CN103392513B - Ferment of aweto mycelia and application of ferment - Google Patents
Ferment of aweto mycelia and application of ferment Download PDFInfo
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- CN103392513B CN103392513B CN201310352684.7A CN201310352684A CN103392513B CN 103392513 B CN103392513 B CN 103392513B CN 201310352684 A CN201310352684 A CN 201310352684A CN 103392513 B CN103392513 B CN 103392513B
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Abstract
The invention relates to a ferment of aweto mycelia and application of the ferment, which can be used for preparing anti-fatigue and immunity-enhancing products. The aweto mycelia are obtained by conducting strain collection, preservation and rejuvenation on wild aweto Cordycepssinensis (Berk.) Sacc. sporocarp, manufacturing a stock culture, a primary culture and a secondary production culture, preparing finished aweto mycelia products and extracting effective ingredients of the finished aweto mycelia products. The method for preparing the aweto mycelia has the advantages that the process is simple and environment-friendly; no waste liquor is discharged; the cost is low. Therefore, the practicability and the maneuverability are high. Experimental results show that the effective ingredients of the aweto mycelia prepared according to the method can remarkably prolong the swimming time of a burdened rat. Therefore, the functions of immunity enhancing and fatigue preventing are favorable; the aweto mycelia can be used for preparing the anti-fatigue and immunity-enhancing products.
Description
Technical field
The present invention relates to a kind of aweto mycelium, relate in particular to a kind of Cordyceps sinensis (Cordyceps sinensis(Berk.) Sace.) mycelial effective ingredient extract and application thereof.
Background technology
Cordyceps sinensis is the complex that section ergot fungus cordyceps sinensis bacterium Cordyceps sinensis (Berk.) Sacc. colonizes in stroma on bat pretty young woman larva and larva corpse.Early summer stroma goes out head, the Dan Sheng of larva, rare 2 ~ 3, bears from host's head, long 4 ~ 11cm, base diameter 1.5 ~ 4mm, and upwards gradually carefully narrow, head enlarges into closely cylindric, brown, long 1 ~ 4.5cm, diameter 2.5 ~ 6mm.The nearly surface of perithecium is raw, and base portion slightly sinks in stroma, oval to avette.Early summer stroma is unearthed, spore takes when not dispersing, and shines to 6 seventy percent dry, removes quasi-fibrous attachment and impurity, dry or dry.Cordyceps known is in the world more than 400 kinds, and Chinese caterpillar fungus can not be equal to and Cordyceps sinensis, not substituting nominally, on medical value, clinical medicine more can not be equal to.The main finger being wherein called Cordyceps sinensis originates in the kind in China Qinghai and Tibet.In general, good than other places of Qinghai, two places, the Tibet Cordyceps sinensis interior quality of producing.Study according to modern pharmacology, Cordyceps of Qinghai Province contains cordycepic acid about 7%, protein about 25%, fat about 8.4%, wherein 82.2% can not synthesize and required unsaturated fatty acid for human body, also containing carbohydrate 28.9%, free amino acid 12 kinds, hydrolyzate amino acid/11 8 kinds, 8 seed amino acids that wherein adult must supply from food all possess, and also have physical growth of children to grow necessary histidine.In addition, still containing multi-vitamin B12, ergot lipidol, hexose alcohol, alkaloid etc.
Support worm, cultivation and educate the necessary condition that grass is Artificial Cultured Cordyceps Sinensis.
Cordyceps militaris is also known as Cordceps militaris, for the stroma (grass) of Clavicipitaceae fungi pupa grass, stroma head is oval, the blunt circle in top, and color is orange or orange red, handle is elongated, cylindrical, host is Noctuidae larva, just dead after often growing pupa, so polypide is oval pupa can't see polypide, differentiate than being easier to.The nineties in 20th century, the scientific and technical personnel of Shanghai academy of agricultural sciences complete the research of the artificial culture technique of Cordceps militaris, certainly, the Cordyceps sinensis of this and really, another kind of Cordyceps, although the Chinese caterpillar fungus composition contained by two kinds is most of identical, content is similar to, medical value and Cordyceps sinensis or incomplete same.The cultivation of current artificial aweto, be mostly inoculation that the wild cordyceps fruit body adopted obtains to peptone be main component medium on, and be different from of the present invention using bat moth larvae as completely bionical medium.
Summary of the invention
In order to solve prior art exist manually bat moth larvae direct infection Cordyceps sinensis fungi cannot be obtained this predicament of the Fermented Cordyceps consistent with wild cordyceps outward appearance efficiently, the invention provides a kind of aweto mycelium fermentation product and application thereof.
Technical scheme of the present invention is: a kind of aweto mycelium, is prepared by following steps:
The first step, carries out tissue and is separated or gathers bacterial classification spore, cultivate, obtain pure bacterial strain, and carry out rejuvenation on Chinese caterpillar fungus culture medium by wild cordyceps Cordyceps sinensis (Berk.) Sacc. fruit body; Described Chinese caterpillar fungus culture medium is the adjust ph that adds water after the making beating of bat moth larvae or dry product bat moth larvae being pulverized is after 6 ~ 6.5, then adds agar and make test tube slant or flat board, solidify, after sterilizing as medium;
Second step, the pure bacterial strain first step obtained accesses in aseptic Chinese caterpillar fungus culture medium, pH value 6-6.5, air humidity 65-85%, oxygen content volume fraction is greater than 20%, and at 10 ~ 15 DEG C, light culture is after 15 ~ 35 days, illumination 4 ~ 6 days, chooses the very fast and bacterial classification that mycelial growth is neat of annesl as mother's kind;
3rd step, plants above-mentioned mother and is seeded in aseptic Chinese caterpillar fungus culture medium after rejuvenation is selected, cultivate 10 ~ 20 days, choose dense, the sturdy mycelium of mycelia as first class inoculum, the same first step of described Chinese caterpillar fungus culture medium;
4th step, inoculates the first class inoculum in the 3rd step in aseptic Chinese caterpillar fungus culture medium and cultivates, and chooses dense, the sturdy mycelium of mycelia and to produce kind as secondary, identical with the first step of described Chinese caterpillar fungus culture medium;
5th step, in an aseptic environment, being produced to plant by secondary inoculates in Chinese caterpillar fungus culture medium, carry out lucifuge cultivate within 9 ~ 14 days, cover with medium to mycelia after, carry out illumination cultivation, light intensity 400 ~ 1000lx, relative air humidity 80% ~ 90%, 10 ~ 15 DEG C, after 7 ~ 12 days, obtain finished product aweto mycelium; After the making beating of bat moth larvae or dry product bat moth larvae are pulverized by described Cordyceps sinensis solid culture medium, adjust ph to 6 ~ 6.5.
Described bat moth larvae is dry product or fresh goods bat moth larvae.
The active ingredient concentrate of described aweto mycelium, method for making is: the finished product aweto mycelium that the 5th step obtains is added its dry product quality 50-100 times of distilled water heating and refluxing extraction 1 ~ 3 time, suction filtration while hot, merging filtrate, is concentrated into 30% ~ 50% of filtrate volume and obtains aweto mycelium active ingredient concentrate.
Described aweto mycelium is preparing the application in develop immunitypty and anti-fatigue product.
The application in develop immunitypty and anti-fatigue product prepared by the effective ingredient concentrate of described aweto mycelium.
beneficial effect
The present invention has given up outward appearance and has unanimously caught composition consistent, combine the similar technique that Cordyceps militaris produces, first bat moth larvae is made into aseptic culture medium, and then inoculate Cordyceps sinensis fungi, improve production efficiency significantly, by " raw material is consistent " this material base with wild cordyceps, produce the aweto mycelium with wild cordyceps " effective ingredient is consistent ", and be applied to prepare develop immunitypty and anti-fatigue product.
And owing to being adopt the bat moth larvae the same with natural cordyceps to be medium, so do not need to add assistant agent in addition again, direct adjust ph is just passable.
Embodiment
Embodiment 1
A kind of preparation method of aweto mycelium:
The first step: the Cordyceps sinensis not yet launching spore gathering Nagqu or Yushu district, Qinghai area, cordyceps sporophore part is entangled with aseptic paper bag, until see the ascospore of aseptic paper bag adduction collection to this Cordyceps sinensis, then at once the paper bag being attached with spore is saved in sterile chamber and seals, storage temperature is no more than 0 DEG C, and the holding time is no more than seven days; If storage temperature is below-18 DEG C, then can be saved to 1 year; Or the whole strain of thecasporous Cordyceps sinensis 75% alcohol will do not launched repeatedly rinse 5 minutes to without after naked eyes visible foreign ,-18-4 DEG C of preservation in sterile chamber is inserted in continuation immersion at once after 5 minutes, the time is no more than seven days;
In the Cordyceps sinensis areal of gathered spore, gather live body bat moth larvae, repeatedly rinse 5 minutes extremely without after naked eyes visible foreign with 75% alcohol immediately after collection, continue immersion and at once insert-18-4 DEG C of preservation in sterile chamber after 5 minutes, the resting period is no more than 20 days; If storage temperature is below-18 DEG C, then can be saved to 1 year;
Obtained spore or whole strain Cordyceps sinensis are thinly sliced or be ground into slurry to be inoculated on Chinese caterpillar fungus culture medium and to cultivate, obtain pure bacterial strain, and carry out rejuvenation; Described Chinese caterpillar fungus culture medium is the adjust ph that adds water after the making beating of bat moth larvae or dry product bat moth larvae being pulverized is after 6 ~ 6.5, then adds agar and make test tube slant or flat board, solidify, after sterilizing as medium;
Second step, the pure bacterial strain first step obtained accesses in aseptic Chinese caterpillar fungus culture medium, and during cultivation, state modulator is temperature 12 DEG C, pH value 6.2, air humidity 75%, oxygen content is greater than 20%, light culture is after 25 days, illumination 5 days, chooses the very fast and bacterial classification that mycelial growth is neat of annesl as mother's kind;
3rd step: above-mentioned mother is planted and is seeded in aseptic Chinese caterpillar fungus culture medium after rejuvenation is selected, cultivate 25 days, choose dense, the sturdy mycelium of mycelia as first class inoculum, the same first step of described Chinese caterpillar fungus culture medium;
4th step, inoculates the first class inoculum in the 3rd step in aseptic Chinese caterpillar fungus culture medium and cultivates, and chooses dense, the sturdy mycelium of mycelia and to produce kind as secondary, identical with the 3rd step of described Chinese caterpillar fungus culture medium;
5th step, in an aseptic environment, produces to plant by secondary and inoculates in Chinese caterpillar fungus culture medium, carry out after lucifuge cultivates and cover with medium to mycelia in 25 days, carrying out illumination cultivation, light intensity 700lx, relative air humidity 85%, obtained finished product aweto mycelium after 9 days by 12 DEG C.Cordyceps sinensis solid culture medium described in this step adds suitable quantity of water, adjust ph to 6.2 after the making beating of bat moth larvae or dry product bat moth larvae being pulverized.
The active ingredient data (Guangdong Province's microorganism detection Spot detection) of aweto mycelium fermentation product of the present invention
Embodiment 2
A kind of preparation method of aweto mycelium:
The first step: the Cordyceps sinensis not yet launching spore gathering Nagqu or Yushu district, Qinghai area, cordyceps sporophore part is entangled with aseptic paper bag, until see the ascospore of aseptic paper bag adduction collection to this Cordyceps sinensis, then at once the paper bag being attached with spore is saved in sterile chamber and seals, storage temperature is no more than 0 DEG C, and the holding time is no more than seven days; If storage temperature is below-18 DEG C, then can be saved to 1 year; Or the whole strain of thecasporous Cordyceps sinensis 75% alcohol will do not launched repeatedly rinse 5 minutes to without after naked eyes visible foreign ,-18-4 DEG C of preservation in sterile chamber is inserted in continuation immersion at once after 5 minutes, the time is no more than seven days;
In the Cordyceps sinensis areal of gathered spore, gather live body bat moth larvae, repeatedly rinse 5 minutes extremely without after naked eyes visible foreign with 75% alcohol immediately after collection, continue immersion and at once insert-18-4 DEG C of preservation in sterile chamber after 5 minutes, the resting period is no more than 20 days; If storage temperature is below-18 DEG C, then can be saved to 1 year;
Obtained spore or whole strain Cordyceps sinensis are thinly sliced or be ground into slurry to be inoculated on Chinese caterpillar fungus culture medium and to cultivate, obtain pure bacterial strain, and carry out rejuvenation; Described Chinese caterpillar fungus culture medium is the adjust ph that adds water after the making beating of bat moth larvae or dry product bat moth larvae being pulverized is after 6, then adds agar and make test tube slant or flat board, solidify, after sterilizing as medium;
Second step, the pure bacterial strain first step obtained accesses in aseptic Chinese caterpillar fungus culture medium, and during cultivation, state modulator is temperature 10 DEG C, pH value 6, air humidity 65%, oxygen content is greater than 20%, light culture is after 35 days, illumination 6 days, chooses the very fast and bacterial classification that mycelial growth is neat of annesl as mother's kind;
3rd step: above-mentioned mother is planted and is seeded in aseptic Chinese caterpillar fungus culture medium after rejuvenation is selected, cultivate 40 days, choose dense, the sturdy mycelium of mycelia as first class inoculum, the same first step of described Chinese caterpillar fungus culture medium;
4th step, inoculates the first class inoculum in the 3rd step in aseptic Chinese caterpillar fungus culture medium and cultivates, and chooses dense, the sturdy mycelium of mycelia and to produce kind as secondary, identical with the 3rd step of described Chinese caterpillar fungus culture medium;
5th step, in an aseptic environment, produces to plant by secondary and inoculates in Chinese caterpillar fungus culture medium, carry out after lucifuge cultivates and cover with medium to mycelia in 40 days, carrying out illumination cultivation, light intensity 400lx, relative air humidity 80%%, obtained finished product aweto mycelium after 12 days by 10 DEG C.Cordyceps sinensis solid culture medium described in this step adds suitable quantity of water, adjust ph to 6 after the making beating of bat moth larvae or dry product bat moth larvae being pulverized.
Embodiment 3
A kind of preparation method of aweto mycelium:
The first step: the Cordyceps sinensis not yet launching spore gathering Nagqu or Yushu district, Qinghai area, cordyceps sporophore part is entangled with aseptic paper bag, until see the ascospore of aseptic paper bag adduction collection to this Cordyceps sinensis, then at once the paper bag being attached with spore is saved in sterile chamber and seals, storage temperature is no more than 0 DEG C, and the holding time is no more than seven days; If storage temperature is below-18 DEG C, then can be saved to 1 year; Or the whole strain of thecasporous Cordyceps sinensis 75% alcohol will do not launched repeatedly rinse 5 minutes to without after naked eyes visible foreign ,-18-4 DEG C of preservation in sterile chamber is inserted in continuation immersion at once after 5 minutes, the time is no more than seven days;
In the Cordyceps sinensis areal of gathered spore, gather live body bat moth larvae, repeatedly rinse 5 minutes extremely without after naked eyes visible foreign with 75% alcohol immediately after collection, continue immersion and at once insert-18-4 DEG C of preservation in sterile chamber after 5 minutes, the resting period is no more than 20 days; If storage temperature is below-18 DEG C, then can be saved to 1 year;
Obtained spore or whole strain Cordyceps sinensis are thinly sliced or be ground into slurry to be inoculated on Chinese caterpillar fungus culture medium and to cultivate, obtain pure bacterial strain, and carry out rejuvenation; Described Chinese caterpillar fungus culture medium is the adjust ph that adds water after the making beating of bat moth larvae or dry product bat moth larvae being pulverized is after 6.5, then adds agar and make test tube slant or flat board, solidify, after sterilizing as medium;
Second step, the pure bacterial strain first step obtained accesses in aseptic Chinese caterpillar fungus culture medium, and during cultivation, state modulator is temperature 15 DEG C, pH value 6.5, air humidity 85%, oxygen content is greater than 20%, light culture is after 15 days, illumination 4 days, chooses the very fast and bacterial classification that mycelial growth is neat of annesl as mother's kind;
3rd step: above-mentioned mother is planted and is seeded in aseptic Chinese caterpillar fungus culture medium after rejuvenation is selected, cultivate 10 days, choose dense, the sturdy mycelium of mycelia as first class inoculum, the same first step of described Chinese caterpillar fungus culture medium;
4th step, inoculates the first class inoculum in the 3rd step in aseptic Chinese caterpillar fungus culture medium and cultivates, and chooses dense, the sturdy mycelium of mycelia and to produce kind as secondary, identical with the 3rd step of described Chinese caterpillar fungus culture medium;
5th step, in an aseptic environment, produces to plant by secondary and inoculates in Chinese caterpillar fungus culture medium, carry out after lucifuge cultivates and cover with medium to mycelia in 10 days, carrying out illumination cultivation, light intensity 1000lx, relative air humidity 90%, obtained finished product aweto mycelium after 7 days by 10 ~ 15 DEG C.Cordyceps sinensis solid culture medium described in this step adds suitable quantity of water, adjust ph to 6.5 after the making beating of bat moth larvae or dry product bat moth larvae being pulverized.
Embodiment 4
A kind of active ingredient concentrate of aweto mycelium, method for making is: finished product aweto mycelium embodiment 1 ~ 3 obtained adds its dry product quality 50-100 times of distilled water heating and refluxing extraction 1 ~ 3 time, suction filtration while hot, merging filtrate, is concentrated into 30% ~ 50% of filtrate volume and obtains aweto mycelium active ingredient concentrate.
Embodiment 5
The SPF level KM male mice of learning from else's experience after quarantining is as laboratory animal, and often to organize 10/cage, group support, ad lib is drunk water for condition is tested.The experimental group concentrate 1.2gkg of the aweto mycelium of embodiment 1
-1d
-1for dosage, every day gives corresponding mouse by 0.2ml/10g body weight gavage; Distilled water control group gives the distilled water of equivalent; Anshen Bunao mixture (Jilin Aodong Yanbian Medicine Industry Co., Ltd) group gives Anshen Bunao mixture by 0.2ml/10g body weight; Once a day, successive administration 30 days.Carry out swimming with a load attached to the body experiment after 30 days, experimental result shows aweto mycelium energy significant prolongation mice burden swimming time of the present invention (result of the test: swimming average time of control group is 25.56 minutes; Swimming average time of Anshen Bunao mixture group is 34 minutes; 45 minutes average times of swimming of experimental group).
Claims (5)
1. an aweto mycelium, is characterized in that, is prepared by following steps:
The first step, carries out tissue and is separated or gathers bacterial classification spore, cultivate, obtain pure bacterial strain, and carry out rejuvenation on Chinese caterpillar fungus culture medium by wild cordyceps Cordyceps sinensis (Berk.) Sacc. fruit body; Described Chinese caterpillar fungus culture medium is that after the making beating of bat moth larvae or dry product bat moth larvae being pulverized, adjust ph is after 6 ~ 6.5, then adds agar and make test tube slant or flat board, solidify, after sterilizing as medium;
Second step, the pure bacterial strain first step obtained accesses in aseptic Chinese caterpillar fungus culture medium, pH value 6-6.5, air humidity 65-85%, oxygen content volume fraction is greater than 20%, and at 10 ~ 15 DEG C, light culture is after 15 ~ 35 days, illumination 4 ~ 6 days, chooses the very fast and bacterial classification that mycelial growth is neat of annesl as mother's kind;
3rd step, plants above-mentioned mother and is seeded in aseptic Chinese caterpillar fungus culture medium after rejuvenation is selected, cultivate 10 ~ 20 days, choose dense, the sturdy mycelium of mycelia as first class inoculum, the same first step of described Chinese caterpillar fungus culture medium;
4th step, inoculates the first class inoculum in the 3rd step in aseptic Chinese caterpillar fungus culture medium and cultivates, and chooses dense, the sturdy mycelium of mycelia and to produce kind as secondary, identical with the first step of described Chinese caterpillar fungus culture medium;
5th step, in an aseptic environment, being produced to plant by secondary inoculates in Chinese caterpillar fungus culture medium, carry out lucifuge cultivate within 9 ~ 14 days, cover with medium to mycelia after, carry out illumination cultivation, light intensity 400 ~ 1000lx, relative air humidity 80% ~ 90%, 10 ~ 15 DEG C, after 7 ~ 12 days, obtain finished product aweto mycelium; After the making beating of bat moth larvae or dry product bat moth larvae are pulverized by described Cordyceps sinensis solid culture medium, adjust ph to 6 ~ 6.5.
2. aweto mycelium as claimed in claim 1, it is characterized in that, described bat moth larvae is dry product or fresh goods bat moth larvae.
3. the active ingredient concentrate of aweto mycelium according to claim 1, it is characterized in that, method for making is: the finished product aweto mycelium that the 5th step obtains is added its dry product quality 50-100 times of distilled water heating and refluxing extraction 1 ~ 3 time, suction filtration while hot, merging filtrate, is concentrated into 30% ~ 50% of filtrate volume and obtains aweto mycelium active ingredient concentrate.
4. aweto mycelium according to claim 1 is preparing the application in develop immunitypty and anti-fatigue product.
5. the application in develop immunitypty and anti-fatigue product prepared by the effective ingredient concentrate of aweto mycelium according to claim 3.
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| CN104145719B (en) * | 2014-09-04 | 2016-06-08 | 重庆市中药研究院 | A kind of Cordyceps fungus mycelium fermentation production method |
| CN104904496A (en) * | 2015-06-18 | 2015-09-16 | 上海善力健生物科技有限公司 | Preparation method for cordyceps sinensis mycelium |
| CN105331543A (en) * | 2015-10-16 | 2016-02-17 | 杨发国 | Production technology of anti-cancer anti-fatigue solid cordyceps sinensis fungal powder |
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| CN102845218A (en) * | 2012-08-23 | 2013-01-02 | 大化金玉草生物科技有限责任公司 | Original ecological separation method of cordyceps sinensis hirsutella sinensis |
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