CN109348983A - A kind of breeding method of cordyceps sinensis - Google Patents
A kind of breeding method of cordyceps sinensis Download PDFInfo
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- CN109348983A CN109348983A CN201811051782.6A CN201811051782A CN109348983A CN 109348983 A CN109348983 A CN 109348983A CN 201811051782 A CN201811051782 A CN 201811051782A CN 109348983 A CN109348983 A CN 109348983A
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- 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
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- 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
- A01G18/20—Culture media, e.g. compost
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- 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
- A01G18/30—Accessories for use before inoculation of spawn, e.g. sterilisers
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- 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
- A01G18/50—Inoculation of spawn
-
- 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
- A01G18/60—Cultivation rooms; Equipment therefor
- A01G18/69—Arrangements for managing the environment, e.g. sprinklers
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- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Mushroom Cultivation (AREA)
Abstract
The present invention relates to cordyceps sinensis technical fields, more particularly to a kind of breeding method of cordyceps sinensis, the breeding method comprises the following steps 1) preparation of culture medium: the high-quality green silkworm chrysalis of selection, is immersed in culture solution after 35min and is fitted into glass culture bottle, obtain culture medium；Wherein, the formula rate of the culture solution are as follows: potato 80-100g, glucose 20-30g, milk powder 15-25g, potassium dihydrogen phosphate 5-7g, magnesium sulfate 3-6g add water constant volume to 1000mL；2) it sterilizes；3) it is inoculated with；4) culture bottle for accessing liquid spawn is moved into culturing room and is protected from light culture, temperature is controlled at 20 DEG C, pH is 4-9, stand 6-8h, it is subsequently put on 25 DEG C, cultivates 10-15min in the oscillation cabinet of 160r/min, magnetic treatment 40min is carried out after taking-up under certain flux condition, being subsequently placed at temperature is 20 DEG C, and pH is cultivated under the conditions of being 6-7；5) it harvests；It is long that technical solution provided by the invention effectively compensates for mycelial growth time existing for the breeding method of existing cordyceps sinensis, cultivates slow-footed defect.
Description
Technical field
The present invention relates to cordyceps sinensis technical fields, and in particular to a kind of breeding method of cordyceps sinensis.
Background technique
Cordyceps are the abbreviations of cordyceps sinensis, belong to one kind of cordyceps sinensis, are Clavicipitaceae fungies, colonize in Hepialidae
The complex of stroma and larva corpse on insect larvae.
Have and adjusts the multiple efficacies such as function of immune system, antitumor, antifatigue, theoretical, the unique formula in forward position,
Exclusive positioning is walked in front end.It is mainly distributed on Tibet, China, based on Qinghai Province, Sichuan and Yunnan, South Korea some areas have
This mushroom.
Silkworm chrysalis cordyceps sinensis is edible and medicinal to be thought to the Research on kinds for having become induction cordyceps militaris sporocarp both at home and abroad
Value can match in excellence or beauty with traditional cordyceps sinensis, and it is wild that the silkworm chrysalis cordyceps sinensis cellulose content that biotechnology is manually cultivated is higher than tradition
Tens times of cordyceps sinensis, it is that wild cordyceps are important, ideal substitute, is more and more favored by market.
The wild cordyceps of scarcity of resources are as traditional precious Chinese medicine and invigorant, and supply falls short of demand always in market.
Chinese patent CN103460981B discloses a kind of novel method for cultivating of Cordceps militaris at present, comprising the following steps: by
Test tube solid spawn access shaking flask switchs to liquid spawn, shaking flask strain access fermentor is carried out numerous, the Cordceps militaris solid of strain expansion
It cultivates cultivation box culture matrix manufacturing, liquid inoculation, cultural hypha, annesl, go out grass management, harvesting.Shake flask culture body strain is matched
Side is as follows: dried silkworm chrysalis meal 35g；Glucose 35g；Maize flour 50g；VB1 2；KH2PO4 5g； MgSO4 1.7g；Water 3500ml；
Fermentation tank culture formula is as follows: pressing the canned amount 70L of the every 100L of fermentor.Soybean powder 200g；Maize flour 1400g；Sucrose 1400g；
KH2PO4 70g； MgSO4 70g；Dried silkworm chrysalis meal 1050g；Add defoaming agent 7ml, salad oil 7-10ml in tank；The training of Cordceps militaris solid
It is as follows to educate cultivation box nutrient solution prescription: peptone 4g；KH2PO4 1g； MgSO4 1g；VB1 1；Bean cake powder 16.67g；Sugar
10g；Water 1000ml.Novel method for cultivating provided by the invention, easy to operate, cultivation period is shorter, and toxigenic capacity is lower, culture
Success rate is higher, and yield is high, also greatly improves at cordycepin content in grass.
But that there is also mycelial growth times is long for the breeding method of existing cordyceps sinensis, cultivates slow-footed defect.
Summary of the invention
In order to overcome the shortcomings of the prior art, the object of the present invention is to provide a kind of breeding methods of cordyceps sinensis, effectively
The breeding method for compensating for existing cordyceps sinensis existing for mycelial growth time it is long, cultivate slow-footed defect.
In order to achieve the above purpose, it is achieved by the following technical programs:
A kind of breeding method of cordyceps sinensis, the breeding method comprise the following steps:
1) preparation of culture medium: the high-quality green silkworm chrysalis of selection, is immersed in culture solution loading glass culture after 35min
In bottle, culture medium is obtained；
Wherein, the formula rate of the culture solution are as follows: potato 80-100g, glucose 20-30g, milk powder 15-25g, phosphorus
Acid dihydride potassium 5-7g, magnesium sulfate 3-6g, add water constant volume to 1000mL；
2) it sterilizes: culture bottle sealing being put into high-pressure sterilizing pot and is sterilized, is warming up to 100 DEG C, closes exhaust valve,
When high-pressure sterilizing pot pressure one periodically keeps 1h, taking-up is cooling；
3) it is inoculated with: aseptically, being sprayed liquid spawn with inoculating gun even in media surface；
4) it manages:
1. the culture bottle for accessing liquid spawn, which is moved to culturing room, is protected from light culture, temperature control is at 20 DEG C, and pH 4-9 is quiet
6-8h is set, 25 DEG C is subsequently put on, cultivates 10-15min in the oscillation cabinet of 160r/min, after taking-up under certain flux condition
Magnetic treatment 40min is carried out, continuing thereafter with and being placed in temperature is 20 DEG C, and pH is cultivated under the conditions of being 6-7；
2. control cultivation temperature is 25 DEG C, pH 6-7, intensity of illumination 0- when mycelia protrusion occurs in culture medium top
10Lx, relative humidity are certain.
5) it harvests: when fructification stroma is rodlike in Chinese red or crocus and no longer grows, with stainless steel pocket knife or cutting
Knife adopts down it from base portion, is put into cleaner and enters freeze-drying, is stored in sealed plastic bag after dry, transports refrigeration
Library is stored refrigerated.
Preferably, the silkworm chrysalis place of production is the pollution-free area of Rizhao inshore in the step 1).
Preferably, step 2) the mesohigh sterilizing pot inner pressure is 1.4-1.5bar.
Preferably, the mass ratio of liquid spawn and culture medium is 1:30-35 in the step 3).
Preferably, it is 6-7 that cultivation stage pH is protected from light in the step 4).
Preferably, the magnetic flux of magnetic treatment is 6mT in the step 4).
Preferably, relative humidity is 78-80% in the step 4).
Using above-mentioned technical solution, beneficial effects of the present invention are as follows:
(1) milk powder can be used as the nitrogen nutrition of cordyceps mycelial growth, and play apparent promote to the growth of cordyceps
Effect.
(2) mycelium is equal energy sprout growth in 4-9 in pH, and most fast, the higher or lower speed of growth is grown in 6-7
Slow down, significant difference.
(3) illumination is not required cordyceps, and some researches show that the mycelial speed of growth is with intensity of illumination
Enhancing and slow down, under illumination weaker condition (illumination be less than 10lx), mycelial growth is most fast.
(4) to strain carry out magnetic treatment, can be improved SOD activity, can also improve peroxidase, catalase and
The activity of a variety of enzymes such as polyphenol oxidase, the metabolism of enhancing body substance and energetic supersession, promote mycelial growth, accelerate winter worm
Careless cultivation process.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, implement below in conjunction with the present invention
Example, technical scheme in the embodiment of the invention is clearly and completely described.Based on the embodiment of the present invention, this field is general
Logical technical staff every other embodiment obtained without creative efforts, belongs to protection of the present invention
Range.
Embodiment 1:
A kind of breeding method of cordyceps sinensis, the breeding method comprise the following steps:
1) preparation of culture medium: the high-quality green silkworm chrysalis of selection, is immersed in culture solution loading glass culture after 35min
In bottle, culture medium is obtained；
Wherein, the formula rate of the culture solution are as follows: potato 80g, glucose 30g, milk powder 15g, potassium dihydrogen phosphate
7g, magnesium sulfate 3g, add water constant volume to 1000mL；
2) it sterilizes: culture bottle sealing being put into high-pressure sterilizing pot and is sterilized, is warming up to 100 DEG C, closes exhaust valve,
1h is kept when high-pressure sterilizing pot pressure is 1.5bar, is taken out, it is cooling；
3) it is inoculated with: aseptically, being sprayed liquid spawn with inoculating gun even in media surface；
4) it manages:
1. the culture bottle for accessing liquid spawn, which is moved to culturing room, is protected from light culture, temperature is controlled at 20 DEG C, pH 4, is stood
8h is subsequently put on 25 DEG C, cultivates 10min in the oscillation cabinet of 160r/min, carries out magnetic after taking-up under the flux condition of 6mT
40min is handled, being subsequently placed at temperature is 20 DEG C, and pH is cultivated under the conditions of being 7；
2. control cultivation temperature is 25 DEG C, pH 6, and intensity of illumination is when mycelia protrusion occurs in culture medium top
10Lx, relative humidity 78%；
5) it harvests: when fructification stroma is rodlike in Chinese red or crocus and no longer grows, with stainless steel pocket knife or cutting
Knife adopts down it from base portion, is put into cleaner and enters freeze-drying, is stored in sealed plastic bag after dry, transports refrigeration
Library is stored refrigerated.
Embodiment 2:
A kind of breeding method of cordyceps sinensis, the breeding method comprise the following steps:
1) preparation of culture medium: the high-quality green silkworm chrysalis of selection, is immersed in culture solution loading glass culture after 35min
In bottle, culture medium is obtained；
Wherein, the formula rate of the culture solution are as follows: potato 100g, glucose 20g, milk powder 25g, potassium dihydrogen phosphate
5g, magnesium sulfate 6g, add water constant volume to 1000mL；
2) it sterilizes: culture bottle sealing being put into high-pressure sterilizing pot and is sterilized, is warming up to 100 DEG C, closes exhaust valve,
1h is kept when high-pressure sterilizing pot pressure is 1.4bar, is taken out, it is cooling；
3) it is inoculated with: aseptically, being sprayed liquid spawn with inoculating gun even in media surface；
4) it manages:
1. the culture bottle for accessing liquid spawn, which is moved to culturing room, is protected from light culture, temperature is controlled at 20 DEG C, pH 9, is stood
6h is subsequently put on 25 DEG C, cultivates 15min in the oscillation cabinet of 160r/min, carries out magnetic after taking-up under the flux condition of 6mT
40min is handled, being subsequently placed at temperature is 20 DEG C, and pH is cultivated under the conditions of being 6；
2. when mycelia protrusion occurs in culture medium top, control cultivation temperature is 25 DEG C, pH 7, intensity of illumination 0Lx,
Relative humidity is 80%；
5) it harvests: when fructification stroma is rodlike in Chinese red or crocus and no longer grows, with stainless steel pocket knife or cutting
Knife adopts down it from base portion, is put into cleaner and enters freeze-drying, is stored in sealed plastic bag after dry, transports refrigeration
Library is stored refrigerated.
Embodiment 3:
A kind of breeding method of cordyceps sinensis, the breeding method comprise the following steps:
1) preparation of culture medium: the high-quality green silkworm chrysalis of selection, is immersed in culture solution loading glass culture after 35min
In bottle, culture medium is obtained；
Wherein, the formula rate of the culture solution are as follows: potato 90g, glucose 25g, milk powder 20g, potassium dihydrogen phosphate
6g, magnesium sulfate 5g, add water constant volume to 1000mL；
2) it sterilizes: culture bottle sealing being put into high-pressure sterilizing pot and is sterilized, is warming up to 100 DEG C, closes exhaust valve,
1h is kept when high-pressure sterilizing pot pressure is 1.4bar, is taken out, it is cooling；
3) it is inoculated with: aseptically, being sprayed liquid spawn with inoculating gun even in media surface；
4) it manages:
1. the culture bottle for accessing liquid spawn, which is moved to culturing room, is protected from light culture, temperature is controlled at 20 DEG C, pH 6, is stood
7h is subsequently put on 25 DEG C, cultivates 15min in the oscillation cabinet of 160r/min, carries out magnetic after taking-up under the flux condition of 6mT
40min is handled, being subsequently placed at temperature is 20 DEG C, and pH is cultivated under the conditions of being 6；
2. when mycelia protrusion occurs in culture medium top, control cultivation temperature is 25 DEG C, pH 6, intensity of illumination 10Lx,
Relative humidity is 80%；
5) it harvests: when fructification stroma is rodlike in Chinese red or crocus and no longer grows, with stainless steel pocket knife or cutting
Knife adopts down it from base portion, is put into cleaner and enters freeze-drying, is stored in sealed plastic bag after dry, transports refrigeration
Library is stored refrigerated.
Embodiment 4:
A kind of breeding method of cordyceps sinensis, the breeding method comprise the following steps:
1) preparation of culture medium: the high-quality green silkworm chrysalis of selection, is immersed in culture solution loading glass culture after 35min
In bottle, culture medium is obtained；
Wherein, the formula rate of the culture solution are as follows: potato 95g, glucose 25g, milk powder 20g, potassium dihydrogen phosphate
7g, magnesium sulfate 4g, add water constant volume to 1000mL；
2) it sterilizes: culture bottle sealing being put into high-pressure sterilizing pot and is sterilized, is warming up to 100 DEG C, closes exhaust valve,
1h is kept when high-pressure sterilizing pot pressure is 1.4bar, is taken out, it is cooling；
3) it is inoculated with: aseptically, being sprayed liquid spawn with inoculating gun even in media surface；
4) it manages:
1. the culture bottle for accessing liquid spawn, which is moved to culturing room, is protected from light culture, temperature is controlled at 20 DEG C, pH 7, is stood
7h is subsequently put on 25 DEG C, cultivates 10min in the oscillation cabinet of 160r/min, carries out magnetic after taking-up under the flux condition of 6mT
40min is handled, being subsequently placed at temperature is 20 DEG C, and pH is cultivated under the conditions of being 7；
2. when mycelia protrusion occurs in culture medium top, control cultivation temperature is 25 DEG C, pH 7, intensity of illumination 0Lx,
Relative humidity is 78%；
5) it harvests: when fructification stroma is rodlike in Chinese red or crocus and no longer grows, with stainless steel pocket knife or cutting
Knife adopts down it from base portion, is put into cleaner and enters freeze-drying, is stored in sealed plastic bag after dry, transports refrigeration
Library is stored refrigerated.
Keep other conditions consistent, only control is protected from light cultivation stage pH difference, tests mycelial growth rate, as a result as follows:
| PH=4 | PH=6 | PH=7 | PH=9 | |
| Mycelia growth coefficient | 15 | 48 | 49 | 20 |
Cordyceps fructification and commercially available Yushu district, Qinghai wild cordyceps fructification prepared by the embodiment of the present invention 1
Warp let-off Chinese Academy of Sciences Shenyang ecology institute's agricultural product security and environmental quality inspection center are detected, and testing result is as follows:
| Inspection project | Embodiment 1 | Wild cordyceps | Unit |
| Asparatate | 2670 | 1899 | mg/100g |
| Threonine | 1689 | 1177 | mg/100g |
| Serine | 1695 | 838 | mg/100g |
| Glutamic acid | 4060 | 14822 | mg/100g |
| Glycine | 1278 | 757 | mg/100g |
| Alanine | 2075 | 861 | mg/100g |
| Cystine | 342 | 319 | mg/100g |
| Valine | 1368 | 798 | mg/100g |
| Isoleucine | 2450 | 2607 | mg/100g |
| Leucine | 2083 | 1754 | mg/100g |
| Tyrosine | 1998 | 731 | mg/100g |
| Phenylalanine | 867 | 665 | mg/100g |
| Lysine | 846 | 862 | mg/100g |
| Histidine | 445 | 190 | mg/100g |
| Arginine | 1678 | 1206 | mg/100g |
| Proline | 1468 | 906 | mg/100g |
| Cordycepin | 1423 | 118 | mg/kg |
| Cordycepic acid | 15.25 | 8.05 | mg/100g |
| Thick many candies | 26.78 | 6.60 | g/100g |
| SOD | 593 | 189 | u/g |
| Protein | 37.7 | 36.2 | g/100g |
As seen from the above table, the silkworm chrysalis cordyceps sinensis that embodiment 1 is cultivated rich in the nutrition such as amino acid, cordycepin, cordycepic acid at
Point, and the content of the physiologically active ingredients such as cordycepic acid and cordycepin will far be more than wild cordyceps, the embodiment of the present invention is cultivated
Cordyceps sinensis have it is antitumor, keep fit and healthy, anti-bacteria and anti-virus, immunological regulation, remove a variety of pharmacology and the health care such as free radical
Effect.
The above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although with reference to the foregoing embodiments
Invention is explained in detail, those skilled in the art should understand that: it still can be to aforementioned each implementation
Technical solution documented by example is modified or equivalent replacement of some of the technical features；And these modification or
Replacement, the spirit and scope for technical solution of various embodiments of the present invention that it does not separate the essence of the corresponding technical solution.
Claims (7)
1. a kind of breeding method of cordyceps sinensis, which is characterized in that the breeding method comprises the following steps:
1) preparation of culture medium: the high-quality green silkworm chrysalis of selection, is immersed in culture solution after 35min and is fitted into glass culture bottle,
Obtain culture medium；
Wherein, the formula rate of the culture solution are as follows: potato 80-100g, glucose 20-30g, milk powder 15-25g, di(2-ethylhexyl)phosphate
Hydrogen potassium 5-7g, magnesium sulfate 3-6g, add water constant volume to 1000mL；
2) it sterilizes: culture bottle sealing being put into high-pressure sterilizing pot and is sterilized, is warming up to 100 DEG C, exhaust valve is closed, works as high pressure
Autoclave pressure one periodically keeps 1h, takes out, cooling；
3) it is inoculated with: aseptically, being sprayed liquid spawn with inoculating gun even in media surface；
4) it manages:
1. the culture bottle for accessing liquid spawn, which is moved to culturing room, is protected from light culture, temperature is controlled at 20 DEG C, pH 4-9, stands 6-
8h is subsequently put on 25 DEG C, cultivates 10-15min in the oscillation cabinet of 160r/min, carries out magnetic after taking-up under certain flux condition
40min is handled, being subsequently placed at temperature is 20 DEG C, and pH is cultivated under the conditions of being 6-7；
2. when mycelia protrusion occurs in culture medium top, control cultivation temperature is 25 DEG C, pH 6-7, intensity of illumination 0-10Lx,
Relative humidity is certain；
5) it harvests:, will with stainless steel pocket knife or scissors when fructification stroma is rodlike in Chinese red or crocus and no longer grows
It is adopted down from base portion, is put into cleaner and is entered freeze-drying, be stored in sealed plastic bag after dry, transports freezer refrigeration
It saves.
2. a kind of breeding method of cordyceps sinensis according to claim 1, which is characterized in that the silkworm chrysalis place of production is in the step 1)
Rizhao inshore is pollution-free area.
3. a kind of breeding method of cordyceps sinensis according to claim 1, which is characterized in that step 2) the mesohigh autoclave
Interior pressure is 1.4-1.5bar.
4. a kind of breeding method of cordyceps sinensis according to claim 1, which is characterized in that in the step 3) liquid spawn with
The mass ratio of culture medium is 1:30-35.
5. a kind of breeding method of cordyceps sinensis according to claim 1, which is characterized in that be protected from light culture rank in the step 4)
Section pH is 6-7.
6. a kind of breeding method of cordyceps sinensis according to claim 1, which is characterized in that the magnetic of magnetic treatment in the step 4)
Flux is 6mT.
7. a kind of breeding method of cordyceps sinensis according to claim 1, which is characterized in that relative humidity is in the step 4)
78-80%.
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