A method for rapidly forming fruiting bodies by inducing blood-ear mycelium through a culture medium

Abstract

The present application relates to edible mushroom culture technical field, specifically disclose a kind of method for inducing fruiting body rapid formation by culture medium of blood ear pure mycelium, comprising the following steps: (1) blood ear mycelium culture: blood ear spore germination mycelium is inoculated to enrichment culture medium P1 and is cultured, and blood ear mycelium after enrichment is obtained;(2) blood ear fruiting body culture: the mycelium after enrichment is inoculated to induction culture medium P2 and is cultured, and blood ear fruiting body is induced to form.This application effectively inhibits abnormal conversion of blood ear mycelium from mycelial type to yeast type by mycelium enrichment culture medium P1, thereby significantly improving the biomass of blood ear mycelium, in addition, the present application proposes blood ear pure mycelium ear forming technology independent of blood mark tenacisporae associated, breaks through traditional double-bacteria co-culture mode, and provides a new method for blood ear breeding work.

Description

Technical Field

[0001] This invention relates to the field of edible fungi cultivation technology, and in particular to a method for inducing the rapid formation of fruiting bodies from pure mycelia of *Hymenochloa chinensis* using a culture medium. Background Technology

[0002] Tremella sanguinea, a rare edible and medicinal fungus belonging to the genus Tremella in the family Tremellaceae, possesses fruiting bodies with rich nutritional value and significant medicinal effects. However, current market demand for Tremella sanguinea primarily relies on the collection of wild resources. Although some artificial cultivation techniques have achieved small-scale fruiting, challenges remain, including long growth cycles, strain degeneration, and low fruiting rates.

[0003] *Hypericum sanguinolentum* is a dimorphic fungus, exhibiting a life cycle that alternates between yeast-like and hyphal forms. During cultivation, *Hypericum sanguinolentum* hyphae readily transform into yeast-like spores, subsequently reproducing asexually via budding, making it difficult to develop into robust fruiting bodies. Furthermore, research indicates that the formation of *Hypericum sanguinolentum* fruiting bodies is highly dependent on the synergistic effect of its symbiotic fungus, *Stereum sanguinolentum*. *Hypericum sanguinolentum* hyphae alone cannot form fruiting bodies in lignocellulose culture media; only the presence of *Stereum sanguinolentum*, which possesses highly efficient lignin and cellulose decomposition capabilities, allows for the formation of *Hypericum sanguinolentum* fruiting bodies.

[0004] Traditional methods for cultivating *Hypericum aegyptium* require the simultaneous cultivation of both *Hypericum aegyptium* and its associated fungi. However, the growth competition between the two fungi during mixed cultivation is difficult to control precisely, resulting in a low fruiting rate. Furthermore, *Hypericum aegyptium* has a long growth cycle; under artificial cultivation conditions, it takes 4-8 months from inoculation to fruiting body maturity, which significantly limits its large-scale production and application. Summary of the Invention

[0005] In order to overcome the shortcomings and deficiencies of the existing technology, the purpose of this invention is to provide a method for inducing pure mycelia of *Hypericum aegyptium* to form fruiting bodies, using a fast-acting nutrient compound formula to achieve the differentiation of pure mycelia of *Hypericum aegyptium* into fruiting bodies.

[0006] This invention breaks through the traditional theory that blood otorrhea depends on the presence of associated bacteria, providing key technical support for its industrial production.

[0007] The objective of this invention is achieved through the following technical solution: a method for inducing pure hyphae of *Hypericum aegyptium* to form fruiting bodies, comprising the following steps:

[0008] (1) Culture of Hemlock mycelium: The mycelium germinating from Hemlock spores was inoculated into enrichment medium P1 and cultured to obtain enriched Hemlock mycelium;

[0009] (2) Culture of Blood Ear fruiting bodies: The enriched mycelium was inoculated into induction medium P2 and cultured to induce the formation of Blood Ear fruiting bodies.

[0010] Furthermore, the enrichment medium P1 in step (1) comprises the following components by weight:

[0011] 5g potato starch, 10g maltose, 10g glucose, 0.5g KH2PO4, 0.5g MgSO4, 1g superphosphate, 10mg vitamin B15, 2g fish peptone, 20g agar, 1L water.

[0012] Furthermore, potato starch provides natural carbon and nitrogen sources and growth factors; maltose and glucose, as readily available carbon sources, synergistically promote fungal mycelial metabolism; KH2Po4 provides phosphorus and potassium ions, supports nucleic acid and ATP synthesis, and helps regulate the enzymatic reaction environment, thereby promoting mycelial growth; magnesium ions in MgSO4 activate enzymes in the EMP pathway and TCA cycle related to energy metabolism, while sulfur is an indispensable component for amino acid synthesis; superphosphate replenishes phosphate and calcium ions, enhances cell wall stability, and regulates pH buffering capacity; fish peptone provides a high-quality nitrogen source and essential amino acids; vitamin B1 significantly enhances mycelial vitality and growth rate by promoting sugar metabolism.

[0013] Specifically, enrichment medium P1 can effectively inhibit the abnormal transformation of hyphae into spores and increase the hyphal growth rate.

[0014] Furthermore, the temperature of the enrichment medium P1 in step (1) is 21–23.5°C.

[0015] Furthermore, in step (1), the number of days of culture in enrichment medium P1 is 20 to 30 days.

[0016] Furthermore, the formulation of the induction medium P2 in step (2) comprises the following components by weight:

[0017] 5g potato starch, 20g glucose, 1g KH2Po4, 1g superphosphate, 100–200mg L-ascorbic acid, 300–600mg cyclic adenosine monophosphate, 10–20mg chitosan, 20g agar, 1L water.

[0018] Furthermore, L-ascorbic acid protects hyphae from oxidative damage and prolongs metabolic activity through its antioxidant effects; cyclic adenosine monophosphate (cAMP), as an intracellular signaling molecule, activates differentiation-related signaling pathways; and chitosan, as a natural polysaccharide inducing factor, can promote hyphal aggregation and differentiation, and contribute to cell wall synthesis.

[0019] Specifically, the induction medium P2 can effectively promote the differentiation of hyphae into fruiting body primordia.

[0020] Furthermore, the specific culture method for inducing mycelium to form fruiting bodies in step (2) includes the following steps:

[0021] The enriched mycelia were inoculated into induction medium P2 at a temperature of 18–20°C and then cultured in the dark for one week. Light stimulation was selectively applied during the culture period. After the light stimulation was completed, the culture was continued in the dark until primordia were formed.

[0022] Furthermore, the period of the light stimulation is 48 hours.

[0023] Furthermore, the 48-hour light stimulation cycle consists of two alternating cycles of 8 hours of light and 16 hours of darkness.

[0024] Furthermore, the light intensity of the light stimulus is 100–200 lux.

[0025] Furthermore, the culture temperature for inducing mycelium to form fruiting bodies is 18–20°C. First, the mycelium is cultured in the dark for one week, followed by selective light stimulation. After the light stimulation is completed, the mycelium continues to be cultured in the dark until primordia are formed.

[0026] Furthermore, the light stimulation cycle is 48 hours, including 8 hours of light and 16 hours of darkness, alternating twice, with the light intensity controlled between 100 and 200 lux.

[0027] Furthermore, in step (2), the induction culture medium P2 is cultured for 30 to 40 days.

[0028] According to the solution provided by the present invention, compared with the prior art, the beneficial effects of the present invention are:

[0029] This invention effectively inhibits the abnormal conversion of *Hypericum haematomyces* hyphae from hyphal morphology to yeast morphology through hyphal enrichment culture medium, thereby increasing the biomass of *Hypericum haematomyces* hyphae.

[0030] This invention transfers enriched pure mycelia of *Hypericum halophilum* to a fruiting body induction medium, enabling the pure mycelia of *Hypericum halophilum* to form robust fruiting bodies within 30–40 days after being separated from its symbiotic fungus *Hypericum halophilum*. This breaks through the current bottleneck of symbiotic dependence between *Hypericum halophilum* and its symbiotic fungus, effectively shortening the production cycle of *Hypericum halophilum*. By replacing wild collection with efficient artificial cultivation technology, it alleviates the ecological pressure on wild *Hypericum halophilum* resources.

[0031] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention and to implement it in accordance with the contents of the specification, and in order to make the above and other objects, features and advantages of the present invention more apparent and understandable, specific embodiments of the present invention are described below. Attached Figure Description

[0032] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly described below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation on the scope of this application.

[0033] Figure 1 This is a schematic diagram showing the growth status of *Hypericum schistosomiasis* hyphae on YCM medium.

[0034] Figure 2 This is a schematic diagram showing the growth status of *Hypericum schistosomiasis* hyphae on PDA medium.

[0035] Figure 3 A schematic diagram showing the growth status of *Hypericum schistosum* hyphae on MMN medium;

[0036] Figure 4 A schematic diagram showing the growth status of *Hypericum schistosum* hyphae on the enrichment medium P1 of this invention;

[0037] Figure 5 This is a schematic diagram of a micrograph of *Hypericum haematomyces* hyphae.

[0038] Figure 6 This is a schematic diagram showing the dark growth of *Hypericum hygroscopicum* hyphae on induction medium P2 in Example 1.

[0039] Figure 7 This is a schematic diagram showing the growth status of *Hypericum hainanense* hyphae on the induction medium P2 in Example 1 after light induction.

[0040] Figure 8 This is a schematic diagram of the growth status of *Hypericum hygroscopicum* hyphae on the induction medium P2 in Example 2 after light induction.

[0041] Figure 9 A schematic diagram comparing the aqueous solution and pure water used to soak the fruiting bodies of *Hypericum esculentum*.

[0042] Figure 10 The third-generation ITS sequencing sequence listing of *Hypericum haematomyces* hyphae and fruiting bodies;

[0043] Figure 11 The results of NCBI alignment of hematuria ITS sequences;

[0044] Figure 12 Statistical analysis of data processing results for third-generation sequencing samples;

[0045] Figure 13 Statistical analysis of species at various levels in third-generation sequencing samples. Detailed Implementation

[0046] To facilitate understanding by those skilled in the art, the present invention will be further described below with reference to embodiments. The content mentioned in the embodiments is not intended to limit the present invention.

[0047] Example 1

[0048] This embodiment provides a method for inducing the rapid formation of fruiting bodies from pure hyphae of *Hypericum halophilum*, including the following steps:

[0049] (1) Culture of Hemlock mycelium: The mycelium germinating from Hemlock spores was inoculated into enrichment medium P1 and cultured to obtain enriched Hemlock mycelium;

[0050] (2) Culture of Blood Ear fruiting bodies: The enriched mycelium was inoculated into induction medium P2 and cultured to induce the formation of Blood Ear fruiting bodies.

[0051] In step (1), the pure mycelium enrichment medium P1 for *Hypericum aegyptium* is prepared. The specific formula is as follows: 5g potato starch, 10g maltose, 10g glucose, 0.5g KH2PO4, 0.5g MgSO4, 1g superphosphate, 5-10mg vitamin B1, 2g fish peptone, 20g agar, and 1L water.

[0052] The method for preparing the enrichment culture medium P1 in step (1) includes the following steps:

[0053] a. Weigh out 5g potato starch, 10g maltose, 10g glucose, 0.5g KH2Po4, 0.5g MgSO4, 1g superphosphate, 2g fish peptone, 20g agar, and 1L water; sterilize at 121℃ and 0.1MPa for 30min.

[0054] b. After the culture medium cools to 50℃–60℃, add the filtered vitamin B1 solution to a final concentration of 5mg / L;

[0055] c. After mixing, dispense the culture medium into 9cm petri dishes, let it cool and solidify before use.

[0056] Furthermore, the enriched *Hypericum hygroscopicum* mycelium was inoculated into enrichment medium P1, specifically as follows:

[0057] In a clean bench, the cooled blade was used to flammate and cut the germinating hyphae of *Heterophyllum hemangiosum* into uniformly sized pieces. These pieces were then transferred to the enrichment medium P1 used in this example or other commonly used fungal culture media and cultured in a constant temperature incubator at 22°C in the dark for 30 days. The results are as follows: Figure 4 As shown.

[0058] Furthermore, the morphological characteristics of the *Hypericum haematomyces* hyphae in the enrichment medium P1 were analyzed:

[0059] During mycelial culture, a portion of mycelial samples was picked from the enrichment medium P1 and observed under an optical microscope; for example... Figure 5 As shown, only the morphology of a single *Hypericum hainanense* hyphae was observed in the field of view, and no other associated bacteria or miscellaneous bacteria were found, indicating that the culture was a pure *Hypericum hainanense* hyphae.

[0060] In step (2), the preparation of the blood ear fruiting body induction medium P2 has the following specific formula:

[0061] 5g potato starch, 20g glucose, 1g KH2Po4, 1g superphosphate, 100mg L-ascorbic acid, 300mg cyclic adenosine monophosphate, 10mg chitosan, 20g agar, 1L water.

[0062] The method for preparing the induction medium P2 in step (2) includes the following steps:

[0063] a. Weigh out 5g potato starch, 20g glucose, 1g KH2Po4, 1g superphosphate, 100mg L-ascorbic acid, 10mg chitosan, 20g agar, and 1L water; sterilize at 121℃ and 0.1MPa for 30min.

[0064] b. After the culture medium cools down to 50℃–60℃, add the filtered cyclic adenosine monophosphate solution to a final concentration of 300 mg / L;

[0065] c. Dispense the culture medium into 9cm petri dishes, let it cool and solidify before use.

[0066] Furthermore, the *Hypericum haematomyces* hyphae in enrichment medium P1 were inoculated into induction culture P2:

[0067] In a clean bench, using a sterilized blade, the enriched *Heterophyllum hematuriae* hyphae were transferred to the induction medium P2 in this embodiment. The culture was then carried out in the dark at 20°C for 35 days. The hyphal growth status was as follows: Figure 6 As shown.

[0068] Preferably, the hyphae of *Hypericum aegyptium* were cultured in induction medium P2 at 20°C in the dark for one week, followed by 48-hour light stimulation cycles (8 hours of light followed by 16 hours of darkness, repeated twice) at a light intensity of 100 lux. After the light stimulation, the mycelium was cultured in the dark at 20°C for another 35 days. The hyphal growth status was as follows: Figure 7 As shown.

[0069] like Figure 6 and Figure 7 The results showed that *Haematomyces cerevisiae* hyphae could be successfully induced to form fruiting bodies under both dark and light stimulation conditions in culture medium P2. The comparison revealed that light effectively promoted the differentiation process of *Haematomyces cerevisiae* hyphae, thereby improving the quality and efficiency of fruiting body formation.

[0070] In this embodiment, the formulation comparison test includes:

[0071] The test culture media were YCM, PDA, MMN, P1, and P2, as shown in Table 1 below.

[0072]

[0073] Table 1

[0074] Furthermore, the preparation of culture media: the preparation methods of YCM, PDA, and MMN culture media can be carried out in accordance with existing conventional techniques, and the preparation methods of P1 and culture media P2 are as described in this invention.

[0075] Specifically, the inoculation and data measurement methods include: cutting the mycelia of germinating *Bacillus hematuriae* into uniformly sized pieces using a sterile blade in a clean bench, and transferring them to YCM, PDA, MMN, P1 and P2 media respectively, and carrying out mycelial enrichment culture at 22°C in the dark for 30 days.

[0076] In addition, the mycelia enriched in culture medium P1 were punched sequentially with a sterile 200 μL pipette tip and then transferred to YCM, PDA, MMN, P1 and P2 culture media respectively with a sterile inoculation needle. They were then induced and cultured in the dark at 20°C for 40 days.

[0077] During the cultivation period, the transformation of mycelium into spores and the formation of fruiting bodies were observed and statistically analyzed. The growth status of mycelium and spores was also recorded. The calculation method is as follows, and the specific results are shown in Table 1.

[0078]

[0079] Specifically, the results and analysis include: Figure 1 – Figure 3 The results showed that while *Heterophyllum hexandrum* hyphae could grow on YCM, PDA, and MMN media, they generally underwent extensive transformation into yeast-like spores, and the hyphal biomass was low; in contrast, Figure 4 The results showed that the culture medium P1 of the present invention exhibited significant advantages, effectively inhibiting the excessive conversion of Hectoria hyphae into spores and significantly increasing the biomass of the hyphae.

[0080] During the mycelial enrichment stage, the yeast-like spore formation rate on YCM medium was 90%, with numerous spores that were wrinkled and off-white. On PDA medium, the yeast-like spore formation rate was 85%, with relatively smooth, light yellow, and viscous spores. On MMN medium, the yeast-like spore formation rate was 75%, with smooth, yellow, viscous spores and a moist surface. In medium P1 of this invention, the yeast-like spore formation rate was 10%, with fewer spores that were off-white and had a relatively dry surface. In medium P2 of this invention, the yeast-like spore formation rate was 15%, with fewer spores that formed smooth small round dots.

[0081] The appeal results fully demonstrate that the enrichment medium P1 of the present invention can efficiently enrich Haematomyces hyphae, effectively solving the problem of low hyphae quantity caused by the large-scale conversion of existing cultured hyphae into spores, and providing a better solution for the efficient amplification and culture of Haematomyces hyphae.

[0082] During the fruiting body induction stage, the fruiting body formation rate on YCM medium was 10%, with sparse and weak hyphae, extremely slow growth, and no pigment production. On PDA medium, the fruiting body formation rate was 30%, with sparse, yellowish-brown hyphae, slow growth, and pigment production. On MMN medium, the fruiting body formation rate was 35%, with dense, yellowish-brown hyphae, relatively fast growth, and pigment production. In the present invention's medium P1, the fruiting body formation rate was 55%, with thicker, yellowish-brown hyphae, rapid growth, and significant pigment production. In the present invention's medium P2, the fruiting body formation rate was 90%, with yellowish-brown, slow-growing, relatively thick hyphae, significant pigment production, and easy fruiting body formation. Therefore, it is evident that the fruiting body formation rate of *Hypericum aegyptium* induction medium P2 of the present invention is significantly higher than other formulations.

[0083] Example 2

[0084] This embodiment provides a method for inducing pure hyphae of *Hypericum aegyptium* to form fruiting bodies, including the following steps:

[0085] (1) Culture of Hemlock mycelium: The mycelium germinating from Hemlock spores was inoculated into enrichment medium P1 and cultured to obtain enriched Hemlock mycelium;

[0086] (2) Culture of Blood Ear fruiting bodies: The enriched mycelium was inoculated into induction medium P2 and cultured to induce the formation of Blood Ear fruiting bodies.

[0087] In step (1), the pure mycelium enrichment medium P1 for *Hypericum aureum* is prepared. The specific formula is as follows: 5g potato starch, 10g maltose, 10g glucose, 0.5g KH2PO4, 0.5g MgSO4, 1g superphosphate, 2g fish peptone, 20g agar, 10mg vitamin B1, and 1L water.

[0088] The method for preparing the enrichment culture medium P1 in step (1) includes the following steps:

[0089] a. Weigh out 5g potato starch, 10g maltose, 10g glucose, 0.5g KH2Po4, 0.5g MgSO4, 1g superphosphate, 2g fish peptone, 20g agar, and 1L water; sterilize at 121℃ and 0.1MPa for 30min.

[0090] b. After the culture medium cools to 50℃–60℃, add the filtered vitamin B1 solution to a final concentration of 10mg / L;

[0091] c. After mixing, dispense the culture medium into 9cm petri dishes, let it cool and solidify before use.

[0092] Furthermore, the enriched *Hypericum hygroscopicum* mycelium was inoculated into enrichment medium P1, specifically as follows:

[0093] In a clean bench, the mycelia germinating from the *Hypericum spp.* were cut into uniformly sized pieces by a blade cooled by flame. These pieces were then transferred to culture medium P1 in this embodiment and other commonly used fungal culture media and cultured at 23°C in a constant temperature incubator to obtain enriched *Hypericum spp.*

[0094] In step (2), the preparation of the blood ear fruiting body induction medium P2 has the following specific formula:

[0095] 5g potato starch, 20g glucose, 1g KH2Po4, 1g superphosphate, 200mg L-ascorbic acid, 600mg cyclic adenosine monophosphate, 20mg chitosan, 20g agar, 1L water.

[0096] The method for preparing the induction medium P2 in step (2) includes the following steps:

[0097] a. Weigh out 5g potato starch, 20g glucose, 1g KH2Po4, 1g superphosphate, 200mg L-ascorbic acid, 20mg chitosan, 20g agar, and 1L water; sterilize at 121℃ and 0.1MPa for 30min.

[0098] b. After the culture medium cools to 50℃–60℃, add the filtered cyclic adenosine monophosphate solution to a final concentration of 600mg / L. Dispense the culture medium into 500mL plastic tissue culture bottles and allow it to cool and solidify before use.

[0099] Furthermore, the *Hypericum haematomyces* hyphae in enrichment medium P1 were inoculated into induction culture P2:

[0100] In a clean bench, the enriched Haematomyces hyphae were transferred to the P2 induction medium in this embodiment using a sterilized blade. The medium was then cultured in the dark at 19°C for one week in a constant temperature incubator. Subsequently, light stimulation was performed for 48 hours, including 8 hours of light and 16 hours of darkness, alternating twice, with a light intensity of 200 lux. After the light stimulation was completed, the medium was cultured in the dark for another 40 days.

[0101] like Figure 8 As shown in the figure, the growth status of fruiting bodies induced by light on culture medium P2 in Example 2 is shown. It can be seen from the figure that under this culture condition, pure hyphae of *Hypericum aegyptium* can efficiently induce the formation of fruiting bodies.

[0102] like Figure 9 As shown in the image, a comparison of the aqueous solution and pure water used to extract the fruiting bodies of *Hypericum aegyptium* is presented. It is clearly visible that the extract from the fruiting bodies induced by mycelium is amber-red, a color highly consistent with that of the extract from wild *Hypericum aegyptium* fruiting bodies. This result indicates that, through induced culture, pure *Hypericum aegyptium* mycelium can successfully produce fruiting bodies with typical characteristics.

[0103] In this embodiment, *Hypericum aureum* hyphae were collected from enrichment medium P1 and labeled A1, and induced *Hypericum aureum* fruiting bodies were collected from medium P2 and labeled A2. A1 and A2 were sent to BioMed Biotechnology Co., Ltd. for third-generation ITS full-length sequencing, with a sequencing data volume of 5Gb.

[0104] In this embodiment, Figure 10 The results of the sample sequencing data processing are statistical. The results show that the average ITS sequence length of samples A1 and A2 is 502bp, and the validity of the sequencing analysis data is 100%, indicating that the sequencing data results are reliable.

[0105] In this embodiment, Figure 11 The results show that the species counts at each level of the sequencing samples are as follows: Kindom, Phylum, Class, Order, Family, Genus, and Species represent the seven taxonomic levels of Kingdom, Phylum, Class, Order, Family, Genus, and Species, respectively. The results show that only one species exists in the *Hygrophora* hyphae enriched in culture medium P1 and the *Hygrophora* fruiting bodies induced in culture medium P2 in this invention.

[0106] In this embodiment, Figure 12 It is the third-generation sequencing ITS sequence of Haematomyces hyphae and fruiting bodies, totaling 502 bp.

[0107] In this embodiment, Figure 13The results of the NCBI alignment of the ITS sequence of *Hymenoplastyces cerevisiae* show that the fragment is 100% identical to the *Tremellasanguinea* sequence in GenBank. Therefore, at the molecular level, it can be determined that the hyphae and fruiting bodies are *Hymenoplastyces cerevisiae*.

[0108] The above embodiments are preferred implementations of the present invention. In addition, the present invention can be implemented in other ways. Any obvious substitutions without departing from the concept of the present invention are within the protection scope of the present invention.

Claims

1. A method for inducing the rapid formation of fruiting bodies from pure hyphae of *Hypericum perforatum* using a culture medium, characterized in that, Includes the following steps: (1) Culture of Hemlock mycelium: The mycelium of Hemlock spores was inoculated into enrichment medium P1 and cultured to obtain enriched Hemlock mycelium; (2) Culture of Blood Ear fruiting bodies: The enriched mycelium was inoculated into induction medium P2 and cultured to induce the formation of Blood Ear fruiting bodies; The enrichment medium P1 in step (1) is composed of the following components by weight: 5g potato starch, 10g maltose, 10g glucose, 0.5g KH2PO4, 0.5g MgSO4, 1g superphosphate, 10mg vitamin B15, 2g fish peptone, 20g agar, 1L water; The formulation of the induction medium P2 in step (2) consists of the following components by weight: 5g potato starch, 20g glucose, 1g KH2Po4, 1g superphosphate, 100-200mg L-ascorbic acid, 300-600mg cyclic adenosine monophosphate, 10-20mg chitosan, 20g agar, 1L water. The specific culture method for inducing mycelium to form fruiting bodies in step (2) includes the following steps: The enriched mycelia were inoculated into induction medium P2 at a temperature of 18-20°C and then cultured in the dark for one week. During the culture period, light stimulation was selectively applied. After the light stimulation was completed, the culture was continued in the dark until primordia were formed.

2. The method for inducing rapid fruiting body formation of pure hyphae of *Hypericum aegyptium* via culture medium according to claim 1, characterized in that, The temperature for mycelial enrichment culture in step (1) is 21–23.5℃.

3. The method for inducing rapid fruiting body formation of pure hyphae of *Hypericum aegyptium* via culture medium according to claim 1, characterized in that, In step (1), the number of days of culture in enrichment medium P1 is 20~30 days.

4. The method for inducing rapid fruiting body formation of pure hyphae of *Hypericum aegyptium* via culture medium according to claim 1, characterized in that, In step (2), the induction culture medium P2 is cultured for 30 to 40 days.

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