CN112715276A - Culture method of cordyceps sobolifera spore powder - Google Patents
Culture method of cordyceps sobolifera spore powder Download PDFInfo
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- CN112715276A CN112715276A CN202110024079.1A CN202110024079A CN112715276A CN 112715276 A CN112715276 A CN 112715276A CN 202110024079 A CN202110024079 A CN 202110024079A CN 112715276 A CN112715276 A CN 112715276A
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
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- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a culture method of cordyceps sobolifera spore powder, which comprises the following steps: step 1, performing amplification culture on cordyceps sobolifera strains; step 2, inoculating the strain obtained by the enlarged culture in the step 1 into a solid culture medium for solid culture; step 3, collecting a culture medium of the cordyceps sobolifera spore powder; wherein, the solid culture process in the step 2 adopts whole illumination culture, and the illumination intensity is 200-500 Lux.
Description
Technical Field
The invention relates to a culture method of cordyceps sobolifera spore powder, in particular to a spherical thin-layer culture method of cordyceps sobolifera spore powder, and belongs to the field of cordyceps sinensis spore powder culture.
Background
The cicada fungus (Isaria cicadae Miquel) is called cicada pupa grass, cicadas, cicada mushrooms and the like, is a nymph of Isaria cicada, which is a fungus of Clavipitaceae and parasitizes Isaria cicada in cicadae, bamboo cicadas, mole cricket, mountain cicadas, black locust and the like, absorbs nutrition of insect bodies and converts the nutrition into mycelium when the climate environment is proper, and the top branch germinates to be similar to a corolla, so the cicada fungus is called as cicada fungus. The cordyceps sobolifera has a history of more than one thousand years as a medicine, ancient classical medical treatises such as Nanbei dynasty (five century of the Gongyuan) 'Legong Baoji treatise, Song dynasty Susong' materia medica of graph Jing, Song dynasty Tang cautiously 'Jing history type emergency materia medica' and Ming dynasty Li Shizhen 'materia medica outline' are recorded, and the cordyceps sobolifera has rich nutritional value.
The cordyceps sobolifera spore powder is used as a propagation organ of cordyceps sobolifera, most of nutrition of a cordyceps sobolifera culture is concentrated in the inoculation period, and the cordyceps sobolifera spore powder is rich in amino acid, fatty acid, dietary fiber and vitamin, wherein the protein is as high as 41.6%, and the dietary fiber is as high as 34.35%. Researches show that the cordyceps sobolifera spore powder has important effects on preventing and treating constipation, preventing rectum cancer and colon cancer, preventing coronary heart disease and the like.
Regarding the culture of spore powder, in the prior art, in the process of culturing cordyceps sobolifera sporophore bundles, methods such as adjusting culture conditions and prolonging culture time are used for improving the yield of spore powder on the cordyceps sobolifera sporophore bundles. The culture period is long, the energy consumption is high, the yield is low, the treatment is complicated, the comprehensive cost of the cordyceps sobolifera spore powder is high, and the market competitiveness is greatly reduced.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides the culture method of the cordyceps sobolifera spore powder, and the cordyceps sobolifera spore powder cultured by the method has high yield and high effective components, is suitable for industrial culture and has obvious application advantages.
As one aspect of the present invention, the present invention provides a method for culturing cordyceps sobolifera spore powder, comprising the steps of:
step 1, performing amplification culture on cordyceps sobolifera strains;
step 2, inoculating the strain obtained by the enlarged culture in the step 1 into a solid culture medium for solid culture;
step 3, collecting a culture medium of the cordyceps sobolifera spore powder;
wherein, the solid culture process in the step 2 adopts whole illumination culture, and the illumination intensity is 200-500 Lux.
Preferably, the illumination intensity is 200Lux-400 Lux; most preferably, the illumination intensity is 300 Lux.
Preferably, the illumination time is more than 18 hours/day; further preferably, the illumination time is more than 20 hours/day; still more preferably, the illumination time is greater than 22 hours/day; most preferably, the illumination time is 24 hours/day.
Preferably, in the solid culture process in the step 2, the thickness of the solid culture medium is 0.2-1.2 cm; further preferably, the thickness of the solid culture medium is 0.3-1.0 cm; further preferably, the thickness of the solid culture medium is 0.4-0.9 cm; most preferably, the thickness of the solid medium is 0.5-0.7 cm.
Preferably, in the solid culture process in the step 2, the culture temperature is 24-26 ℃, the culture humidity is 50-80%, and CO is added2The concentration is not more than 2000 ppm. Further preferably, the humidity of the culture is controlled to be 60% -80% when the culture starts until the upper surface of the grain is full of spores; when the upper surface of the grain is full of spores and the whole surface of the grain is full of spores, namely the grains are collected, the humidity is controlled to be 50% -60%. Since the grain is granular, the initial sporulation starts from the upper surface of the grain and eventually spores are produced on both the sides and bottom of the whole grain. The method adopts a method for controlling humidity in stages, wherein the first stage is from the beginning of culture to the overgrowth of spores on the upper surface of grains, and the humidity is controlled to be 60-80%, preferably 70%; and in the second stage, the overgrowth of the spores on the upper surface of the grain to the full surface of the grain is carried out, and the humidity is controlled to be 50-60%, and the humidity is preferably 50%.
Preferably, the inoculation amount in the step 2 is 5% -25%; further preferably, the inoculation amount is 10% -20%; still more preferably, the inoculum size is 15%.
Preferably, the solid medium in step 2 is a cereal medium, the cereal is selected from any one or more of barley, wheat, oat, buckwheat, wheat kernel, rye, bran, corn, soybean meal and rice, and the solid medium is any one or more of the above cereals after simple crushing; further preferably, the solid culture medium has a feed-water ratio of 1: (0.8-1.2); still further preferably, the solid medium has a feed-water ratio of 1: 1.
preferably, the strain expansion culture of the cordyceps sobolifera in the step 1 aims at expanding the strain number, and comprises any one or more culture modes of slant culture, shake flask culture, seeding tank culture and fermentation tank culture. The culture medium used for the scale-up culture is a medium conventional in the art. Preferably, the solid culture medium of the slant test tube is a PDA culture medium, and the liquid culture medium is 3.5 percent of white granulated sugar, 1 percent of yeast extract powder and 0.02 percent of soybean protein concentrate. The culture temperature of the liquid culture is 23-27 ℃, and preferably 25 ℃.
The feed-water ratio is the weight/volume ratio (kg/L) of the grains to the water in the preparation process of the solid culture medium.
The inoculation amount is XX%' in the invention refers to the percentage of the volume (L) of the inoculated seed liquid in the weight (kg) of the solid culture medium; the weight of the solid medium is the weight of the grains.
In the present invention, unless otherwise stated, the operations performed are preferably performed under substantially aseptic conditions, as is well known in the art, in order to avoid contamination with germs, and the various tools and materials used are sterilized.
The invention has the beneficial effects that: the invention adopts a specific culture method, the culture is mainly spore powder, only a small amount of cordyceps sobolifera sporocarp is generated or no cordyceps sobolifera sporocarp is generated, and the yield of the spore powder is greatly improved.
Drawings
FIG. 1 is an external view of a culture bag.
FIG. 2 is an internal view of the culture bag.
FIG. 3 is an external view of the culture using the culture cassette.
FIG. 4 is an internal view of the case culture.
FIG. 5 is an external view of the cultivation in an incubator.
FIG. 6 is an internal view of cultivation in an incubator.
Detailed Description
EXAMPLE 1 bag culture
1, liquid culture:
1.1 stock/cultivar preparation
1.1.1 preparation of the culture Medium
1.1.1.1 culture Medium formulation:
| serial number | Name of raw and auxiliary materials | Percentage of | 1L dosage |
| 1 | Yeast extract (FM802) | 1％ | 10g |
| 2 | Soy protein concentrate | 0.02％ | 0.2g |
| 3 | White granulated sugar | 3.5％ | 35g |
| 4 | Filtered water | - | To add to 1L |
Note: the seed liquid loading in the 2L triangular flask is 500-800 ml.
1.1.1.2 preparation method of spore suspension
Preparing an inoculation tool, and subpackaging the filtered water into test tubes, wherein each test tube is filled with 12ml of water. Sterilizing at 121 + -1 deg.C for 40min, and cooling to room temperature.
Wiping an ultra-clean workbench with 75% ethanol, transferring sterilized water, a triangular flask and a culture medium into the workbench, performing ultraviolet sterilization for 30min, performing aseptic operation, pouring 10ml of sterile water into 1 slant strain test tube, scraping conidia with an inoculating loop or an inoculating shovel, and shaking uniformly to obtain a spore suspension for later use.
1.1.2 preparation of first-class stock seed solution
Weighing the raw materials for preparing the culture medium according to the proportion of 1.1.1.1, subpackaging according to the amount of 500 plus one ml of culture medium in a 2L triangular bottle, plugging a silica gel plug, sterilizing at 121 +/-1 ℃ for 40min, naturally cooling, directly adding spore suspension into the culture medium in an ultraclean workbench under the aseptic operation, inoculating 1 slant strain into a triangular bottle, covering the silica gel plug, and carrying out constant-temperature shaking culture at the temperature of 25 +/-1 ℃ and 150rpm for 48-55h to obtain a primary stock seed solution.
1.1.3 preparation of seed liquid for fermentation tank cultivation
1.1.3.1 preparation of culture Medium
Weighing the raw materials for preparing the culture medium according to the proportion of 1.1.1.1, and fixing the volume. The liquid filling amount of the fermentation tank does not exceed 70 percent of the total volume.
1.1.3.2 Sterilization
Before sterilization, whether all valves are loosened or not, whether water leakage or air leakage occurs or not needs to be checked, and sterilization can be carried out if no looseness exists. The sterilization method is carried out according to the operation specification of the fermentation system, the temperature is reduced to 25 +/-1 ℃ for standby after sterilization, the pressure of the tank body is adjusted to be about 0.05MPa, and positive pressure is kept.
1.1.3.3 cultivation of cultivars in fermentors
Inoculating by flame method, wherein the inoculation amount of the seed liquid is 1.5-3% (the actual inoculation amount can be slightly adjusted according to the concentration of the seed liquid). Before inoculation, the micro positive pressure of the tank body is maintained, the inoculation port is disinfected by 75% ethanol, the cotton ring is ignited and placed in the inoculation port, the inoculation cover is slowly opened, and the first-class original seed bottle is inoculated into a fermentation tank after being disinfected by flame for about half a minute. After inoculation, the inoculation cover is covered.
The culture temperature is 25 + -1 deg.C, aeration ratio is 0.71-1.2vvm (15-25m/h), and tank pressure is maintained at 0.05-0.1 MPa.
1.1.4 can
The culture time is 19-28h, and the strain can be placed in the pot if the strain meets the release requirement of the cordyceps sobolifera cultivated species. Before the tank is placed, the material discharging port needs to be sterilized by steam for at least 20 min. Before the inoculation silicone tube is in butt joint with the discharging tube, the mouth of the discharging tube needs to be disinfected by 75% ethanol, after the butt joint, the silicone tube needs to be clamped, and steam is introduced again for sterilization for more than 20 min.
2, solid culture:
the culture bag is used as a culture container, the size of the culture bag is 950mm by 550mm, and the size of the bottom support of the culture bag is 540mm by 270mm by 60 mm.
2.1 Medium formulation
The solid culture medium comprises wheat and purified water, wherein the charging amount of each bag is 0.3kg, and the ratio of the cleaned wheat to the purified water is 1: 1.
2.1.1 cleaning and sub-packaging of cereals
And starting the cleaning machine, pouring the grains into a feeding hole of the cleaning machine, operating the equipment according to the specification, and cleaning the grains for later use. According to the established specification, subpackaging by using a subpackaging machine, subpackaging in a culture bag, binding the bag opening, placing in a sterilization tray, and placing the sterilization tray on a sterilization vehicle for sterilization;
2.1.2 Sterilization of solid Medium
And (4) pushing the sterilization vehicle into the sterilization cabinet, closing the cabinet door, and operating the equipment according to the specification of the sterilization cabinet to perform sterilization. Sterilizing at 0.146 + -0.002 MPa and 127 + -2 deg.C for 45-50 min.
After sterilization, the culture medium is transferred into a forced cooling chamber and cooled to room temperature for inoculation.
2.2 solid inoculation
2.2.1 Sterilization of inoculation tools
The silicone tube and the inoculating gun are connected in series, two ends are wrapped with more than 8 layers of gauze, the bag is placed into a PP bag, the bag opening is tightened, and the sterilization is carried out for 45-50min at 127 +/-2 ℃.
2.2.2 inoculation control
An inoculating worker holds the inoculating gun by hand, and the other person provides a solid culture medium for inoculating operation; when the inoculating gun is not used, the gun head needs to be placed on flame to prevent bacteria contamination. The seed liquid of the fermentation tank is inoculated into the solid culture medium in a proportion of 15 percent, and the fermentation tank is placed in a tank for inoculation.
2.2.3 Strain blending treatment
After inoculation, the culture bag is kneaded or reversed back and forth to uniformly mix the bacteria liquid and the culture medium, then the culture bag is transferred to a culture rack and paved, the whole bottom support of the culture bag is uniformly paved to enable the height of the culture material to be 0.5cm (after wheat paving is finished, the height of the culture material is equivalent to the thickness of a layer of wheat), and at least 5cm of gap is reserved between the edge of the bag and the wheat to facilitate stretching of the bag during illumination culture.
2.3 spore powder culture
2.3.1 culture conditions
The illumination culture intensity is 300Lux, the temperature is 25 ℃, the concentration of carbon dioxide is controlled not to exceed 2000ppm, and the humidity is controlled to be 70% when the culture starts until the upper surface of the grain is just full of spores; when the upper surface of the grain is full of spores until the whole surface of the grain is full of spores, namely the grains are collected, the humidity is controlled to be 50%; the illumination time is 24h/d, and the culture time is 20-28 days.
2.3.2 bundling and opening bags
The bag tying and opening are carried out after 2-5 days of culture: the bag is pricked by sterilized stainless steel pricks, the number of the pricked holes is 50-60, the aperture is less than or equal to 3mm, and the pricked holes are uniformly distributed. The culture bag which is perforated is placed into the culture bag bottom support to be supported, the bag corners are folded inwards and supported by using two hands, and the bag opening is folded upwards, so that the culture bag is rectangular, is similar to a small greenhouse, and allows a culture to grow under the set conditions, and is shown in figure 1.
2.4 Chamber management
When the culture starts to grow spores on the upper surface of grains, the bottom of the culture is still adhered to the culture bag, the bottom of the culture is separated from the bag in a manual intervention mode, the culture is placed back to the original position after separation, the culture can not be adhered to the bag, at the moment, the bottom of the culture can also obtain more spaces and oxygen, the rapid growth of spore powder is facilitated, and the culture is continued.
3, harvesting:
3.1 harvesting treatment
The harvesting table is connected with a power supply, the dust removal switch is turned on, ventilation is carried out, the movable window of the harvesting cover is opened, the culture is taken out of the culture bag and placed on the harvesting table, the culture is broken off and placed in a baking pan. The culture results are shown in FIG. 2.
3.2 dehumidification, drying and sieving
Directly dehumidifying for 18-42 h after enough quantity required by drying each batch is collected; the dehumidification condition is that the temperature of the dehumidification room is controlled below 30 ℃, and the humidity is controlled below 60%; the drying time is 12-18 h; the drying conditions were 80. + -. 1 ℃.
3.3 sieving
And (3) putting the dried culture with the spore powder into a shaking sieve for sieving (the sieve aperture is 100 meshes), detecting the appearance and the moisture of the spore powder, and checking to be qualified for later use.
The culture results are shown in FIGS. 1 and 2.
EXAMPLE 2 cassette culture
1, liquid culture:
1.1 stock/cultivar preparation
1.1.1 preparation of the culture Medium
1.1.1.1 culture Medium formulation:
| serial number | Name of raw and auxiliary materials | Percentage of | 1L dosage |
| 1 | Yeast extract (FM802) | 1％ | 10g |
| 2 | Soy protein concentrate | 0.02％ | 0.2g |
| 3 | White granulated sugar | 3.5％ | 35g |
| 4 | Filtered water | - | To add to 1L |
Note: the seed liquid loading in the 2L triangular flask is 500-800 ml.
1.1.1.2 preparation method of spore suspension
Preparing an inoculation tool, and subpackaging the filtered water into test tubes, wherein each test tube is filled with 12ml of water. Sterilizing at 121 + -1 deg.C for 40min, and cooling to room temperature.
Wiping an ultra-clean workbench with 75% ethanol, transferring sterilized water, a triangular flask and a culture medium into the workbench, performing ultraviolet sterilization for 30min, performing aseptic operation, pouring 10ml of sterile water into 1 slant strain test tube, scraping conidia with an inoculating loop or an inoculating shovel, and shaking uniformly to obtain a spore suspension for later use.
1.1.2 preparation of first-class stock seed solution
Weighing the raw materials for preparing the culture medium according to the proportion of 1.1.1.1, and filling 10-20L of purified water into a seeding tank. Placing a certain amount of culture medium raw materials into a seeding tank, sterilizing for 30 minutes at the temperature of 121 +/-1 ℃, cooling, and inoculating a certain amount of spore suspension prepared in advance into a fermentation tank by using a flame inoculation method. The culture temperature is 25 + -1 deg.C, and aeration ratio is 0.71-1.2 vvm. Maintaining the pressure in the tank at 0.05-0.1MPa, and culturing for 44-48h to obtain the first-stage stock seed solution.
1.1.3 preparation of seed liquid for fermentation tank cultivation
1.1.3.1 preparation of culture Medium
Weighing the raw materials for preparing the culture medium according to the proportion of 1.1.1.1, and fixing the volume. The liquid filling amount of the fermentation tank does not exceed 70 percent of the total volume.
1.1.3.2 Sterilization
Before sterilization, whether all valves are loosened or not, whether water leakage or air leakage occurs or not needs to be checked, and sterilization can be carried out if no looseness exists. The sterilization method is carried out according to the operation specification of the fermentation system, the temperature is reduced to 25 +/-1 ℃ for standby after sterilization, the pressure of the tank body is adjusted to be about 0.05MPa, and positive pressure is kept.
1.1.3.3 cultivation of cultivars in fermentors
Sterilizing the transfer line between the seed tank and the fermentation tank with steam for 30-40min, and increasing the pressure of the seed tank to 0.15-0.18MPa. The pressure of the fermentation tank is adjusted to 0.02-0.05 MPa. And opening a valve required for liquid transfer, and closing the valve after liquid in the seed tank is completely transferred to the fermentation tank and liquid transfer is finished.
The culture temperature is 25 + -1 deg.C, aeration ratio is 0.71-1.2vvm (15-25m/h), and tank pressure is maintained at 0.05-0.1 MPa.
1.1.4 can
The culture time is 19-28h, and the strain can be placed in the pot if the strain meets the release requirement of the cordyceps sobolifera cultivated species. Before the tank is placed, the material discharging port needs to be sterilized by steam for at least 20 min. Before the inoculation silicone tube is in butt joint with the discharging tube, the mouth of the discharging tube needs to be disinfected by 75% ethanol, after the butt joint, the silicone tube needs to be clamped, and steam is introduced again for sterilization for more than 20 min.
2, solid culture:
the procedure is essentially the same as in example 1, except that the culture bag is replaced with a culture box, and the hole pricking and the bag opening are omitted.
Harvesting: the procedure is as in example 1.
The culture results are shown in FIGS. 3 and 4.
EXAMPLE 3 incubator culture
Liquid culture: the procedure is as in example 1.
Solid culture: the procedure is essentially the same as in example 1, the bag is replaced with an incubator, eliminating the need for puncturing and "bag bracing".
Harvesting: the procedure is as in example 1.
The culture results are shown in FIGS. 5 and 6.
Example 4 Effect of illumination on spore powder production
Wheat is used as a culture medium, a culture bag is used as a culture container, the charging amount is 300 g/bag (the thickness of the culture medium is 0.5cm), the ratio of feed to water is 1:1, the thickness of the culture medium is about 0.5cm, the inoculation amount is 15%, the culture temperature is 25 ℃, and the culture time is 20 days. 6 experimental groups were set, 3 replicates per group, and the effect of different light intensities on spore powder yield was examined (other operating steps and parameters refer to example 1).
The results are shown in Table 1.
The spore powder yield is found to be optimal by the statistics of the spore powder yield with the light intensity of 300 Lux.
TABLE 1 Effect of illumination on spore powder production
Note: the yield of spore powder is defined as dry weight of spore powder/amount of powder charged per bag x 100%, as follows.
Example 5 Effect of culture humidity on spore powder production
Wheat is used as a culture medium, a culture bag is used as a culture container, the charging amount is 300 g/bag (the thickness of the culture medium is 0.5cm), the illumination intensity is 300Lux, the inoculation amount is 15%, the culture temperature is 25 ℃, and the culture time is 20 days. 2 experimental groups were set, 3 replicates each, and the effect of the humidity of the culture environment on the spore powder yield was examined (other operating steps and parameters refer to example 1).
The results are shown in Table 2.
Spore powder yield statistics show that the ratio of material to water is 1: the yield of the spore powder is optimal when the yield is 1 hour.
TABLE 2 Effect of cultivation humidity on spore powder yield
EXAMPLE 6 Effect of Medium thickness on spore powder production
Wheat is used as a culture medium, a culture bag is used as a culture container, and the ratio of material to water is 1:1, the illumination intensity is 300Lux, the inoculation amount is 15%, the culture temperature is 25 ℃, and the culture time is 20 days. 5 experimental groups were set, 3 replicates per group, and the effect of different media charges on spore powder production was examined (other operating steps and parameters refer to example 1).
The results are shown in Table 3.
Spore powder yield statistics show that the yield of the spore powder is optimal when the charging amount is 300g and the thickness of the culture medium is 0.5 cm.
TABLE 3 Effect of Medium Loading on spore powder production
Example 7 Effect of solid Medium Water ratio on spore powder production
Wheat is used as a culture medium, a culture bag is used as a culture container, the charging amount is 300 g/bag (the thickness of the culture medium is 0.5cm), the illumination intensity is 300Lux, the inoculation amount is 15%, the culture temperature is 25 ℃, and the culture time is 20 days. 5 experimental groups were set, 3 replicates per group, and the effect of different feed water ratios on spore powder yield was examined (other operating steps and parameters refer to example 1).
The results are shown in Table 4.
Spore powder yield statistics show that the ratio of material to water is 1: the yield of the spore powder is optimal when the yield is 1 hour.
TABLE 4 influence of Medium Water ratio on spore powder yield
| Serial number | Ratio of material to water | Spore powder Dry weight (g) | Yield of spore powder (%) |
| 1 | 1:0.6 | 55.17 | 6.13 |
| 2 | 1:0.8 | 65.25 | 7.25 |
| 3 | 1:1.0 | 115.11 | 12.79 |
| 4 | 1:1.2 | 67.5 | 7.50 |
| 5 | 1:1.4 | 57.23 | 6.35 |
Claims (10)
1. A culture method of cordyceps sobolifera spore powder is characterized by comprising the following steps:
step 1, performing amplification culture on cordyceps sobolifera strains;
step 2, inoculating the strain obtained by the enlarged culture in the step 1 into a solid culture medium for solid culture;
step 3, collecting a culture medium of the cordyceps sobolifera spore powder;
wherein, the solid culture process in the step 2 adopts whole illumination culture, and the illumination intensity is 200-500 Lux.
2. The culture method according to claim 1, wherein the illumination intensity is 200Lux to 400 Lux; preferably, the illumination intensity is 300 Lux.
3. The culture method according to claim 2, wherein the light irradiation time is more than 18 hours/day; preferably, the illumination time is more than 20 hours/day; further preferably, the illumination time is more than 22 hours/day; most preferably, the illumination time is 24 hours/day.
4. The culture method according to claim 1, wherein in the solid culture process of step 2, the culture temperature is 24 ℃ to 26 ℃, the culture humidity is 50% to 80%, and CO is present2The concentration is not more than 2000 ppm.
5. The culture method according to claim 4, wherein the culture humidity is: when the culture is started and the upper surface of the grain is just full of spores, the humidity is controlled to be 60-80%; when the upper surface of the grain is full of spores, the humidity is controlled to be 50% -60%.
6. The culture method according to claim 5, wherein the culture humidity is: controlling the humidity to be 70% when the culture is started until the upper surface of the grain is just full of spores; when the upper surface of the grain is full of spores, the humidity is controlled to be 50%.
7. The culture method according to claim 1, wherein in the solid culture process of step 2, the thickness of the solid medium is 0.2 to 1.2 cm; preferably, the thickness of the solid culture medium is 0.3-1.0 cm; further preferably, the thickness of the solid culture medium is 0.4-0.9 cm; most preferably, the thickness of the solid medium is 0.5-0.7 cm.
8. The culture method of claim 1, wherein the amount of inoculation in step 2 is 5% to 25%; preferably, the inoculation amount is 10% -20%; further preferably, the inoculum size is 15%.
9. The culture method according to claim 1, wherein the solid medium in step 2 is a grain medium, and the grain is selected from one or more of barley, wheat, oat, buckwheat, wheat kernel, rye, bran, corn, soybean meal and rice.
10. The culture method according to claim 9, wherein the solid medium has a feed-water ratio of 1: (0.8-1.2); preferably, the solid culture medium has a feed-water ratio of 1: 1.
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