CN115553179A - Artificial cordyceps militaris and preparation method thereof - Google Patents
Artificial cordyceps militaris and preparation method thereof Download PDFInfo
- Publication number
- CN115553179A CN115553179A CN202211223208.0A CN202211223208A CN115553179A CN 115553179 A CN115553179 A CN 115553179A CN 202211223208 A CN202211223208 A CN 202211223208A CN 115553179 A CN115553179 A CN 115553179A
- Authority
- CN
- China
- Prior art keywords
- culture medium
- inoculation
- culture
- sterilization
- temperature
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 35
- 241001264174 Cordyceps militaris Species 0.000 title claims abstract description 31
- 239000001963 growth medium Substances 0.000 claims abstract description 78
- 238000011081 inoculation Methods 0.000 claims abstract description 59
- 230000001954 sterilising effect Effects 0.000 claims abstract description 41
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 41
- 238000000034 method Methods 0.000 claims abstract description 22
- 238000003306 harvesting Methods 0.000 claims abstract description 19
- 238000001816 cooling Methods 0.000 claims abstract description 14
- 238000004519 manufacturing process Methods 0.000 claims abstract description 14
- 244000025254 Cannabis sativa Species 0.000 claims abstract description 8
- 241000233866 Fungi Species 0.000 claims abstract description 8
- 230000008569 process Effects 0.000 claims abstract description 7
- 238000003860 storage Methods 0.000 claims description 32
- 239000007788 liquid Substances 0.000 claims description 26
- 241000894006 Bacteria Species 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 239000000047 product Substances 0.000 claims description 18
- 238000009423 ventilation Methods 0.000 claims description 17
- 238000004806 packaging method and process Methods 0.000 claims description 15
- 241000190633 Cordyceps Species 0.000 claims description 14
- 230000000694 effects Effects 0.000 claims description 13
- 241000255789 Bombyx mori Species 0.000 claims description 12
- -1 polypropylene Polymers 0.000 claims description 12
- 238000012258 culturing Methods 0.000 claims description 11
- 238000001035 drying Methods 0.000 claims description 11
- 235000015097 nutrients Nutrition 0.000 claims description 11
- 238000002955 isolation Methods 0.000 claims description 10
- 229920001817 Agar Polymers 0.000 claims description 9
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 9
- 239000004743 Polypropylene Substances 0.000 claims description 9
- 239000008272 agar Substances 0.000 claims description 9
- 238000005286 illumination Methods 0.000 claims description 9
- 238000009630 liquid culture Methods 0.000 claims description 9
- 229920001155 polypropylene Polymers 0.000 claims description 9
- 239000007787 solid Substances 0.000 claims description 9
- 239000000126 substance Substances 0.000 claims description 9
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 8
- 241000382353 Pupa Species 0.000 claims description 8
- 241001248610 Ophiocordyceps sinensis Species 0.000 claims description 7
- 238000007789 sealing Methods 0.000 claims description 7
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 6
- 229910002091 carbon monoxide Inorganic materials 0.000 claims description 6
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- 229910052760 oxygen Inorganic materials 0.000 claims description 6
- 239000001301 oxygen Substances 0.000 claims description 6
- 239000005022 packaging material Substances 0.000 claims description 6
- 230000002035 prolonged effect Effects 0.000 claims description 6
- 238000005303 weighing Methods 0.000 claims description 6
- 229920000742 Cotton Polymers 0.000 claims description 5
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 5
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 5
- 241000238631 Hexapoda Species 0.000 claims description 4
- 230000006866 deterioration Effects 0.000 claims description 4
- 229910052742 iron Inorganic materials 0.000 claims description 4
- 230000008018 melting Effects 0.000 claims description 4
- 238000002844 melting Methods 0.000 claims description 4
- 239000000243 solution Substances 0.000 claims description 4
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 240000001624 Espostoa lanata Species 0.000 claims description 3
- 235000009161 Espostoa lanata Nutrition 0.000 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 239000004698 Polyethylene Substances 0.000 claims description 3
- 244000061456 Solanum tuberosum Species 0.000 claims description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 3
- 238000009825 accumulation Methods 0.000 claims description 3
- 230000032683 aging Effects 0.000 claims description 3
- 238000007605 air drying Methods 0.000 claims description 3
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 3
- 229910052782 aluminium Inorganic materials 0.000 claims description 3
- 230000001580 bacterial effect Effects 0.000 claims description 3
- 244000052616 bacterial pathogen Species 0.000 claims description 3
- 238000009835 boiling Methods 0.000 claims description 3
- 239000002775 capsule Substances 0.000 claims description 3
- 230000008859 change Effects 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- 239000003034 coal gas Substances 0.000 claims description 3
- 239000002131 composite material Substances 0.000 claims description 3
- 238000005520 cutting process Methods 0.000 claims description 3
- 230000007850 degeneration Effects 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 239000008157 edible vegetable oil Substances 0.000 claims description 3
- 238000005562 fading Methods 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 239000011888 foil Substances 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 239000012535 impurity Substances 0.000 claims description 3
- 239000004615 ingredient Substances 0.000 claims description 3
- 230000005764 inhibitory process Effects 0.000 claims description 3
- 239000002655 kraft paper Substances 0.000 claims description 3
- 238000011068 loading method Methods 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 239000000123 paper Substances 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000002985 plastic film Substances 0.000 claims description 3
- 229920006255 plastic film Polymers 0.000 claims description 3
- 229920000573 polyethylene Polymers 0.000 claims description 3
- 239000012286 potassium permanganate Substances 0.000 claims description 3
- 238000000746 purification Methods 0.000 claims description 3
- 238000005057 refrigeration Methods 0.000 claims description 3
- 150000003839 salts Chemical class 0.000 claims description 3
- 238000007711 solidification Methods 0.000 claims description 3
- 230000008023 solidification Effects 0.000 claims description 3
- 238000005507 spraying Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 2
- 238000004140 cleaning Methods 0.000 claims 1
- 230000002354 daily effect Effects 0.000 claims 1
- 230000004069 differentiation Effects 0.000 claims 1
- 230000003203 everyday effect Effects 0.000 claims 1
- 238000011049 filling Methods 0.000 claims 1
- 235000016709 nutrition Nutrition 0.000 claims 1
- 230000035764 nutrition Effects 0.000 claims 1
- 230000009467 reduction Effects 0.000 claims 1
- 230000001502 supplementing effect Effects 0.000 claims 1
- 238000007726 management method Methods 0.000 abstract description 21
- 238000005516 engineering process Methods 0.000 abstract description 5
- 238000013070 change management Methods 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 4
- 238000002845 discoloration Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The artificial cordyceps militaris and the preparation method thereof are a fungus production technology, which make up the gap of the prior cordyceps militaris production technology, and the preparation method comprises the following processes: the preparation of strains → the preparation of culture medium → bottling → sterilization → cooling → inoculation → spawn running management → color change management → grass growing management → harvesting → processing.
Description
Technical Field
The invention relates to a fungus production technology, in particular to artificial cordyceps militaris and a preparation method thereof.
Background
Cordyceps militaris, also known as Cordyceps militaris and Cordyceps militaris, is a complex of insect and fungus formed after Cordyceps militaris is infected by insect pupae, and has similar medical and health promotion effects to natural Cordyceps militaris. At present, the production technology of the cordyceps militaris in the market is rare.
Disclosure of Invention
The invention aims to provide a novel artificial cordyceps militaris and a preparation method thereof. The preparation method makes up the gap of the prior cordyceps militaris production technology, and comprises the following steps: preparing strains → preparing culture medium → bottling → sterilizing → cooling → inoculating → managing spawn running → color changing managing → managing grass → harvesting → processing;
first line of bacterial preparation
1.1 Production of mother seeds
The process flow for preparing the artificial cordyceps mother seeds comprises the following steps: preparation of culture medium → selection and collection of seed source of artificial cordyceps sinensis → tissue separation (spore isolation) → purification culture → mother strain. The preparation method comprises the following steps:
1.1.1 culture medium formula:
150-250 g of potato, 15-25 g of glucose, 1-5 g of potassium dihydrogen phosphate, 1-2 g of magnesium sulfate, 15-25 g of agar, 15: 4 tablets of Vb1 and 800-1200 ml of water.
1.1.2 The preparation method comprises the following steps:
(1) Weighing: accurately weighing various substances according to the proportion of the various substances in the formula.
(2) Extracting juice: the preparation method comprises boiling the above materials in water, and maintaining with slow fire for 15-20 min; then filtered through gauze to leave the juice. Filtering and then making up the water content to 800-1200 ml.
(3) Dissolution of other substances: adding agar into the filtrate, heating to melt the agar, stirring continuously during melting, and adding other nutrients to dissolve.
(4) Subpackaging: subpackaging the prepared culture medium into test tubes when the culture medium is hot; during subpackaging, the loading amount is preferably 1/5-1/4 of the height of the test tube; the culture medium is not adhered to the tube opening during subpackage, so as to avoid pollution.
(5) Tampon and package
After the culture medium is subpackaged, a cotton plug is plugged at the opening of the test tube; after the cotton plugs are plugged, every 7 test tubes are bundled, and a layer of kraft paper is wrapped outside the test tubes for fastening.
(6) And (3) sterilization: the mother culture medium is generally sterilized by autoclaving for 25-35 minutes; the test tube culture medium is vertically placed in a wide-mouth bottle or an iron frame during sterilization, then is put in a pressure cooker for sterilization, and is naturally cooled after 25-35 minutes.
(7) Cooling and swinging the inclined plane: after sterilization, naturally reducing the pressure to zero, opening the pressure cooker, taking out the test tube, and placing the test tube on an inclined plane when the test tube is hot; the length of the inclined plane is 1/2-2/3 of that of the test tube; after solidification, the culture medium is a slant culture medium.
1.1.3 Strain isolation
Tissue isolation:
(1) Cutting a small piece of tissue with the length of 0.1-0.2 cm from the surface-sterilized and sterile-cleaned cordyceps militaris stroma under the aseptic condition, and connecting the tissue piece to the center of a culture medium by using a sterile inoculating needle.
(2) The inoculated medium was placed at 22 ℃ and cultured in the dark.
(3) And (4) about 15 days, when new hyphae extend on the culture medium, selecting the hyphae which are free of pollution and strong in growth, and storing the hyphae in a refrigerator for later use.
1.2 Preparation of liquid spawn
1.2.1 The formula of the culture medium is as follows:
30 g of glucose, 20 g of peptone, 0.5 g of monopotassium phosphate, 2.5 g of salt and 1000 ml of water.
1.2.2 Bottling: the prepared liquid culture medium is respectively filled into wide-mouth bottles, each bottle is 3500-4500 ml, a few drops of edible oil are dropped into the bottle bottles to play a defoaming role, a polypropylene film is used for sealing and tightening, and sterilization is carried out.
1.2.3 And (3) sterilization: and placing the wide-mouth bottle filled with the liquid culture medium into an autoclave for sterilization for 35-45 minutes.
1.2.4 Inoculation: placing the sterilized culture medium, mother seeds and inoculation tools on a super clean workbench or in an inoculation box, and then sterilizing the inoculation chamber, the super clean workbench or the inoculation box; when inoculating, inoculating the mother strain into liquid culture medium by aseptic technique, inoculating 1-2 pieces of Cordyceps militaris mother strain into a bottle of culture medium; each parent seed is 2 times 2 cm in size, i.e. 4 cm square.
1.2.5 Culturing: placing in dark environment after inoculation, standing for 2-3 days to recover growth of mycelium, and then introducing oxygen to perform shake culture; shaking for 2-3 days to finish the preparation of liquid strain; the prepared liquid strain is stored in a refrigerator, the temperature of the refrigerator is kept at 2-5 ℃ for standby use, and the liquid strain is used up in 10-20 days as much as possible.
Preparing the second culture medium and bottling
2.1 Silkworm chrysalis culture medium
The silkworm pupa is selected from high-quality silkworm pupas according to the selection standard of no impurity, no mildew, no worm damage, no peculiar smell, normal color, uniform size, 2-4 cm long and 8-10 mm thick. Air drying fresh pupa Bombycis.
2.2 Bottling: bottling in 400-600 ml wide-mouth bottle, each bottle contains 25-35 g silkworm chrysalis, and the mouth is wrapped with polypropylene film.
The method for binding comprises the following steps: and (5) tying two times at the bottle mouth by using a rubber band to ensure that the bottle mouth is tightly tied.
Third sterilization and inoculation
3.1 And (3) sterilization: maintaining the pressure at 0.15 MPa for 1 hour or at 100 deg.c for 10 hours; when sterilizing, the water in the pot is sufficiently added, the bottles are compactly placed, and the pot is taken out after naturally reducing the pressure.
And (4) directly placing the material bottle into an inoculation room after sterilization, cooling, and cooling to normal temperature.
3.2 Inoculation:
3.2.1 disinfection:
the inoculation chamber (box) needs to be thoroughly disinfected by formaldehyde, the dosage of the formaldehyde is 10 ml and 5 g potassium permanganate in each cubic meter of space, then the door and the window are closed and closed for 30 minutes, and simultaneously the ultraviolet lamp is turned on for irradiation disinfection; when in use, the culture medium, the inoculation tool, the alcohol lamp, the alcohol cotton ball, the lighter and the like which are sterilized and cooled are put into an inoculation chamber (box), and the inoculation chamber is started to irradiate and sterilize for 30 minutes by the ultraviolet lamp.
3.2.2 liquid spawn treatment:
the liquid strain cultured by shaking can be directly used for inoculation; but for the condition that the concentration of the bacteria balls is large (the concentration of the bacteria balls is more than 50 percent) and the culture production amount is large, water is needed to be added into the liquid bacteria for dilution; the concentration of the bacteria balls reaches about 50 percent, and the method is suitable for production inoculation.
3.2.3 inoculation:
directly inoculating liquid strains into a solid culture medium in an aseptic state; directly injecting the mixture on the surface of the solid medium in a spraying mode.
The inoculation operation process is carried out in an inoculation chamber (box), and the inoculated culture chamber is moved into a culture chamber; after inoculation, vertically placing the bottle in a dark environment for culturing to ensure that hyphae germinate, and then placing the bottle on a shelf for culturing after field planting; otherwise, the bacteria liquid or nutrient solution and culture medium will fall to one side, resulting in uneven grass; the culture chamber is sterilized before use, and is used after ventilation.
Fourth spawn running period management
After hyphae germinate, the hyphae can be put on a culture frame to enter the spawn running period for management; the spawn running period is easily polluted by mixed bacteria, so closed culture is required.
4.1 temperature: the culture temperature of the mycelium can not be lower than 17 ℃ at least and can not be higher than 24 ℃ at most; preferably, incubation is carried out at constant temperature.
4.2 Humidity: the relative humidity of air is required to be kept to be 65% -70% in the spawn running period, and the air can be properly sprayed on the indoor ground or in the air for humidification.
4.3 Ventilating: the workshop is properly ventilated during the hypha growth stage.
If the culture chamber is heated by a stove, attention needs to be paid to discharge coal gas, namely carbon monoxide, in time; because carbon monoxide has a certain inhibition effect on the growth of hyphae.
4.4 Illumination: the culture should be protected from light at this stage.
4.5 Daily management:
4.5.1 examining the bacteria vial: the fungus bottles are frequently checked to find that hyphae grow slowly, reasons need to be searched in time so as to take proper measures to remedy the hyphae, and polluted hyphae need to be treated to minimize the loss.
4.5.2 constant temperature culture: the mycelium culture stage is preferably carried out at constant temperature, and the temperature is not higher or lower.
About 15 days later, the next stage of management can be carried out.
Fifth color cycle management
5.1 Ventilating: the indoor ventilation is required to be carried out once a day, 35-45 minutes each time, and the air is kept fresh.
5.2 Humidity: and controlling the concentration to be about 75%.
5.3 Illumination: scattered light irradiation is required for 12 to 14 hours per day.
5.4 Temperature: the temperature in the grass-growing period is controlled between 20 ℃ and 25 ℃.
Sixth management of emergence period
6.1 Temperature: the temperature of the primordium, namely the prototypical bulge growing on the surface of the culture medium is controlled to be 18-22 ℃ before the occurrence; the temperature may be suitably raised after the primordia have appeared, and the temperature does not exceed 28 ℃.
6.2 Ventilating: ventilating after the primordium appears, wherein the ventilation is required to be carried out once a day in the room, and the air is kept fresh every 30-60 minutes; the indoor ventilation time is increased to two to three times per day as the fruiting body differentiates and grows.
6.3 Humidity: and keeping the relative humidity of the air between 80 and 90 percent.
6.4 Illumination: the natural scattered light irradiates for about 12 hours in the daytime.
Usually, according to the above management conditions, after about 10-20 days, the height of the fruiting body can reach more than 7 cm, and the fruiting body can be harvested when there appear small red-orange spots on the upper part of the stroma, i.e. the shell of the seed capsule.
Seventh harvest management
7.1 Timing of harvesting
When the height of the stroma is as high as about 7-9 cm, yellow protrusions appear on the upper part, and a plurality of small thorns grow on the top end, the whole stroma is orange red and does not grow for a long time, which indicates that the stroma is mature, and then the stroma can be harvested.
The harvest is too early, the nutrient accumulation does not reach the highest point, and the yield is influenced; after late harvest, the ascospores begin to emit, consuming nutrients and reducing the effective ingredients.
7.2 Harvesting method
During collection, the cordyceps militaris is collected integrally by using sterile tweezers, and then the collected cordyceps militaris is placed together in order, preferably directly on a clean drying sieve, so that the cordyceps militaris can be dried or aired in time.
Note that: and thousands of people do not expose to the sun to prevent the color fading of the fruiting body.
7.3 Drying process
The vacuum freeze drying is to dry the product by utilizing the principle that water in the product is directly sublimated into steam from solid ice under the condition of quick freezing and vacuum environment and is pumped away by a vacuum pump, and the product can be basically recovered to the state before drying after rehydration.
The dried artificial cordyceps stroma has the weight of 1/6 of the fresh weight, darker color than the fresh product and orange color.
Eighth strip of packaging and storage
8.1 Packaging requirements
(1) The packaging material has good pressure resistance, moisture resistance and sealing property; the necessary moisture protection should be taken in the package to prevent moisture ingress.
(2) The packaging material should meet the sanitary requirements and should not cause degeneration, deterioration, discoloration, or flavor change of Cordyceps.
(3) The packing bag should be marked with the registration, quantity, processing date of the cordyceps sinensis and the mark of 'damp-proof and rain-proof'.
8.2 Packaging method
The storage in a short time is generally made of polyethylene or polypropylene plastic films into small bags, the small bags are bagged and sealed, and then the small bags are packaged into a carton every 10 bags.
When the storage time is long, the glass paper bag and the composite film packaging bag can be used; the characteristics of light tightness, oxygen impermeability and moisture impermeability of the aluminum foil in the film bag are met, so that the dry artificial cordyceps product is stored under the anoxic condition, and the storage effect is further improved.
8.3 Storage of
(1) Low temperature storage
Under the condition of low temperature, the activity of biological enzyme is reduced, and the activity of pathogenic bacteria is inhibited, so that the artificial cordyceps sinensis is stored at low temperature and is stored under the refrigeration condition of 0-4 ℃, the storage period can be prolonged, and the aging can be prevented.
(2) Ventilated storage
The product is stored in a ventilation storage warehouse with good air fluidity or placed in a ventilation shade place, relatively small air humidity is kept, and the storage time limit can be prolonged; some drying and dehumidifying agent can be put into the sealed storage container to prevent mildew.
Detailed Description
The preparation method comprises the following steps: the preparation of strains → the preparation of culture medium → bottling → sterilization → cooling → inoculation → spawn running management → color changing management → grass growing management → harvesting → processing;
first line of bacterial preparation
1.1 Production of mother seeds
The process flow for preparing the artificial cordyceps mother seeds comprises the following steps: preparation of culture medium → selection and collection of seed source of artificial cordyceps → tissue isolation (spore isolation) → purification culture → mother seed. The preparation method comprises the following steps:
1.1.1 culture medium formula:
150-250 g of potato, 15-25 g of glucose, 1-5 g of monopotassium phosphate, 1-2 g of magnesium sulfate, 15-25 g of agar, 1:4 tablets of Vb and 800-1200 ml of water.
1.1.2 The preparation method comprises the following steps:
(1) Weighing: accurately weighing various substances according to the proportion of the various substances in the formula.
(2) Extracting juice: the preparation method comprises boiling the above materials in water, and maintaining with slow fire for 15-20 min; then filtered through gauze, leaving the juice. Filtering and then making up the water content to 800-1200 ml.
(3) Dissolution of other substances: adding agar into the filtrate, heating to melt agar, stirring continuously during melting, and adding other nutrients to dissolve.
(4) Subpackaging: subpackaging the prepared culture medium into test tubes when the culture medium is hot; during subpackaging, the loading amount is preferably 1/5-1/4 of the height of the test tube; the culture medium is not adhered to the tube opening during subpackage, so as to avoid pollution.
(5) Tampon and package
After the culture medium is subpackaged, a cotton plug is plugged at the opening of the test tube; after the cotton plugs are plugged, every 7 test tubes are bundled, and a layer of kraft paper is wrapped outside the test tubes for fastening.
(6) And (3) sterilization: the mother culture medium is generally sterilized by autoclaving for 25-35 minutes; the test tube culture medium is vertically placed in a wide-mouth bottle or an iron frame during sterilization, then is put in a pressure cooker for sterilization, and is naturally cooled after 25-35 minutes.
(7) Cooling and swinging the inclined plane: after sterilization, naturally reducing the pressure to zero, opening the pressure cooker, taking out the test tube, and placing the test tube on an inclined plane when the test tube is hot; the length of the inclined plane is 1/2-2/3 of that of the test tube; after solidification, the culture medium is a slant culture medium.
1.1.3 Strain isolation
Tissue isolation:
(1) Cutting a small piece of tissue with the length of 0.1-0.2 cm from the surface-sterilized and sterile-cleaned cordyceps militaris stroma under the aseptic condition, and connecting the tissue piece to the center of a culture medium by using a sterile inoculating needle.
(2) The inoculated medium was placed at 22 ℃ and cultured in the dark.
(3) And (4) about 15 days, when new hyphae extend on the culture medium, selecting the hyphae which are free of pollution and strong in growth, and storing the hyphae in a refrigerator for later use.
1.2 Preparation of liquid spawn
1.2.1 The formula of the culture medium is as follows:
30 g of glucose, 20 g of peptone, 0.5 g of monopotassium phosphate, 2.5 g of salt and 1000 ml of water.
1.2.2 Bottling: the prepared liquid culture medium is respectively filled into wide-mouth bottles, each bottle is 3500-4500 ml, a few drops of edible oil are dropped into the bottle bottles to play a defoaming role, a polypropylene film is used for sealing and tightening, and sterilization is carried out.
1.2.3 And (3) sterilization: and placing the wide-mouth bottle filled with the liquid culture medium into an autoclave for sterilization for 35-45 minutes.
1.2.4 Inoculation: placing the sterilized culture medium, mother seeds and inoculation tools on a super clean workbench or in an inoculation box, and then sterilizing the inoculation chamber, the super clean workbench or the inoculation box; when inoculating, inoculating the mother strain into liquid culture medium by aseptic technique, inoculating 1-2 pieces of Cordyceps militaris mother strain into a bottle of culture medium; each parent seed is 2 times 2 cm in size, i.e. 4 cm square.
1.2.5 Culturing: placing in dark environment after inoculation, standing for 2-3 days to recover hypha growth, and then performing shake culture by introducing oxygen; shaking for 2-3 days to finish the preparation of liquid strain; the prepared liquid strain is stored in a refrigerator, the temperature of the refrigerator is kept at 2-5 ℃ for standby use, and the liquid strain is used up in 10-20 days as much as possible.
Preparing the second culture medium and bottling
2.1 Silkworm chrysalis culture medium
The silkworm pupa is selected from high-quality silkworm pupas according to the selection standard of no impurity, no mildew, no worm damage, no peculiar smell, normal color, uniform size, length of 2-4 cm and thickness of 8-10 mm. Air drying fresh pupa Bombycis.
2.2 Bottling: bottling in 400-600 ml wide-mouth bottle, each bottle contains 25-35 g silkworm chrysalis, and the mouth is wrapped with polypropylene film.
The method for binding comprises the following steps: and (4) tying the bottle mouth twice by using a rubber band to ensure that the bottle mouth is tightly tied.
Third sterilization and inoculation
3.1 And (3) sterilization: maintaining the pressure of 0.15 MPa for 1 hour or maintaining the temperature of 100 ℃ at normal pressure for 10 hours; when sterilizing, the water in the pot is sufficiently added, the bottles are compactly placed, and the pot is taken out after naturally reducing the pressure.
And (4) directly placing the material bottle into an inoculation room after sterilization, cooling, and cooling to normal temperature.
3.2 Inoculation:
3.2.1 disinfection:
the inoculation chamber (box) needs to be thoroughly disinfected by formaldehyde, the dosage of the formaldehyde is 10 ml and 5 g potassium permanganate in each cubic meter of space, then the door and the window are closed and closed for 30 minutes, and simultaneously the ultraviolet lamp is turned on for irradiation disinfection; when in use, the culture medium, the inoculation tool, the alcohol lamp, the alcohol cotton ball, the lighter and the like which are sterilized and cooled are put into an inoculation chamber (box) and the inoculation chamber is turned on to be irradiated and sterilized by the ultraviolet lamp for 30 minutes.
3.2.2 liquid spawn treatment:
the liquid strain cultured by shaking can be directly used for inoculation; but for the condition that the concentration of the bacteria balls is large (the concentration of the bacteria balls is more than 50 percent) and the culture production amount is large, water is needed to be added into the liquid bacteria for dilution; the concentration of the bacteria balls reaches about 50 percent, and the method is suitable for production inoculation.
3.2.3 inoculation:
directly inoculating liquid strains into a solid culture medium in an aseptic state; directly injecting the mixture on the surface of the solid medium in a spraying mode.
The inoculation operation process is carried out in an inoculation chamber (box), and the inoculated culture chamber is moved into a culture chamber; after inoculation, vertically placing the bottle in a dark environment for culturing to ensure that hyphae germinate, and then placing the bottle on a shelf for culturing after field planting; otherwise, the bacteria solution or nutrient solution and culture medium will fall to one side, resulting in uneven grass growth; the culture chamber is sterilized before use, and is used after ventilation.
Fourth spawn running period management
After hyphae germinate, the hyphae can be put on a culture frame to enter the spawn running period for management; the spawn running period is easily polluted by mixed bacteria, so closed culture is required.
4.1 temperature: the culture temperature of the mycelium can not be lower than 17 ℃ at least and can not be higher than 24 ℃ at most; preferably, incubation is carried out at constant temperature.
4.2 Humidity: the relative humidity of air is required to be kept to be 65% -70% in the spawn running period, and the air can be properly sprayed on the indoor ground or in the air for humidification.
4.3 Ventilating: the workshop is properly ventilated during the hypha growth stage.
If the culture chamber is heated by a stove, attention needs to be paid to discharge coal gas, namely carbon monoxide, in time; because carbon monoxide has a certain inhibition effect on the growth of hyphae.
4.4 Illumination: the culture should be protected from light at this stage.
4.5 Daily management:
4.5.1 examining the bacteria flasks: the fungus bottles need to be checked frequently, hyphae grow slowly, reasons need to be found in time so as to take appropriate measures to remedy, and polluted fungi need to be treated, so that the loss is reduced to the lowest point.
4.5.2 constant temperature culture: the mycelium culture stage is preferably carried out at constant temperature, and the temperature is not higher or lower.
About 15 days later, the next stage of management can be carried out.
Fifth color cycle management
5.1 Ventilating: the indoor ventilation is required to be carried out once a day, 35-45 minutes each time, and the air is kept fresh.
5.2 Humidity: the control is about 75 percent.
5.3 Illumination: scattered light irradiation is required for 12 to 14 hours per day.
5.4 Temperature: the temperature in the grass-growing period is controlled between 20 ℃ and 25 ℃.
Sixth management of emergence period
6.1 Temperature: the temperature of the primordium, namely the prototypical bulge growing on the surface of the culture medium is controlled to be 18-22 ℃ before the occurrence; the temperature may be suitably raised after the primordia have appeared, and the temperature does not exceed 28 ℃.
6.2 Ventilating: ventilating after the primordium appears, wherein the ventilation is required to be carried out once a day in the room, and the air is kept fresh every 30-60 minutes; the indoor ventilation time is increased to two to three times per day as the fruiting body differentiates and grows.
6.3 Humidity: and keeping the relative humidity of the air between 80 and 90 percent.
6.4 Illumination: the natural scattered light irradiates for about 12 hours in the daytime.
Generally, according to the management conditions, after about 10-20 days, the height of the sporocarp can reach more than 7 cm, and small orange-red dots, namely the seed capsule shells, appear on the upper part of the stroma.
Seventh harvest management
7.1 Timing of harvesting
When the height of the stroma is as high as about 7-9 cm, yellow protrusions appear on the upper part, and a plurality of small thorns grow on the top end, the whole stroma is orange red and does not grow for a long time, which indicates that the stroma is mature, and then the stroma can be harvested.
The harvest is too early, the nutrient accumulation does not reach the highest point, and the yield is influenced; after late harvest, the ascospores begin to emit, consuming nutrients and reducing the effective ingredients.
7.2 Harvesting method
During collection, the cordyceps militaris is collected integrally by using sterile tweezers, and then the collected cordyceps militaris is placed together in order, preferably directly on a clean drying sieve, so that the cordyceps militaris can be dried or aired in time.
Note that: and thousands of people do not expose to the sun to prevent the color fading of the fruiting body.
7.3 Dry processing
The vacuum freeze drying is to dry the product by utilizing the principle that water in the product is directly sublimated into steam from solid ice in a quick freezing and vacuum environment and is pumped away by a vacuum pump, and the product can basically recover to the state before drying after rehydration.
The dried artificial cordyceps stroma has the weight of 1/6 of the fresh weight, darker color than the fresh product and orange color.
Eighth strip of packaging and storage
8.1 Packaging requirements
(1) The packaging material has good pressure resistance, moisture resistance and sealing property; the necessary moisture protection should be taken in the package to prevent moisture ingress.
(2) The packaging material should meet the sanitary requirements and should not cause degeneration, deterioration, discoloration and taste deterioration of Cordyceps.
(3) The packing bag should be marked with the registration, quantity, processing date of the cordyceps sinensis and the mark of 'damp-proof and rain-proof'.
8.2 Packaging method
The storage in a short time is generally made of polyethylene or polypropylene plastic films into small bags, the small bags are bagged and sealed, and then the small bags are packaged into a carton every 10 bags.
When the storage time is long, the glass paper bag and the composite film packaging bag can be used; the characteristics of light tightness, oxygen impermeability and moisture impermeability of the aluminum foil in the film bag are met, so that the dry artificial cordyceps product is stored under the anoxic condition, and the storage effect is further improved.
8.3 Storage of
(1) Low temperature storage
Under the condition of low temperature, the activity of biological enzyme is reduced, and the activity of pathogenic bacteria is inhibited, so that the artificial cordyceps sinensis is stored at low temperature and is stored under the refrigeration condition of 0-4 ℃, the storage period can be prolonged, and the aging can be prevented.
(2) Ventilated storage
The product is stored in a ventilation storage warehouse with good air fluidity or placed in a ventilation shade place, relatively small air humidity is kept, and the storage time limit can be prolonged; some drying and dehumidifying agent can be put into the sealed storage container to prevent mildew.
Claims (1)
1. The artificial cordyceps militaris and the preparation method are characterized in that: the artificial cordyceps militaris is an insect and fungus complex formed after cordyceps militaris of insects is infected by cordyceps militaris, and the preparation method comprises the following steps: preparing strains → preparing culture medium → bottling → sterilizing → cooling → inoculating → managing spawn running → color changing managing → managing grass → harvesting → processing;
first line of bacterial preparation
1.1 Production of mother seeds
The process flow for preparing the artificial cordyceps mother seeds comprises the following steps: preparation of culture medium → selection and collection of seed source of artificial cordyceps → tissue separation (spore isolation) → purification culture → mother seed, the preparation method is as follows:
1.1.1 culture medium formula:
150-250 g of potato, 15-25 g of glucose, 1-5 g of monopotassium phosphate, 1-2 g of magnesium sulfate, 15-25 g of agar, 1:4 tablets of Vb and 800-1200 ml of water;
1.1.2 The preparation method comprises the following steps:
(1) Weighing: accurately weighing various substances according to the proportion of the various substances in the formula;
(2) Extracting juice: the preparation method comprises boiling the above materials in water, and maintaining with slow fire for 15-20 min; then filtered through gauze to leave the juice. Filtering and then supplementing the water to 800-1200 ml;
(3) Dissolve in other substances: adding agar into the filtrate, heating to melt the agar, stirring continuously during melting, and adding other nutrients to dissolve after completely melting;
(4) Subpackaging: subpackaging the prepared culture medium into test tubes when the culture medium is hot; when subpackaging, the loading amount is preferably 1/5-1/4 of the height of the test tube; the culture medium is not required to be adhered to the pipe orifice during subpackaging so as to avoid pollution;
(5) Tampon and package
After the culture medium is subpackaged, plugging a tampon on a test tube opening; after the cotton plugs are plugged, bundling 7 test tubes into a bundle, and wrapping a layer of kraft paper outside the test tubes for fastening;
(6) And (3) sterilization: the mother culture medium is generally sterilized by autoclaving for 25-35 minutes; vertically placing the test tube culture medium in a wide-mouth bottle or an iron frame during sterilization, then placing the wide-mouth bottle or the iron frame into a pressure cooker for sterilization, and naturally cooling after 25-35 minutes;
(7) Cooling and swinging the inclined plane: after sterilization, naturally reducing the pressure to zero, opening the pressure cooker, taking out the test tube, and placing the test tube on an inclined plane when the test tube is hot; the length of the inclined plane is 1/2-2/3 of that of the test tube; after solidification, the culture medium is a slant culture medium;
1.1.3 Strain isolation
Tissue isolation:
(1) Cutting a small piece of cordyceps militaris stroma which is subjected to surface sterilization and sterile cleaning under the sterile condition, wherein the small piece of tissue is 0.1-0.2 cm, and taking a tissue block by using a sterile inoculating needle to connect the tissue block at the center of a culture medium;
(2) Placing the inoculated culture medium at 22 ℃, and culturing under a dark condition;
(3) About 15 days, when new hyphae extend on the culture medium, selecting the hyphae which are pollution-free and strong in growth, and storing the hyphae in a refrigerator for later use;
1.2 Preparation of liquid Strain
1.2.1 The formula of the culture medium is as follows:
30 g of glucose, 20 g of peptone, 0.5 g of monopotassium phosphate, 2.5 g of salt and 1000 ml of water;
1.2.2 And (4) bottling: respectively filling the prepared liquid culture medium into wide-mouth bottles, wherein each bottle is 3500-4500 ml, a few drops of edible oil are dripped into the bottle bottles to play a defoaming role, and sealing and tightening by using a polypropylene film for sterilization;
1.2.3 And (3) sterilization: placing the wide-mouth bottle filled with the liquid culture medium into an autoclave for sterilization for 35-45 minutes;
1.2.4 Inoculation: placing the sterilized culture medium, mother seeds and inoculation tools on a super clean workbench or in an inoculation box, and then sterilizing the inoculation chamber, the super clean workbench or the inoculation box; when inoculating, inoculating the mother strain into liquid culture medium by aseptic technique, inoculating 1-2 pieces of Cordyceps militaris mother strain into a bottle of culture medium; each mother seed is 2 multiplied by 2 cm, namely 4 square centimeters;
1.2.5 Culturing: placing in dark environment after inoculation, standing for 2-3 days to recover hypha growth, and then performing shake culture by introducing oxygen; shaking for 2-3 days to finish the preparation of liquid strain; storing the prepared liquid strain in a refrigerator, wherein the temperature of the refrigerator is kept at 2-5 ℃ for standby use, and the liquid strain is used up in 10-20 days as much as possible;
preparing the second culture medium and bottling
2.1 Silkworm chrysalis culture medium
Selecting high-quality silkworm pupas according to the selection standard, wherein the selection standard is that no impurity, no mildew, no worm damage, no peculiar smell, normal color, uniform size, length of 2-4 cm and thickness of 8-10 mm are contained; air-drying fresh silkworm pupa;
2.2 Bottling: bottling in 400-600 ml wide-mouth bottle, wherein each bottle contains 25-35 g silkworm chrysalis, and the opening is wrapped with polypropylene film;
the buckling method comprises the following steps: two times of tying are carried out at the bottle mouth by using a rubber band to ensure that the bottle mouth is tightly tied;
third sterilization and inoculation
3.1 And (3) sterilization: maintaining the pressure at 0.15 MPa for 1 hour or at 100 deg.c for 10 hours; when in sterilization, the water in the pot is sufficiently added, the bottles are compactly placed, and the bottles are taken out of the pot after natural pressure reduction;
directly placing the material bottle into an inoculation room for cooling after sterilization, and cooling to normal temperature;
3.2 Inoculation:
3.2.1 disinfection:
the inoculation chamber (box) needs to be thoroughly disinfected by formaldehyde, the dosage of the formaldehyde is 10 ml and 5 g potassium permanganate in each cubic meter of space, then the door and the window are closed and closed for 30 minutes, and simultaneously the ultraviolet lamp is turned on for irradiation disinfection; when in use, the culture medium, the inoculation tool, the alcohol lamp, the alcohol cotton ball, the lighter and the like which are sterilized and cooled are put into an inoculation chamber (box), and the inoculation chamber is started to be irradiated by the ultraviolet lamp for disinfection for 30 minutes;
3.2.2 liquid spawn treatment:
the liquid strain cultured by shaking can be directly used for inoculation; but for the condition that the concentration of the bacteria balls is large (the concentration of the bacteria balls is more than 50 percent) and the culture production amount is large, water is needed to be added into the liquid bacteria for dilution; the concentration of the bacteria balls reaches about 50 percent, and the method is suitable for production inoculation;
3.2.3 inoculation:
directly inoculating liquid strains into a solid culture medium in an aseptic state; directly injecting the mixture on the surface of a solid culture medium in a spraying mode;
the inoculation operation process is carried out in an inoculation chamber (box), and the inoculated culture chamber is moved into a culture chamber; after inoculation, vertically placing the bottle in a dark environment for culturing to ensure that hyphae germinate, and then placing the bottle on a shelf for culturing after field planting; otherwise, the bacteria liquid or nutrient solution and culture medium will fall to one side, resulting in uneven grass; the culture room is sterilized before use and is used after ventilation;
fourth spawn running period management
After hyphae germinate, putting the hyphae on a culture frame for management in a spawn running period; the spawn running period is easily polluted by mixed bacteria, so closed culture is required;
4.1 temperature: the culture temperature of the mycelium can not be lower than 17 ℃ at least and can not be higher than 24 ℃ at most; preferably, constant temperature culture is carried out;
4.2 Humidity: the relative humidity of air is required to be kept to be 65% -70% in the spawn running period, and the air can be properly sprayed on the indoor ground or in the air for humidification;
4.3 Ventilating: ventilating the workshop properly in the hypha growth stage;
if the culture chamber is heated by a stove, attention needs to be paid to discharge coal gas, namely carbon monoxide, in time; carbon monoxide has certain inhibition effect on the growth of hyphae;
4.4 Illumination: culturing in dark place;
4.5 Daily management:
4.5.1 examining the bacteria vial: checking the fungus bottles frequently, finding that hyphae grow slowly, searching for reasons in time so as to take appropriate measures to remedy, and treating polluted bacteria to minimize loss;
4.5.2 constant temperature culture: the mycelium culture stage is preferably carried out at constant temperature, and the temperature is forbidden to be high or low;
about 15 days later, the next stage of management can be carried out;
fifth color cycle management
5.1 Ventilating: ventilating for 35-45 min every day to keep air fresh;
5.2 Humidity: controlling the concentration to be about 75%;
5.3 Illumination: the irradiation of scattered light is required for 12-14 hours each day;
5.4 Temperature: the temperature in the grass growing period is controlled between 20 ℃ and 25 ℃;
sixth management of emergence period
6.1 Temperature: the temperature of the primordium, namely the prototypical bulge growing on the surface of the culture medium is controlled to be 18-22 ℃ before the occurrence; the temperature can be properly raised after the primordium appears, and the temperature does not exceed 28 ℃;
6.2 Ventilating: ventilating after the primordium appears, wherein the ventilation is required to be carried out once a day in the room, and the air is kept fresh every 30-60 minutes; increasing the indoor ventilation time to two to three times per day along with the differentiation and growth of the sporocarp;
6.3 Humidity: keeping the relative humidity of air at 80% -90%;
6.4 Illumination: naturally scattering light in the daytime for about 12 hours;
generally, according to the management conditions, after about 10-20 days, the height of the sporocarp can reach more than 7 cm, and small orange-red dots appear on the upper part of the stroma, namely the seed capsule shell can be harvested;
seventh harvest management
7.1 Timing of harvesting
When the stroma is as high as about 7-9 cm, yellow protrusions appear on the upper part, and a plurality of small thorns grow on the top end, the whole stroma is orange and does not grow for a long time, which indicates that the stroma is mature, and then the stroma can be harvested;
the harvest is too early, the nutrient accumulation does not reach the highest point, and the yield is influenced; after late harvest, the ascospores begin to emit, so that nutrition is consumed and effective ingredients are reduced;
7.2 Harvesting method
During collection, the cordyceps militaris is collected integrally by using sterile tweezers, and then the collected cordyceps militaris is placed together in order, preferably directly on a clean drying sieve, so as to be dried or aired in time;
note that: thousands of people do not need to be exposed to the sun to prevent the color of the fruiting body from fading;
7.3 Drying process
The vacuum freeze drying is to dry the product by utilizing the principle that water in the product is directly sublimated into steam from solid ice in a quick-freezing and vacuum environment and is pumped away by a vacuum pump, and the product can basically recover to the state before drying after rehydration;
the weight of the dried artificial cordyceps stroma is 1/6 of the fresh weight, the color is darker than that of the fresh product, and the artificial cordyceps stroma is orange yellow;
eighth strip of packaging and storage
8.1 Packaging requirements
(1) The packaging material has good pressure resistance, moisture resistance and sealing property; necessary moisture-proof measures are adopted in the packaging box to prevent moisture from entering;
(2) The packaging material is required to meet the sanitary requirement and cannot cause the degeneration, the deterioration, the color change and the taste change of the cordyceps sinensis;
(3) The packaging bag should be marked with the registration, quantity, processing date and the mark of 'damp-proof and rainproof';
8.2 Packaging method
Storing for a short time, making into small bags with polyethylene or polypropylene plastic film, bagging, sealing, and packaging into a carton every 10 bags;
when the storage time is long, the glass paper bag and the composite film packaging bag can be used; the characteristics of light tightness, oxygen impermeability and moisture impermeability of the aluminum foil in the film bag are met, so that the dry artificial cordyceps product is stored under the anoxic condition, and the storage effect is further improved;
8.3 Storage of
(1) Low temperature storage
Under the condition of low temperature, the activity of the biological enzyme is reduced, and the activity of pathogenic bacteria is inhibited, so that the artificial cordyceps sinensis is stored at low temperature and is stored under the refrigeration condition of 0-4 ℃, the storage period can be prolonged, and the aging can be prevented;
(2) Ventilated storage
The product is stored in a ventilation storage warehouse with good air fluidity or placed in a ventilation shade place, relatively small air humidity is kept, and the storage time limit can be prolonged; some drying and dehumidifying agent can be put into the sealed storage container to prevent mildew.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211223208.0A CN115553179A (en) | 2022-10-08 | 2022-10-08 | Artificial cordyceps militaris and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211223208.0A CN115553179A (en) | 2022-10-08 | 2022-10-08 | Artificial cordyceps militaris and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115553179A true CN115553179A (en) | 2023-01-03 |
Family
ID=84745224
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211223208.0A Pending CN115553179A (en) | 2022-10-08 | 2022-10-08 | Artificial cordyceps militaris and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115553179A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101720627A (en) * | 2009-12-16 | 2010-06-09 | 江苏江南生物科技有限公司 | Method for culturing cordyceps militaris by living silkworm chrysalises |
| CN107810781A (en) * | 2017-11-10 | 2018-03-20 | 广西沙田仙人滩农业投资有限公司 | A kind of method that silkworm living cultivates Cordyceps militaris |
| WO2022088280A1 (en) * | 2020-10-27 | 2022-05-05 | 苏州家和蚕业生物科技有限公司 | Cultivating method for silkworm pupa-cultured cordyceps militaris |
-
2022
- 2022-10-08 CN CN202211223208.0A patent/CN115553179A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101720627A (en) * | 2009-12-16 | 2010-06-09 | 江苏江南生物科技有限公司 | Method for culturing cordyceps militaris by living silkworm chrysalises |
| CN107810781A (en) * | 2017-11-10 | 2018-03-20 | 广西沙田仙人滩农业投资有限公司 | A kind of method that silkworm living cultivates Cordyceps militaris |
| WO2022088280A1 (en) * | 2020-10-27 | 2022-05-05 | 苏州家和蚕业生物科技有限公司 | Cultivating method for silkworm pupa-cultured cordyceps militaris |
Non-Patent Citations (3)
| Title |
|---|
| 徐秀华编著: "北虫草栽培新技术", 28 February 2010, 华中科技大学出版社, pages: 49 - 78 * |
| 曾文虎等: "蛹虫草高产栽培技术", 蛹虫草高产栽培技术, no. 13, 14 July 2014 (2014-07-14), pages 106 - 108 * |
| 李昊编著: "虫草人工栽培技术", 31 May 2001, 金盾出版社, pages: 36 - 39 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102668878B (en) | Method for cultivating flammulina velutipes by using aquilaria-sinensis-chip edible mushroom culture medium | |
| CN107125023B (en) | Edible fungus culture medium and edible fungus culture method using same | |
| CN109392592B (en) | Phellinus igniarius cultivation method | |
| CN103416225A (en) | Method for preparing high-quality ganoderma lucidum compost | |
| CN110367039A (en) | A kind of production method reducing Neurospora infection Dictyophora rubrovalvata cultivar | |
| CN1939115A (en) | Cultivation of brown crab-flavor mushroom | |
| CN108633614A (en) | A kind of greenhouse cultivation method of oyster mushroom | |
| CN107396730A (en) | A kind of method that rhizoma Gastrodiae is cultivated using tea branch | |
| KR20200012367A (en) | How to grow flower mushrooms | |
| CN1939116A (en) | Cultivation of white crab-flavor mushroom | |
| CN111527991A (en) | Process for making edible reed fungus sticks | |
| CN108207484A (en) | A kind of breeding method of silkworm chrysalis cordyceps sinensis | |
| CN115553179A (en) | Artificial cordyceps militaris and preparation method thereof | |
| CN109122024A (en) | A kind of method of the efficient bag cultivating of Pleurotus eryngii | |
| CN115152537A (en) | Method for manufacturing Armillaria mellea substitute bacteria stick | |
| CN109874598A (en) | A kind of standardized planting method of red sesame | |
| CN108812063A (en) | A method of cultivation of agaricus bisporus kind is made using synthetic substrate | |
| CN104737775A (en) | Organic pleurotus eryngii production process | |
| CN113079946A (en) | Edible fungus cultivation method and culture medium | |
| CN112931046A (en) | Bag-cultivated mushroom compost formula | |
| CN106665316A (en) | Organic stereoscopic tank for efficiently planting bell peppers and method for planting same in organic stereoscopic tank | |
| CN111480511A (en) | Cultivation and production process of fresh reed fungi | |
| CN113940231A (en) | High-yield cultivation technology and management method for oyster mushrooms | |
| CN110999718A (en) | Planting method of pleurotus geesteranus | |
| KR20160087512A (en) | Cultivating method of tree ear and the composition of cultur medium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |