demand. Fortunately, artificially cultivated Cordyceps militaris and its products are now available. In current scientific research, there are two main methods for cultivating Cordyceps militaris: one is to use crops such as wheat and rice as substrates to cultivate fruiting bodies, and the other is to obtain fruiting bodies by parasitizing silkworm pupae with Cordyceps militaris fungus. In recent years, due to the high economic value of Cordyceps militaris, research on liquid fermentation of Cordyceps militaris has gradually increased. Liquid fermentation can produce key bioactive compounds such as cordycepin and polysaccharides.
[0003] However, in the existing technology, the liquid fermentation of Cordyceps militaris can only obtain mycelium, and cannot directly cultivate Cordyceps militaris fruiting bodies with complete morphology in liquid culture medium. In order to obtain fruiting bodies, it is necessary to combine it with other methods such as solid cultivation. This not only increases the complexity of the production process and the cost input, but also makes it difficult to give full play to the advantages of liquid fermentation in terms of production efficiency, cost control and large-scale production, thus forming a significant technical barrier. Summary of the Invention
[0004] The purpose of this invention is to provide a method for obtaining Cordyceps militaris fruiting bodies through liquid culture in order to solve the problems mentioned above.
[0005] The technical solution adopted in this invention is as follows: A method for obtaining Cordyceps militaris fruiting bodies through liquid culture, the method comprising the following steps: S1: Under aseptic conditions, the preserved Cordyceps militaris slant culture was inoculated onto PDA solid medium and cultured at 22°C in the dark for 7 days to complete the activation of the culture, providing a robust germplasm base for subsequent liquid inoculation.
[0006] S2: Take the activated strain and inoculate it into the culture medium at a ratio of 4 5mm diameter mycelial blocks per 100mL of liquid culture medium. After inoculation, let it stand in the dark for 24 hours to allow the strain to adapt to the environment. Then transfer it to a constant temperature shaker and culture it at 150r / min and 25℃ for 3 days to obtain uniformly sized mycelial balls, which will provide sufficient and consistent mycelial material for subsequent mycelial growth culture.
[0007] S3: Transfer the culture flasks containing bacterial balls to an environment of 18℃ and keep them in the dark for 7-10 days to allow the bacterial balls to grow further and intertwine to form an aerial mycelium film on the surface of the culture medium, creating a structural basis for subsequent color change upon exposure to light.
[0008] S4: After the culture medium is covered with aerial mycelium film, move the culture bottle to an artificial climate chamber with a temperature of 22℃, humidity of 70%-75% and light intensity of 1000Lx. Maintain 12 hours of light per day for light culture until the white film completely turns orange-yellow, which promotes the mycelium on the film to twist and form primordia, thus preparing the morphology for the growth of fruiting bodies.
[0009] S5: When the primordia on the mycelium grow to 0.5-1cm, adjust the culture temperature to 20℃, increase the humidity to 85%-95%, continue to maintain 12 hours of light per day, and continue to culture for 30-35 days to finally obtain mature Cordyceps militaris fruiting bodies.
[0010] In a preferred embodiment, in step S1, the nutrient composition of the PDA solid culture medium is as follows: 200g of potato, peeled and chopped, is added to 1000mL of water, boiled for 30min, filtered through 4 layers of gauze to obtain the filtrate, and then 20g of glucose, 1.5g of fish peptone, 1.5g of KH2PO4, 0.5g of MgSO4, 0.5g of CaCl2, and 20g of agar are added, and the volume is adjusted to 1000mL, maintaining the natural pH.
[0011] In a preferred embodiment, in step S1, the prepared culture medium is sterilized by steam at 0.1 MPa pressure and 121°C for 30 minutes. After cooling to room temperature, the preserved Cordyceps militaris slant culture is inoculated onto the surface of the culture medium. The inoculated culture medium is then transferred to a light-protected environment at 22°C and cultured for 7 days to ensure that the culture fully recovers its vitality and provides a robust and high-quality seed source for subsequent liquid culture.
[0012] In a preferred embodiment, in step S2, the nutrient composition of the liquid culture medium is as follows: 200g of peeled and diced potatoes and 20g of silkworm pupae are added to 1000mL of water, boiled for 30min, filtered through 4 layers of gauze to obtain the filtrate, and 20g of sucrose, 1.5g of fish peptone, 1.5g of KH2PO4, 0.5g of MgSO4, and 0.5g of CaCl2 are added, and the volume is adjusted to 1000mL, maintaining the natural pH value.
[0013] In a preferred embodiment, in step S2, the liquid culture medium is sterilized with steam at 0.1 MPa pressure and 121°C for 30 minutes. After cooling to room temperature, inoculation is completed at a ratio of 4 activated bacterial blocks with a diameter of 5 mm per 100 ml of culture medium. After inoculation, the culture container is sealed with a polypropylene breathable film and then placed in a light-proof environment for 24 hours to allow the strain to gradually adapt to the nutritional environment of the liquid culture medium, thus preparing for the rapid growth of mycelium.
[0014] In a preferred embodiment, in step S3, after the static adaptation is completed, the sealed culture container is transferred to a constant temperature shaker, the shaker speed is set to 150 r / min, the culture temperature is controlled at 25°C, and the culture is continued for 3 days. The culture is a closed culture, and the sealing method is to use a polypropylene breathable film for sealing.
[0015] Under the continuous vibration of the shaker, the inoculum disperses evenly in the liquid culture medium and grows and multiplies rapidly, eventually forming uniformly sized mycelial balls. The mycelium inside these mycelial balls is vigorous and stable, providing a large amount of high-quality mycelial material for the subsequent mycelial growth stage, effectively ensuring the continuity and stability of the entire culture process.
[0016] In a preferred embodiment, in step S4, during the mycelial growth stage, the culture bottle containing a large number of mycelial balls needs to be transferred to an environment at 18°C and kept in a static, light-protected state for 7 to 10 days. The culture is a closed culture, and the sealing method is to use a polypropylene breathable membrane for sealing. Under relatively low temperature and light-free conditions, the mycelium will germinate and grow further, gradually extending towards the surface of the liquid culture medium and intertwining and aggregating on the surface, eventually forming a tightly structured aerial mycelial film. This film is the core foundation for subsequent color change upon exposure to light and the growth of fruiting bodies, providing the necessary structural support and nutrient supply for the differentiation and development of primordia.
[0017] In a preferred embodiment, in step S5, the culture is a closed culture, and the closed method is to seal it with a polypropylene breathable film; the culture bottle is transferred to an artificial climate chamber, the ambient temperature is set at 22°C, the air humidity is controlled between 70% and 75%, and the light intensity is adjusted to 1000 Lx.
[0018] In a preferred embodiment, step S5 involves maintaining 12 hours of light exposure daily. Under the combined effects of temperature, humidity, and light, the initially white bacterial film gradually transforms into orange-yellow, and the hyphae on the film begin to twist and differentiate, gradually forming primordia primordia. Controlling the environmental parameters at this stage directly affects the quantity and quality of primordia formation, requiring strict maintenance of the stability of all conditions.
[0019] In a preferred embodiment, in step S6, when the primordia on the mycelium grow to 0.5 to 1 cm, the fruiting body maturation stage begins. At this time, the culture environment parameters are adjusted: the temperature is lowered to 20°C, while the air humidity is increased to 85% to 95%, and 12 hours of light per day is maintained for 30 to 35 days. Under the combined effects of suitable temperature, high humidity, and stable light, the primordia will continuously elongate and thicken, gradually developing into fully formed Cordyceps militaris fruiting bodies. Throughout the entire culture process, close monitoring of environmental changes is necessary to avoid sudden temperature changes, excessive humidity fluctuations, etc., to ensure the normal growth and maturation of the fruiting bodies, ultimately obtaining a high-quality Cordyceps militaris fruiting body product.
[0020] In summary, due to the adoption of the above technical solution, the beneficial effects of the present invention are: 1. This invention enables the direct cultivation of complete Cordyceps militaris fruiting bodies in a liquid culture medium, overcoming the limitations of traditional liquid fermentation which can only obtain mycelium. Compared with solid substrate cultivation methods, liquid culture offers advantages such as speed, low cost, and small footprint, allowing for the rapid production of a large number of propagules with more stable culture quality and effectively reducing the risk of contamination. Furthermore, the harvesting process in liquid culture is simpler, allowing direct separation of mycelium from the culture medium, and the fruiting bodies can be harvested together with the mycelium after maturity, significantly increasing biomass harvesting and providing a new pathway for the large-scale, standardized production of Cordyceps militaris.
[0021] 2. In this invention, by optimizing the culture medium formula and culture conditions, the accumulation of antioxidant active substances such as flavonoids and polyphenols in the fruiting bodies of Cordyceps militaris is significantly increased, and the cordycepin content reaches its peak in the later stage of fruiting body growth, effectively enhancing the medicinal and economic value of the product. The waste liquid culture medium generated by liquid culture has low nutrient content, is easier to treat subsequently, and has less environmental impact, which is in line with the concept of green and environmentally friendly production. It provides technical support for the sustainable development of the Cordyceps militaris industry and can significantly increase the content of functional components in Cordyceps militaris. Attached Figure Description
[0022] Figure 1 This is a schematic diagram illustrating the process principle of the present invention; Figure 2This is a comparison chart of cordycepin and adenosine content at different growth stages of Cordyceps militaris in this invention, where MY: mycelial stage, MM: mycelial stage, CP: color change stage, PR: primordium stage, and FB: fruiting body stage. Detailed Implementation
[0023] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.
[0024] Reference Figure 1-2 , Example 1: A method for obtaining Cordyceps militaris fruiting bodies through liquid culture, comprising the following steps: (1) Activation of the strain: Under aseptic conditions, the preserved Cordyceps militaris slant culture was inoculated into PDA solid medium and cultured at 22℃ in the dark for 7 days. The PDA solid medium contained the following nutrients: 200g of peeled and diced potatoes were added to 1000mL of water, boiled for 30min, filtered through 4 layers of gauze, and the filtrate was collected. 20g of glucose, 1.5g of fish peptone, 1.5g of KH2PO4, 0.5g of MgSO4, 0.5g of CaCl2, and 20g of agar were added to make up to 1000mL, and the pH was adjusted to natural. The medium was then autoclaved at 0.1MPa and 121℃.
[0025] (2) Fermentation culture and mycelial growth: A 300mL tissue culture flask was filled with 100mL of liquid culture medium and sealed with a polypropylene breathable film. The nutrient composition of the liquid culture medium was as follows: 200g of peeled and diced potatoes and 20g of silkworm pupae were added to 1000mL of water and boiled for 30min. The filtrate was filtered through four layers of gauze, and 20g of sucrose, 1.5g of fish peptone, 1.5g of KH2PO4, 0.5g of MgSO4, and 0.5g of CaCl2 were added. The volume was adjusted to 1000mL, and the pH was set to natural. The flask was then autoclaved at 0.1MPa and 121℃. After cooling to room temperature, the activated mycelium was inoculated into the liquid culture medium under aseptic conditions. Four mycelial blocks with a diameter of 5mm were inoculated into each 100mL of culture medium. After inoculation, the flask was allowed to stand in the dark for 24h. The culture flask was then incubated in a constant temperature shaker at 150r / min and 25℃ for 3 days to obtain uniformly sized mycelial balls.
[0026] (3) Mycelium growth culture: Place the culture bottle from which a large number of mycelium balls were generated in step (2) in a 18℃ environment and keep it in the dark for 7-10 days.
[0027] (4) Light cultivation: When the culture medium in step (3) is covered with aerial mycelium film, place it in an artificial climate chamber with an ambient temperature of 22℃, humidity of 70%-75% and light intensity of 1000Lx for light cultivation, and expose it to light for 12 hours a day until the white film turns orange-yellow.
[0028] (5) Fruiting body culture: When the mycelial primordia in step (4) grow to 0.5-1cm, adjust the culture temperature to 20℃. The humidity is 85%-95%, and the plant is exposed to light for 12 hours a day. The culture time is 30-35 days to obtain Cordyceps militaris fruiting bodies.
[0029] Example 2: (1) Activation of the strain: Under aseptic conditions, the preserved Cordyceps militaris slant culture was inoculated into PDA solid medium and cultured at 22℃ in the dark for 7 days. The PDA solid medium contained the following nutrients: 200g of peeled and diced potatoes were added to 1000mL of water, boiled for 30min, filtered through 4 layers of gauze, and the filtrate was collected. 20g of glucose, 1.5g of fish peptone, 1.5g of KH2PO4, 0.5g of MgSO4, 0.5g of CaCl2, and 20g of agar were added to make up to 1000mL, and the pH was adjusted to natural. The medium was then autoclaved at 0.1MPa and 121℃.
[0030] (2) Fermentation culture and mycelial growth: A 300mL tissue culture flask was filled with 100mL of liquid culture medium and sealed with a polypropylene breathable film. The nutrient composition of the liquid culture medium was as follows: 200g of peeled and diced potatoes and 20g of silkworm pupae were added to 1000mL of water and boiled for 30min. The filtrate was filtered through four layers of gauze, and 20g of glucose, 1.5g of fish peptone, 1.5g of KH2PO4, 0.5g of MgSO4, and 0.5g of CaCl2 were added to a final volume of 1000mL. The pH was adjusted to 0.1MPa and 121℃ for autoclaving. After cooling to room temperature, the activated mycelium was inoculated into the liquid culture medium under aseptic conditions. Three mycelial blocks with a diameter of 5mm were inoculated into each 100mL of culture medium. After inoculation, the flask was allowed to stand in the dark for 24h. The culture flask was then incubated in a constant temperature shaker at 150r / min and 25℃ for 3 days to obtain uniformly sized mycelial balls.
[0031] (3) Mycelium growth culture: Place the culture bottle from which a large number of mycelium balls were generated in step (2) in a 18℃ environment and keep it in the dark for 7-10 days.
[0032] (4) Light cultivation: When the culture medium in step (3) is covered with aerial mycelium film, place it in an artificial climate chamber with an ambient temperature of 22℃, humidity of 70%-75% and light intensity of 1000Lx for light cultivation, and expose it to light for 12 hours a day until the white film turns orange-yellow.
[0033] (5) Fruiting body culture: When the mycelial primordia in step (4) grow to 0.5-1cm, adjust the culture temperature to 20℃. The humidity is 85%-95%, and the plant is exposed to light for 12 hours a day. The culture time is 30-35 days to obtain Cordyceps militaris fruiting bodies.
[0034] Example 3 (1) Activation of the strain: Under aseptic conditions, the preserved Cordyceps militaris slant culture was inoculated into PDA solid medium and cultured at 22℃ in the dark for 7 days. The PDA solid medium contained the following nutrients: 200g of peeled and diced potatoes were added to 1000mL of water, boiled for 30min, filtered through 4 layers of gauze, and the filtrate was collected. 20g of glucose, 1.5g of fish peptone, 1.5g of KH2PO4, 0.5g of MgSO4, 0.5g of CaCl2, and 20g of agar were added to make up to 1000mL, and the pH was adjusted to natural. The medium was then autoclaved at 0.1MPa and 121℃.
[0035] (2) Fermentation culture and mycelial growth: A 300mL tissue culture flask was filled with 100mL of liquid culture medium and sealed with a polypropylene breathable film. The nutrient composition of the liquid culture medium was as follows: 200g of peeled and diced potatoes and 20g of silkworm pupae were added to 1000mL of water and boiled for 30min. The filtrate was filtered through four layers of gauze, and 20g of glucose, 1.5g of beef extract, 1.5g of KH2PO4, 0.5g of MgSO4, and 0.5g of CaCl2 were added to bring the volume to 1000mL. The pH was set to 0.1MPa and 121℃ for autoclaving. After cooling to room temperature, the activated mycelium was inoculated into the liquid culture medium under aseptic conditions. Four mycelial blocks with a diameter of 5mm were inoculated into each 100mL of culture medium. After inoculation, the culture flask was left to stand in the dark for 24h. The culture flask was then incubated in a constant temperature shaker at 150r / min and 25℃ for 3 days to obtain uniformly sized mycelial balls.
[0036] (3) Mycelium growth culture: Place the culture bottle from which a large number of mycelium balls were generated in step (2) in a 18℃ environment and keep it in the dark for 7-10 days.
[0037] (4) Light cultivation: When the culture medium in step (3) is covered with aerial mycelium film, place it in an artificial climate chamber with an ambient temperature of 22℃, humidity of 70%-75% and light intensity of 1000Lx for light cultivation, and expose it to light for 12 hours a day until the white film turns orange-yellow.
[0038] (5) Fruiting body culture: When the mycelial primordia in step (4) grow to 0.5-1cm, adjust the culture temperature to 20℃. The humidity is 85%-95%, and the plant is exposed to light for 12 hours a day. The culture time is 30-35 days to obtain Cordyceps militaris fruiting bodies.
[0039] The effects of different carbon and nitrogen sources on the content of phytosporum flavonoids and polyphenols are shown in the table below: Flavonoids (mg / g) Polyphenols (mg / g) ABTS free radical scavenging (mg / mL) Example 1 3.93 13.93 8.05 Example 2 3.67 11.64 8.42 Example 3 2.71 10.67 9.78 The effects of different carbon and nitrogen sources on the flavonoid and polyphenol content of Cordyceps militaris fruiting bodies are shown in the table below: Flavonoids Polyphenols Example 1 2.89 21.04 Example 2 2.73 17.37 As shown in Tables 1 and 2, the Cordyceps militaris substrate and fruiting bodies grown using the culture medium formulation of Example 1 have higher contents of flavonoids and polyphenols. Furthermore, it can be seen that the Cordyceps militaris substrate of Example 1 achieved an ABTS free radical scavenging rate of 50% at a sample concentration of 8.05 mg / mL. These comparative data fully demonstrate that the cultivation method provided by this invention can significantly enhance the accumulation of antioxidant active substances such as flavonoids and polyphenols in Cordyceps militaris.
[0040] like Figure 2 As shown, adenosine content is higher during the MY, MM, and CP stages. During the FB stage, adenosine content increases to meet the needs of cordycepin synthesis, acting as a precursor to cordycepin. Cordycepin content shows a trend of first decreasing and then increasing across the five stages (MY, MM, CP, PR, and FB), finally reaching its peak during the FB stage. If the goal is to obtain cordycepin, harvesting is most suitable when the fruiting body apex is swollen.
[0041] From the above, we can conclude that: This invention enables the direct cultivation of complete Cordyceps militaris fruiting bodies in a liquid culture medium, overcoming the limitations of traditional liquid fermentation which can only obtain mycelium. Compared to solid substrate cultivation methods, liquid culture offers advantages such as speed, low cost, and small footprint, allowing for the rapid production of a large number of propagules with more stable culture quality and effectively reducing the risk of contamination. Furthermore, the harvesting process in liquid culture is simpler, allowing direct separation of mycelium from the culture medium, and the mature fruiting bodies can be harvested together with the mycelium, significantly increasing biomass harvesting and providing a new pathway for the large-scale, standardized production of Cordyceps militaris.
[0042] In this invention, by optimizing the culture medium formula and cultivation conditions, the accumulation of antioxidant active substances such as flavonoids and polyphenols in the fruiting bodies of Cordyceps militaris is significantly increased, and the cordycepin content reaches its peak in the later stage of fruiting body growth, effectively enhancing the medicinal and economic value of the product. The waste liquid culture medium generated from liquid culture has low nutrient content, is easier to treat subsequently, and has less environmental impact, conforming to the concept of green and environmentally friendly production. This provides technical support for the sustainable development of the Cordyceps militaris industry and can significantly increase the content of functional components in Cordyceps militaris.
[0043] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.
[0044] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims
1. A method for obtaining Cordyceps militaris fruiting bodies through liquid culture, characterized in that: The method includes the following steps: S1: Under aseptic conditions, the preserved Cordyceps militaris slant culture was inoculated onto PDA solid medium and cultured at 22°C in the dark for 7 days to complete the activation of the culture, providing a robust seed source for subsequent liquid inoculation; S2: Take the activated strain and inoculate it into the culture medium at a ratio of 4 5mm diameter mycelial blocks per 100mL of liquid culture medium. After inoculation, let it stand in the dark for 24 hours to allow the strain to adapt to the environment. Then transfer it to a constant temperature shaker and culture it at 150r / min and 25℃ for 3 days to obtain uniformly sized mycelial balls, which will provide sufficient and consistent mycelial material for subsequent mycelial growth culture. S3: Transfer the culture flask containing the bacterial balls after cultivation to a 18°C environment and incubate in the dark for 7-10 days to allow the bacterial balls to grow further and intertwine on the surface of the culture medium to form an aerial mycelium film, creating a structural basis for subsequent color change upon exposure to light; S4: After the culture medium is covered with aerial mycelium film, move the culture bottle to an artificial climate chamber with a temperature of 22℃, humidity of 70%-75% and light intensity of 1000Lx. Maintain 12 hours of light per day for light culture until the white film completely turns orange-yellow, which promotes the mycelium on the film to twist and form primordia, thus preparing the morphology for the growth of fruiting bodies. S5: When the primordia on the mycelium grow to 0.5-1cm, adjust the culture temperature to 20℃, increase the humidity to 85%-95%, continue to maintain 12 hours of light per day, and continue to culture for 30-35 days to finally obtain mature Cordyceps militaris fruiting bodies.
2. The method for obtaining Cordyceps militaris fruiting bodies by liquid culture as described in claim 1, characterized in that: In step S1, the nutrient composition of the PDA solid culture medium is as follows: 200g of potato, peeled and chopped, is added to 1000mL of water, boiled for 30min, filtered through 4 layers of gauze to obtain the filtrate, and 20g of glucose, 1.5g of fish peptone, 1.5g of KH2PO4, 0.5g of MgSO4, 0.5g of CaCl2, and 20g of agar are added, and the volume is adjusted to 1000mL, maintaining the natural pH value.
3. The method for obtaining Cordyceps militaris fruiting bodies by liquid culture as described in claim 1, characterized in that: In step S1, the prepared culture medium is sterilized by steam for 30 minutes at 0.1 MPa pressure and 121°C. After cooling to room temperature, the preserved Cordyceps militaris slant culture is inoculated onto the surface of the culture medium. Then, the inoculated culture medium is transferred to a light-proof environment at 22°C and cultured for 7 days.
4. The method for obtaining Cordyceps militaris fruiting bodies by liquid culture as described in claim 1, characterized in that: In step S2, the nutrient composition of the liquid culture medium is as follows: 200g of potato (peeled and chopped) and 20g of silkworm pupae are added to 1000mL of water, boiled for 30min, filtered through 4 layers of gauze to obtain the filtrate, 20g of sucrose, 1.5g of fish peptone, 1.5g of KH2PO4, 0.5g of MgSO4, and 0.5g of CaCl2 are added, and the volume is adjusted to 1000mL, maintaining the natural pH value.
5. The method for obtaining Cordyceps militaris fruiting bodies by liquid culture as described in claim 1, characterized in that: In step S2, the liquid culture medium is sterilized with steam at 0.1 MPa and 121°C for 30 minutes. After cooling to room temperature, the culture medium is inoculated at a ratio of 4 activated bacterial blocks with a diameter of 5 mm per 100 ml of culture medium. After inoculation, the culture container is sealed with a polypropylene breathable film and then placed in a light-proof environment for 24 hours.
6. The method for obtaining Cordyceps militaris fruiting bodies by liquid culture as described in claim 1, characterized in that: In step S3, after the static adaptation is completed, the sealed culture container is transferred to a constant temperature shaker. The shaker speed is set to 150 r / min, the culture temperature is controlled at 25℃, and the culture is continued for 3 days. The culture is a closed culture, and the sealing method is to use a polypropylene breathable film for sealing.
7. The method for obtaining Cordyceps militaris fruiting bodies by liquid culture as described in claim 1, characterized in that: In step S4, during the mycelial growth stage, the culture bottle containing a large number of mycelial balls needs to be transferred to an environment of 18°C and kept in a static, light-protected state for 7 to 10 days. The culture is a closed culture, and the sealing method is to use a polypropylene breathable film for sealing.
8. The method for obtaining Cordyceps militaris fruiting bodies by liquid culture as described in claim 1, characterized in that: In step S5, the culture is a closed culture, which is sealed by using a polypropylene breathable film; the culture bottle is transferred to an artificial climate chamber, the ambient temperature is set at 22°C, the air humidity is controlled between 70% and 75%, and the light intensity is adjusted to 1000 Lx.
9. The method for obtaining Cordyceps militaris fruiting bodies by liquid culture as described in claim 1, characterized in that: In step S5, the light exposure time is maintained at 12 hours per day.
10. The method for obtaining Cordyceps militaris fruiting bodies by liquid culture as described in claim 1, characterized in that: In step S6, when the primordia on the bacterial film grow to 0.5 to 1 cm, the fruiting body matures and enters the cultivation stage. At this time, the cultivation environment parameters are adjusted, the temperature is lowered to 20°C, the air humidity is increased to 85% to 95%, and the light time is maintained for 12 hours a day for 30 to 35 days.