CN102405763B - Cultivating method of aweto - Google Patents
Cultivating method of aweto Download PDFInfo
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Abstract
The invention discloses a cultivating method of aweto, which is characterized in that: wild aweto in Qinghai Province of China is adopted as an initial strain, aweto liquid spawn is obtained through separation cultivation, amplification cultivation and liquid cultivation of the initial strain, and the liquid spawn is inoculated onto a two-year-old bat larval body to obtain the aweto through a bedstead-type cultivation way or a bottle-type cultivation way. Due to the adoption of the cultivating method, the technical problem of the prior art that the aweto cannot be artificially cultivated is solved.
Description
Technical field the invention belongs to medicine breed of edible fungus technical field, relates to a kind of breeding method of Cordyceps sinensis.
Although background technology Cordyceps sinensis has worm and careless profile concurrently, the non-grass of non-worm, belong to the Homonemeae biology.Cordyceps sinensis and northern Chinese caterpillar Fungus belong to the Eumycota Cordyceps, but are different kinds.Their growing environment and medicinal efficacy are completely different.Cordyceps sinensis is Clavicipitaceae, and fungi Cordyceps sinensis is to colonize in stroma on the Hepialidae insect larvae and the complex of larva corpse, is a kind of traditional famous and precious tonic Chinese herbal medicine material, and the multiple efficacies such as the function of immune system of adjusting, antitumor, antifatigue are arranged.Cordyceps sinensis have protect the liver beneficial lung, supplying vital essence and marrow, stop blooding, reduce phlegm, antitumor, can also grow lung, each bacterioid, hypotensive is resisted in kidney tonifying.The cordycepin of one of its main component, it is a kind of material that there is antibiosis and suppress the cell division effect, medically with foreign medical science, by the isotope detection technology, Cordyceps sinensis to be studies confirm that, being grass element and being that oxalic acid studies confirm that it is that cordycepic acid in grass also has very high kidney tonifying, establishing-Yang, benefit essence, Dingchuan effect in addition in Chinese caterpillar fungus, and can resist significantly cell ageing.In pharmacology modern study result, Cordyceps of Qinghai Province contains cordycepic acid approximately 7%, carbohydrate 28.9%, fat approximately 8.4%, protein approximately 25%, in fat, 82.2% is unsaturated fatty acid, in addition, still contain cobalamin, ergot lipidol, hexose alcohol, alkaloid etc.Because narrow, the natural parasitic rate of wild cordyceps Distribution Area is low, to living environment conditional request harshness, so resource own is more limited.Because Cordyceps sinensis main product ground ecotope suffers the people, be heavy damage again in recent years, a large amount of blindly unreasonable excavating cause resource increasingly to reduce, and output descends year by year.And Cordyceps sinensis is due to can medicine-food two-purpose, and people constantly find the pharmacological action that it is new, thus a plurality of country to its multiplication that needs, price goes up year by year, international market is day by day in short supply.In order to meet the demand of world market, the research work to Cordyceps sinensis has all been carried out in succession as provinces such as Sichuan, Hubei, Henan, Shaanxi, Yunnan and Hunan in current many areas.People relate to phorozoon conclusive evidence, ecotope investigation, chemical composition and the aspects such as pharmacological action analysis, artificial cultivation to the research of Cordyceps sinensis.
Taiwan has started the technical research of Developing Cultivate Cordyceps sinensis in recent years, still, relevant report is not yet arranged at present.Cordyceps sinensis, as one of raw material of health food, because the property of medicine of Cordyceps sinensis is gentle, is not easy to have side effects, so can among subtle, change physique.But Cordyceps sinensis also is not easy to collect, so price is naturally high.Carry out tame method, utilize mycelium to cultivate and can reduce a large amount of production costs, for the mankind benefit!
Summary of the invention the purpose of this invention is to provide a kind of breeding method of Cordyceps sinensis, to overcome the deficiencies in the prior art.
To achieve these goals, the breeding method of Cordyceps sinensis of the present invention, its main points are to comprise the following steps:
The first step, the female inclined-plane of planting of Cordyceps sinensis one-level are cultivated:
1, culture medium prescription:
Potato 200 grams, wheat skin 50 grams, potassium dihydrogen phosphate 2 grams, glucose 20 grams, peptone 10 grams, streptomycin 0.5 gram, agar 20 grams, 4 of Cobastabs, 1000 milliliters, water;
2, compound method:
The potato of cleaning peeling is thinly sliced and added 1000 milliliters, water and boil 30 minutes, soft and when not rotten to potato, consider and remove the potato residue with 4 layers of gauze, taking juice adds agar 20 grams in pot, peptone 10 grams, potassium dihydrogen phosphate 2 grams, vitamin B4 tablet, streptomycin 0.5 gram, glucose 20 grams, wheat skin 50 grams also stir, continuing to be heated to agar dissolves fully, rapid packing test tube while hot, the amount of each cuvette cartridge medium and the volume ratio of test tube are 1: 5, tampon beyond the Great Wall after installing, then every 10 of test tube is bundled into to a bundle, wrap and put into the pressure cooker sterilizing with brown paper, be heated to pressure and reach 0.5Kg/m
2the time, the air valve that begins to rehearse, discharge cold air, makes pointer get back to 0 point, when then shutting air bleeding valve and continuing to be heated to 121 ℃, pressurize takes the dish out of the pot in 40 minutes, and while hot test tube is put into to inclined-plane, after cooling the solid test tube slant is standby,
3, Isolation and Culture:
The stroma of fresh stalwartness is cleaned to the rear medicinal alcohol with 0.1% mercuric chloride solution or 70% to its disinfection at the Qinghai Wild Cordyceps sinensis of 8 centimeter length, remove its surperficial mercuric chloride or alcohol with sterile water again, after the Cordyceps sinensis surface sterilization is processed, cut the skin of Cordyceps sinensis stroma by sterile razor blade, the fritter that cuts 1 cubic millimeter of its interior tissue with vaccinating lancet moves rapidly to be received on test tube slant, and tampon beyond the Great Wall rapidly, then put into the insulating box of 20-28 ℃ and cultivate and within 15-20 days, obtain the female inclined-plane of planting of Cordyceps sinensis one-level;
Second step, the female expansion of planting inclined-plane of Cordyceps sinensis secondary are cultivated:
The female slant tube of planting of the one-level of separation is enlarged to cultivation again, obtain the female slant tube of planting of secondary, the medium of the female kind of secondary slant tube is identical with the medium of the female kind of one-level slant tube, move strict sterile working while connecing, one-level is female plants slant tube and moves and meet 20-30 and prop up the female slant tube of planting of secondary, puts into the insulating box of 20-28 ℃ after inoculation and cultivates and within 10-15 days, obtain the female inclined-plane of planting of Cordyceps sinensis secondary;
The cultivation of the 3rd step, Cordyceps sinensis liquid spawn:
1, culture medium prescription:
2500 milliliters of corn 250 grams, peptone 25 grams, dusty yeast 2.5 grams, potassium dihydrogen phosphate 2.5 grams, pangamic acid sheet, streptomycin 2.5 grams, magnesium sulfate 1 gram, glucose 40 grams, water;
2, compound method:
Corn is cleaned and to be put into pot and add 2500 milliliters, water and boil 30 minutes with clear water, and four layers of filtered through gauze of cooling rear use remove slag, and get its juice in pot, add peptone 25 grams, dusty yeast 2.5 grams, potassium dihydrogen phosphate 2.5 grams, Cobastab
15, streptomycin 2.5 grams, magnesium sulfate 1 gram, glucose 40 grams, then be heated to all dissolve, after cooling, be divided in 500 mL of saline bottles, 250 milliliters of every bottled liquid measures, with sterilizing in pressure cooker after the sealing of tampon jam-pack, when temperature reaches 121 ℃, pressurize takes the dish out of the pot in 2 hours, together puts into sterile board together with the female inclined-plane of planting of Cordyceps sinensis secondary after cooling, prepares switching;
3, the cultivation of liquid spawn:
Strictly observe the sterile working rules in switching process, switching process is that female kind of Cordyceps sinensis will grown up in slant tube is transferred in liquid nutrient medium, during inoculation, remove female agar of planting on piece, female piece of planting is swum in above the liquid medium in saline bottle, after inoculation, be put in insulating box at 20-28 ℃, constant temperature culture 2-3 days, when the mother on the strain cultivation liquid level plants the bacterium piece that block length is 2.5cm to diameter, just saline bottle is transferred on bottle swingging machine at 20-28 ℃ of shaken cultivation 7-14 days, obtained the Cordyceps sinensis liquid spawn;
The cultivation of the 4th step, Cordyceps sinensis:
1, the selection in cultivation place:
Select leeward on the sunny side, clean, the pollution-free source of Gao Kuo, surrounding environment, have clean water source, draining to build easily receptacle and cultivation canopy, orientation sits in the north facing the south, receptacle is wanted and the cultivation canopy connects together, and is convenient to management;
2, the raising of bat moth worm:
Raising bat moth worm will raise at aseptic receptacle, require: the temperature of receptacle is 25 ℃-28 ℃, humidity is 90%, and keep dark or low-light according to condition, the food of bat moth worm is Chinese sorghum, carrot and potato, or manually use wheat bran 50 grams, potato starch 10 grams, defatted soy flour 25 grams, dried mulberry leaf powder 20 grams, sucrose 5 grams, aureomycin 0.1 gram, sorbic acid 0.3 gram, ascorbic acid 1.5 grams, 250 milliliters of manufactured feeds that prepare in proportion of aquae destillata, receptacle is wanted regular sterilizing, sterilization, do sanitation and hygiene well, raise after 5-8 month bat moth worm both produced two generation the bat larva, two generation the bat larval growth within 30 days, obtain 2 age the bat larva,
3, the cultivation of Cordyceps sinensis:
The cultivation canopy will select the good soil of earth, venting capability that the silt content mass ratio is 50% to set up green house, and green house sits in the north facing the south, and use windowpane, and planting type has the cultivation of bedstead formula and bottle formula to cultivate two kinds:
1), bedstead formula cultivation:
(1), the preparation of cultivation bedstead rib: be provided with multilayer in booth and cultivate bedstead, every layer height 12-20 centimetre, width 1.2-1.5 rice, length and green house are corresponding, the top layer of cultivating bedstead is high apart from 1 meter of ceiling, leaves 0.8 meter wide passage between each bedstead, is convenient to well management; Load compost on each layer of bedstead, require 8 centimetres of soil thickness, loosen the soil 10Kg, artifical compound feed 10Kg of compost effluent husky 50Kg, vegetable garden soil 30Kg, yellow mud mixes together in proportion, and the adjusting soil pH value is 6-7, humidity is 40-60%, then compost is made to ridge;
(2), inoculation: cultured Cordyceps sinensis liquid spawn, pack in sterile little sprayer, during 7 thirty thirty to 9, uniformly liquid spawn is sprayed onto on 2 age bat larvas at night, every spray in 8-10 hour once, connect and spray 2-3 time, success ratio of inoculation is higher, in order to improve success ratio of inoculation, sneak into aweto mycelium in the food of larva, after inoculation, again by 2 age the bat larva put on the rib of cultivating bedstead, allow its food limit, limit bore in soil, put bat larva 150-200 of 2 age for every square metre, above putting after worm, with leaf or film, build,
(3), postvaccinal management: after inoculation, to note observing 2 age the bat larva active situation, and to prevent ant, the harm of cockroach, after 7-10 days, larva just starts to infect Cordyceps Militaris, infected larva, body colour is faded to faint yellow by dark brown, and creep slowly, last whole body is covered with the canescence mycelia and ossifys dead, inoculating in latter 1 month regulates the room temperature is 10 ℃-20 ℃, relative air humidity is 50%-60%, 1-1.5 in individual month, regulate temperature at 14 ℃-25 ℃, air humidity 60%-70%, after 1 first quarter moon, when the larva head is sprouted fruit body, regulate temperature at 26 ℃-32 ℃, humidity is 70%-80%, every day, door window ventilated 2 hours, be beneficial to growing of cordyceps militaris sporocarp, again after 1-2 month, stroma can grow to 10-12 centimetre high, thick 1.5-5 millimeter club-like, color is yellowish-brown, ripe, just can gather,
2), bottle formula cultivation:
(1), the preparation of culture bottle: use Cans as culture bottle, the composition of compost is identical with the bedstead cultivation, and the compost bottling requires: dress soil will loosen, and is conducive to larva and pierces in soil, 2 centimeters of soil thickness of dress, for well, install by sterilization and put into and cultivate canopy;
(2), inoculation: strict sterile working, cultured Cordyceps sinensis liquid spawn, pack in sterile little sprayer, during 7 thirty thirty to 9, uniformly liquid spawn is sprayed onto on 2 age bat larvas at night, every spray in 8-10 hour once, connect and spray 2-3 time, after having inoculated, put into by every bottle of 8 insects on the soil of bottle, then cover tightly and get final product with leaf or film;
(3), postvaccinal management: manage with bedstead.
The present invention be take Qinghai Wild Cordyceps sinensis as starting strain, through separating the Cordyceps sinensis liquid spawn of cultivating, enlarging cultivation, liquid culture system, liquid-spawn inoculation is obtained to Cordyceps sinensis through the cultivation of bedstead formula or the cultivation of bottle formula to 2 ages on bat larva body.The invention solves prior art and manually can not cultivate the technical problem of Cordyceps sinensis.
Bat moth worm be the cultivation Cordyceps sinensis first-selected medium, with 2 age bat larva the best.The optimum growing condition of Cordyceps sinensis: temperature is 20 ℃-28 ℃, form stroma 10 ℃-25 ℃ of temperature, in the time of 26 ℃-32 ℃, stroma grows native face, the growth temperature of spore is 28 ℃-℃ 32 ℃, stages all will keep certain temperature difference, utilize the temperature difference to stimulate, enhancing development, the temperature difference should be controlled in the scope of 5 ℃-10 ℃.Humidity: dry condition need to be filled out in the larva microbiological contamination of burying, relative air humidity remains on 50%-60%, mycelia grows in polypide, and to require relative temperature be 60%-70%, being split into of sclerotium requires relative moisture 10%-20%, the relative moisture that the needs of the formation of stroma and snake grass are higher, best with 80%-90%.Illumination: do not need illumination in spore germination and vegetative stage, but transforming from the asexual stage to the perfect stage, namely, after inoculation is cultivated 80-90 days, just need to increase scattered light to a certain degree, to stimulate formation and the growth of stroma.Air: Cordyceps sinensis grows little to the requirement of oxygen, only needs a small amount of air just.Acid-base value: Cordyceps sinensis likes the ambient growth of meta-acid to grow, and the pH value of medium is advisable with 6-7.
Embodiment
Embodiment 1: the breeding method of Cordyceps sinensis, carry out in the steps below:
The first step, the female inclined-plane of planting of Cordyceps sinensis one-level are cultivated:
1, culture medium prescription:
Potato 200 grams, wheat skin 50 grams, potassium dihydrogen phosphate 2 grams, glucose 20 grams, peptone 10 grams, streptomycin 0.5 gram, agar 20 grams, 4 of Cobastabs, 1000 milliliters, water;
2, compound method:
The potato of cleaning peeling is thinly sliced and added 1000 milliliters, water and boil 30 minutes, soft and when not rotten to potato, consider and remove the potato residue with 4 layers of gauze, taking juice adds agar 20 grams in pot, peptone 10 grams, potassium dihydrogen phosphate 2 grams, vitamin B4 tablet, streptomycin 0.5 gram, glucose 20 grams, wheat skin 50 grams also stir, continuing to be heated to agar dissolves fully, rapid packing test tube while hot, the amount of each cuvette cartridge medium and the volume ratio of test tube are 1: 5, tampon beyond the Great Wall after installing, then every 10 of test tube is bundled into to a bundle, wrap and put into the pressure cooker sterilizing with brown paper, be heated to pressure and reach 0.5Kg/m
2the time, the air valve that begins to rehearse, discharge cold air, makes pointer get back to 0 point, when then shutting air bleeding valve and continuing to be heated to 121 ℃, pressurize takes the dish out of the pot in 40 minutes, and while hot test tube is put into to inclined-plane, after cooling the solid test tube slant is standby,
3, Isolation and Culture:
By the stroma of fresh stalwartness after the Qinghai Wild Cordyceps sinensis of 8 centimeter length cleans with 0.1% mercuric chloride solution to its disinfection, remove its surperficial mercuric chloride with sterile water again, after the Cordyceps sinensis surface sterilization is processed, cut the skin of Cordyceps sinensis stroma by sterile razor blade, the fritter that cuts 1 cubic millimeter of its interior tissue with vaccinating lancet moves rapidly to be received on test tube slant, and tampon beyond the Great Wall rapidly, then put into the insulating box of 25 ℃ and cultivate and within 18 days, obtain the female inclined-plane of planting of Cordyceps sinensis one-level;
Second step, the female expansion of planting inclined-plane of Cordyceps sinensis secondary are cultivated:
The female slant tube of planting of the one-level of separation is enlarged to cultivation again, obtain the female slant tube of planting of secondary, the medium of the female kind of secondary slant tube is identical with the medium of the female kind of one-level slant tube, move strict sterile working while connecing, the female kind of an one-level slant tube moves and connects the female slant tubes of planting of 25 secondarys, and the insulating box cultivation of putting into 25 ℃ after inoculation obtains the female kind of Cordyceps sinensis secondary inclined-plane in 12 days;
The cultivation of the 3rd step, Cordyceps sinensis liquid spawn:
1, culture medium prescription:
Corn 250 grams, peptone 25 grams, dusty yeast 2.5 grams, potassium dihydrogen phosphate 2.5 grams, Cobastab
12500 milliliters of 5, streptomycin 2.5 grams, magnesium sulfate 1 gram, glucose 40 grams, water;
2, compound method:
Corn is cleaned and to be put into pot and add 2500 milliliters, water and boil 30 minutes with clear water, and four layers of filtered through gauze of cooling rear use remove slag, and get its juice in pot, add peptone 25 grams, dusty yeast 2.5 grams, potassium dihydrogen phosphate 2.5 grams, Cobastab
15, streptomycin 2.5 grams, magnesium sulfate 1 gram, glucose 40 grams, then be heated to all dissolve, after cooling, be divided in 500 mL of saline bottles, 250 milliliters of every bottled liquid measures, with sterilizing in pressure cooker after the sealing of tampon jam-pack, when temperature reaches 121 ℃, pressurize takes the dish out of the pot in 2 hours, together puts into sterile board together with the female inclined-plane of planting of Cordyceps sinensis secondary after cooling, prepares switching;
3, the cultivation of liquid spawn:
Strictly observe the sterile working rules in switching process, switching process is that female kind of Cordyceps sinensis will grown up in slant tube is transferred in liquid nutrient medium, during inoculation, remove female agar of planting on piece, female piece of planting is swum in above the liquid medium in saline bottle, after inoculation, be put in insulating box at 25 ℃, constant temperature culture 3 days, when the mother on the strain cultivation liquid level plants the bacterium piece that block length is 2.5cm to diameter, just saline bottle is transferred on bottle swingging machine 25 ℃ of shaken cultivation 10 days, obtained the Cordyceps sinensis liquid spawn;
The cultivation of the 4th step, Cordyceps sinensis:
1, the selection in cultivation place:
Select leeward on the sunny side, clean, the pollution-free source of Gao Kuo, surrounding environment, have clean water source, draining to build easily receptacle and cultivation canopy, orientation sits in the north facing the south, receptacle is wanted and the cultivation canopy connects together, and is convenient to management;
2, the raising of bat moth worm:
Raising bat moth worm will raise at aseptic receptacle, require: the temperature of receptacle is 25 ℃, humidity is 90%, and keep dark, the food of bat moth worm is manually to use wheat bran 50 grams, potato starch 10 grams, defatted soy flour 25 grams, dried mulberry leaf powder 20 grams, sucrose 5 grams, aureomycin 0.1 gram, sorbic acid 0.3 gram, ascorbic acid 1.5 grams, 250 milliliters of manufactured feeds that prepare in proportion of aquae destillata, receptacle is wanted regular sterilizing, sterilization, do sanitation and hygiene well, raise after 7 months bat moth worm both produced two generation the bat larva, two generation the bat larval growth within 30 days, be 2 age the bat larva,
3, the cultivation of Cordyceps sinensis:
The cultivation canopy will select the good soil of earth, venting capability that the silt content mass ratio is 50% to set up green house, and green house sits in the north facing the south, and use windowpane, adopts bedstead formula cultivation Cordyceps sinensis:
(1), the preparation of cultivation bedstead rib: be provided with multilayer in booth and cultivate bedstead, 16 centimetres of every layer heights, 1.3 meters of width, length and green house are corresponding, the top layer of cultivating bedstead is high apart from 1 meter of ceiling, leaves 0.8 meter wide passage between each bedstead, is convenient to well management; Load compost on each layer of bedstead, require 8 centimetres of soil thickness, loosen the soil 10Kg, artifical compound feed 10Kg of compost effluent husky 50Kg, vegetable garden soil 30Kg, yellow mud mixes together in proportion, and regulating soil pH value is 6, humidity is 50%, then compost is made to ridge;
(2), inoculation: cultured Cordyceps sinensis liquid spawn, pack in sterile little sprayer, 8 are sprayed onto liquid spawn on 2 age bat larvas uniformly at night, every spray in 9 hours once, connect and spray 3 times, after inoculation, again by 2 age the bat larva put on the rib of cultivating bedstead, allow its food limit, limit bore in soil, every square metre put 2 age 180 of bat larvas, with leaf, build above putting after worm;
(3), postvaccinal management: after inoculation, to note observing 2 age the bat larva active situation, and to prevent ant, the harm of cockroach, after 8 days, larva just starts to infect Cordyceps Militaris, infected larva, body colour is faded to faint yellow by dark brown, and creep slowly, last whole body is covered with the canescence mycelia and ossifys dead, inoculate that to regulate the room temperature in latter 1 month be 15 ℃, relative air humidity is 55%, 1.5 in individual month, regulate temperature at 18 ℃, air humidity 65%, after 1 first quarter moon, when the larva head is sprouted fruit body, regulate temperature at 28 ℃, humidity is 75%, every day, door window ventilated 2 hours, be beneficial to growing of cordyceps militaris sporocarp, again after 1.5 months, stroma can grow to 11 centimetres high, thick 2 millimeters club-likes, color is yellowish-brown, ripe, just can gather.
Embodiment 2: according to embodiment 1, adopt bottle formula cultivation Cordyceps sinensis:
(1), the preparation of culture bottle: use Cans as culture bottle, the composition of compost is identical with the bedstead cultivation, and the compost bottling requires: dress soil will loosen, and is conducive to larva and pierces in soil, 2 centimeters of soil thickness of dress, for well, install by sterilization and put into and cultivate canopy;
(2), inoculation: strict sterile working, cultured Cordyceps sinensis liquid spawn, pack in sterile little sprayer, during 8 thirty, uniformly liquid spawn is sprayed onto on 2 age bat larvas at night, every spray in 10 hours once, connect and spray 3 times, after having inoculated, put into by every bottle of 8 insects on the soil of bottle, then cover tightly and get final product with leaf;
(3), postvaccinal management: manage with bedstead.
Claims (1)
1. the breeding method of a Cordyceps sinensis is characterized in that comprising the following steps:
The first step, the female inclined-plane of planting of Cordyceps sinensis one-level are cultivated:
(1), culture medium prescription:
1000 milliliters of potato 200 grams, wheat skin 50 grams, potassium dihydrogen phosphate 2 grams, glucose 20 grams, peptone 10 grams, streptomycin 0.5 gram, agar 20 grams, vitamin B4 tablet, water;
(2), compound method:
The potato of cleaning peeling is thinly sliced and added 1000 milliliters, water and boil 30 minutes, soft and when not rotten to potato, with 4 layers of gauze elimination potato residue, taking juice adds agar 20 grams in pot, peptone 10 grams, potassium dihydrogen phosphate 2 grams, vitamin B4 tablet, streptomycin 0.5 gram, glucose 20 grams, wheat skin 50 grams also stir, continuing to be heated to agar dissolves fully, rapid packing test tube while hot, the amount of each cuvette cartridge medium and the volume ratio of test tube are 1: 5, tampon beyond the Great Wall after installing, then every 10 of test tube is bundled into to a bundle, wrap and put into the pressure cooker sterilizing with brown paper, be heated to pressure and reach 0.5Kg/m
2the time, the air valve that begins to rehearse, discharge cold air, makes pointer get back to 0 point, when then shutting air bleeding valve and continuing to be heated to 121 ℃, pressurize takes the dish out of the pot in 40 minutes, and while hot test tube is put into to inclined-plane, after cooling the solid test tube slant is standby,
(3), Isolation and Culture:
By the stroma of fresh stalwartness after the Qinghai Wild Cordyceps sinensis of 8 centimeter length cleans with 0.1% mercuric chloride solution to its disinfection, remove its surperficial mercuric chloride with sterile water again, after the Cordyceps sinensis surface sterilization is processed, cut the skin of Cordyceps sinensis stroma by sterile razor blade, the fritter that cuts 1 cubic millimeter of its interior tissue with vaccinating lancet moves rapidly to be received on test tube slant, and tampon beyond the Great Wall rapidly, then put into the insulating box of 25 ℃ and cultivate and within 18 days, obtain the female inclined-plane of planting of Cordyceps sinensis one-level;
Second step, the female expansion of planting inclined-plane of Cordyceps sinensis secondary are cultivated:
The female slant tube of planting of the one-level of separation is enlarged to cultivation again, obtain the female slant tube of planting of secondary, the medium of the female kind of secondary slant tube is identical with the medium of the female kind of one-level slant tube, move strict sterile working while connecing, the female kind of an one-level slant tube moves and connects the female slant tubes of planting of 25 secondarys, and the insulating box cultivation of putting into 25 ℃ after inoculation obtains the female kind of Cordyceps sinensis secondary inclined-plane in 12 days;
The cultivation of the 3rd step, Cordyceps sinensis liquid spawn:
(1), culture medium prescription:
Corn 250 grams, peptone 25 grams, dusty yeast 2.5 grams, potassium dihydrogen phosphate 2.5 grams, Cobastab
12500 milliliters of 5, streptomycin 2.5 grams, magnesium sulfate 1 gram, glucose 40 grams, water;
(2), compound method:
Corn is cleaned and to be put into pot and add 2500 milliliters, water and boil 30 minutes with clear water, and four layers of filtered through gauze of cooling rear use remove slag, and get its juice in pot, add peptone 25 grams, dusty yeast 2.5 grams, potassium dihydrogen phosphate 2.5 grams, Cobastab
15, streptomycin 2.5 grams, magnesium sulfate 1 gram, glucose 40 grams, then be heated to all dissolve, are divided in 500 mL of saline bottles after cooling,
250 milliliters of every bottled liquid measures, with sterilizing in pressure cooker after the sealing of tampon jam-pack, when temperature reaches 121 ℃, pressurize takes the dish out of the pot in 2 hours, together puts into sterile board together with the female inclined-plane of planting of Cordyceps sinensis secondary after cooling, prepares switching;
(3), the cultivation of liquid spawn:
Strictly observe the sterile working rules in switching process, switching process is that female kind of Cordyceps sinensis will grown up in slant tube is transferred in liquid nutrient medium, during inoculation, remove female agar of planting on piece, female piece of planting is swum in above the liquid medium in saline bottle, after inoculation, be put in insulating box at 25 ℃, constant temperature culture 3 days, when the mother on the strain cultivation liquid level plants the bacterium piece that block length is 2.5cm to diameter, just saline bottle is transferred on bottle swingging machine 25 ℃ of shaken cultivation 10 days, obtained the Cordyceps sinensis liquid spawn;
The cultivation of the 4th step, Cordyceps sinensis:
(1), the selection in cultivation place:
Select leeward on the sunny side, clean, the pollution-free source of Gao Kuo, surrounding environment, have clean water source, draining to build easily receptacle and cultivation canopy, orientation sits in the north facing the south, receptacle is wanted and the cultivation canopy connects together, and is convenient to management;
(2), the raising of bat moth worm:
Raising bat moth worm will raise at aseptic receptacle, require: the temperature of receptacle is 25 ℃, humidity is 90%, and keep dark, the food of bat moth worm is manually to use wheat bran 50 grams, potato starch 10 grams, defatted soy flour 25 grams, dried mulberry leaf powder 20 grams, sucrose 5 grams, aureomycin 0.1 gram, sorbic acid 0.3 gram, ascorbic acid 1.5 grams, 250 milliliters of manufactured feeds that prepare in proportion of aquae destillata, receptacle is wanted regular sterilizing, sterilization, do sanitation and hygiene well, raise after 7 months bat moth worm both produced two generation the bat larva, two generation the bat larval growth within 30 days, be 2 age the bat larva,
(3), the cultivation of Cordyceps sinensis:
The cultivation canopy will select the good soil of earth, venting capability that the silt content mass ratio is 50% to set up green house, and green house sits in the north facing the south, and use windowpane, adopts bedstead formula cultivation Cordyceps sinensis:
1., the preparation of cultivation bedstead rib: be provided with multilayer in booth and cultivate bedstead, 16 centimetres of every layer heights, 1.3 meters of width, length and green house are corresponding, the top layer of cultivating bedstead is high apart from 1 meter of ceiling, leaves 0.8 meter wide passage between each bedstead, is convenient to well manage; Load compost on each layer of bedstead, require 8 centimetres of soil thickness, loosen the soil 10Kg, artifical compound feed 10Kg of compost effluent husky 50Kg, vegetable garden soil 30Kg, yellow mud mixes together in proportion, and regulating soil pH value is 6, humidity is 50%, then compost is made to ridge;
2., inoculation: cultured Cordyceps sinensis liquid spawn, pack in sterile little sprayer, 8 are sprayed onto liquid spawn on 2 age bat larvas uniformly at night, every spray in 9 hours once, connect and spray 3 times, after inoculation, again by 2 age the bat larva put on the rib of cultivating bedstead, allow its food limit, limit bore in soil, every square metre put 2 age 180 of bat larvas, with leaf, build above putting after worm;
3., postvaccinal management: after inoculation, to note observing 2 age the bat larva active situation, and to prevent ant, the harm of cockroach, after 8 days, larva just starts to infect Cordyceps Militaris, infected larva, body colour is faded to faint yellow by dark brown, and creep slowly, last whole body is covered with the canescence mycelia and ossifys dead, inoculate that to regulate the room temperature in latter 1 month be 15 ℃, relative air humidity is 55%, 1.5 in individual month, regulate temperature at 18 ℃, air humidity 65%, after 1 first quarter moon, when the larva head is sprouted fruit body, regulate temperature at 28 ℃, humidity is 75%, every day, door window ventilated 2 hours, be beneficial to growing of cordyceps militaris sporocarp, again after 1.5 months, stroma grows to 11 centimetres high, thick 2 millimeters club-likes, color is yellowish-brown, ripe, just can gather.
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