Cultivation method of selenium-enriched cordyceps sinensis sporocarp

Abstract

The invention belongs to the field of biological fermentation of cordyceps militaris, and particularly relates to a culture method of selenium-enriched cordyceps militaris sporocarp. The method comprises the following steps: inoculating a seed solution of a cordyceps militaris strain which is transferred and propagated for multiple times into a fermentation culture medium to carry out mycelium static fermentation, when the mycelia are full of the mycelia and the height of the lawn of aerial mycelia is 3-6mm, carrying out stimulation treatment on the mycelia, and then carrying out coloring and fruiting culture, so as to obtain the selenium-enriched cordyceps sinensis sporocarp. The culture method provided by the invention can effectively control the content of organic selenium, the content of cordycepin and the height of the sporocarp of the selenium-enriched cordyceps sinensis, so that ideal medicinal quality is achieved. Besides, by adopting a liquid fermentation culture method, the culture efficiency can be improved, the culture period can be shortened, the controllability is high, and the method is suitable for large-scale production of the selenium-enriched cordyceps sinensis sporocarp and has a good industrial application prospect.

Description

Technical Field

[0001] The invention belongs to the field of Cordyceps militaris biological fermentation, and particularly relates to a method for culturing selenium-enriched Cordyceps militaris fruiting bodies. Background Art

[0002] In recent years, with the increasing demand for Cordyceps products at home and abroad, the artificial cultivation technology and scale of Cordyceps militaris have been greatly improved, providing a large amount of high-quality Cordyceps raw materials for the medical and health care product markets at home and abroad. Solid cultivation is generally used in the production of Cordyceps militaris, which has a high mushroom yield and good fruiting body growth. However, the long cultivation cycle of solid cultivation, unstable yield and quality during production, and low biological efficiency still restrict the large-scale production of Cordyceps militaris. Therefore, the establishment of a new and efficient cultivation technology is an urgent problem to be solved in Cordyceps production.

[0003] In addition to growth efficiency, biological activity is also a focus of Cordyceps militaris cultivation. Selenium (Se) is a non-metallic element and one of the 15 nutrients necessary for the human body. Years of research have confirmed that selenium has multiple functions such as anti-cancer, anti-aging, anti-oxidation, enhancing human immunity, regulating the absorption and utilization of vitamins, regulating protein synthesis, and enhancing reproductive function. It is also an important component of muscle function. Selenium-rich Cordyceps is mostly found in solid cultivation, but the biological content is low and the conversion cycle is long.

[0004] Therefore, it is of great significance to develop a new method for culturing selenium-enriched Cordyceps fruiting bodies based on liquid fermentation technology. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a method for cultivating selenium-rich Cordyceps fruiting bodies in view of the shortcomings of low production efficiency of Cordyceps militaris and low cordycepin and organic selenium content in the prior art.

[0006] In order to solve the above technical problems, the present invention discloses a method for culturing selenium-enriched Cordyceps fruiting bodies, which is characterized in that the seed liquid of the Cordyceps militaris strain is inoculated into a fermentation medium for static fermentation of the mycelium, and when the mycelium is full and the height of the aerial mycelium is more than 3 mm, the mycelium is stimulated and then color-changing and mushroom-producing culture is carried out to obtain the selenium-enriched Cordyceps fruiting bodies.

[0007] The Cordyceps militaris strains include but are not limited to the flat plate bacteria CGMCC No. 18581 of the present invention, and strains that can be fermented to prepare Cordyceps militaris fruiting bodies in the prior art are all within the scope of protection of the present invention.

[0008] The seed liquid of the Cordyceps militaris strain is a seed liquid obtained by inoculation and expansion of a multi-stage seed culture medium.

[0009] Specifically, the formula of the multi-stage seed culture medium is: 25-40 g / L glucose, 10-20 g / L peptone, 5-10 g / L yeast powder, 0.1-2 g / L potassium dihydrogen phosphate, 0.1-2 g / L magnesium sulfate, 20-100 mg / L selenate and / or selenite, pH 5.8.

[0010] Furthermore, the concentration of selenate and / or selenite in the multi-stage seed culture medium is increased in a gradient of 10 to 100%; preferably, in a gradient of 40 to 60%.

[0011] Specifically, the inoculation and expansion are performed 2 to 4 times, preferably 3 times.

[0012] Specifically, the inoculation amount is 5-30% v / v, and the volume ratio of the plate area of ​​the Cordyceps militaris strain to the seed culture medium is 10-30 cm 2 : 100~300cm 3 Preferably, the inoculation volume is 25% v / v, and the volume ratio of the plate area of ​​the Cordyceps militaris strain to the seed culture medium is 15 cm 2 : 200cm 3 .

[0013] Specifically, the culture conditions for the expansion culture are: culture at a rotation speed of 150-220 rpm and a temperature of 26-29° C. until the mycelium wet weight is greater than 180 g / L.

[0014] Wherein, the inoculation amount is 5-30% v / v; the preferred inoculation amount is 20-25% v / v.

[0015] The fermentation medium has a formula of: glucose 10-20 g / L, sucrose 5-15 g / L, corn flour 2-10 g / L, peptone 1-15 g / L, yeast powder 1-6 g / L, VB1 0.05-0.5 g / L, potassium dihydrogen phosphate 0.5-2 g / L, magnesium sulfate 0.1-0.5 g / L, adenine 0.1-5 g / L, glycine 2-20 g / L, uracil nucleotide 1-5 g / L, selenate and / or selenite 0.5-100 mg / L, pH 6.0. Preferably, adenine 1-2 g / L, glycine 6-10 g / L, uracil nucleotide 2-3 g / L, selenate and / or sodium selenite 20-40 mg / L.

[0016] More preferably, the fermentation medium has a formula of: 15 g / L glucose, 15 g / L sucrose, 5 g / L corn flour, 12 g / L peptone, 6 g / L yeast powder, 0.08 g / L VB1, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate, 1 g / L adenine, 10 g / L glycine, 3 g / L uracil nucleotide, 40 mg / L selenate and / or selenite, pH 6.0.

[0017] The mycelium is statically fermented under the following fermentation conditions: light-shading culture under a positive micro-pressure environment of 0.1-0.5 MPa, ventilation volume of 0.05-0.2 vvm, temperature of 22-29° C., and humidity of 65-70%.

[0018] Preferably, the mycelium is statically fermented, and the fermentation conditions are: shading culture under a positive micro-pressure environment of 0.3 MPa, a ventilation volume of 0.1 vvm, a temperature of 26° C., and a humidity of 70%.

[0019] Wherein, the stimulation treatment is contact scribing or non-contact laser irradiation.

[0020] Specifically, the contact streaking is: streaking the mycelium in the culture bottle containing the fermentation medium, with the parallel lines spaced 1 to 3 cm apart; the non-contact laser irradiation is: irradiating the laser for 5 to 20 minutes in the dark.

[0021] The color changing and fruiting mushroom culture is divided into an induction phase and a growth phase.

[0022] Specifically, the culture conditions during the induction period are: temperature 20-26° C., light intensity 100-200 lx, humidity 55-65%, and light color change culture. When the mycelium turns orange, small brown water droplets appear on the surface of the fermentation medium, and fruiting body buds appear, continue culturing for 4-6 days.

[0023] Preferably, the culture conditions during the induction period are: temperature 25°C, light intensity 120lx, humidity 60%, light color change culture, and culture until the skin turns orange, small brown water droplets appear on the surface of the culture medium, and fruiting body buds appear, and continue to culture for 4 to 6 days.

[0024] Specifically, during the growth stage, the culture conditions are: 20-26°C, light intensity 200-500lx, humidity 80-95% for 6-8h; 12-18°C, no light, humidity 80-95% for 16-18h, and continuous culture for 3-6 days; then the temperature is controlled at 24°C, the humidity is 80-95%, and the light hours are maintained at 16h per day to promote the growth of fruiting bodies, and the culture is carried out for 10-16 days.

[0025] Preferably, during the growth stage, the culture conditions are: 24°C, light intensity 200-500lx, humidity 80-95% for 8h; 16°C, no light, humidity 80-95% for 16h, and continuous culture for 3-6 days; then the temperature is controlled at 24°C, the humidity is 80-95%, and the light hours are maintained at 16h per day to promote the growth of fruiting bodies, and the culture is carried out for 10-16 days.

[0026] Beneficial effects:

[0027] (1) The present invention provides a method for culturing selenium-rich Cordyceps fruiting bodies, which can effectively control the organic selenium content, cordycepin content and fruiting body height of the selenium-rich Cordyceps fruiting bodies to achieve ideal medicinal quality.

[0028] (2) The present invention adopts a liquid fermentation culture method, which can improve the culture efficiency, shorten the culture cycle, and has strong controllability, and is suitable for large-scale industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments, and the above and / or other advantages of the present invention will become more clear.

[0030] Figure 1 This is a picture of the fruiting bodies cultured on different substrates in Example 3. DETAILED DESCRIPTION

[0031] The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials described are commercially available unless otherwise specified.

[0032] In the following examples, the plate bacteria CGMCC No.18581, its detailed information has been disclosed in patent CN111117896A.

[0033] Example 1: Effect of selenite on the culture of Cordyceps militaris fruiting bodies

[0034] Scrape the plate bacteria CGMCC No.18581 into a shake flask containing 150 mL of primary seed culture medium containing different selenite concentrations. The ratio of the plate bacteria area to the shake flask liquid volume is 15 cm 2 : 200cm 3 , the first-level seed solution was obtained by culturing at 180 rpm and 26°C. The results showed (Table 1) that high concentration of selenite (60 mg / L) had a certain inhibitory effect on the growth of mycelium, and the wet weight of the mycelium in seed culture was far less than 180 g / L, so low concentration of selenite was used for first-level seed culture.

[0035] The first-level seed liquid obtained by culturing with the first-level seed medium containing 30 mg / L selenite concentration is transferred to the second-level seed medium at an inoculation amount of 20% v / v, and cultured at the same rotation speed and temperature until the wet weight of the bacteria is not less than 180 g / L. Then, the second-level seed liquid is transferred to the third-level seed medium at the same inoculation amount for 3 generations, and cultured at the same rotation speed and temperature until the wet weight of the bacteria is not less than 180 g / L to obtain the third-level seed liquid. From the first-level seed culture to the third-level seed culture, the selenite concentration is gradually increased as the expansion level increases.

[0036] The formula of the primary seed culture medium is as follows: 25 g / L glucose, 12 g / L peptone, 10 g / L yeast powder, 0.5 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate, 30 mg / L or 60 mg / L selenite, pH 5.8.

[0037] The secondary seed culture medium has a formula of: 25 g / L glucose, 12 g / L peptone, 10 g / L yeast powder, 0.5 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate, 45 mg / L selenite, and a pH of 5.8.

[0038] The formula of the tertiary seed culture medium is: 25 g / L glucose, 12 g / L peptone, 10 g / L yeast powder, 0.5 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate, 68 mg / L selenite, pH 5.8.

[0039] The cultured tertiary seed liquid was transferred to 200 mL of fermentation medium containing different selenite concentrations at 10-20% v / v, placed in a 0.01 MPa micro-positive pressure environment under shade culture, and cultured for about 5 days at a ventilation volume of 0.1 vvm, a temperature of 26°C, and a humidity of 70%, until the mycelium was fully covered and the aerial mycelium moss was more than 3 mm high.

[0040] The fermentation medium has a formula of: 15 g / L glucose, 15 g / L sucrose, 5 g / L corn flour, 12 g / L peptone, 6 g / L yeast powder, 0.08 g / L VB1, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate, 20-50 mg / L selenite, pH 6.0.

[0041] The culture bottle containing the fermentation medium is streaked in the clean bench, with parallel lines spaced about 2 cm apart, and placed in a plant incubator for mushroom cultivation management. Cultivation conditions: temperature 25°C, light intensity 120lx, humidity 60%, light color change cultivation, and continue cultivation for 5 days when the mushroom skin turns orange, small brown water droplets appear on the surface of the culture medium, and fruiting body buds appear.

[0042] Fruiting body growth process: 24℃, light intensity 200-500lx, humidity 80-95% for 8h; 16℃, no light, humidity 80-95% for 16h, and continue for 5 days. Then control the temperature to 24℃, humidity 80-95%, maintain 16h of light hours per day to promote fruiting body growth, and cultivate for 16 days.

[0043] The results of the effects of adding selenite at the seed stage and fermentation stage on the growth and product of Cordyceps are shown in Table 1.

[0044] Table 1 Effects of adding selenite at the seed stage and fermentation stage on the growth and product of Cordyceps

[0045]

[0046] From the fermentation results in Table 1, it can be found that during the fermentation process, within the lower selenite concentration range, as the amount of selenite added increased, the selenium content in the mycelium increased significantly; but when the selenite concentration exceeded 50 mg / L, it had a significant inhibitory effect on the growth of the fruiting body.

[0047] Example 2: Effect of uracil nucleotides on selenium content and cordycepin content in Cordyceps

[0048] The culture methods of the primary (selenite concentration is 30 mg / L), secondary and tertiary seed solutions are the same as those in Example 1.

[0049] The cultured tertiary seed liquid was transferred to 200 mL of fermentation medium containing different uracil nucleotide concentrations at 20% v / v, placed in a 0.03 MPa micro-positive pressure environment with shade culture, and cultured for about 4 days at a ventilation volume of 0.1 vvm, a temperature of 26°C, and a humidity of 70%, until the mycelium was fully covered and the aerial mycelium moss was more than 3 mm high.

[0050] The fermentation medium has a formula of: 15 g / L glucose, 15 g / L sucrose, 5 g / L corn flour, 12 g / L peptone, 6 g / L yeast powder, 0.08 g / L VB1, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate, 1-5 g / L uracil nucleotide, 40 mg / L selenite, pH 6.0.

[0051] The culture bottle containing the fermentation medium is streaked in the clean bench, with parallel lines spaced about 2 cm apart, and placed in a plant incubator for mushroom cultivation management. Cultivation conditions: temperature 25°C, light intensity 120lx, humidity 60%, light color change cultivation, and continue cultivation for 5 days when the mushroom skin turns orange, small brown water droplets appear on the surface of the culture medium, and fruiting body buds appear.

[0052] Fruiting body growth process: 24℃, light intensity 200-500lx, humidity 80-95% for 8h; 16℃, no light, humidity 80-95% for 16h, and continue for 5 days. Then control the temperature to 24℃, humidity 80-95%, maintain 16h of light hours per day to promote fruiting body growth, and cultivate for 16 days.

[0053] The results of the effect of adding uracil nucleotides on the growth and product of Cordyceps sinensis during the above fermentation stage are shown in Table 2.

[0054] Table 2 Effects of uracil nucleotides on selenium content and cordycepin content in Cordyceps

[0055] Uracil nucleotide g / L Fruiting body height cm Selenium content ug / g Cordycepin content mg / g 1 3.5±0.1 26.2±0.9 5.9±0.1 2 3.7±0.1 62.3±1.1 6.4±0.2 3 4.3±0.1 73.5±1.6 6.7±0.2 4 4.4±0.2 76.3±1.4 6.3±0.3 5 4.4±0.1 81.2±2.3 6.5±0.2

[0056] The results are shown in Table 2. The greater the amount of uracil nucleotide added, the more beneficial it is to the growth of the fruiting body, and the selenium content in the fruiting body is significantly increased, but the increase in cordycepin content is not obvious. Among them, when the concentration of uracil nucleotide is higher than 3g / L, the rate of increase in selenium content in the fruiting body begins to slow down. Considering economy and efficiency improvement, 3g / L of uracil nucleotide is selected as the optimal addition amount.

[0057] Example 3: Effect of substrate on fruiting body culture

[0058] The culture methods of the primary (selenite concentration is 30 mg / L), secondary and tertiary seed solutions are the same as those in Example 1.

[0059] The cultured tertiary seed liquid was transferred to 200 mL of fermentation medium containing different adenine concentrations at 20% v / v, placed in a 0.03 MPa micro-positive pressure environment with shade culture, and cultured for about 4 days at a ventilation volume of 0.1 vvm, a temperature of 26°C, and a humidity of 70%, until the mycelium was fully covered and the aerial mycelium moss was more than 3 mm high.

[0060] The formula of the fermentation medium is as follows: 15 g / L glucose, 15 g / L sucrose, 5 g / L corn flour, 12 g / L peptone, 6 g / L yeast powder, 0.08 g / L VB1, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate, 3 g / L uracil nucleotide, 1-5 g / L adenine, 40 mg / L selenite, pH 6.0.

[0061] The other subsequent culture processes are the same as in Example 2.

[0062] The results of the effect of adding adenine in the above fermentation stage on the growth and product of Cordyceps are shown in Table 3 and Figure 1As shown in A, B, and C, the increase in the amount of adenine added can increase the content of cordycepin, but it does not promote the absorption of selenium. When the addition amount exceeds 3g / L, the inhibitory effect on the growth of fruiting bodies is more obvious, and when the addition amount is 5g / L, the cordycepin content decreases. Therefore, the fermentation medium containing 1-2g / L adenine was used for the next study.

[0063] On the basis of the above-mentioned fermentation medium containing 1-2 g / L adenine, different concentrations of glycine are further added to the fermentation medium to replace part of the adenine. The formula of the fermentation medium is: 15 g / L glucose, 15 g / L sucrose, 5 g / L corn flour, 12 g / L peptone, 6 g / L yeast powder, 0.08 g / L VB1, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate, 3 g / L uracil nucleotide, 1-2 g / L adenine, 6-10 g / L glycine, 40 mg / L selenite, pH 6.0.

[0064] The results of the effect of adding glycine during the above fermentation stage on the growth and product of Cordyceps are shown in Tables 3 and Figure 1 As shown in D in Figure 1, replacing part of adenine with glycine can significantly increase the cordycepin content, accompanied by a 10% increase in selenium content. In particular, when the glycine addition amount is 10g / L, the fruiting body grows stronger.

[0065] Table 3 Effect of substrate on fruiting body culture

[0066] Adenine g / L Glycine g / L Fruiting body height cm Selenium content ug / g Cordycepin content mg / g Remark 1 —— 4.4±0.1 71.3±1.2 8.90±0.25 Figure 1 A 2 —— 4.1±0.2 75.2±1.9 14.41±0.22 3 —— 3.3±0.1 55.8±1.3 16.75±0.14 Figure 1 B 4 —— 2.4±0.1 42.3±0.9 18.33±0.30 5 —— 2.2±0.1 20.8±1.3 17.54±0.23 Figure 1 C 1 6 4.5±0.2 78.9±1.8 18.38±0.65 1 10 4.7±0.2 81.4±2.1 25.22±1.15 Figure 1 D 2 6 3.8±0.1 84.2±2.4 26.03±1.02 2 10 3.9±0.2 83.2±1.1 28.82±1.11

[0067] Example 4: Effects of ventilation and pressure maintenance on the cultivation of Cordyceps fruiting bodies

[0068] The culture methods of the primary (selenite concentration is 30 mg / L), secondary and tertiary seed solutions are the same as those in Example 1.

[0069] The cultured tertiary seed liquid was transferred to 200 mL fermentation medium at 20% v / v, placed in a 0.1-0.5 MPa micro-positive pressure environment under shade culture, and cultured for about 4 days at a ventilation volume of 0.05-0.2 vvm, a temperature of 26°C, and a humidity of 70%, until the mycelium was fully covered and the aerial mycelium moss was more than 3 mm high.

[0070] The fermentation medium has a formula of: 15 g / L glucose, 15 g / L sucrose, 5 g / L corn flour, 12 g / L peptone, 6 g / L yeast powder, 0.08 g / L VB1, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate, 3 g / L uracil nucleotide, 1 g / L adenine, 10 g / L glycine, 40 mg / L selenite, pH 6.0.

[0071] The culture bottle containing the fermentation medium is streaked in the clean bench, with parallel lines spaced about 2 cm apart, and placed in a plant incubator for mushroom cultivation management. Cultivation conditions: temperature 25°C, light intensity 120lx, humidity 60%, light color change cultivation, and continue cultivation for 4 to 5 days when the mushroom skin turns orange, small brown water droplets appear on the surface of the culture medium, and fruiting body buds appear.

[0072] Fruiting body growth process: 24℃, light intensity 200-500lx, humidity 80-95% for 8h; 16℃, no light, humidity 80-95% for 16h, and continue for 3-6 days. Then control the temperature at 24℃, humidity 80-95%, and maintain 16h of light hours per day to promote fruiting body growth, and cultivate for 10-15 days.

[0073] The results of the effects of changes in air pressure and ventilation volume in the micro-positive pressure environment during the fermentation stage on the growth and product of Cordyceps are shown in Table 4.

[0074] Table 4 Effects of ventilation and pressure maintenance on Cordyceps fruiting body culture

[0075]

[0076]

[0077] As shown in Table 4, ventilation and pressure maintenance have a significant promoting effect on the growth of mycelium and fruiting body, and can significantly shorten the culture time of the mycelium growth stage and the fruiting body growth stage. Under the conditions of pressure of 0.3Mpa, 0.1vvm, the culture time of the fermentation stage was shortened from 30 days in the control group (pressure of 0.1Mpa, 0.1vvm) to 24 days, and the cycle was reduced by 20%.

[0078] The present invention provides a method for cultivating a selenium-enriched Cordyceps fruiting body. There are many methods and approaches to implement the technical solution. The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention. All components not specified in this embodiment can be implemented using existing technologies.

Claims

1. A method for cultivating selenium-enriched Cordyceps sinensis fruiting bodies, characterized in that: The seed liquid of the Cordyceps militaris strain is inoculated into the fermentation medium for static fermentation of the mycelium. When the mycelium is fully covered and the aerial mycelium is 3 to 6 mm high, the mycelium is stimulated and then color change and mushroom production culture is carried out to obtain selenium-enriched Cordyceps militaris fruiting bodies.

2. The culture method according to claim 1, characterized in that The seed liquid of the Cordyceps militaris strain is a seed liquid obtained by inoculation and expansion of a multi-stage seed culture medium; The multi-stage seed culture medium has the following formulas: 25-40 g / L glucose, 10-20 g / L peptone, 5-10 g / L yeast powder, 0.1-2 g / L potassium dihydrogen phosphate, 0.1-2 g / L magnesium sulfate, 20-100 mg / L selenate and / or selenite, pH 5.

8.

3. The culture method according to claim 2, characterized in that The number of inoculation and expansion is 2 to 4 times; the inoculation volume is 5 to 30% v / v, and the volume ratio of the plate area of ​​the Cordyceps militaris strain to the seed culture medium is 10 to 30 cm 2 : 100~300cm 3 The culture conditions of the expansion culture are as follows: the rotation speed is 150-220 rpm and the temperature is 26-29° C. until the wet weight of the mycelium is greater than 180 g / L; the concentration of the selenate and / or selenite is increased in a gradient of 10-100% in the multi-stage seed culture medium.

4. The culture method according to claim 1, characterized in that The inoculation has an inoculation volume of 5 to 30% v / v.

5. The culture method according to claim 1, characterized in that The fermentation medium has a formula of: 10-20 g / L glucose, 5-15 g / L sucrose, 2-10 g / L corn flour, 1-15 g / L peptone, 1-6 g / L yeast powder, 0.05-0.5 g / L VB1, 0.5-2 g / L potassium dihydrogen phosphate, 0.1-0.5 g / L magnesium sulfate, 0.1-5 g / L adenine, 2-20 g / L glycine, 1-5 g / L uracil nucleotide, 0.5-100 mg / L selenate and / or selenite, and a pH of 6.

0.

6. The culture method according to claim 1, characterized in that The mycelium static fermentation has the following fermentation conditions: light-shading culture under a positive micro-pressure environment of 0.1-0.5 MPa, a ventilation volume of 0.05-0.2 vvm, a temperature of 22-29° C., and a humidity of 65-70%.

7. The culture method according to claim 1, characterized in that: The stimulation treatment is contact streaking or non-contact laser irradiation; wherein the contact streaking is: streaking the mycelium in the culture bottle containing the fermentation medium, with parallel lines spaced 1 to 3 cm apart; the non-contact laser irradiation is: laser irradiation for 5 to 20 minutes in dark conditions.

8. The culture method according to claim 1, characterized in that: The color changing and fruiting mushroom culture is divided into an induction phase and a growth phase.

9. The culture method according to claim 8, characterized in that: The culture conditions during the induction period are: temperature 20-26°C, light intensity 100-200lx, humidity 55-65%, and light color change culture. When the mycelium turns orange, small brown water droplets appear on the surface of the fermentation medium, and fruiting body buds appear, continue to culture for 4-6 days.

10. The culture method according to claim 8, characterized in that: During the growth stage, the culture conditions are as follows: 20-26°C, light intensity of 200-500lx, and humidity of 80-95% for 6-8h; 12-18°C, no light, and humidity of 80-95% for 16-18h, and continuous culture for 3-6 days; then the temperature is controlled at 24°C, the humidity is 80-95%, and the light hours are maintained at 16h per day to promote the growth of fruiting bodies, and the culture is carried out for 10-16 days.

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