CN103749149A - Method for cultivating artificial worm grass with mealworm pupa as carrier - Google Patents
Method for cultivating artificial worm grass with mealworm pupa as carrier Download PDFInfo
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- CN103749149A CN103749149A CN201310728836.9A CN201310728836A CN103749149A CN 103749149 A CN103749149 A CN 103749149A CN 201310728836 A CN201310728836 A CN 201310728836A CN 103749149 A CN103749149 A CN 103749149A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
Provided is a method for cultivating artificial worm grass with mealworm pupa as a carrier. Bacterial suspension is prepared by using wild natural cordyceps sinensis bacterial strains produced in the Tibet plateau and inoculated on the live mealworm pupa, and mealworm pupa worm grass is produced through artificial cultivation. The method includes the following steps of (1) screening of worm grass bacterial strains; (2) preparation of mealworm pupa; (3) preparation of the bacterial suspension; (4) inoculation; (5) cultivation of stroma; (6) collection and storage of the worm grass. The mealworm pupa is used as a host, the cordyceps sinensis bacterial suspension is inoculated artificially, and the artificial worm grass is cultivated. Inoculation success rate and ossification after inoculation reach above 85%, and growing rate of sporocarp is more than 100%. The period from inoculation to maturity of the sporocarp is about 50-60 days.
Description
Technical field
The invention belongs to medicinal fungus culture technique field, be specifically related to a kind ofly take yellow meal worm and be that carrier cultivates the method for artificial aweto, relate in particular to wild natural cordyceps bacterial strain that a kind of use originates in Qinghai-Tibet Platean and prepare bacterial suspension inoculation to form the breeding method of artificial aweto on live body yellow meal worm through artificial culture.
Background technology
Along with the raising of people's living standard with to healthy attention, Cordyceps sinensis and auxiliary product thereof become one and develop industry rapidly.Cordyceps sinensis is classified national second class protection species as, and various places government also appearance policy taboo is dug, limit and dig, but not excavating of Cordyceps sinensis is curbed.Cordyceps sinensis is a kind of rare Chinese medicine of China, because it needs special growth conditions, vegetative period is longer, thereby very rare in occurring in nature quantity, along with the change of environmental condition, output is downward trend gradually, and social demand is increasing, therefore very famous and precious, price is also high, reach in the market every kilogram of 40,000 yuan of left and right.
Cordyceps militaris (Cordyceps militaris) is compared with Cordyceps sinensis (Cordyceps): Chinese caterpillar fungus is famous and precious traditional Chinese medicine, has very high tonic and is worth and medical value.Kind China Chinese caterpillar fungus has kind more than 200, and what wherein using value was the highest is to originate in the Cordyceps sinensis in Tibet and the Cordyceps militaris that originates in Chang Zishan.The two belongs to Ascomycetes Hypocreales Clavicipitaceae Cordyceps in biological classification.Cordyceps sinensis originates from natural, is that aweto parasitizes bat moth larvae and forms.Yellow meal worm Chinese caterpillar fungus is that cordyceps is inoculated in to yellow mealworm pupal cell, through artificial culture, forms.The cultivation of yellow meal worm Chinese caterpillar fungus at present has formed scale.Yellow meal worm Chinese caterpillar fungus and Cordyceps sinensis medicine change into minute and pharmacological function all very close.Research shows, in yellow meal worm Chinese caterpillar fungus and the multiple grass of winter worm, inorganic elements kind is basically identical, and the content of the two selenium is all higher, yellow luxuriant higher 3.3 times and 5.8 times than selenium-rich anticancer Chinese medicine respectively.
At present, the Pan Limei of domestic Beihua University passes through inoculation Cordyceps Militaris by Yellow meal worm larva, under the conditions such as certain temperature, humidity, illumination, through taking pains to foster of a couple of days, successfully cultivated yellow meal worm Chinese caterpillar fungus, but infection rate is closely 30%, does not reach the demand of large production.
The people such as professor Li Chunru of microbial control province of Agricultural University Of Anhui key lab utilize Taiwan Chinese caterpillar fungus (Cordyceps formosana) to carry out infection experiment to the insect yellow mealworm of feeding, result shows: yellow mealworm is at bacterium colony contact infection, just cast off a skin percutaneous puncture-inoculation and 1 * 105/ml of spore concentration of larva dips under processing, obtained relatively high infection rate, reach respectively 30%, 25% and 25%, it is higher that the larva that just casted off a skin is generally organized its infection rate than dark larva.
Application number is 200510101348.0, and denomination of invention is the patent of " fruiting bodies of cordyceps militaris and breeding method thereof that the yellow meal worm of take is host ", its method be comprise bacterial classification preparation, to yellow meal worm inoculation with go out the incubation steps such as grass.The present invention be take family's yellow meal worm as host, artificial infection cordyceps, and simulation Cordyceps militaris natural ecological environment, artificial culture forms.And the bacterial classification of this invention is spore suspension, the cultivation in later stage is to cultivate under natural environment, and drying and processing adopts high temperature treatment, and this is very large on the impact of the quality of Cordyceps militaris and active ingredient.
Summary of the invention
The object of this invention is to provide the wild natural cordyceps bacterial strain spore inoculating that a kind of use originates in Qinghai-Tibet Platean and to artificial culture on organic live body yellow meal worm, form the breeding method of yellow meal worm Chinese caterpillar fungus.
For achieving the above object, the breeding method of a kind of yellow meal worm Chinese caterpillar fungus of the present invention, is characterized in that, comprises the following steps:
1) screening of Cordyceps Militaris bacterial strain
The wild fresh Cordyceps strain of Qinghai-Tibet Platean will be originated in, first be inoculated in yellow meal worm powder (removing the cured matter of epidermis) as the dull and stereotyped growth of nitrogenous source, treat that mycelium covers with flat board, repeated screening is counted strain until obtain growing way than the pure bacterial strain of Cordyceps sinensis very fast, proterties is stable, usings above-mentioned bacterial strains as object bacterial strain of the present invention;
2) preparation of yellow meal worm
By Yellow meal worm larva in the indoor worm pupa of cultivating to obtain, for cultivating fruiting bodies of cordyceps militaris.In aseptic operating platform, live body pupa, with about 75% alcohol disinfecting 6-9 minute, is used to aseptic water washing 5-6 time, dry and do host;
3) making of bacteria suspension
The Chinese caterpillar fungus strain of screening is inoculated in and is usingd on the culture medium flat plate that yellow meal worm powder (remove epidermis cured matter) makes as main nitrogen, put at 20 ℃ and cultivate, after there are a large amount of mycelia, a picking part is inoculated in aforesaid liquid medium and cultivates, cultivate 4-5 days, use high speed discretizer, with 10000r/min, homogenate 1min had both obtained bacteria suspension;
4) inoculation
In aseptic operating platform, with asepsis injector, draw bacteria suspension and inject above-mentioned yellow meal worm afterbody or mouthpart, by 0.1-0.15 milliliter/only inject bacterium liquid;
5) cultivation of stroma
Yellow meal worm after processing is put in to aseptic vial or large glass dish, protects to pupal cell at 20 ℃ stiff; Then the Sterile culture room of cultivating stiff pupal cell and be placed in the natural lighting that 15-25 ℃, relative air humidity are 85-95% is cultivated;
6) Collection and conservation of Chinese caterpillar fungus
After stroma maturation, be placed under 60 ℃ of environment and dry 1-5 hour, put shady and cool dry place sealing and preserve, or carry out low tempertaure storage after adopting vacuum cooling drying.
Chinese caterpillar fungus of the present invention be take yellow meal worm as host, artificial infection bacteria suspension.Success ratio of inoculation, postvaccinal rigid rate reach more than 85%, and the rate that grows of fruit body reaches more than 100%.From being inoculated into the about 50-60 days of maturation of fruit body.
Embodiment
Below in conjunction with specific embodiment, the invention will be further described:
Live body yellow meal worm in the present invention independently provides for our company laboratory.
Yellow meal worm Chinese caterpillar fungus of the present invention is cultivated technical scheme: during artificial culture yellow meal worm Chinese caterpillar fungus, the Chinese caterpillar fungus strain of screening is inoculated in and usings yellow meal worm powder as on the organic medium of main nitrogen, put at 20 ℃ and cultivate, after there are a large amount of mycelia, a picking part is placed in liquid nutrient medium and makes cultivation, cultivates 4-5 days, smashes mycelium pellet both obtained bacteria suspension by high speed discretizer, by bacterial suspension inoculation in above-mentioned yellow meal worm afterbody or mouthpart, to pupal cell until stiff.Then stiff yellow mealworm pupal cell is placed under the condition that is applicable to Cordyceps militaris cultivation and cultivates.Technology concrete scheme is as follows:
1) screening of Cordyceps Militaris bacterial strain
The wild fresh Cordyceps strain of Qinghai-Tibet Platean will be originated in, first be inoculated in yellow meal worm powder as the dull and stereotyped growth of nitrogenous source, treat that mycelium covers with flat board, repeated screening is counted strain until obtain growing way than the pure bacterial strain of Cordyceps sinensis very fast, proterties is stable, usings above-mentioned bacterial strains as object bacterial strain of the present invention;
2) preparation of yellow meal worm
By Yellow meal worm larva in the indoor worm pupa of cultivating to obtain, for cultivating fruiting bodies of cordyceps militaris.In aseptic operating platform, live body pupa, with about 75% alcohol disinfecting 6-9 minute, is used to aseptic water washing 5-6 time, dry and do host;
3) making of bacteria suspension
The Chinese caterpillar fungus strain of screening is inoculated in and is usingd on the culture medium flat plate that yellow meal worm powder makes as main nitrogen, put at 20 ℃ and cultivate, after there are a large amount of mycelia, a picking part is inoculated in aforesaid liquid medium and cultivates, cultivate 4-5 days, use high speed discretizer, with 10000r/min, homogenate 1min had both obtained bacteria suspension;
4) inoculation
In aseptic operating platform, with asepsis injector, draw bacteria suspension and inject above-mentioned yellow meal worm afterbody or mouthpart, by 0.1-0.15 milliliter/only inject bacterium liquid;
5) cultivation of stroma
Yellow meal worm after processing is put in to aseptic vial or large glass dish, protects to pupal cell at 20 ℃ stiff; Then the Sterile culture room of cultivating stiff pupal cell and be placed in the natural lighting that 15-25 ℃, relative air humidity are 85-95% is cultivated;
6) Collection and conservation of Chinese caterpillar fungus
After stroma maturation, be placed under 60 ℃ of environment and dry 1-5 hour, put shady and cool dry place sealing and preserve, or carry out low tempertaure storage after adopting vacuum cooling drying.
Embodiment mono-:
The artificial aweto breeding method that the live body yellow meal worm of take is medium.
Concrete technical scheme is as follows:
(1) screening of Cordyceps Militaris bacterial strain
1, bacterial classification is made
Choose high yield, high-quality, precocity, originate in Qinghai-Tibet wild fresh cordyceps sporophore, alcohol with 75% carries out surface sterilization, with sterile water wash several (general 5-6 time), the polypide in Cordyceps sinensis is removed, remainder is positioned in medium and cultivates.Repurity after mycelia is covered with flat board.Through repeatedly obtaining the female kind of number strain after purifying.
Mother is planted and expanded after cultivation, be placed in and using yellow meal worm powder as on the organic medium of nitrogenous source, at 18 ℃-25 ℃, cultivate 20-30d, observe growing state, have living contaminants if see, the female kind of reply is further purified; If without living contaminants, can continue to cultivate, after one month, there is orange red fruitbody production, female kind is reliably described.
Liquid nutrient medium is sub-packed in 500ml triangular flask, and 121 ℃, sterilizing 20 minutes, plants mother with oese, to receive in triangular flask under aseptic condition.Inoculation is placed under 18-22 ℃ of condition, shakes kind of a cultivation 6d left and right, after even bead to be formed, with high speed disperser high speed 10000r/min, processes 1min and has both obtained bacteria suspension.
2, host prepares
By Yellow meal worm larva in the indoor worm pupa of cultivating to obtain, for cultivating fruiting bodies of cordyceps militaris.In aseptic operating platform, live body pupa, with about 75% alcohol disinfecting 6-9 minute, is used to aseptic water washing 1 minute, dry and do host;
3, inoculation
In aseptic operating platform, with asepsis injector, draw bacteria suspension and inject above-mentioned yellow meal worm afterbody or mouthpart, by 0.1-0.15 milliliter/only inject bacteria suspension;
4, the cultivation of stroma
Yellow meal worm after processing is put in to aseptic vial or large glass dish, protects to pupal cell at 20 ℃ stiff; Then the Sterile culture room of cultivating stiff pupal cell and be placed in the natural lighting that 15-25 ℃, relative air humidity are 85-95% is cultivated;
5, the Collection and conservation of yellow meal worm Chinese caterpillar fungus
After stroma maturation, be placed under 60 ℃ of environment and dry 1-5 hour, put shady and cool dry place sealing and preserve, or in order to prevent that high temperature from destroying the part nutrients in Chinese caterpillar fungus, can adopt after vacuum cooling drying low tempertaure storage.
Claims (7)
1. the method that the yellow meal worm of take is cultivated artificial aweto as carrier, is characterized in that, comprises the following steps: the screening of Cordyceps Militaris bacterial strain, the preparation of yellow meal worm, the making of bacteria suspension, inoculation, the cultivation of stroma, the steps such as the Collection and conservation of Chinese caterpillar fungus.
2. based on claimed in claim 1, take the method that yellow meal worm cultivates artificial aweto as carrier, it is characterized in that the screening of described Cordyceps Militaris bacterial strain, to originate in the wild fresh Cordyceps strain of Qinghai-Tibet Platean, first be inoculated in the flat board growth that yellow meal worm powder is made as main nitrogen, treat that mycelium covers with flat board, repeated screening is counted strain until obtain growing way than the pure bacterial strain of Cordyceps sinensis very fast, proterties is stable, usings above-mentioned bacterial strains as object bacterial strain of the present invention.
3. based on claimed in claim 1, take the method that yellow meal worm cultivates artificial aweto as carrier, it is characterized in that the preparation of described yellow meal worm, in aseptic operating platform, by 10-15 yellow meal worm in age larva with about 75% alcohol disinfecting 6-9 minute, with aseptic water washing 5-6 time, be placed in aseptic easily interior ventilation and dry, do host standby.
4. based on claimed in claim 1, take the method that yellow meal worm cultivates artificial aweto as carrier, it is characterized in that the making of described bacteria suspension: be that the Chinese caterpillar fungus strain of screening is inoculated in and is usingd on the culture medium flat plate that yellow meal worm powder makes as main nitrogen, put at 20 ℃ and cultivate, after there are a large amount of mycelia, a picking part is inoculated in aforesaid liquid medium and cultivates, and cultivates 4-5 days, uses high speed discretizer, with 10000r/min, homogenate 1min had both obtained bacteria suspension.
5. based on claimed in claim 1, take the method that yellow meal worm cultivates artificial aweto as carrier, it is characterized in that described inoculation: in aseptic operating platform, with asepsis injector, draw bacteria suspension and inject above-mentioned yellow meal worm pupa afterbody or mouthpart, by 0.1-0.15 milliliter/only inject bacterium liquid.
6. based on claimed in claim 1, take yellow meal worm and be that carrier cultivates the method for artificial aweto, it is characterized in that the cultivation of described stroma: the yellow meal worm after processing is put in to aseptic vial or large glass dish, protects to pupal cell at 20 ℃ stiff; Then the Sterile culture room of cultivating stiff pupal cell and be placed in the natural lighting that 15-25 ℃, relative air humidity are 85-95% is cultivated.
7. based on claimed in claim 1, take the method that yellow meal worm cultivates artificial aweto as carrier, the Collection and conservation that it is characterized in that described Chinese caterpillar fungus: be by stroma after maturation, be placed under 60 ℃ of environment and dry 1-5 hour, put shady and cool dry place sealing and preserve, or carry out low tempertaure storage after adopting vacuum cooling drying.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105861314A (en) * | 2015-01-24 | 2016-08-17 | 保山学院 | Method for culturing Isaria cicadae Miquel by using Tenebrio molitor pupae |
| CN108184540A (en) * | 2018-01-29 | 2018-06-22 | 泰安市农业科学研究院 | A kind of method that Cordyceps militaris is cultivated using live body Yellow meal worm larva as host |
| CN116250510A (en) * | 2023-04-26 | 2023-06-13 | 伊犁哈萨克自治州林业科学研究院 | Artificial rearing method for leg-swelling bees |
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| CN116250510A (en) * | 2023-04-26 | 2023-06-13 | 伊犁哈萨克自治州林业科学研究院 | Artificial rearing method for leg-swelling bees |
| CN116250510B (en) * | 2023-04-26 | 2024-06-04 | 伊犁哈萨克自治州林业科学研究院 | Artificial rearing method for leg-swelling bees |
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Application publication date: 20140430 |