CN106797804A - A kind of method that silkworm using abnormal early stage cultivates Cordyceps militaris - Google Patents
A kind of method that silkworm using abnormal early stage cultivates Cordyceps militaris Download PDFInfo
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- KQLDDLUWUFBQHP-UHFFFAOYSA-N Cordycepin Natural products C1=NC=2C(N)=NC=NC=2N1C1OCC(CO)C1O KQLDDLUWUFBQHP-UHFFFAOYSA-N 0.000 description 1
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- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- OFEZSBMBBKLLBJ-BAJZRUMYSA-N cordycepin Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)C[C@H]1O OFEZSBMBBKLLBJ-BAJZRUMYSA-N 0.000 description 1
- OFEZSBMBBKLLBJ-UHFFFAOYSA-N cordycepine Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(CO)CC1O OFEZSBMBBKLLBJ-UHFFFAOYSA-N 0.000 description 1
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- 235000013399 edible fruits Nutrition 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
- A01K67/04—Silkworms
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Mycology (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Mushroom Cultivation (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to a kind of method that silkworm using abnormal early stage cultivates Cordyceps militaris, the method be the silkworm with live body metamorphosis early stage as host, cordyceps are inoculated with using the method for injection, because the silkworm body of abnormal early stage is big, body fluid is few, the features such as love, so inoculation survival rate is greatly improved.Inorganic salt concentration in bacterium culture medium is doubled and injected again by the present invention before inoculation, and the inorganic salt environment for providing and being more suitable for grow in silkworm chrysalis for bacterium, will hair bacterium time shortening 18%, growth time shortening 15.6%.After using the method for the present invention, the rigid rate of the silkworm of abnormal early stage reaches 94%~100%, and fructification extracting rate long reaches 90%~100%；Cordyceps militaris breeding method of the invention has easy to operate, cultivation low cost and experiment high repeatability and other advantages, it is adaptable to extensive cultivation, has broad application prospects.
Description
Technical field
The present invention relates to field of edible fungus culture, a kind of method for being related to silkworm using abnormal early stage to cultivate Cordyceps militaris.
Background technology
Cordyceps militaris shows also known as Cordceps militaris, research, species and content and the Cordyceps sinensis phase of Cordyceps militaris active chemical
It is same or similar, also there is same or analogous function to human body, they contain cordycepic acid, cordycepin, Cordyceps sinensis polysaccharide, SO days enzymes
With the bioactivator such as ergosterol, with enhance immunity, antitumor, anti-oxidant, antifatigue, radioresistance, hypoglycemic, drop
Blood fat and anti-aging etc. are acted on.
2009, Ministry of Health of the People's Republic of China announced No. 3, and approval Cordyceps militaris is new resource food, amount≤2
Gram/day.2013, Ministry of Health of the People's Republic of China announced No. 1, now ratified Tea Flower, Suaeda salsa seed oil, U.S. rattan really
Oil, rhus glabra oil, Guangdong Cordyceps militaris fruiting body, A Sayi fruit and the lobate layer bacterium fermentation mycelium of currant are new resource food.
Production technology summary is Guangdong Cordyceps militaris strain through inoculation, culture, dries and is made after harvesting fructification, amount≤3 gram/day.Together
When Cordyceps militaris by social recognition, its promotion prospect is huge.
Silkworm chrysalis is posted as Cordyceps militaris, it is desirable to which silkworm chrysalis is full fresh, such as silkworm pupate after the time it is long, shell soften, in vivo
Nutrients is consumed, and contamination rate is high.The full bacterium amount of injection well of silkworm chrysalis is restricted height, and strain injection volume is small, and the hair bacterium time is long, microbiological contamination
Probability is high, and success rate declines.
5 age silkworms are posted as Cordyceps militaris, and silkworm plumpness is good, and strain injection volume is restricted, and 5 age silkworm activities are very big, strain
Flow out a part again after injection.The Chinese patent literature of publication number CN102440149 is using ice cube or in 4~6 DEG C of environment
Middle 20~30min of hypothermic anesthesia, reduces by 5 age silkworm activities, but the method increases operation, shifts environment, increases contamination probability, takes
When arduously take the energy.
The content of the invention
It is an object of the invention to provide a kind of method that silkworm using abnormal early stage cultivates Cordyceps militaris, it is used to solve existing
The inefficient problem of Cordyceps militaris is cultivated with silkworm chrysalis or silkworm.
To achieve the above object, the invention discloses following technical scheme：
The method that a kind of silkworm using abnormal early stage disclosed by the invention cultivates Cordyceps militaris, methods described includes：Cordyceps militaris
The preparation of strain：Cordyceps militaris inclined-plane master clock is inoculated on slant medium, activated spawn, is scraped after mycelium is shown in that light turns Huang
Mycelium on slant medium, is inoculated into fluid nutrient medium；Strain spreads cultivation：By liquid Cordyceps militaris spawn obtained above after
Continuous lucifuge culture, first static gas wave refrigerator 48h is shaken 2 times daily with 140r/min later, and 5min is shaken every time, continues to cultivate 4-5 days；
Inoculation：Liquid spawn addition dipotassium hydrogen phosphate and the magnesium sulfate obtained after spreading cultivation, make the dipotassium hydrogen phosphate and sulfuric acid in bacterium solution
The concentration of magnesium is changed into original 1.5-2 times, then is shaken 10 minutes with 140r/min, and the bacterium solution after desinfection chamber will shake up is injected through disappearing
In poison and the host's silkworm body after draining, from silkworm postmedian stomata inserting needle, side withdraw of the needle marginal not is penetrated, until untill injection is full；Secretly
Training orientation：The silkworm lucifuge culture that will be cultivated after inoculation, 11~13 giant silkworms are thoroughly ossify, and the silkworm body of 16~20 days 2/3 is covered with white
Color mycelia, bacterium completes hair；Optical culture is managed：The silkworm completed through light culture hair bacterium is placed in the container of culture Cordyceps militaris in illumination
Under conditions of cultivate 20~25 days.
Above-mentioned a kind of silkworm using abnormal early stage disclosed by the invention is cultivated in the method for Cordyceps militaris, host in the inoculation
Selection finishes cocoon and the silkworm of the abnormal early stage for becoming pupa of not casting off a skin.
Above-mentioned a kind of silkworm using abnormal early stage disclosed by the invention is cultivated in the method for Cordyceps militaris, the silkworm be tussah or
Silkworm.
Above-mentioned a kind of silkworm using abnormal early stage disclosed by the invention is cultivated in the method for Cordyceps militaris, the Cordyceps militaris spawn
Preparation in fluid nutrient medium include following components in percentage by weight：Potato 15-25%, glucose 1%-2%, sucrose
0.5%-1%, peptone 0.5%, potassium dihydrogen phosphate 0.15-0.3%, magnesium sulfate 0.1-0.15% and vitamin B1 0.001-
0.01%, remaining is distilled water.
Above-mentioned a kind of silkworm using abnormal early stage disclosed by the invention is cultivated in the method for Cordyceps militaris, is expanding in the inoculation
Liquid spawn addition dipotassium hydrogen phosphate and the magnesium sulfate obtained after training, make the quality of dipotassium hydrogen phosphate described in bacterium solution and magnesium sulfate
Percentage is changed into 0.25-0.6% and 0.15-0.3% respectively.
Above-mentioned a kind of silkworm using abnormal early stage disclosed by the invention is cultivated in the method for Cordyceps militaris, during the strain spreads cultivation
21~23 DEG C of the cultivation temperature of lucifuge culture.
Above-mentioned a kind of silkworm using abnormal early stage disclosed by the invention is cultivated in the method for Cordyceps militaris, the light culture management
In cultivation temperature be 15~17 DEG C, culture humidity be 45%~55%.
Above-mentioned a kind of silkworm using abnormal early stage disclosed by the invention is cultivated in the method for Cordyceps militaris, the optical culture management
In intensity of illumination be 200~400Lx, cultivation temperature be 20~23 DEG C, culture humidity be 85%~95%.
The invention has the advantages that：
The method be with live body metamorphosis early stage silkworm as host, using injection method be inoculated with cordyceps because abnormal
The silkworm body of early stage is big, and body fluid is few, the features such as love, so inoculation survival rate is greatly improved.The present invention is before inoculation by bacterium
Inorganic salt concentration is doubled and injected again in kind of culture medium, and the inorganic salt environment for providing and being more suitable for, general grows in silkworm chrysalis for bacterium
The hair bacterium time shortens 26.7%, and growth time shortens 15.6%.After using the method for the present invention, the rigid rate of the silkworm of abnormal early stage
94%~100% is reached, fructification extracting rate long reaches 90%~100%；Cordyceps militaris breeding method of the invention has operation side
Just low cost and experiment high repeatability and other advantages, are cultivated, it is adaptable to extensive cultivation, is had broad application prospects.
Specific embodiment
Following examples are used to illustrate the present invention, but are not limited to the scope of the present invention.
Embodiment 1
With same strain, same method selects 50 silkworms and 50 mulberry silkworm chrysalis to be host respectively, specific to utilize
The method and step that the silkworm of abnormal early stage cultivates Cordyceps militaris is as follows：
(1) selection of host：Cocoon is finished in selection and the silkworm of change pupa of not casting off a skin is host, selects the mulberry silkworm chrysalis just changed in quality and pupated
It is host；
(2) preparation of Cordyceps militaris spawn：The preparation of Cordyceps militaris spawn：Cordyceps militaris inclined-plane parent species are inoculated in slant medium
On, activated spawn, after mycelium is shown in that light turns Huang, the mycelium on scraping slant medium is inoculated into fluid nutrient medium, institute
The fluid nutrient medium stated in the preparation of Cordyceps militaris spawn includes following components in percentage by weight：Potato 20%, glucose 2%,
Sucrose 0.5%, peptone 0.5%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.1% and vitamin B1 0.01%, remaining is steaming
Distilled water；
(3) strain spreads cultivation：The liquid spawn that step (2) is obtained continues lucifuge culture, and 21~23 DEG C of cultivation temperature is static
Culture 48h, is shaken 2 times, each 5min daily with 140r/min later, continues to cultivate 4-5 days；
(4) it is inoculated with：Liquid spawn addition dipotassium hydrogen phosphate and the magnesium sulfate that step (3) is obtained, make its concentration be changed into phosphorus
Sour hydrogen dipotassium 0.3%, magnesium sulfate 0.15% shakes 10min with 140r/min, and the cocoon before choosing metamorphosis cuts off cocoon and takes out silkworm, disappears
Poison, drain, in and host's silkworm body after draining sterilized in the bacterium solution injection after desinfection chamber will shake up；Injecting method：From silkworm
Postmedian stomata inserting needle, side withdraw of the needle marginal not is penetrated, until untill injection is full；
(5) light culture management：The silkworm that will be cultivated through step (4) and mulberry silkworm chrysalis lucifuge culture, cultivation temperature are 15~17
DEG C, culture humidity is 45%~55%, is checked in time, and choose the silkworm being infected by bacteria, 11~13 giant silkworms are thoroughly ossify, 16~20
Its 2/3 silkworm body is covered with white hypha, and bacterium completes hair；
(6) optical culture management：The silkworm and mulberry silkworm chrysalis that are completed through step (5) culture hair bacterium are placed in the appearance of culture Cordyceps militaris
Device is cultivated under conditions of illumination, intensity of illumination be 200~400Lx, cultivation temperature be 20~23 DEG C, culture humidity be 85%~
95%, incubation time is 20~25 days.
The mulberry silkworm chrysalis of table 1 and silkworm hair bacterium end time contrast table
| Number of days | 15 | 16 | 17 | 18 | 19 | 20 | 21 |
| Mulberry silkworm chrysalis hair bacterium terminates number | 0 | 0 | 2 | 5 | 14 | 19 | 2 |
| Silkworm hair bacterium terminates number | 0 | 5 | 35 | 9 | 0 | 0 | 0 |
Send out bacterium in above-mentioned table 1 to terminate white hair occur by silkworm body 2/3rds to be defined, as can be seen from Table 1 the hair bacterium of mulberry silkworm chrysalis
Time is considerably longer than the hair bacterium time of silkworm.
The mulberry silkworm chrysalis of table 2 and silkworm harvesting end time contrast table
| Number of days | 19 | 20 | 21 | 22 | 23 | 24 | 25 | 26 |
| Mulberry silkworm chrysalis harvesting terminates number | 0 | 0 | 0 | 1 | 4 | 14 | 13 | 6 |
| Silkworm harvesting terminates number | 0 | 8 | 21 | 17 | 1 | 0 | 0 | 0 |
Harvested in above-mentioned table 2 and terminate to be defined by fructification spore long, the collecting time of mulberry silkworm chrysalis is obvious as can be seen from Table 2
It is longer than the collecting time of silkworm.
The mulberry silkworm chrysalis of table 3 and silkworm cultivate Cordyceps militaris interpretation of result contrast table
| Project | The hair bacterium time (my god) | Ossify rate (%) | Collecting time (my god) | Go out careless (%) |
| Mulberry silkworm chrysalis | 19.3 | 84 | 24.5 | 90 |
| Silkworm | 17.1 | 98 | 21.2 | 96 |
In above-mentioned table 3, collecting time is counted from after optical culture management, and rigid rate is ossify number and total ratio；Go out
Carelessness is ratio of the number with sum for grass.As can be seen from Table 3 mulberry silkworm chrysalis can be drawn from hair bacterium time, collecting time
Silkworm is considerably longer than, but the rate that ossifys saves 14.9%, harvesting with careless silkworm apparently higher than mulberry silkworm chrysalis, the hair bacterium time of silkworm is gone out
Time reduces 15.6%, so silkworm is adapted to be host than mulberry silkworm chrysalis.
Embodiment 2
With same strain, same method selects 50 tussahs and 50 tussah chrysalis to be host respectively, specific to utilize
The method and step that the tussah of abnormal early stage cultivates Cordyceps militaris is as follows：
(1) selection of host：Cocoon is finished in selection and the tussah of change pupa of not casting off a skin is host, selects the tussah chrysalis just changed in quality and pupated
It is host；
(2) preparation of Cordyceps militaris spawn：The preparation of Cordyceps militaris spawn：Cordyceps militaris inclined-plane parent species are inoculated in slant medium
On, activated spawn, after mycelium is shown in that light turns Huang, the mycelium on scraping slant medium is inoculated into fluid nutrient medium, institute
The fluid nutrient medium stated in the preparation of Cordyceps militaris spawn includes following components in percentage by weight：Potato 19%, glucose 1%,
Sucrose 1%, peptone 0.5%, potassium dihydrogen phosphate 0.2%, magnesium sulfate 0.12% and vitamin B1 0.005%, remaining is distillation
Water.
(3) strain spreads cultivation：The liquid spawn that step (2) is obtained continues lucifuge culture, and 21~23 DEG C of cultivation temperature is static
Culture 48h, is shaken 2 times, each 5min daily with 140r/min later, continues to cultivate 4-5 days；
(4) it is inoculated with：Liquid spawn addition dipotassium hydrogen phosphate and the magnesium sulfate that step (3) is obtained, make its concentration be changed into phosphorus
Sour hydrogen dipotassium 0.4%, magnesium sulfate 0.24% shakes 10min with 140r/min, and the cocoon before choosing metamorphosis cuts off cocoon and takes out tussah, disappears
Poison, drain, in and host's silkworm body after draining sterilized in the bacterium solution injection after desinfection chamber will shake up；Injecting method：From tussah
Postmedian stomata inserting needle, side withdraw of the needle marginal not is penetrated, until untill injection is full；
(5) light culture management：The silkworm that will be cultivated through step (4) and mulberry silkworm chrysalis lucifuge culture, cultivation temperature are 15~17
DEG C, culture humidity is 45%~55%, is checked in time, and choose the silkworm being infected by bacteria, 11~13 giant silkworms are thoroughly ossify, 16~20
Its 2/3 silkworm body is covered with white hypha, and bacterium completes hair；
(6) optical culture management：The tussah and tussah chrysalis that are completed through step (5) culture hair bacterium are placed in the appearance of culture Cordyceps militaris
Device is cultivated under conditions of illumination, intensity of illumination be 200~400Lx, cultivation temperature be 20~23 DEG C, culture humidity be 85%~
95%, incubation time is 20~25 days.
The tussah chrysalis of table 4 and tussah hair bacterium end time contrast table
| Number of days | 16 | 17 | 18 | 19 | 20 | 21 | 22 |
| Tussah chrysalis hair bacterium terminates number | 6 | 19 | 15 | 4 | |||
| Tussah hair bacterium terminates number | 3 | 34 | 10 | 1 |
Send out bacterium in above-mentioned table 4 to terminate white hair occur by silkworm body 2/3rds to be defined, as can be seen from Table 4 the hair bacterium of tussah chrysalis
Time is considerably longer than the hair bacterium time of tussah.
The tussah chrysalis of table 5 and tussah harvesting end time contrast table
| Number of days | 19 | 20 | 21 | 22 | 23 | 24 | 25 | 26 |
| Tussah chrysalis harvesting terminates number | 2 | 3 | 13 | 18 | 4 | |||
| Tussah harvesting terminates number | 3 | 18 | 18 | 8 |
Harvested in above-mentioned table 5 and terminate to be defined by fructification spore long, the collecting time of tussah chrysalis is obvious as can be seen from Table 5
It is longer than the collecting time of tussah.
The tussah chrysalis of table 6 and tussah cultivate Cordyceps militaris interpretation of result contrast table
| Project | The hair bacterium time (my god) | Ossify rate (%) | Collecting time (my god) | Go out careless (%) |
| Tussah chrysalis | 20.4 | 88 | 24.5 | 90 |
| Tussah | 17.2 | 96 | 21.7 | 98 |
In above-mentioned table 6, collecting time is counted from after optical culture management, and rigid rate is ossify number and total ratio；Go out
Carelessness is ratio of the number with sum for grass.As can be seen from Table 6 tussah chrysalis can be drawn from hair bacterium time, collecting time
Tussah is considerably longer than, but the rate that ossifys saves 18%, during harvesting with careless tussah apparently higher than tussah chrysalis, the hair bacterium time of tussah is gone out
Between reduce 12.9%, so tussah is adapted to be host than tussah chrysalis.
Although the present invention is described in detail above to have used general explanation and specific embodiment, at this
On the basis of invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Therefore,
These modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (8)
1. a kind of method that silkworm using abnormal early stage cultivates Cordyceps militaris, it is characterised in that methods described includes：
The preparation of Cordyceps militaris spawn：Cordyceps militaris inclined-plane parent species are inoculated on slant medium, activated spawn treats that mycelium is shown in light
After turning Huang, the mycelium on scraping slant medium is inoculated into fluid nutrient medium；
Strain spreads cultivation：By liquid Cordyceps militaris spawn obtained above continue lucifuge culture, static gas wave refrigerator 48h first, later with
140r/min is shaken 2 times daily, and 5min is shaken every time, continues to cultivate 4-5 days；
Inoculation：Liquid spawn addition dipotassium hydrogen phosphate and the magnesium sulfate obtained after spreading cultivation, make dipotassium hydrogen phosphate in bacterium solution and
The concentration of magnesium sulfate is changed into original 1.5-2 times, then is shaken 10 minutes with 140r/min, the bacterium solution injection after desinfection chamber will shake up
In sterilized and host's silkworm body after draining, from silkworm postmedian stomata inserting needle, side withdraw of the needle marginal not is penetrated, until injection is full being
Only；
Light culture is managed：The silkworm lucifuge culture that will be cultivated after inoculation, 11~13 giant silkworms are thoroughly ossify, the silkworm body of 16~20 days 2/3
It is covered with white hypha, bacterium completes hair；
Optical culture is managed：The silkworm completed through light culture hair bacterium is placed in the container of culture Cordyceps militaris and is cultivated under conditions of illumination
20~25 days.
2. the method that a kind of silkworm using abnormal early stage according to claim 1 cultivates Cordyceps militaris, it is characterised in that described
Tropical area finishes cocoon and the silkworm of the abnormal early stage for becoming pupa of not casting off a skin in inoculation.
3. the method that a kind of silkworm using abnormal early stage according to claim 2 cultivates Cordyceps militaris, it is characterised in that described
Silkworm is tussah or silkworm.
4. the method that a kind of silkworm using abnormal early stage according to claim 1 cultivates Cordyceps militaris, it is characterised in that described
Fluid nutrient medium in the preparation of Cordyceps militaris spawn includes following components in percentage by weight：Potato 15-25%, glucose 1%-
2%th, sucrose 0.5%-1%, peptone 0.5%, potassium dihydrogen phosphate 0.15-0.3%, magnesium sulfate 0.1-0.15% and vitamin B1
0.001-0.01%, remaining is distilled water.
5. the method that a kind of silkworm using abnormal early stage according to claim 1 cultivates Cordyceps militaris, it is characterised in that described
Liquid spawn addition dipotassium hydrogen phosphate and the magnesium sulfate obtained after spreading cultivation in inoculation, make dipotassium hydrogen phosphate described in bacterium solution and sulphur
The mass percent of sour magnesium is changed into 0.25-0.6% and 0.15-0.3% respectively.
6. the method that a kind of silkworm using abnormal early stage according to claim 1 cultivates Cordyceps militaris, it is characterised in that described
Strain spreads cultivation 21~23 DEG C of the cultivation temperature of middle lucifuge culture.
7. the method that a kind of silkworm using abnormal early stage according to claim 1 cultivates Cordyceps militaris, it is characterised in that described
Cultivation temperature in light culture management is 15~17 DEG C, and culture humidity is 45%~55%.
8. the method that a kind of silkworm using abnormal early stage according to claim 1 cultivates Cordyceps militaris, it is characterised in that described
Intensity of illumination in optical culture management is 200~400Lx, and cultivation temperature is 20~23 DEG C, and culture humidity is 85%~95%.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107810781A (en) * | 2017-11-10 | 2018-03-20 | 广西沙田仙人滩农业投资有限公司 | A kind of method that silkworm living cultivates Cordyceps militaris |
| CN109328858A (en) * | 2018-08-02 | 2019-02-15 | 河北晶懋生物医药科技有限公司 | The method of bag tea, candy, drink and wine and its bag tea of preparation, candy, drink and wine are prepared using Cordyceps militaris |
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| CN112772282A (en) * | 2021-03-11 | 2021-05-11 | 齐齐哈尔大学 | Cultivation method of cordyceps militaris |
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1094091A (en) * | 1994-02-05 | 1994-10-26 | 李春燕 | The seed selection of cordyceps militaris excellent species and high yield cultivating method |
| CN102100152A (en) * | 2010-11-17 | 2011-06-22 | 鲁东大学 | Artificial culture method and culture medium for fruiting bodies of cordyceps militaris |
| CN102440149A (en) * | 2011-10-10 | 2012-05-09 | 江苏科技大学 | Method for cultivating cordyceps militaris by using silkworm larvae |
| CN104164367A (en) * | 2014-08-01 | 2014-11-26 | 沁阳市西向食用菌研究所 | Dried silkworm cordyceps militaris and culture method thereof |
| CN104472208A (en) * | 2014-11-11 | 2015-04-01 | 冀宏 | Liquid culture method for cordyceps militaris stroma |
| CN105733957A (en) * | 2016-03-01 | 2016-07-06 | 鲁东大学 | Method for culturing cordyceps militaris with slant solid strains effectively instead of liquid strains |
| CN106244463A (en) * | 2016-08-01 | 2016-12-21 | 鲁东大学 | The cultural method of the Cordyceps mycelium of selenium-rich high cordycepin content |
-
2017
- 2017-01-24 CN CN201710062706.4A patent/CN106797804A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1094091A (en) * | 1994-02-05 | 1994-10-26 | 李春燕 | The seed selection of cordyceps militaris excellent species and high yield cultivating method |
| CN102100152A (en) * | 2010-11-17 | 2011-06-22 | 鲁东大学 | Artificial culture method and culture medium for fruiting bodies of cordyceps militaris |
| CN102440149A (en) * | 2011-10-10 | 2012-05-09 | 江苏科技大学 | Method for cultivating cordyceps militaris by using silkworm larvae |
| CN104164367A (en) * | 2014-08-01 | 2014-11-26 | 沁阳市西向食用菌研究所 | Dried silkworm cordyceps militaris and culture method thereof |
| CN104472208A (en) * | 2014-11-11 | 2015-04-01 | 冀宏 | Liquid culture method for cordyceps militaris stroma |
| CN105733957A (en) * | 2016-03-01 | 2016-07-06 | 鲁东大学 | Method for culturing cordyceps militaris with slant solid strains effectively instead of liquid strains |
| CN106244463A (en) * | 2016-08-01 | 2016-12-21 | 鲁东大学 | The cultural method of the Cordyceps mycelium of selenium-rich high cordycepin content |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107810781A (en) * | 2017-11-10 | 2018-03-20 | 广西沙田仙人滩农业投资有限公司 | A kind of method that silkworm living cultivates Cordyceps militaris |
| CN109328858A (en) * | 2018-08-02 | 2019-02-15 | 河北晶懋生物医药科技有限公司 | The method of bag tea, candy, drink and wine and its bag tea of preparation, candy, drink and wine are prepared using Cordyceps militaris |
| CN109845580A (en) * | 2019-02-27 | 2019-06-07 | 湖南省蚕桑科学研究所 | A method of with 5 instar larvae of silkworm for host's artificial culture snowy peak cordyceps sinensis |
| CN110692434A (en) * | 2019-10-15 | 2020-01-17 | 吉林省蚕业科学研究院 | Method for culturing cordyceps sinensis of dichotomous rhinoceros |
| CN112166954A (en) * | 2020-10-27 | 2021-01-05 | 苏州家和蚕业生物科技有限公司 | Culture method of cordyceps militaris |
| WO2022088280A1 (en) * | 2020-10-27 | 2022-05-05 | 苏州家和蚕业生物科技有限公司 | Cultivating method for silkworm pupa-cultured cordyceps militaris |
| CN112772282A (en) * | 2021-03-11 | 2021-05-11 | 齐齐哈尔大学 | Cultivation method of cordyceps militaris |
| CN112772282B (en) * | 2021-03-11 | 2022-12-02 | 齐齐哈尔大学 | Cultivation method of cordyceps militaris |
| CN114793755A (en) * | 2022-04-18 | 2022-07-29 | 常熟理工学院 | Method for cultivating silkworm cordyceps militaris with high rigidification rate |
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