CN112840947A - Method for cultivating cordyceps sinensis by taking hawthorn as main matrix - Google Patents
Method for cultivating cordyceps sinensis by taking hawthorn as main matrix Download PDFInfo
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- 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
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- 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
Abstract
A method for culturing Cordyceps with fructus crataegi as main matrix comprises inoculating multi-stage domesticated Cordyceps strain into culture medium composed of fructus crataegi, fructus crataegi branches and leaves and Oryza Glutinosa as main raw materials, and culturing high-quality Cordyceps fruiting body suitable for good growth in fructus crataegi matrix. Through the implementation of the invention, the hawthorn fruit residues, the crushed hawthorn branches and leaves and the glutinous rice form the culture medium with rich and comprehensive nutrition, cordyceps mycelia grow rapidly and robustly under the nutrition, cordyceps fruiting body individuals also grow robustly and biotransformation rate is high. By implementing the method, the cordyceps sinensis test tube strain, the cordyceps sinensis liquid strain and the procedure of cultivating cordyceps sinensis sporocarp are trained and domesticated in several stages, and the domestication is mainly carried out by gradually increasing the content of the hawthorn in a culture medium and gradually reducing the pH value, so that cordyceps sinensis hyphae are gradually adapted to be completely adapted to grow in the hawthorn culture medium, and finally, the high-quality and high-yield cordyceps sinensis sporocarp is grown in the hawthorn culture medium.
Description
Technical Field
The invention belongs to the technical field of edible fungus cultivation, and particularly relates to a method for cultivating cordyceps sinensis by taking hawthorn as a main matrix.
Background
The hawthorn fruit is sour and sweet and tasty, can generate body fluid to quench thirst, can be used as a medicine, enters spleen, stomach and liver meridians, has the effects of promoting digestion to eliminate stagnation, activating blood circulation and dissipating blood stasis, and can be eaten by 76 percent.
According to the determination, the hawthorn contains 18 kinds of amino acids, particularly the content of vitamin C is higher, which reaches 89mg/100 g, 17 times higher than that of apples, and 30 times higher than that of pears. In addition, per 100g contains 397kJ of energy, 73g of water, 0.5g of protein, 0.6g of fat, 3.1g of dietary fiber, 22g of carbohydrate, 100. mu.g of carotene, 17. mu.g of vitamin A, 0.02mg of thiamine, 0.02mg of riboflavin, 0.4mg of nicotinic acid, 53mg of vitamin C, 7.32mg of vitamin E, 299mg of potassium, 5.4mg of sodium, 52mg of calcium, 19mg of magnesium, 0.9mg of iron, 0.24mg of manganese, 0.28mg of zinc, 0.11mg of copper, 24mg of phosphorus and 1.22. mu.g of selenium, bioactive substances and nutritional ingredients such as maslinic acid, flavone and SOD, and components having inhibitory effects on Escherichia coli, Pseudomonas aeruginosa and Bacillus dysenteriae. The pectin content in hawthorn is the first of all fruits and reaches 6.4%, and according to the latest research, pectin has the function of a radiation-proof substance and can take away half of radioactive elements (strontium, cobalt, palladium and the like) from the body. Pectin also has adsorptive and antibacterial properties, and can remove bacteria and toxins from the intestine and bind water, thereby treating diarrhea. The compounds such as vitexin contained in fructus crataegi have anticancer effect, and fructus crataegi is useful for preventing cancer. The hawthorn also contains abundant calcium and carotene, the calcium content is first of fruits, the carotene content is second to that of the Chinese dates and the Chinese gooseberries, and the hawthorn is most suitable for children to eat. The hawthorn can be singly used or combined with other foods and medicines to improve the food therapy effect. The hawthorn contains various organic acids, so that the vitamin C in the hawthorn can be kept, and the hawthorn cannot be damaged even under the condition of heating, so that the vitamin C can be still preserved after the hawthorn is prepared into products such as hawthorn cakes and the like. The hawthorn is also rich in triterpenoid olefine acids such as carotene, calcium, oleanolic acid, ornithine, crataetin and the like and beneficial components such as flavonoids (a plurality of chemical components such as flavonoid polyflavan, trimeric flavan, tannin and the like), can relax blood vessels, strengthen and regulate cardiac muscle, increase the amplitude of ventricular and cardiac motion and the blood flow of coronary artery, reduce serum cholesterol and lower blood pressure; in addition, fructus crataegi has adjuvant treatment effect on heart activity dysfunction, vascular neurosis, and fibrillation arrhythmia; the hawthorn is an ideal health food for preventing and treating cardiovascular diseases and a food with better curative effect, and can be used for preventing and treating hypertension, hyperlipidemia, coronary heart disease and the like.
The hawthorn has important medicinal value, and becomes a good medicine for tonifying spleen, stimulating appetite, promoting digestion, removing food stagnation, promoting blood circulation and reducing phlegm since ancient times. The hawthorn fruit is used as a medicine, has mild nature and sour and sweet taste, enters spleen, stomach and liver channels, and has the functions of promoting digestion, invigorating stomach, promoting blood circulation, removing blood stasis, astringing and stopping dysentery. It can be used for treating phlegm retention, abdominal distention, acid regurgitation, dysentery, intestinal wind, lumbago, hernia, puerperal infantile occipital pain, lochiorrhea, and infantile dyspepsia.
In addition, the yield of the hawthorn leaves and the yield of the fruits are equal, and one hawthorn tree strain capable of producing 100 jin of hawthorn fruits can also produce 100 jin of hawthorn leaves. Therefore, the raw material resources of the hawthorn leaves are also very rich. At present, the hawthorn leaves are only burned as firewood except for being naturally rotten when falling to the ground, and the hawthorn leaves are not effectively developed and utilized. Authors such as Lixiaoling, Zhang Xiaomin and the like at the institute of Life sciences and technology of Shanxi university in 1 month in 2005 published the article "nutrition and development value of Hawthorn leaf" in Shanxi forestry. The nutrient components and the effects of the hawthorn leaves are explained in detail, and the main contents are as follows:
the amino acid composition of the hawthorn leaves is as follows: the hawthorn leaves contain 17 kinds of amino acids, wherein the content of aspartic acid, glutamic acid, alanine, cystine and isoleucine is far higher than that of hawthorn fruits, and the total content of various amino acids is 36.38% higher than that of the hawthorn leaves. Different from the same hawthorn leaf sample, the content of amino acid is approximately in the following sequence: grain > light, preserved fruit, cheese > group, egg > cysts. The total of 20 amino acids constituting human proteins, 8 of which are not synthesized in the human body and must be supplied from food, are called essential amino acids. The hawthorn fruit contains 17 amino acids, which comprise 8 amino acids necessary for human body. Generally, the content of amino acids in the leaves is higher, the young leaves are higher than the mature leaves, and the newly collected leaves are higher than those stored for a longer time. Amino acids are essential nutrients for the composition of human and animal body proteins and for maintaining normal metabolism, and have important physiological effects.
② trace elements of hawthorn leaves: mr. Sun Zhang has determined 25 kinds of elements in haw leaf and haw fruit, and the results show that Ca, Mg, K and P contained in haw leaf and Fe and A1 contained in haw leaf are constant and the rest elements are trace. The content of most elements in the hawthorn leaves is higher than that of hawthorn fruits. Be. The contents of Bi, Pb, Cd, As and Hg are all very small. The contents of residual harmful components of the hawthorn leaves, namely hexachloro cyclohexane, DDT, arsenic, copper and lead are all lower than the national food sanitation standard value, and the hawthorn leaves are harmless to human bodies. Recently, the death rate of 60 types of cardiovascular diseases is analyzed and discovered that sr, Ca, Mg, Li, Si and the like can reduce the death rate of the diseases, the action mechanism of the elements possibly competes with Na in intestines to absorb sites, so that the absorption of Na is reduced and the excretion of Na is increased, and excessive Na in vivo is related to the cardiovascular diseases such as hypertension. Therefore, the hawthorn fruits and the hawthorn leaves have therapeutic effects on cardiovascular diseases such as hypertension, coronary heart disease, angina and the like, and can be related to high contents of Sr, Ca, Mg and K and low contents of Na besides flavonoid components. The hawthorn leaves contain a certain amount of Mn (45.5 mg/kg). Manganese deficiency has been reported to be associated with arteriosclerosis. Manganese can improve lipid metabolism of atherosclerosis patients, and has effect of removing lipid. Therefore, the traditional Chinese medicine composition is probably one of the reasons for reducing blood fat and preventing and treating atherosclerosis of hawthorn fruits and hawthorn leaves. The hawthorn leaves contain considerable amounts of P (2150mg/kg), Fe (420mg/kg) and Zn (682 mg/kg), which are all required by human bodies, so that the content of beneficial elements in the hawthorn leaves is higher than or close to that in the hawthorn fruits, in addition, the total content of amino acid is also higher than that of the hawthorn leaves (2.40 percent) by 10.30 percent and simultaneously contains various flavonoid components, and the hawthorn leaves have obvious curative effects on cardiovascular diseases.
③ Vc content of hawthorn leaves: the Vc content of hawthorn leaves in the mature period is 820 mg-1224 mg/1009, which is about 10 times of that of fruits (89 mg/1009). The Vc content of the leaves before the leaves fall from 5 months to 11 months tends to rise, and the Vc content of the brown leaves falling to the ground is extremely low. The Vc content is highest at the beginning of 11 months, and at the moment, the Vc content is the harvest season of the hawthorn fruits, so that the growth and the yield of the hawthorn fruits are not influenced, and leaves can be harvested after the fruits are harvested. Vitamin c promotes collagen synthesis. Collagen is an important protein constituting connective tissue, thereby accelerating the healing of wounds and preventing bleeding from small blood vessels. At the same time, Vc promotes cortisone synthesis, iron absorption and antibody production. Vc is easy to combine with nitrosamine, a carcinogen, and has an anticancer effect. In the absence of Vc, capillary vessel embrittlement and permeability increase are caused, which causes subcutaneous, mucosal, muscular, gingival bleeding, fracture and the like.
VB of hawthorn leaveslAnd VB2The content is as follows: folium crataegi containing VBl(0.04mg/100g)、VB2(0.08mg/100g) and nicotinic acid (0.06 mg/1009). VB1Is an important part for maintaining the normal activity of the nervous system and is also one of the coenzymes of sugar metabolism. Lack of VB1Susceptible to beriberi, heavy lower limbs, numb skin of hands and feet, accelerated heartbeat, enlarged heart, heart failure and other diseases. VB2Can promote wound and infection recovery, is an essential component for maintaining eye health, and plays a role of coenzyme in oxidation. VB2In the absence, the main symptoms are cheilitis, glossitis, angular stomatitis, keratitis, scrotitis, etc. Nicotinic acid supplies energy during injury, fever, vomiting, diarrhea, bowel disease and pregnancy while helping the skin maintain normal function and activate energy in many biological reactions.
Fifthly, the content of the flavone in the hawthorn leaves is as follows: the content of total flavone in the hawthorn leaves is 3.18-9.17%, wherein the content of vitexin is 8.705 mg/g. Vitexin is one of effective natural bioactive components in flavonoids compounds, and has good anticancer effect. The folium crataegi contains various flavonoids such as rutin, hyperoside, quercetin, vitexin, etc. Pharmacological and clinical experiments show that: the hawthorn leaf flavonoid compound has the effects of reducing blood pressure, reducing blood fat, increasing coronary flow, improving myocardial blood and oxygen supply and the like for cardiovascular diseases, and has good curative effects on the cardiovascular diseases such as hypertension, hyperlipidemia, coronary heart disease, angina pectoris and the like. In addition, the hawthorn leaf flavonoid compound has higher antioxidant and anti-free radical activity, and is an ideal additive for producing natural health-care food. The hawthorn leaves in autumn not only have the highest flavone content, but also have low water content, and are convenient to store. The hawthorn leaves are picked preferably after the hawthorn fruits are picked in autumn, so that the normal growth of the hawthorn trees and the yield of the hawthorn fruits are not influenced. The folium crataegi contains much flavone compounds, and its content is similar to that of folium Eucalypti Robustae (E.macrocorhyncha) and Eucalyptus (E.yomani), and the extraction of flavone compounds from these leaves has been industrialized. The hawthorn leaf flavone has good pharmacological action and is widely applied to clinical treatment, so the hawthorn leaf can be used as a promising natural medicine extraction raw material and has production value.
In conclusion, the whole body of the hawthorn is treasure, the hawthorn fruits and the hawthorn branches and leaves are fully utilized and processed into products with higher added values, and the significance is very important.
Disclosure of Invention
The invention provides a brand-new method for culturing cordyceps sinensis strains by taking hawthorn as a matrix based on the abundant components of hawthorn fruits and hawthorn branches and leaves and fully and effectively utilizing the hawthorn fruits and the hawthorn branches and leaves. The method specifically comprises the following steps: a culture medium which is mainly composed of hawthorn fruits, hawthorn branches and leaves and glutinous rice is inoculated with multi-stage domesticated cordyceps sinensis strains to culture high-quality cordyceps sinensis fruiting bodies which are suitable for good growth in hawthorn substrates.
The invention is realized by the following technical scheme:
a method for culturing cordyceps sinensis by taking hawthorn as a main matrix adopts the technical scheme that the method comprises the following steps:
1. the culture medium comprises the following components in percentage by weight: 50-60 parts of hawthorn fruit residues, 15-20 parts of hawthorn branch and leaf crushed materials, 15-25 parts of glutinous rice and 3-5 parts of silkworm chrysalis powder.
2. Raw material treatment: mixing the above fructus crataegi fruit residue, pulverized fructus crataegi branches and leaves, Oryza Glutinosa and pupa Bombycis, stirring, and adding nutrient solution to maintain water content of the mixture at 50-60% to obtain culture medium.
3. And (3) sterilizing a culture medium: and placing the culture medium in a sterilization container for sterilization and disinfection.
4. Inoculation: inoculating multi-stage domesticated Cordyceps strain to culture medium under sterile environment.
5. And (3) bacteria cultivation: culturing the culture medium inoculated with Cordyceps strain in dark environment at 15-20 deg.C and humidity of 60-75% until the culture medium is full of mycelia.
6. Color conversion: culturing the culture medium with mycelia at 20-25 deg.C under 70-80% humidity in light until the mycelia turn orange.
7. Culturing cordyceps sinensis fruiting bodies: placing the medium which is changed into orange color at 20-25 deg.C, humidity of 80-90% and illumination environment, and culturing until Cordyceps fruiting body grows.
Further, the preparation method of the hawthorn fruit residues comprises the following steps:
(1) collecting raw materials: collecting fresh hawthorn fruits with moderate maturity and good quality as raw materials;
(2) raw material treatment: picking out insect-resistant fruits, rotten fruits and impurities, and cleaning the hawthorn fruits;
(3) boiling: putting the hawthorn fruits into a pot, adding clear water to submerge the hawthorn fruits, and then boiling the hawthorn fruits until the hawthorn fruits are soft and rotten;
(4) separation: breaking the hawthorn fruits to remove kernels and pedicles;
(5) pulping: beating the pulp into jam;
(6) separating dregs and juice: squeezing the jam to obtain juice to obtain fructus crataegi pomace.
Further, the preparation method of the hawthorn branch and leaf crushed material comprises the following steps:
(1) collecting raw materials: collecting thoroughly dried and moldless hawthorn branches and leaves as raw materials;
(2) raw material treatment: pulverizing the branches and leaves of fructus crataegi, and sieving with 50-60 mesh sieve to obtain fructus crataegi branch and leaf powder.
Further, the preparation method of the nutrient solution comprises the following steps:
(1) the raw materials comprise the following components in percentage by weight: 1000 ml of fresh hawthorn juice, 20-30 g of glucose, 2-3 g of monopotassium phosphate, 1-2 g of magnesium sulfate and 5-6 g of peptone;
(2) raw material treatment: putting the hawthorn juice, glucose, potassium dihydrogen phosphate, magnesium sulfate and peptone into a heating container, and then heating until all materials are completely dissolved in water to obtain the nutrient solution.
Further, the sterilization method comprises the following steps:
(1) and (3) high-pressure sterilization: sterilizing in a pressure sterilizing container at 121 deg.C under 0.15 MPa for 30-50 min;
(2) normal pressure sterilization: the mixture is sterilized in a normal pressure sterilization container at 100 ℃ for 240-300 minutes.
Further, the multi-stage domestication method of the cordyceps sinensis strains comprises the following steps:
(1) first stage test tube strain cultivation: preparing 850-900 ml of clear water, 100-150 ml of fresh hawthorn juice, 20-30 g of polygonatum odoratum powder, 20-30 g of corn flour, 20-30 g of glucose and 15-20 g of agar powder, putting the above materials into a heating container together, heating until the materials are completely dissolved in water to obtain a matrix solution, adjusting the pH value of the matrix solution to 7.5-8.5, then subpackaging the matrix solution into test tubes, sterilizing and placing a slant to obtain a slant test tube, inoculating commercially available cordyceps militaris strains into the slant test tube in an aseptic environment, culturing the cordyceps militaris strains in a dark light environment at 15-20 ℃ until the cordyceps militaris strains are full of tubes, and placing the test tube full of cordyceps militaris strains in a dark light environment at 20-25 ℃ until the cordyceps militaris strains are turned orange to obtain first-stage test tube strains;
(2) the first stage liquid strain cultivation: preparing 800 ml of clear water of 750 plus materials, 250 ml of fresh hawthorn juice of 200 plus materials, 50-60 g of rhizoma polygonati odorati powder, 20-30 g of glucose and 5-6 g of peptone, putting the above materials into a heating container, heating until the materials are completely dissolved in water to obtain a nutrient solution, adjusting the pH of the nutrient solution to 6.5-7.5, subpackaging the nutrient solution into a liquid strain culture container, sterilizing to obtain a liquid strain culture medium, inoculating the first-stage test tube strain obtained by culturing in the step (1) into the liquid strain culture medium in an aseptic environment, and finally carrying out submerged fermentation culture in a dark environment at 15-20 ℃ until the liquid strain culture medium is full of mycelia or hyphae to obtain a first-stage liquid strain;
(3) first stage sporophore cultivation: preparing 700 ml of clean water of 650 plus materials, 350 ml of fresh hawthorn juice of 300 plus materials, 50-60 g of rhizoma polygonati odorati powder, 20-30 g of glucose and 5-6 g of peptone, putting the above materials into a heating container together, heating until the materials are completely dissolved in water to obtain a nutrient solution, and adjusting the pH value of the nutrient solution to 6-7 for later use; taking 50-70 parts of glutinous rice, 15-20 parts of hawthorn fruit residues, 10-25 parts of hawthorn branch and leaf crushed materials and 3-5 parts of silkworm chrysalis powder, mixing the glutinous rice, the hawthorn fruit residues, the hawthorn branch and leaf crushed materials and the silkworm chrysalis powder to obtain a mixed material, mixing a nutrient solution into the mixed material, adjusting the water content of the mixed material to 55-60% to obtain a culture medium, subpackaging the culture medium into culture containers and sterilizing, inoculating the first-stage liquid strain obtained by the culture in the step (2) into the culture medium in an aseptic environment, placing the inoculated culture medium in a dark environment at a temperature of 15-20 ℃, a humidity of 70-75% until cordyceps mycelia overgrow with a culture medium, placing the culture medium full of cordyceps mycelia in a dark environment at a temperature of 20-25 ℃, a humidity of 80-90% and a light environment until the cordyceps is turned to orange, finally culturing at 20-25 deg.C and humidity of 80-90% under visible light environment until Cordyceps fruiting body grows;
(4) and (3) second-stage test tube strain cultivation: 600 ml of clear water with 550 ml, 450 ml of fresh hawthorn juice with 400 ml of waste water, 20-30 g of polygonatum odoratum powder, 20-30 g of glucose, 2-3 g of monopotassium phosphate, 1-2 g of magnesium sulfate, 5-6 g of peptone and 15-20 g of agar powder, putting the above materials into a heating container together, heating until various materials are completely dissolved in water to obtain matrix solution, adjusting pH of the matrix solution to 5.5-6, subpackaging in test tubes, sterilizing, placing on slant to obtain slant test tube, separating the first stage Cordyceps militaris fruiting body obtained by culturing in step (3) into slant test tubes under aseptic environment, culturing at 15-20 deg.C in dark light until the Cordyceps mycelia overgrow the tube, and culturing in 20-25 deg.C in light environment until the Cordyceps mycelia turn orange to obtain second stage test tube strain;
(5) and (3) liquid strain cultivation in the second stage: 500 ml of clear water of 450-;
(6) second stage sporocarp cultivation: 400 ml of clear water of 350-; taking 50-70 parts of hawthorn fruit residues, 20-25 parts of hawthorn branch and leaf crushed materials, 10-20 parts of glutinous rice and 3-5 parts of silkworm chrysalis powder, mixing the hawthorn fruit residues, the hawthorn branch and leaf crushed materials, the glutinous rice and the silkworm chrysalis powder to obtain a mixed material, mixing a nutrient solution into the mixed material, adjusting the water content of the mixed material to 55-60% to obtain a culture medium, subpackaging the culture medium into culture containers and sterilizing, inoculating the second-stage liquid strain obtained by the culture in the step (5) into the culture medium in an aseptic environment, placing the inoculated culture medium in a dark environment at the temperature of 15-20 ℃, the humidity of 70-75% until cordyceps mycelia overgrow with a culture medium, placing the culture medium full of cordyceps mycelia in a dark environment at the temperature of 20-25 ℃, the humidity of 80-90% and a light environment until cordyceps is turned to orange, finally culturing at 20-25 deg.C and humidity of 80-90% under visible light environment until Cordyceps fruiting body grows;
(7) third-stage test tube strain cultivation: 700 ml of fresh hawthorn juice with 650 plus materials, 350 ml of clear water with 300 plus materials, 20-30 g of rhizoma polygonati odorati powder, 20-30 g of glucose, 2-3 g of monopotassium phosphate, 1-2 g of magnesium sulfate, 5-6 g of peptone and 15-20 g of agar powder, putting the above materials into a heating container together, then heating until various materials are completely dissolved in water to obtain matrix solution, then subpackaging the matrix solution in test tubes, sterilizing the test tubes, placing the test tubes on an inclined plane to obtain inclined test tubes, separating the fruiting body of the second stage Cordyceps militaris obtained by culturing in step (6) into slant test tubes under aseptic environment, culturing at 15-20 deg.C in dark light until the cordyceps mycelia overgrow the tube, and culturing in a test tube at 20-25 deg.C in light environment until the cordyceps mycelia turn orange to obtain third stage test tube strain.
(8) Liquid strain cultivation in the third stage: 850-900 ml of fresh hawthorn juice, 100-150 ml of clear water, 50-60 g of polygonatum odoratum powder, 20-30 g of glucose and 5-6 g of peptone, all the materials are put into a heating container, then the heating container is heated until all the materials are completely dissolved in water to obtain nutrient solution, the nutrient solution is respectively filled into a liquid strain culture container and sterilized to obtain a liquid strain culture medium, the third-stage test tube strain obtained by the culture in the step (7) is inoculated into the liquid strain culture medium in an aseptic environment, and finally the third-stage liquid strain is obtained by deep fermentation culture under the environment of 15-20 ℃ and dark light until the liquid strain culture medium is full of bacteria balls or hyphae.
Further, the illumination environment is given 100-200lux illumination.
Further, the illumination environment is given 300-600lux illumination.
The Cordyceps militaris is rich in cordycepin, cordycepic acid, polysaccharide and other substances. The traditional Chinese medicine considers that the cordyceps militaris can tonify both lung yin and kidney yang, is mainly used for treating kidney deficiency, impotence and spermatorrhea, waist and knee soreness, weakness after illness, chronic cough and weakness, cough with phlegm and blood caused by overexertion, spontaneous perspiration and night sweat and the like, and is a traditional Chinese medicine capable of balancing and regulating yin and yang simultaneously. The health-care functional components of the cordyceps militaris not only contain cordycepin, but also contain cordyceps polysaccharide, which is a human body immunopotentiator recognized by international medicine. The traditional Chinese medicine considers that the traditional Chinese medicine has the effects of strengthening body resistance and consolidating constitution, has obvious curative effects on senile chronic bronchitis and pulmonary heart disease, can improve the detoxifying capability of the liver, has the effect of protecting the liver, and improves the antiviral and anti-radiation capability of the body. Cordycepic acid, D-mannitol, is a basic medicine for treating cardiovascular and cerebrovascular diseases, and has effects of scavenging free radicals, dilating blood vessel, and lowering blood pressure. The nucleotide has effects of inhibiting blood platelet aggregation, preventing heart and brain thrombosis, eliminating chloasma, senile plaque and acne, resisting aging and wrinkle, caring skin, etc. Cordyceps militaris is a kind of ascomycete, and sexual reproduction is carried out by using the heterothallic cooperation. The non-sexual type is paecilomyces chrysalis. The mature sporocarp can form ascospore (propagation unit), the spore spreads with wind, the spore falls on proper insect body, and starts to germinate to form mycelium. The mycelium continuously develops and begins to spread into the pupa body, so that the pupa body can be infected by fungi, the tissue in the pupa body is decomposed, the nutrition in the pupa body is used as a material and an energy source for the growth and development of the pupa body, and finally, the interior of the pupa body is completely decomposed. When the mycelium of Cordyceps militaris decomposes various tissues and organs in pupa, the development of mycelium also enters a new stage. Form rod-like stroma (fruit body) with orange or red-orange top part slightly enlarged. The cordyceps militaris can also be bred into a complete stroma under artificial conditions, the culture medium for cultivating the cordyceps militaris is not limited to insect bodies, and the most widely used culture medium for artificially cultivating the cordyceps militaris at present is rice, wheat, corn and the like.
The glutinous rice is rich in protein, fat, saccharides, calcium, phosphorus, iron, vitamin B1, vitamin B2, nicotinic acid, starch and the like, is rich in nutrition, is a warm-tonifying and body-strengthening food, and has the effects of tonifying middle-jiao and Qi, strengthening spleen and nourishing stomach, and stopping sweating due to debility.
The invention has the following advantages:
1. the method for cultivating the cordyceps sinensis by taking the hawthorn as the matrix has novel conception, simple production process and easy implementation.
2. Through the implementation of the invention, the culture medium consisting of hawthorn fruit residues, hawthorn branch and leaf crushed materials and sticky rice is used, hawthorn contains 18 kinds of amino acids, multiple vitamins, multiple trace elements such as potassium, sodium, calcium, magnesium, selenium and the like, and bioactive substances such as maslinic acid, flavone, SOD and the like; the hawthorn leaves contain 17 amino acids, wherein the content of aspartic acid, glutamic acid, alanine, cystine and isoleucine is far higher than that of hawthorn fruits, the total content of various amino acids is 36.38% of the hawthorn leaves compared with the hawthorn fruits, Ca, Mg, K and P contained in the hawthorn leaves and Fe and A1 contained in the hawthorn leaves belong to constants, the content of most elements in the hawthorn leaves is higher than that of the hawthorn fruits, the hawthorn leaves contain various flavonoid compounds such as rutin, hyperin, quercetin, vitexin and the like, the content of the total flavonoids is 3.18-9.17%, the content of the vitexin is 8.705Mg/g, and the vitexin is one of effective natural bioactive components in the flavonoid compounds and has a good anticancer effect; the glutinous rice is rich in protein, fat, sugar, calcium, phosphorus, iron, vitamin B1, vitamin B2, nicotinic acid, starch and the like, the three substances form a culture medium with rich and comprehensive nutrition, cordyceps mycelia grow rapidly and robustly under the nutrition, cordyceps fruiting body individuals also grow robustly, and the biotransformation rate is high.
3. Because hawthorn fruits are sour, the pH of the hawthorn fruits is about 3-4, and cordyceps sinensis strains are difficult to grow on a hawthorn culture medium. By implementing the method, the cordyceps sinensis test tube strain, the cordyceps sinensis liquid strain and the procedure of cultivating cordyceps sinensis sporocarp are trained and domesticated in several stages, and the domestication is mainly carried out by gradually increasing the content of the hawthorn in a culture medium and gradually reducing the pH value, so that cordyceps sinensis hyphae are gradually adapted to be completely adapted to grow in the hawthorn culture medium, and finally, the high-quality and high-yield cordyceps sinensis sporocarp is grown in the hawthorn culture medium.
4. The hawthorn fruit and branches and leaves have high fiber content, and the hawthorn fruit contains a large amount of tannin and flavonoid substances, so the hawthorn fruit has bitter taste. By implementing the invention, the cordyceps mycelia germinate and grow in the hawthorn matrix, and the cordyceps mycelia can effectively decompose and differentiate hawthorn, so that the hawthorn becomes small molecules, and the hawthorn product is more beneficial to human body absorption after people eat the hawthorn product. The bitter and astringent taste of hawthorn is also decomposed. In addition, heavy metal and pesticide residues in hawthorn fruits and branches and leaves can be removed through fermentation, which is different from the era of drug trisection.
5. By implementing the invention, the current situation that deep-processed hawthorn products are rare in the current market is changed, the postpartum way of hawthorn is solved, the development of related industries of hawthorn fruits is driven, a new economic growth point is formed, and great economic benefits and social benefits are achieved.
Detailed Description
The process of the present invention is further illustrated below with reference to examples.
A method for culturing cordyceps sinensis by taking hawthorn as a main matrix adopts the technical scheme that the method comprises the following steps: a culture medium which is mainly composed of hawthorn fruits, hawthorn branches and leaves and glutinous rice is inoculated with multi-stage domesticated cordyceps sinensis strains to culture high-quality cordyceps sinensis fruiting bodies which are suitable for good growth in hawthorn substrates.
The specific implementation mode is as follows:
1. collecting high-quality test-tube cordyceps militaris strains sold in the market as a strain source.
2. The preparation method of the hawthorn fruit residue and the hawthorn fruit juice comprises the following steps:
(1) collecting raw materials: collecting fresh hawthorn fruits with moderate maturity and good quality as raw materials;
(2) raw material treatment: picking out insect-resistant fruits, rotten fruits and impurities, and cleaning the hawthorn fruits;
(3) boiling: putting the hawthorn fruits into a pot, adding clear water to submerge the hawthorn fruits, and then boiling the hawthorn fruits until the hawthorn fruits are soft and rotten;
(4) pulping: beating the pulp into jam;
(5) separating juice and residue: squeezing the jam according to conventional method to obtain fruit residue and fruit juice.
3. The preparation method of the crushed hawthorn branches and leaves comprises the following steps:
(1) collecting raw materials: collecting thoroughly dried and moldless hawthorn branches and leaves as raw materials;
(2) raw material treatment: pulverizing branches and leaves of fructus crataegi to obtain 50-60 mesh powder.
4. Collecting glutinous rice without insect pest and mildew as raw material.
5. The preparation method of the polygonatum powder comprises the following steps:
(1) collecting raw materials: collecting dried and moldy-free polygonatum odoratum as a raw material;
(2) raw material treatment: pulverizing rhizoma Polygonati Odorati to obtain rhizoma Polygonati Odorati powder of 120 meshes.
6. The preparation method of the corn flour comprises the following steps:
(1) collecting raw materials: collecting the dried and mildewless corns as a raw material;
(2) raw material treatment: pulverizing semen Maydis to obtain semen Maydis powder with 120 mesh sieve.
7. The preparation method of the strain comprises
(1) First stage test tube strain cultivation: preparing 900 ml of clear water, 100 ml of fresh hawthorn juice, 30 g of polygonatum powder, 20 g of glucose, 3g of potassium dihydrogen phosphate, 1g of magnesium sulfate, 5g of peptone and 16 g of agar powder, putting the above materials into a heating container, heating until all the materials are completely dissolved in water to obtain a matrix solution, adjusting the pH of the matrix solution to 8.5, subpackaging the matrix solution into test tubes, putting the test tubes into a pressure sterilization container, sterilizing at 121 ℃ and 0.15 MPa for 30 minutes, taking out the test tubes, placing the test tubes on a slope to obtain a slope test tube, inoculating cordyceps militaris strains into the slope test tube in a sterile environment, culturing at 15-20 ℃ in a dark light environment until the cordyceps militaris hypha is full of the tube, placing the test tube full of cordyceps militaris hypha at 20-25 ℃, and culturing under 200lux light environment until the cordyceps militaris turns to orange yellow to obtain first-stage test tube strains.
(2) The first stage liquid strain cultivation: preparing 800 ml of clear water, 200 ml of fresh hawthorn juice, 60 g of polygonatum odoratum powder, 20 g of glucose and 5g of peptone, putting all the materials into a heating container, heating until all the materials are completely dissolved in water to obtain a nutrient solution, adjusting the pH of the nutrient solution to 7.5, then subpackaging the nutrient solution into a liquid strain culture container, sterilizing at 121 ℃ and 0.15 MPa for 30 minutes to obtain a liquid strain culture medium, inoculating the first-stage test tube strain obtained by culturing in the step 1 into the liquid strain culture medium in an aseptic environment, and finally carrying out submerged fermentation culture in a dark light environment at 15-20 ℃ until the liquid strain culture medium is full of bacteria balls or hyphae to obtain a first-stage liquid strain.
(3) First stage sporophore cultivation: preparing 700 ml of clear water, 300 ml of fresh hawthorn juice, 60 g of polygonatum powder, 30 g of glucose and 6g of peptone, putting the above materials into a heating container, heating until the materials are completely dissolved in water to obtain a nutrient solution, and adjusting the pH value of the nutrient solution to 6.5 for later use; taking 60 parts of glutinous rice, 20 parts of hawthorn fruit residues, 15 parts of hawthorn branch and leaf crushed materials and 5 parts of silkworm chrysalis powder, mixing the glutinous rice, the hawthorn fruit residues, the hawthorn branch and leaf crushed materials and the silkworm chrysalis powder to obtain a mixed material, mixing a nutrient solution into the mixed material, adjusting the water content of the mixed material to 55-60% to obtain a culture medium, subpackaging the culture medium into culture containers, placing the culture medium in a normal-pressure sterilization container, sterilizing at 100 ℃ for 300 minutes, inoculating the first-stage liquid strain obtained in the step 2 in an aseptic environment into the culture medium, placing the inoculated culture medium in a dark light environment at the temperature of 15-20 ℃, the humidity of 70-75% to culture the cordyceps mycelia until the cordyceps mycelia overgrow with the culture medium, placing the culture medium overgrowing with the cordyceps mycelia at the temperature of 20-25 ℃, the humidity of 80-90%, and culturing the cordyceps mycelia in a 300lux light environment until the cordyceps mycelia turns orange, and finally culturing at 20-25 deg.C and humidity of 80-90% under 500lux illumination until Cordyceps fruiting body grows.
(4) And (3) second-stage test tube strain cultivation: putting the above materials into a heating container together with clear water of 600 ml, fresh hawthorn juice of 400 ml, polygonatum powder of 25 g, glucose of 25 g, potassium dihydrogen phosphate of 2.5 g, magnesium sulfate of 1.5 g, peptone of 5.5 g and agar powder of 16 g, then heating until the materials are completely dissolved in water to obtain a matrix solution, adjusting the pH of the matrix solution to 6, then packaging the matrix solution into test tubes, sterilizing the test tubes, placing the test tubes on a slant to obtain slant test tubes, separating the first stage cordyceps militaris cultured in the step 3 into the slant test tubes under an aseptic environment, culturing the test tubes at 15-20 ℃ under a dark light environment until cordyceps mycelia are full of the tubes, finally placing the test tubes full of cordyceps mycelia at 20-25 ℃ and culturing under a 200lux light environment until the cordyceps mycelia are turned to orange yellow to obtain second stage test tube strains.
(5) And (3) liquid strain cultivation in the second stage: 500 ml of clear water, 500 ml of fresh hawthorn juice, 50 g of polygonatum powder, 20 g of glucose and 5g of peptone are put into a heating container together, then the materials are heated until the materials are completely dissolved in the water to obtain nutrient solution, the pH value of the nutrient solution is adjusted to 5.5, the nutrient solution is subpackaged into liquid strain culture containers, the liquid strain culture medium is obtained by sterilizing the nutrient solution at 121 ℃ and 0.15 MPa for 30 minutes, the second-stage test tube strain obtained by culturing in the step 4 is inoculated into the liquid strain culture medium in an aseptic environment, and finally, the second-stage liquid strain is obtained by deep fermentation culture in a dark light environment at 15-20 ℃ until the liquid strain culture medium is full of bacteria balls or hyphae.
(6) Second stage sporocarp cultivation: putting 400 ml of clear water, 600 ml of fresh hawthorn juice, 60 g of polygonatum powder, 30 g of glucose and 6g of peptone into a heating container, heating until the materials are completely dissolved in water to obtain a nutrient solution, and adjusting the pH value of the nutrient solution to 5 for later use; taking 65 parts of hawthorn fruit residues, 20 parts of hawthorn branch and leaf crushed materials, 10 parts of glutinous rice and 5 parts of silkworm chrysalis powder, mixing the hawthorn fruit residues, the hawthorn branch and leaf crushed materials, the glutinous rice and the silkworm chrysalis powder to obtain a mixed material, mixing a nutrient solution into the mixed material, adjusting the water content of the mixed material to 55-60% to obtain a culture medium, subpackaging the culture medium into culture containers, placing the culture medium into a normal-pressure sterilization container, sterilizing at 100 ℃ for 300 minutes, inoculating the second-stage liquid strain obtained by the culture in the step 5 into the culture medium in an aseptic environment, placing the inoculated culture medium in a dark light environment at the temperature of 15-20 ℃, the humidity of 70-75% to culture the mycelia to overgrow the cordyceps sinensis culture medium, placing the culture medium full of cordyceps sinensis mycelia at the temperature of 20-25 ℃, the humidity of 80-90%, and culturing the cordyceps sinensis in a 300lux light environment until the cordyceps sinensis turns orange, and finally culturing at 20-25 deg.C and humidity of 80-90% under 600lux illumination until Cordyceps fruiting body grows.
(7) Third-stage test tube strain cultivation: 700 ml of fresh hawthorn juice, 300 ml of clear water, 30 g of polygonatum powder, 30 g of glucose, 3g of potassium dihydrogen phosphate, 2g of magnesium sulfate, 6g of peptone and 18 g of agar powder, putting the above materials into a heating container, heating until all the materials are completely dissolved in water to obtain a matrix solution, then subpackaging the matrix solution into test tubes, sterilizing the test tubes, placing the test tubes on an inclined plane to obtain inclined test tubes, separating the second-stage cordyceps militaris sporocarp obtained by culturing in the step 6 into the inclined plane test tubes under an aseptic environment, culturing under a dark light environment at 15-20 ℃ until cordyceps mycelium is full of the tubes, and finally placing the test tubes full of cordyceps mycelium at 20-25 ℃ and culturing under a 200lux light environment until the cordyceps mycelium is turned into orange yellow to obtain a third-stage test tube strain.
(8) Liquid strain cultivation in the third stage: 850-900 ml of fresh hawthorn juice, 100-150 ml of clear water, 50-60 g of polygonatum odoratum powder, 20-30 g of glucose and 5-6 g of peptone, all the materials are put into a heating container, then the heating container is heated until all the materials are completely dissolved in water to obtain nutrient solution, the nutrient solution is respectively filled into a liquid strain culture container and sterilized to obtain a liquid strain culture medium, the third-stage test tube strain obtained by the culture in the step (7) is inoculated into the liquid strain culture medium in an aseptic environment, and finally the third-stage liquid strain is obtained by deep fermentation culture under the environment of 15-20 ℃ and dark light until the liquid strain culture medium is full of bacteria balls or hyphae.
8. The method for cultivating the cordyceps sinensis fruiting bodies comprises the following steps:
(1) the culture medium comprises the following components in percentage by weight: 60 parts of hawthorn fruit residues, 20 parts of hawthorn branch and leaf crushed materials, 15 parts of glutinous rice and 5 parts of silkworm chrysalis powder.
(2) Raw material treatment: mixing the above fructus crataegi fruit residue, pulverized fructus crataegi branches and leaves, Oryza Glutinosa and pupa Bombycis, stirring, and adding nutrient solution to maintain water content of the mixture at 50-60% to obtain culture medium.
(3) And (3) sterilizing a culture medium: the medium was placed in a normal pressure sterilization vessel and sterilized at 100 ℃ for 300 minutes to sterilize.
(4) Inoculation: inoculating the third stage liquid strain into the culture medium under sterile condition.
(5) And (3) bacteria cultivation: culturing the culture medium inoculated with Cordyceps strain in dark environment at 15-20 deg.C and humidity of 60-75% until the culture medium is full of mycelia.
(6) Color conversion: placing the culture medium full of mycelia at 20-25 deg.C and humidity of 70-80%, and culturing under 200lux illumination until the mycelia turn orange.
(7) Culturing cordyceps sinensis fruiting bodies: placing the medium which is changed into orange color at 20-25 deg.C and humidity of 80-90%, and culturing under 600lux illumination until the Cordyceps fruiting body grows.
As described above, it will be apparent to those skilled in the art that other various modifications can be made based on the technical solution and the technical idea of the present invention, and all such modifications shall fall within the scope of the appended claims.
Claims (8)
1. A method for culturing cordyceps sinensis by taking hawthorn as a main matrix is characterized by comprising the following steps:
(1) the culture medium comprises the following components in percentage by weight: 50-60 parts of hawthorn fruit residues, 15-20 parts of hawthorn branch and leaf crushed materials, 15-25 parts of glutinous rice and 3-5 parts of silkworm chrysalis powder;
(2) raw material treatment: mixing the above fructus crataegi fruit residue, pulverized fructus crataegi branches and leaves, Oryza Glutinosa and pupa Bombycis, stirring, and adding nutrient solution to maintain water content of the mixture at 50-60% to obtain culture medium;
(3) and (3) sterilizing a culture medium: placing the culture medium in a sterilization container for sterilization and disinfection;
(4) inoculation: inoculating multi-stage domesticated Cordyceps strain into culture medium under sterile environment;
(5) and (3) bacteria cultivation: culturing the culture medium inoculated with Cordyceps strain in dark environment at 15-20 deg.C and humidity of 60-75% until the culture medium is full of mycelia;
(6) color conversion: placing the culture medium full of mycelia in 20-25 deg.C, humidity of 70-80% and light environment, and culturing until the mycelia turns orange;
(7) culturing cordyceps sinensis fruiting bodies: placing the medium which is changed into orange color at 20-25 deg.C, humidity of 80-90% and illumination environment, and culturing until Cordyceps fruiting body grows.
2. The method for cultivating cordyceps sinensis by taking hawthorn as a main matrix according to claim 1, wherein the preparation method of hawthorn pomace in the step (1) is as follows:
(1) collecting raw materials: collecting fresh hawthorn fruits with moderate maturity and good quality as raw materials;
(2) raw material treatment: picking out insect-resistant fruits, rotten fruits and impurities, and cleaning the hawthorn fruits;
(3) boiling: putting the hawthorn fruits into a pot, adding clear water to submerge the hawthorn fruits, and then boiling the hawthorn fruits until the hawthorn fruits are soft and rotten;
(4) separation: breaking the hawthorn fruits to remove kernels and pedicles;
(5) pulping: beating the pulp into jam;
(6) separating dregs and juice: squeezing the jam to obtain juice to obtain fructus crataegi pomace.
3. The method for cultivating cordyceps sinensis by taking hawthorn as a main matrix according to claim 1, wherein the preparation method of the hawthorn branch and leaf crushed material in the step (1) is as follows:
(1) collecting raw materials: collecting thoroughly dried and moldless hawthorn branches and leaves as raw materials;
(2) raw material treatment: pulverizing the branches and leaves of fructus crataegi, and sieving with 50-60 mesh sieve to obtain fructus crataegi branch and leaf powder.
4. The method for cultivating cordyceps sinensis by taking hawthorn as a main matrix according to claim 1, wherein the preparation method of the nutrient solution in the step (2) is as follows:
(1) the raw materials comprise the following components in percentage by weight: 1000 ml of fresh hawthorn juice, 20-30 g of glucose, 2-3 g of monopotassium phosphate, 1-2 g of magnesium sulfate and 5-6 g of peptone;
(2) raw material treatment: putting the hawthorn juice, glucose, potassium dihydrogen phosphate, magnesium sulfate and peptone into a heating container, and then heating until all materials are completely dissolved in water to obtain the nutrient solution.
5. The method for cultivating cordyceps sinensis by taking hawthorn as a main substrate according to claim 1, wherein the sterilization method in the step (3) is as follows:
(1) and (3) high-pressure sterilization: sterilizing in a pressure sterilizing container at 121 deg.C under 0.15 MPa for 30-50 min;
(2) normal pressure sterilization: the mixture is sterilized in a normal pressure sterilization container at 100 ℃ for 240-300 minutes.
6. The method for cultivating cordyceps sinensis by taking hawthorn as a main substrate according to claim 1, wherein the domestication method of the multistage domesticated cordyceps sinensis strain in the step (4) is as follows:
(1) first stage test tube strain cultivation: preparing 850-900 ml of clear water, 100-150 ml of fresh hawthorn juice, 20-30 g of polygonatum odoratum powder, 20-30 g of corn flour, 20-30 g of glucose and 15-20 g of agar powder, putting the above materials into a heating container together, heating until the materials are completely dissolved in water to obtain a matrix solution, adjusting the pH value of the matrix solution to 7.5-8.5, then subpackaging the matrix solution into test tubes, sterilizing and placing a slant to obtain a slant test tube, inoculating commercially available cordyceps militaris strains into the slant test tube in an aseptic environment, culturing the cordyceps militaris strains in a dark light environment at 15-20 ℃ until the cordyceps militaris strains are full of tubes, and placing the test tube full of cordyceps militaris strains in a dark light environment at 20-25 ℃ until the cordyceps militaris strains are turned orange to obtain first-stage test tube strains;
(2) the first stage liquid strain cultivation: preparing 800 ml of clear water of 750 plus materials, 250 ml of fresh hawthorn juice of 200 plus materials, 50-60 g of rhizoma polygonati odorati powder, 20-30 g of glucose and 5-6 g of peptone, putting the above materials into a heating container, heating until the materials are completely dissolved in water to obtain a nutrient solution, adjusting the pH of the nutrient solution to 6.5-7.5, subpackaging the nutrient solution into a liquid strain culture container, sterilizing to obtain a liquid strain culture medium, inoculating the first-stage test tube strain obtained by culturing in the step (1) into the liquid strain culture medium in an aseptic environment, and finally carrying out submerged fermentation culture in a dark environment at 15-20 ℃ until the liquid strain culture medium is full of mycelia or hyphae to obtain a first-stage liquid strain;
(3) first stage sporophore cultivation: preparing 700 ml of clean water of 650 plus materials, 350 ml of fresh hawthorn juice of 300 plus materials, 50-60 g of rhizoma polygonati odorati powder, 20-30 g of glucose and 5-6 g of peptone, putting the above materials into a heating container together, heating until the materials are completely dissolved in water to obtain a nutrient solution, and adjusting the pH value of the nutrient solution to 6-7 for later use; taking 50-70 parts of glutinous rice, 15-20 parts of hawthorn fruit residues, 10-25 parts of hawthorn branch and leaf crushed materials and 3-5 parts of silkworm chrysalis powder, mixing the glutinous rice, the hawthorn fruit residues, the hawthorn branch and leaf crushed materials and the silkworm chrysalis powder to obtain a mixed material, mixing a nutrient solution into the mixed material, adjusting the water content of the mixed material to 55-60% to obtain a culture medium, subpackaging the culture medium into culture containers and sterilizing, inoculating the first-stage liquid strain obtained by the culture in the step (2) into the culture medium in an aseptic environment, placing the inoculated culture medium in a dark environment at a temperature of 15-20 ℃, a humidity of 70-75% until cordyceps mycelia overgrow with a culture medium, placing the culture medium full of cordyceps mycelia in a dark environment at a temperature of 20-25 ℃, a humidity of 80-90% and a light environment until the cordyceps is turned to orange, finally culturing at 20-25 deg.C and humidity of 80-90% under visible light environment until Cordyceps fruiting body grows;
(4) and (3) second-stage test tube strain cultivation: 600 ml of clear water with 550 ml, 450 ml of fresh hawthorn juice with 400 ml of waste water, 20-30 g of polygonatum odoratum powder, 20-30 g of glucose, 2-3 g of monopotassium phosphate, 1-2 g of magnesium sulfate, 5-6 g of peptone and 15-20 g of agar powder, putting the above materials into a heating container together, heating until various materials are completely dissolved in water to obtain matrix solution, adjusting pH of the matrix solution to 5.5-6, subpackaging in test tubes, sterilizing, placing on slant to obtain slant test tube, separating the first stage Cordyceps militaris fruiting body obtained by culturing in step (3) into slant test tubes under aseptic environment, culturing at 15-20 deg.C in dark light until the Cordyceps mycelia overgrow the tube, and culturing in 20-25 deg.C in light environment until the Cordyceps mycelia turn orange to obtain second stage test tube strain;
(5) and (3) liquid strain cultivation in the second stage: 500 ml of clear water of 450-;
(6) second stage sporocarp cultivation: 400 ml of clear water of 350-; taking 50-70 parts of hawthorn fruit residues, 20-25 parts of hawthorn branch and leaf crushed materials, 10-20 parts of glutinous rice and 3-5 parts of silkworm chrysalis powder, mixing the hawthorn fruit residues, the hawthorn branch and leaf crushed materials, the glutinous rice and the silkworm chrysalis powder to obtain a mixed material, mixing a nutrient solution into the mixed material, adjusting the water content of the mixed material to 55-60% to obtain a culture medium, subpackaging the culture medium into culture containers and sterilizing, inoculating the second-stage liquid strain obtained by the culture in the step (5) into the culture medium in an aseptic environment, placing the inoculated culture medium in a dark environment at the temperature of 15-20 ℃, the humidity of 70-75% until cordyceps mycelia overgrow with a culture medium, placing the culture medium full of cordyceps mycelia in a dark environment at the temperature of 20-25 ℃, the humidity of 80-90% and a light environment until cordyceps is turned to orange, finally culturing at 20-25 deg.C and humidity of 80-90% under visible light environment until Cordyceps fruiting body grows;
(7) third-stage test tube strain cultivation: 700 ml of fresh hawthorn juice with 650 plus materials, 350 ml of clear water with 300 plus materials, 20-30 g of rhizoma polygonati odorati powder, 20-30 g of glucose, 2-3 g of monopotassium phosphate, 1-2 g of magnesium sulfate, 5-6 g of peptone and 15-20 g of agar powder, putting the above materials into a heating container together, then heating until various materials are completely dissolved in water to obtain matrix solution, then subpackaging the matrix solution in test tubes, sterilizing the test tubes, placing the test tubes on an inclined plane to obtain inclined test tubes, separating the fruiting body of the second stage Cordyceps militaris obtained by culturing in step (6) into slant test tubes under aseptic environment, culturing at 15-20 deg.C in dark light until the cordyceps mycelia overgrow the tube, and culturing in a test tube at 20-25 deg.C in light environment until the cordyceps mycelia turn orange to obtain third stage test tube strain.
(8) Liquid strain cultivation in the third stage: 850-900 ml of fresh hawthorn juice, 100-150 ml of clear water, 50-60 g of polygonatum odoratum powder, 20-30 g of glucose and 5-6 g of peptone, all the materials are put into a heating container, then the heating container is heated until all the materials are completely dissolved in water to obtain nutrient solution, the nutrient solution is respectively filled into a liquid strain culture container and sterilized to obtain a liquid strain culture medium, the third-stage test tube strain obtained by the culture in the step (7) is inoculated into the liquid strain culture medium in an aseptic environment, and finally the third-stage liquid strain is obtained by deep fermentation culture under the environment of 15-20 ℃ and dark light until the liquid strain culture medium is full of bacteria balls or hyphae.
7. The method for cultivating cordyceps sinensis by using hawthorn as a main substrate as claimed in claim 1, wherein the illumination environment in the step (6) is 100-200lux illumination.
8. The method for cultivating cordyceps sinensis by using hawthorn as a main substrate as claimed in claim 1, wherein the illumination environment in the step (7) is given 300-600lux of illumination.
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