CN102405763A - Cultivating method of aweto - Google Patents
Cultivating method of aweto Download PDFInfo
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- CN102405763A CN102405763A CN2010102970998A CN201010297099A CN102405763A CN 102405763 A CN102405763 A CN 102405763A CN 2010102970998 A CN2010102970998 A CN 2010102970998A CN 201010297099 A CN201010297099 A CN 201010297099A CN 102405763 A CN102405763 A CN 102405763A
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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.
Though background technology Cordyceps sinensis has the profile of worm and grass concurrently, the non-grass of non-worm belongs 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 stroma and the complex of larva corpse that colonizes on the Hepialidae insect larvae, is a kind of traditional famous and precious tonic Chinese herbal medicine material, and 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, kidney tonifying opposing each bacterioid, hypotensive.The cordycepin of one of its main component; Be a kind of have antibiosis and material that suppresses the cell division effect; Be that medically research confirms to Cordyceps sinensis with the isotope detection technology with foreign medical science; In the Chinese caterpillar fungus be grass plain be that oxalic acid also has research to confirm to be the cordycepic acid in the grass also to have very high kidney tonifying, establishing-Yang, smart, the Dingchuan effect of benefit, and can resist cell ageing significantly.Among the pharmacology modern study result, it is about 7% that Qinghai Cordyceps sinensis contains cordycepic acid, carbohydrate 28.9%; Fat is about 8.4%, and protein is about 25%, and 82.2% is unsaturated fatty acid in the fat; In addition, still contain cobalamin, ergot lipidol, hexose alcohol, alkaloid etc.Narrow, the natural parasitic rate in area is low, harsh to the living environment conditional request because wild cordyceps distributes, so resource own is more limited.Again because Cordyceps sinensis main product ground ecotope suffers that the people is heavy damage, a large amount of blindly unreasonable excavating cause resource to reduce day by day in recent years, and output descends year by year.And Cordyceps sinensis is because 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, the international market is in short supply day by day.In order to satisfy the demand of world market, present many areas such as provinces such as Sichuan, Hubei, Henan, Shaanxi, Yunnan and Hunan have all carried out the research work to Cordyceps sinensis in succession.People relate to phorozoon conclusive evidence, ecotope investigation, chemical composition and aspects such as pharmacological action analysis, artificial cultivation to the research of Cordyceps sinensis.
Taiwan has begun the technical research of development cultivation Cordyceps sinensis in recent years, still, relevant report is not arranged as yet at present.Cordyceps sinensis because the property of medicine of Cordyceps sinensis is gentle, is not easy to have side effects, so can among subtle, change physique as one of raw material of health food.But Cordyceps sinensis also is not easy to collect, so price is high naturally.Carry out tame method, utilize mycelium to cultivate and to 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 deficiency of prior art.
To achieve these goals, the breeding method of Cordyceps sinensis of the present invention, its main points are may further comprise the steps:
The first step, the female slant culture of planting of Cordyceps sinensis one-level:
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 in water;
2, compound method:
The potato of cleaning peeling thinly sliced add 1000 milliliters in water and boiled 30 minutes, soft and when mashed, remove the potato residue to potato with 4 layers of gauzes worry; Taking juice adds agar 20 grams, peptone 10 grams, potassium dihydrogen phosphate 2 grams, vitamin B4 tablet, streptomycin 0.5 gram, glucose 20 grams, wheat skin 50 grams and stirs in pot; Continue to be heated to agar and dissolve fully, rapid while hot packing test tube, the amount of each cuvette cartridge medium and the volume ratio of test tube are 1: 5; Tampon beyond the Great Wall after having adorned; Be bundled into a bundle to per 10 of test tube then, wrap with brown paper and put into the pressure cooker sterilization, be heated to pressure and reach 0.5Kg/m
2The time, the air valve that begins to rehearse is discharged cold air, makes pointer get back to 0 point, pressurize took the dish out of the pot in 40 minutes when shutting air bleeding valve then and continuing to be heated to 121 ℃, and put into the inclined-plane to test tube while hot, after the cooling the solid test tube slant is subsequent use;
3, strain separating is cultivated:
With the stroma of fresh stalwartness the Qinghai Wild Cordyceps sinensis of 8 centimeter length clean the back with the medicinal alcohol of 0.1% mercuric chloride solution or 70% to its disinfection; Remove its surperficial mercuric chloride or alcohol with sterile water again; After the Cordyceps sinensis surface sterilization is handled; Cut the skin of Cordyceps sinensis stroma with sterile razor blade; The fritter that cuts 1 cubic millimeter of its interior tissue with vaccinating lancet moves rapidly to be received on the test tube slant, and tampon beyond the Great Wall rapidly, puts into 20-28 ℃ insulating box then and cultivates and promptly get the female kind of Cordyceps sinensis one-level inclined-plane in 15-20 days;
Second step, the female enlarged culture of planting the inclined-plane of Cordyceps sinensis secondary:
The female slant tube of planting of the one-level of separating is carried out enlarged culture again; Get 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 when connecing the Strict aseptic operation, 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 20-28 ℃ insulating box after inoculation finishes and cultivates and promptly get the female kind of Cordyceps sinensis secondary inclined-plane in 10-15 days;
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 cleaned with clear water put into pot and add 2500 milliliters in water and boiled 30 minutes, removing slag with four layers of filtered through gauze in the cooling back, gets its juice in pot, adds that peptone 25 grams, dusty yeast 2.5 grams, potassium dihydrogen phosphate 2.5 restrain, Cobastab
15, streptomycin 2.5 grams, magnesium sulfate 1 gram, glucose 40 grams; Be heated to off-bottom then, be divided in after the cooling in the 500 mL of saline bottles, 250 milliliters of every bottled liquid measures; Sealing the back with the tampon jam-pack sterilizes in pressure cooker; Pressurize took the dish out of the pot in 2 hours when temperature reached 121 ℃, and sterile board is together put into together with the female inclined-plane of planting of Cordyceps sinensis secondary in the cooling back, prepares switching;
3, the cultivation of liquid spawn:
Strictly observe the sterile working rules in the switching process; Switching process is that female kind of the Cordyceps sinensis that has grown up in the slant tube is transferred in the liquid nutrient medium, during inoculation, remove female agar of planting on the piece; Female piece of planting is swum in above the liquid medium in the saline bottle; Be put into after the inoculation in the insulating box at 20-28 ℃, constant temperature culture 2-3 days, treat that mother on the strain cultivation liquid level plants block length when diameter is the bacterium piece of 2.5cm; Just saline bottle was transferred on the bottle swingging machine at 20-28 ℃ of shaken cultivation 7-14 days, 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, Gao Kuo, surrounding environment cleaning, pollution-free source, have cleaning water source, draining to build receptacle and cultivation canopy easily, the orientation sits in the north facing the south, receptacle is wanted and the cultivation canopy connects together, and is convenient to manage;
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 keeps dark or low-light according to condition, and the food of bat moth worm is Chinese sorghum, carrot and potato; Or it is artificial with 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; Sanitation and hygiene will regularly be sterilized, sterilized, do well to receptacle, raise after 5-8 month bat moth worm both produced two generation the bat larva, two generation the bat larval growth promptly got in 30 days 2 age the bat larva;
3, the cultivation of Cordyceps sinensis:
It is that green house is set up in 50% the good soil of earth, venting capability that the cultivation canopy will be selected the silt content mass ratio, and green house sits in the north facing the south, and use windowpane, and planting type has cultivation of bedstead formula and two kinds of bottle formula cultivations:
1), bedstead formula cultivation:
(1), the preparation of cultivation bedstead rib: be provided with multilayer in the 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 management well; On each layer of bedstead, load compost; Require 8 centimetres of soil thickness, loosen the soil 10Kg, artifical compound feed 10Kg of the husky 50Kg of compost effluent, vegetable garden soil 30Kg, yellow mud mixes together in proportion, and the adjusting soil pH value is 6-7; Humidity is 40-60%, makes the ridge to compost then;
(2), inoculation:, in the sterile little sprayer of packing into, be sprayed onto liquid spawn on the 2 age bat larvas uniformly during 7 thirty thirty to 9 at night cultured Cordyceps sinensis liquid spawn; Spray in every separated 8-10 hour once connects and sprays 2-3 time, and success ratio of inoculation is higher; In order to improve success ratio of inoculation, in the food of larva, sneak into aweto mycelium, after inoculation finishes; Again with 2 age the bat larva put on the rib of cultivating bedstead; Let it in soil, bore while eating food, every square metre put 2 ages bat larva 150-200 only, build with leaf or film above putting behind the worm;
(3), postvaccinal management: to note after the inoculation observing 2 age the bat larva active situation, and to prevent the harm of ant, cockroach, after 7-10 days, larva just begins to infect Cordyceps Militaris; Infected larva, body colour is faded to faint yellow by dark brown, and creeps slowly, and last whole body is covered with the canescence mycelia and ossifys dead; Inoculate that to regulate the room temperature in back 1 month be 10 ℃-20 ℃, relative air humidity is 50%-60%, in 1-1.5 month, regulates temperature at 14 ℃-25 ℃; Air humidity 60%-70% after 1 first quarter moon, when the larva head is sprouted fruit body, regulates temperature at 26 ℃-32 ℃; Humidity is 70%-80%, and every day, door window ventilated 2 hours, was beneficial to growing of cordyceps militaris sporocarp; After 1-2 month, stroma can be grown 10-12 centimetre high again, thick 1.5-5 millimeter club-like; Color is a yellowish-brown, and is promptly ripe, just can gather;
2), bottle formula cultivation:
(1), the preparation of culture bottle: as culture bottle, the composition of compost is identical with the bedstead cultivation with Cans, and the compost bottling requires: dress soil will loosen, and helps larva and pierces in the soil, and 2 centimeters of dress soil thickness have been adorned after sterilization is put into and cultivated canopy for well;
(2), the inoculation: strict sterile working, cultured Cordyceps sinensis liquid spawn, in the sterile little sprayer of packing into; Uniformly liquid spawn is sprayed onto on the 2 age bat larvas during 7 thirty thirty to 9 at night; Whenever once, connect and spray 2-3 time, after having inoculated at a distance from spray in 8-10 hour; Put into by every bottle of 8 insects on the soil of bottle, cover completely with leaf or film again and get final product;
(3), postvaccinal management: manage with bedstead.
The present invention is a starting strain with Qinghai Wild Cordyceps sinensis, through the Cordyceps sinensis liquid spawn of separation and Culture, enlarged culture, liquid culture system, liquid-spawn inoculation is obtained Cordyceps sinensis through cultivation of bedstead formula or the cultivation of bottle formula to 2 ages on the bat larva body.The invention solves the prior art manual work and can not cultivate the technical problem of Cordyceps sinensis.
Bat moth worm is the first-selected medium of cultivation Cordyceps sinensis, with 2 age the bat larva best.The optimum growing condition of Cordyceps sinensis: temperature is 20 ℃-28 ℃; Form stroma for 10 ℃-25 ℃ in temperature, stroma grows native face in the time of 26 ℃-32 ℃, and the growth temperature of spore is 28 ℃-℃ 32 ℃; Each stage all will keep certain temperature difference; Utilize the temperature difference to stimulate, promote to grow, the temperature difference should be controlled in 5 ℃-10 ℃ the scope.Humidity: dried condition need 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%, and the higher relative moisture of needs of the formation of stroma and snake grass is best with 80%-90%.Illumination: do not need illumination in spore germination and vegetative stage, but transforming to the perfect stage, just after inoculation is cultivated 80-90 days, just need to increase scattered light to a certain degree, to stimulate the formation and the growth of stroma from the asexual stage.Air: Cordyceps sinensis grows little to the requirement of oxygen, only needs little 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, undertaken by following step:
The first step, the female slant culture of planting of Cordyceps sinensis one-level:
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 in water;
2, compound method:
The potato of cleaning peeling thinly sliced add 1000 milliliters in water and boiled 30 minutes, soft and when mashed, remove the potato residue to potato with 4 layers of gauzes worry; Taking juice adds agar 20 grams, peptone 10 grams, potassium dihydrogen phosphate 2 grams, vitamin B4 tablet, streptomycin 0.5 gram, glucose 20 grams, wheat skin 50 grams and stirs in pot; Continue to be heated to agar and dissolve fully, rapid while hot packing test tube, the amount of each cuvette cartridge medium and the volume ratio of test tube are 1: 5; Tampon beyond the Great Wall after having adorned; Be bundled into a bundle to per 10 of test tube then, wrap with brown paper and put into the pressure cooker sterilization, be heated to pressure and reach 0.5Kg/m
2The time, the air valve that begins to rehearse is discharged cold air, makes pointer get back to 0 point, pressurize took the dish out of the pot in 40 minutes when shutting air bleeding valve then and continuing to be heated to 121 ℃, and put into the inclined-plane to test tube while hot, after the cooling the solid test tube slant is subsequent use;
3, strain separating is cultivated:
With the stroma of fresh stalwartness the Qinghai Wild Cordyceps sinensis of 8 centimeter length clean the back 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 handled, cut the skin of Cordyceps sinensis stroma with sterile razor blade, the fritter that cuts 1 cubic millimeter of its interior tissue with vaccinating lancet moves rapidly to be received on the test tube slant; And tampon beyond the Great Wall rapidly, put into 25 ℃ insulating box then and cultivate and promptly get the female kind of Cordyceps sinensis one-level inclined-plane in 18 days;
Second step, the female enlarged culture of planting the inclined-plane of Cordyceps sinensis secondary:
The female slant tube of planting of the one-level of separating is carried out enlarged culture again; Get 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 aseptic operation when connecing, an one-level mother kind slant tube moves and connects 25 secondarys mother kind slant tubes, and the insulating box cultivation of putting into 25 ℃ after inoculation finishes promptly got Cordyceps sinensis secondary mother kind 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 cleaned with clear water put into pot and add 2500 milliliters in water and boiled 30 minutes, removing slag with four layers of filtered through gauze in the cooling back, gets its juice in pot, adds that peptone 25 grams, dusty yeast 2.5 grams, potassium dihydrogen phosphate 2.5 restrain, Cobastab
15, streptomycin 2.5 grams, magnesium sulfate 1 gram, glucose 40 grams; Be heated to off-bottom then, be divided in after the cooling in the 500 mL of saline bottles, 250 milliliters of every bottled liquid measures; Sealing the back with the tampon jam-pack sterilizes in pressure cooker; Pressurize took the dish out of the pot in 2 hours when temperature reached 121 ℃, and sterile board is together put into together with the female inclined-plane of planting of Cordyceps sinensis secondary in the cooling back, prepares switching;
3, the cultivation of liquid spawn:
Strictly observe the sterile working rules in the switching process; Switching process is that female kind of the Cordyceps sinensis that has grown up in the slant tube is transferred in the liquid nutrient medium, during inoculation, remove female agar of planting on the piece; Female piece of planting is swum in above the liquid medium in the saline bottle; Be put into after the inoculation in the insulating box at 25 ℃, constant temperature culture 3 days treats that mother on the strain cultivation liquid level plants block length when diameter is the bacterium piece of 2.5cm; Just saline bottle is transferred on the bottle swingging machine 25 ℃ of shaken cultivation 10 days, 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, Gao Kuo, surrounding environment cleaning, pollution-free source, have cleaning water source, draining to build receptacle and cultivation canopy easily, the orientation sits in the north facing the south, receptacle is wanted and the cultivation canopy connects together, and is convenient to manage;
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, and the food of bat moth worm is artificial with 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, sanitation and hygiene will regularly be sterilized, sterilized, do well to receptacle; Raise after 7 months bat moth worm both produced two generation the bat larva, two generation the bat larval growth be in 30 days 2 age the bat larva;
3, the cultivation of Cordyceps sinensis:
It is that green house is set up in 50% the good soil of earth, venting capability that the cultivation canopy will be selected the silt content mass ratio, 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 the 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 management well; On each layer of bedstead, load compost; Require 8 centimetres of soil thickness, loosen the soil 10Kg, artifical compound feed 10Kg of the husky 50Kg of compost effluent, vegetable garden soil 30Kg, yellow mud mixes together in proportion, and regulating soil pH value is 6; Humidity is 50%, makes the ridge to compost then;
(2), inoculation: cultured Cordyceps sinensis liquid spawn, in the sterile little sprayer of packing into, 8 are sprayed onto liquid spawn on the 2 age bat larvas uniformly at night; Whenever once, connect and spray 3 times, after inoculation finishes at a distance from spray in 9 hours; Again with 2 age the bat larva put on the rib of cultivating bedstead; Let it in soil, bore while eating food, every square metre put 2 age 180 of bat larvas, build with leaf above putting behind the worm;
(3), postvaccinal management: to note after the inoculation observing 2 age the bat larva active situation, and to prevent the harm of ant, cockroach, after 8 days, larva just begins to infect Cordyceps Militaris, infected larva; Body colour is faded to faint yellow by dark brown, and creeps slowly, and last whole body is covered with the canescence mycelia and ossifys deadly, inoculates that to regulate the room temperature in back 1 month be 15 ℃; Relative air humidity is in 55%, 1.5 month, regulates 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, was beneficial to growing of cordyceps militaris sporocarp, and after 1.5 months, stroma can be grown to 11 centimetres high again; Thick 2 millimeters club-likes, color is a yellowish-brown, and is promptly ripe, just can gather.
Embodiment 2: according to embodiment 1, adopt bottle formula cultivation Cordyceps sinensis:
(1), the preparation of culture bottle: as culture bottle, the composition of compost is identical with the bedstead cultivation with Cans, and the compost bottling requires: dress soil will loosen, and helps larva and pierces in the soil, and 2 centimeters of dress soil thickness have been adorned after sterilization is put into and cultivated canopy for well;
(2), the inoculation: strict sterile working, cultured Cordyceps sinensis liquid spawn, in the sterile little sprayer of packing into; Be sprayed onto liquid spawn on the 2 age bat larvas uniformly during 8 thirty at night; Whenever once, connect and spray 3 times, after having inoculated at a distance from spray in 10 hours; Put into by every bottle of 8 insects on the soil of bottle, cover completely with leaf again and get final product;
(3), postvaccinal management: manage with bedstead.
Claims (2)
1. the breeding method of a Cordyceps sinensis is characterized in that may further comprise the steps:
The first step, the female slant culture of planting of Cordyceps sinensis one-level:
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 in water;
2, compound method:
The potato of cleaning peeling thinly sliced add 1000 milliliters in water and boiled 30 minutes, soft and when mashed, remove the potato residue to potato with 4 layers of gauzes worry; Taking juice adds agar 20 grams, peptone 10 grams, potassium dihydrogen phosphate 2 grams, vitamin B4 tablet, streptomycin 0.5 gram, glucose 20 grams, wheat skin 50 grams and stirs in pot; Continue to be heated to agar and dissolve fully, rapid while hot packing test tube, the amount of each cuvette cartridge medium and the volume ratio of test tube are 1: 5; Tampon beyond the Great Wall after having adorned; Be bundled into a bundle to per 10 of test tube then, wrap with brown paper and put into the pressure cooker sterilization, be heated to pressure and reach 0.5Kg/m
2The time, the air valve that begins to rehearse is discharged cold air, makes pointer get back to 0 point, pressurize took the dish out of the pot in 40 minutes when shutting air bleeding valve then and continuing to be heated to 121 ℃, and put into the inclined-plane to test tube while hot, after the cooling the solid test tube slant is subsequent use;
3, strain separating is cultivated:
With stroma use mass percent after the Qinghai Wild Cordyceps sinensis of 8 centimeter length cleans of fresh stalwartness is that 0.1% mercuric chloride solution is to its disinfection; Remove its surperficial mercuric chloride with sterile water again; After the Cordyceps sinensis surface sterilization is handled; Cut the skin of Cordyceps sinensis stroma with sterile razor blade; The fritter that cuts 1 cubic millimeter of its interior tissue with vaccinating lancet moves rapidly to be received on the test tube slant, and tampon beyond the Great Wall rapidly, puts into 20-28 ℃ insulating box then and cultivates and promptly get the female kind of Cordyceps sinensis one-level inclined-plane in 15-20 days;
Second step, the female enlarged culture of planting the inclined-plane of Cordyceps sinensis secondary:
The female slant tube of planting of the one-level of separating is carried out enlarged culture again; Get 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 when connecing the Strict aseptic operation, 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 20-28 ℃ insulating box after inoculation finishes and cultivates and promptly get the female kind of Cordyceps sinensis secondary inclined-plane in 10-15 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 cleaned with clear water put into pot and add 2500 milliliters in water and boiled 30 minutes, removing slag with four layers of filtered through gauze in the cooling back, gets its juice in pot, adds that peptone 25 grams, dusty yeast 2.5 grams, potassium dihydrogen phosphate 2.5 restrain, Cobastab
15, streptomycin 2.5 grams, magnesium sulfate 1 gram, glucose 40 grams; Be heated to off-bottom then, be divided in after the cooling in the 500 mL of saline bottles, 250 milliliters of every bottled liquid measures; Sealing the back with the tampon jam-pack sterilizes in pressure cooker; Pressurize took the dish out of the pot in 2 hours when temperature reached 121 ℃, and sterile board is together put into together with the female inclined-plane of planting of Cordyceps sinensis secondary in the cooling back, prepares switching;
3, the cultivation of liquid spawn:
Strictly observe the sterile working rules in the switching process; Switching process is that female kind of the Cordyceps sinensis that has grown up in the slant tube is transferred in the liquid nutrient medium, during inoculation, remove female agar of planting on the piece; Female piece of planting is swum in above the liquid medium in the saline bottle; Be put into after the inoculation in the insulating box at 20-28 ℃, constant temperature culture 2-3 days, treat that mother on the strain cultivation liquid level plants block length when diameter is the bacterium piece of 2.5cm; Just saline bottle was transferred on the bottle swingging machine at 20-28 ℃ of shaken cultivation 7-14 days, 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, Gao Kuo, surrounding environment cleaning, pollution-free source, have cleaning water source, draining to build receptacle and cultivation canopy easily, the orientation sits in the north facing the south, receptacle is wanted and the cultivation canopy connects together, and is convenient to manage;
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 keeps dark or low-light according to condition, and the food of bat moth worm is Chinese sorghum, carrot and potato; Or it is artificial with 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; Sanitation and hygiene will regularly be sterilized, sterilized, do well to receptacle, raise after 5-8 month bat moth worm both produced two generation the bat larva, two generation the bat larval growth promptly got in 30 days 2 age the bat larva;
3, the cultivation of Cordyceps sinensis:
It is that green house is set up in 50% the good soil of earth, venting capability that the cultivation canopy will be selected the silt content mass ratio, and green house sits in the north facing the south, and use windowpane, and planting type has cultivation of bedstead formula and two kinds of bottle formula cultivations,
The processing step of said bedstead formula cultivation is following:
(1), the preparation of cultivation bedstead rib: be provided with multilayer in the 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 management well; On each layer of bedstead, load compost; Require 8 centimetres of soil thickness, loosen the soil 10Kg, artifical compound feed 10Kg of the husky 50Kg of compost effluent, vegetable garden soil 30Kg, yellow mud mixes together in proportion, and the adjusting soil pH value is 6-7; Humidity is 40-60%, makes the ridge to compost then;
(2), inoculation:, in the sterile little sprayer of packing into, be sprayed onto liquid spawn on the 2 age bat larvas uniformly during 7 thirty thirty to 9 at night cultured Cordyceps sinensis liquid spawn; Spray in every separated 8-10 hour once connects and sprays 2-3 time, and success ratio of inoculation is higher; In order to improve success ratio of inoculation, in the food of larva, sneak into aweto mycelium, after inoculation finishes; Again with 2 age the bat larva put on the rib of cultivating bedstead; Let it in soil, bore while eating food, every square metre put 2 ages bat larva 150-200 only, build with leaf or film above putting behind the worm;
(3), postvaccinal management: to note after the inoculation observing 2 age the bat larva active situation, and to prevent the harm of ant, cockroach, after 7-10 days, larva just begins to infect Cordyceps Militaris; Infected larva, body colour is faded to faint yellow by dark brown, and creeps slowly, and last whole body is covered with the canescence mycelia and ossifys dead; Inoculate that to regulate the room temperature in back 1 month be 10 ℃-20 ℃, relative air humidity is 50%-60%, in 1-1.5 month, regulates temperature at 14 ℃-25 ℃; Air humidity 60%-70% after 1 first quarter moon, when the larva head is sprouted fruit body, regulates temperature at 26 ℃-32 ℃; Humidity is 70%-80%, and every day, door window ventilated 2 hours, was beneficial to growing of cordyceps militaris sporocarp; After 1-2 month, stroma can be grown 10-12 centimetre high again, thick 1.5-5 millimeter club-like; Color is a yellowish-brown, and is promptly ripe, just can gather.
2. the breeding method of Cordyceps sinensis according to claim 1 is characterized in that the processing step of said bottle formula cultivation is following:
(1), the preparation of culture bottle: as culture bottle, the composition of compost is identical with the bedstead cultivation with Cans, and the compost bottling requires: dress soil will loosen, and 2 centimeters of dress soil thickness have been adorned after sterilize and put into the cultivation canopy;
(2), inoculation: strict sterile working, inoculation are operated identical with the bedstead cultivation, after having inoculated, put into by every bottle of 8 insects on the soil of bottle, cover completely with leaf or film to get final product again;
(3), postvaccinal management: manage with bedstead.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1348988A (en) * | 2000-10-17 | 2002-05-15 | 刑桂兰 | Artificial cultivation of cordyceps |
| TW200833836A (en) * | 2007-02-15 | 2008-08-16 | Comsun Internat Corp | An improved method of cultivating the Cordyceps Militaris by adding sugar cane fiber |
| CN101412967A (en) * | 2007-10-20 | 2009-04-22 | 朱斗锡 | Novel method for cultivating Chinese caterpillar fungus |
| CN101773052A (en) * | 2010-03-16 | 2010-07-14 | 孟庆学 | Preparation and industrial culture of aweto strain |
-
2010
- 2010-09-26 CN CN 201010297099 patent/CN102405763B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1348988A (en) * | 2000-10-17 | 2002-05-15 | 刑桂兰 | Artificial cultivation of cordyceps |
| TW200833836A (en) * | 2007-02-15 | 2008-08-16 | Comsun Internat Corp | An improved method of cultivating the Cordyceps Militaris by adding sugar cane fiber |
| CN101412967A (en) * | 2007-10-20 | 2009-04-22 | 朱斗锡 | Novel method for cultivating Chinese caterpillar fungus |
| CN101773052A (en) * | 2010-03-16 | 2010-07-14 | 孟庆学 | Preparation and industrial culture of aweto strain |
Non-Patent Citations (1)
| Title |
|---|
| 李昊等: "《虫草人工栽培与深度开发》", 31 December 2003, 科学技术文献出版社 * |
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