CN104909869B - Culture medium and preparation method thereof and the method using its cultivation Cordyceps militaris - Google Patents
Culture medium and preparation method thereof and the method using its cultivation Cordyceps militaris Download PDFInfo
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Abstract
A kind of culture medium and preparation method thereof and the method using its cultivation Cordyceps militaris.Culture medium includes culture solution and substrate；Culture solution includes：A great number of elements aqueous solution, trace element water-soluble liquid, organic aqueous solution, the aqueous solution of salt and glucose solution；Substrate is uncooked rice.Production method：Culture solution impregnates substrate 8 12 hours, sterilizes at 18 22 DEG C.The method for cultivating Cordyceps militaris：By culture medium inoculated Cordyceps militaris kind bacterium, cultivated in darkroom；It is put into middle cultivation between the cultivation of 24 hours full spectral illuminations；Cut is delineated in Spawn incubation primary surface；18 22 DEG C, the full spectrum of 80 100 lux of intensity of illumination cultivate between 35 days；When vaccine grows into 0.2 0.4cm by vaccine be transferred to 18 22 DEG C, the full spectrum of 190 210 lux of intensity of illumination cultivate between 23 27 days.The culture medium and method provided using the application, growth cycle is short, nutritive value is high, strong antibacterial.
Description
Technical field
The present invention relates to cordyceps sinensis to cultivate field, in particular to a kind of culture medium and preparation method thereof and uses it
The method for cultivating Cordyceps militaris.
Background technology
Cordyceps militaris is Ascomycotina, ergot Zoopagales, Clavicipitaceae, Cordyceps type sepecies.Latin literary fame is known as
Cordyceps militaris also known as northern Chinese caterpillar Fungus, Cordceps militaris etc. have some areas to be also known as the cordyceps sinensis that leaps up, worldwide distribution
Natural resources quantity is seldom.By stroma (i.e. careless part) and sclerotium (i.e. the corpse part of worm) complex dimerous.Winter
Season, larva was lived in seclusion in soil, and mushroom parasitism wherein, draws nutrition, and larva body is interior dead full of mycelia.To summer, from larva corpse
On bear seedling, likeness in form grass, the Summer Solstice front and rear acquire and obtain.The traditional Chinese medical science thinks that cordyceps sinensis enters lung kidney two warp, can tonifying lung it is cloudy and
Kidney-replenishing cures mainly kidney deficiency, and impotence and seminal emission, soreness of waist and knee joint, eak after being ill, chronic cough is weak, phthisical cough phlegm blood, and spontaneous sweating etc., is only
A kind of one Chinese medicine that can be balanced simultaneously, adjust negative and positive.
The natural yield of Cordyceps militaris far can not meet the needs of people.However, current artificial culture method there is also
Problems, such as：Culture medium entirety unbalanced nutrition, survival rate are low, and sterilizing is insufficient, disease resistance is weak, and the production cycle is long
(prior art needs 80-90 days), low output, condition are poor, and extensive inoculation, nutritive value are low etc..
In view of this, it is special to propose the present invention.
Invention content
The first object of the present invention is to provide a kind of culture medium, and the culture medium has full of nutrition, antibacterial ability
The advantages that strong.
The second object of the present invention is to provide a kind of production method of the culture medium, and this method has operation letter
The advantages that single, at low cost.
The third object of the present invention is the method for providing the use culture medium cultivation Cordyceps militaris, and this method has life
The advantages that long period is short, finished product condition is good, nutritive value is high.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
A kind of culture medium, including culture solution and substrate；The culture solution with volume percent, including：A great number of elements water
Solution 10-20%, trace element water-soluble liquid 10-20%, organic aqueous solution 10-20%, salt aqueous solution 10-20% and grape
Sugar aqueous solution 15-25%, surplus are distilled water；
In a great number of elements aqueous solution, 7-9g potassium nitrate, 13-17g ammonium nitrate, 15-20g are corresponded to per 1000ml distilled water
Potassium dihydrogen phosphate, 1-3g magnesium sulfate and 18-22g calcium nitrate；
In the trace element water-soluble liquid, 12-18g boric acid, 17-25g zinc sulfate, 1-3g sulphur are corresponded to per 1000ml distilled water
Sour manganese and 5-10g copper sulphate；
In the organic aqueous solution, 0.49-0.52g vitamin B1s, 0.18-0.22g dimensions are corresponded to per 1000ml distilled water
Raw element B6,18-22g niacin and 15-20g glycine；
In the aqueous solution of the salt, 15-20g iron sulfites and 27-31g ethanedioic acid tetraacethyls are corresponded to per 1000ml distilled water
Disodium；
In the glucose solution, 450-550g glucose is corresponded to per 1000ml distilled water；
The substrate is uncooked rice；The dosage of the culture solution and the substrate is that culture solution corresponds to use described in per 1ml
Substrate described in 0.8-1.2g.
A great number of elements aqueous solution main purpose is to provide for the nutrients such as nitrogen, phosphorus, potassium, calcium, magnesium；Trace element water-soluble
Liquid main purpose is to provide for the trace elements such as boron, zinc, manganese, copper；Organic aqueous solution main purpose is to provide for amino
Acid and vitamin；The aqueous solution main purpose of salt is to provide for sodium and ferro element；Glucose solution main purpose be in order to
Necessary nutrient is provided.
First, the material demand that can be added in solution is screened, it is ensured that Cordyceps militaris can survive on culture medium；
Secondly, the dosage of various substances has to appropriate and balanced.Therefore, each solution in the culture solution of the application offer is suitable
Volume fraction and each solution in active principle and its appropriate content, can be grown for Cordyceps militaris provide it is sufficient,
The nutrient of weighing apparatus.
The suitable proportioning of culture solution and substrate so that the sufficient utilization that raw material can be suitably is unlikely to subalimentation
It is or superfluous.
Preferably, the culture solution is with volume percent, including：A great number of elements aqueous solution 15%, trace element water-soluble
Liquid 15%, organic aqueous solution 15%, the aqueous solution 15% of salt and glucose solution 20%, surplus are distilled water.
Under above-mentioned volume fraction ratio, the effect of the medium culture Cordyceps militaris of acquisition is preferable.
It is further preferred that in a great number of elements aqueous solution, 8g potassium nitrate, 15g nitric acid are corresponded to per 1000ml distilled water
Ammonium, 18g potassium dihydrogen phosphates, 2g magnesium sulfate and 20g calcium nitrate；
In the trace element water-soluble liquid, 15g boric acid, 21g zinc sulfate, 2g manganese sulfates and 8g are corresponded to per 1000ml distilled water
Copper sulphate；
In the organic aqueous solution, 0.51g vitamin B1s, 0.20g vitamin B6s, 20g are corresponded to per 1000ml distilled water
Niacin and 18g glycine；
In the aqueous solution of the salt, 18g iron sulfites and 29g ethanedioic acid tetraacethyl disodiums are corresponded to per 1000ml distilled water；
Ethanedioic acid tetraacethyl disodium is matched with the other compositions in culture solution, enables to culture medium that can keep for a long time
Cordyceps militaris is fresh and alive.
In the glucose solution, 500g glucose is corresponded to per 1000ml distilled water.
Make culture medium using each solution under said ratio, the effect of the medium culture Cordyceps militaris of acquisition compared with
It is good.Growth cycle is most short, antibacterial ability is most strong, finished product condition is best.
The application also provides a kind of production method of the culture medium, and the method is：
It under the conditions of 18-22 DEG C, is impregnated described substrate 8-12 hours using the culture solution, then sterilized, obtained described
Culture medium.
It is impregnated under the conditions of suitable temperature and time so that the active principle in culture solution can adequately immerse substrate
It is interior, ensure the quality of culture medium.Sterilizing can to avoid affect Cordyceps militaris growth bacterium；While sterilizing and substrate
The process that rice cooks, the process are also the process that the active principle in culture solution is fully merged with substrate rice.
Preferably, the sterilization steps are specially：
Culture medium before sterilizing is put into high-pressure sterilizing pot, heats the 35-45 minutes temperature made in the high-pressure sterilizing pot
Degree reaches 100-105 DEG C；
Then it opens air valve to depressurize 8-12 minutes, makes the high-pressure sterilizing pot external and internal pressure consistent；
Close the air valve continue heat the 18-22 minute temperature made in the high-pressure sterilizing pot reach 125-130 DEG C, press
Power reaches 0.145-0.150mpa, is kept for 18-22 minutes；
Then it opens the air valve to depressurize 8-12 minutes, makes the high-pressure sterilizing pot external and internal pressure consistent.
Control heating time is in order to avoid too fast heating makes the active principle in culture solution can not be fully by rice base
Bottom absorbs；Heating-decompression-reheating-repeatedly is depressurized again, is on the one hand to make rice substrate well-done, on the other hand in base
On the basis of bottom is well-done, the active principle in fusion culture solution is fully absorbed；It is in order to avoid subtracting because too fast to control decompression time
Pressure causes the unbalanced variation even substrate of generation in entire culture medium system half-cooked.
The application also provides a kind of method that Cordyceps militaris is cultivated using the culture medium, and the method includes following steps
Suddenly：
A. Cordyceps militaris kind bacterium is inoculated with by the medium sterilization and after cooling down, is sent into stand in darkroom and cultivate；Then will
Bacterium culture medium is put into middle cultivation between the cultivation of 24 hours full spectral illuminations；
B. when mycelia is changed into golden yellow by white, by manually delineating cut in the Spawn incubation primary surface；
C. the bacterium culture medium after scratching is transferred to 18-22 DEG C, the full light of intensity of illumination 80-100 luxs
3-5 days between spectrum cultivation；Daytime period to light, is protected from light at night；
D. when vaccine grows into 0.2-0.4cm, by vaccine be transferred to 18-22 DEG C, intensity of illumination 190-210 luxs it is complete
23-27 days between spectrum cultivation；Daytime period to light, is protected from light at night.
Rational incubation step cultivates temperature and time, intensity of illumination, light application time, be for improving cultivate quality,
Shorten cultivation period.Period, each factor was coordinated.Delineation cut is to accurately manage, and is left to Cordyceps militaris growth
On the one hand suitable space ensures cultivation amount, on the other hand improves the condition and quality of finished product.
The variation of each stage intensity of illumination agrees with each stage grown with Cordyceps militaris, is adapted with growth.In early days
Intensity of illumination can increase by force very much the inadaptability of Cordyceps militaris, easily so that the vaccine just generated is dead；Later stage intensity of illumination is too
It is weak, it is unfavorable for its fast-growth.
Preferably, in the step a, the cultivation temperature in the darkroom is 18-22 DEG C, and it is 6-8 days to cultivate the time；The training
Cultivation temperature between educating is 18-22 DEG C, and it is 4-7 days to cultivate the time, and intensity of illumination is 45-55 luxs.
It is suitable to cultivate temperature and time and intensity of illumination, it can effectively ensure that cultivation period and cultivate the flat of quality
Weighing apparatus, does not reduce cultivation quality due to the shortening of cultivation period.
It is further preferred that in the step b, the spacing of the adjacent cut is maintained at 0.2-0.8cm.
Suitable spacing, which can effectively ensure that, cultivates appropriate number of Cordyceps militaris on a culture medium, suitable to Cordyceps militaris
Growing space and sufficient nutrition avoid quality being caused to decline due to quantity is excessive.After delineating cut, Cordyceps militaris can be according to people
The cut growth of work delineation, shape, the specification (market price of different grade specifications is different) of Cordyceps militaris finished product can be controlled manually
System, obtains desired finished product Cordyceps militaris.
It is further preferred that need to divulge information daily in the step d, between cultivation twice, it is 25-35 minutes each.
Ventilation is to ensure to cultivate interior suitable oxygen content, and the carbon dioxide that Cordyceps militaris growth generates is discharged, is protected
Demonstrate,prove normal demand of the Cordyceps militaris to oxygen.Suitable ventilation time and ventilation number are effectively ensured Cordyceps militaris cultivation and are constantly in
In excellent environment.
Preferably, ventilation 5-6 points in the morning for the first time, second of ventilation 8-10 points at night.
The ventilation of morning 5-6 points can be to bring ozone between cultivating；Sooner or later appropriate ventilation time can close very much
Interior oxygen content and carbon dioxide content are cultivated in suitable control, and the two is made to remain balanced as possible.Gravity-flow ventilation is not easy always
Temperature between control is cultivated, cultivation cost can be significantly greatly increased by being adjusted and being divulged information using air-conditioning.
In addition, applicant it is emphasized that the application provide culture medium and cultivate Cordyceps militaris method, acquisition into
Product may not necessarily be picked immediately, and culture medium and finished product are transported together to the place of needs, ensure the fresh of finished product；At 3-5 DEG C
Under the conditions of, freshness-retained 3-6 months；Under the conditions of 20 DEG C, freshness-retained 15-20 days.
Compared with prior art, beneficial effects of the present invention are：
(1) culture medium is full of nutrition, balanced；
(2) culture matrix manufacturing is simple, at low cost；
(3) culture medium and cultural method can effectively shorten cultivation period, improve antibacterial disease resistance, increase yield；
(4) the Cordyceps militaris condition cultivated is good, quality is high, good in economic efficiency.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person, the condition suggested according to normal condition or manufacturer carry out.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
The application is mainly the production method and use culture medium cultivation Cordyceps militaris by providing culture medium, culture medium
Method, improve Cordyceps militaris yield and quality, shorten the production cycle, reduce production cost, increase economic efficiency.
Embodiment 1
7g potassium nitrate, 17g ammonium nitrate, 15g potassium dihydrogen phosphates, 3g magnesium sulfate and 18g calcium nitrate are dissolved in 1000ml distillations
In water, fully dissolving obtains a great number of elements aqueous solution；18g boric acid, 17g zinc sulfate, 3g manganese sulfates and 5g copper sulphate are dissolved in
In 1000ml distilled water, fully dissolving obtains trace element water-soluble liquid；By 0.52g vitamin B1s, 0.18g vitamin B6s, 22g cigarettes
Acid and 15g glycine are dissolved in 1000ml distilled water, and fully dissolving obtains organic aqueous solution；By 20g iron sulfites and 27g second
Diacid tetraacethyl disodium is dissolved in 1000ml distilled water, and fully dissolving obtains the aqueous solution of salt；550g edible glucoses are dissolved in
In 1000ml distilled water, fully dissolving obtains glucose solution.
Measure the water of a great number of elements aqueous solution 100ml, trace element water-soluble liquid 200ml, organic aqueous solution 100ml, salt
Solution 200ml and glucose solution 150ml, culture solution is obtained after being sufficiently mixed with 250ml distilled water.
Above-mentioned culture solution 500ml is taken, 400g uncooked rices is weighed and adds in culture solution, under the conditions of 18 DEG C, after impregnating 8 hours
Sterilization treatment is carried out, obtains culture medium.
Inoculation Cordyceps militaris kind bacterium after culture medium cooling, is sent into darkroom at 18 DEG C, stands and cultivate 6 days；Then by strain
Culture medium is put into middle between 18 DEG C, cultivation of the intensity of illumination for 24 hours full spectral illuminations of 45 luxs cultivate 4 days；Treat mycelia by
When white is changed into golden yellow, by manually delineating cut in Spawn incubation primary surface, the spacing of cut is maintained at 0.2cm；Then
Be transferred to 18 DEG C, the full spectrum of 80 lux of intensity of illumination cultivate between 3 days；Daytime period to light, is protected from light at night；Treat that vaccine is grown into
During 0.2cm, by vaccine be transferred to 18 DEG C, the full spectrum of 190 lux of intensity of illumination cultivate between 23 days；Daytime period is to light, evening
It is protected from light, need to divulge information daily twice, 25 minutes every time, ventilation for the first time was in the morning between 5-6 points, and divulge information 8-10 at night for the second time
Between point.
Embodiment 2
9g potassium nitrate, 13g ammonium nitrate, 20g potassium dihydrogen phosphates, 1g magnesium sulfate and 22g calcium nitrate are dissolved in 1000ml distillations
In water, fully dissolving obtains a great number of elements aqueous solution；12g boric acid, 25g zinc sulfate, 1g manganese sulfates and 10g copper sulphate are dissolved in
In 1000ml distilled water, fully dissolving obtains trace element water-soluble liquid；By 0.49g vitamin B1s, 0.22g vitamin B6s, 18g cigarettes
Acid and 20g glycine are dissolved in 1000ml distilled water, and fully dissolving obtains organic aqueous solution；By 15g iron sulfites and 31g second
Diacid tetraacethyl disodium is dissolved in 1000ml distilled water, and fully dissolving obtains the aqueous solution of salt；450g edible glucoses are dissolved in
In 1000ml distilled water, fully dissolving obtains glucose solution.
Measure the water of a great number of elements aqueous solution 200ml, trace element water-soluble liquid 100ml, organic aqueous solution 200ml, salt
Solution 100ml and glucose solution 250ml, culture solution is obtained after being sufficiently mixed with 150ml distilled water.
Above-mentioned culture solution 500ml is taken, 600g uncooked rices is weighed and adds in culture solution, under the conditions of 22 DEG C, is impregnated 12 hours
After be put into high-pressure sterilizing pot, heating 35 minutes temperature made in high-pressure sterilizing pot reach 100 DEG C；Then air valve decompression 8 is opened
Minute, make high-pressure sterilizing pot external and internal pressure consistent；Closing air valve continues 18 minutes temperature made in high-pressure sterilizing pot of heating and reaches
125 DEG C, pressure reach 0.145mpa, kept for 18 minutes；Then it opens air valve to depressurize 8 minutes, makes high-pressure sterilizing pot external and internal pressure
It is consistent.
Inoculation Cordyceps militaris kind bacterium after culture medium cooling, is sent into darkroom at 22 DEG C, stands and cultivate 8 days；Then by strain
Culture medium is put into middle between 22 DEG C, cultivation of the intensity of illumination for 24 hours full spectral illuminations of 55 luxs cultivate 7 days；Treat mycelia by
When white is changed into golden yellow, by manually delineating cut in Spawn incubation primary surface, the spacing of cut is maintained at 0.8cm；Then
Be transferred to 22 DEG C, the full spectrum of 100 lux of intensity of illumination cultivate between 5 days；Daytime period to light, is protected from light at night；Treat that vaccine is grown
During to 0.4cm, by vaccine be transferred to 22 DEG C, the full spectrum of 210 lux of intensity of illumination cultivate between 27 days；Daytime period is to light, evening
On be protected from light, need to divulge information daily twice, 35 minutes every time, ventilation for the first time was in the morning between 5-6 points, second of ventilation 8- at night
Between 10 points.
Embodiment 3
8g potassium nitrate, 15g ammonium nitrate, 18g potassium dihydrogen phosphates, 2g magnesium sulfate and 20g calcium nitrate are dissolved in 1000ml distillations
In water, fully dissolving obtains a great number of elements aqueous solution；15g boric acid, 21g zinc sulfate, 2g manganese sulfates and 8g copper sulphate are dissolved in
In 1000ml distilled water, fully dissolving obtains trace element water-soluble liquid；By 0.51g vitamin B1s, 0.20g vitamin B6s, 20g cigarettes
Acid and 18g glycine are dissolved in 1000ml distilled water, and fully dissolving obtains organic aqueous solution；By 18g iron sulfites and 29g second
Diacid tetraacethyl disodium is dissolved in 1000ml distilled water, and fully dissolving obtains the aqueous solution of salt；500g edible glucoses are dissolved in
In 1000ml distilled water, fully dissolving obtains glucose solution.
Measure the water of a great number of elements aqueous solution 150ml, trace element water-soluble liquid 150ml, organic aqueous solution 150ml, salt
Solution 150ml and glucose solution 200ml, culture solution is obtained after being sufficiently mixed with 200ml distilled water.
Above-mentioned culture solution 500ml is taken, 500g uncooked rices is weighed and adds in culture solution, under the conditions of 20 DEG C, is impregnated 10 hours
After be put into high-pressure sterilizing pot, heating 45 minutes temperature made in high-pressure sterilizing pot reach 105 DEG C；Then air valve decompression 12 is opened
Minute, make high-pressure sterilizing pot external and internal pressure consistent；Closing air valve continues 22 minutes temperature made in high-pressure sterilizing pot of heating and reaches
130 DEG C, pressure reach 0.150mpa, kept for 22 minutes；Then it opens air valve to depressurize 12 minutes, makes high-pressure sterilizing pot external and internal pressure
It is consistent.
Inoculation Cordyceps militaris kind bacterium after culture medium cooling, is sent into darkroom at 20 DEG C, stands and cultivate 7 days；Then by strain
Culture medium is put into middle between 20 DEG C, cultivation of the intensity of illumination for 24 hours full spectral illuminations of 50 luxs cultivate 5 days；Treat mycelia by
When white is changed into golden yellow, by manually delineating cut in Spawn incubation primary surface, the spacing of cut is maintained at 0.5cm；Then
Be transferred to 20 DEG C, the full spectrum of 90 lux of intensity of illumination cultivate between 4 days；Daytime period to light, is protected from light at night；Treat that vaccine is grown into
During 0.3cm, by vaccine be transferred to 20 DEG C, the full spectrum of 200 lux of intensity of illumination cultivate between 25 days；Daytime period is to light, evening
It is protected from light, need to divulge information daily twice, 30 minutes every time, ventilation for the first time between 5-6 points, was divulged information 9 points at night for the second time in the morning
Left and right.
Embodiment 4
8g potassium nitrate, 16g ammonium nitrate, 19g potassium dihydrogen phosphates, 2g magnesium sulfate and 20g calcium nitrate are dissolved in 1000ml distillations
In water, fully dissolving obtains a great number of elements aqueous solution；14g boric acid, 21g zinc sulfate, 2g manganese sulfates and 7g copper sulphate are dissolved in
In 1000ml distilled water, fully dissolving obtains trace element water-soluble liquid；By 0.50g vitamin B1s, 0.21g vitamin B6s, 21g cigarettes
Acid and 17g glycine are dissolved in 1000ml distilled water, and fully dissolving obtains organic aqueous solution；By 18g iron sulfites and 29g second
Diacid tetraacethyl disodium is dissolved in 1000ml distilled water, and fully dissolving obtains the aqueous solution of salt；520g edible glucoses are dissolved in
In 1000ml distilled water, fully dissolving obtains glucose solution.
Measure the water of a great number of elements aqueous solution 150ml, trace element water-soluble liquid 150ml, organic aqueous solution 150ml, salt
Solution 150ml and glucose solution 200ml, culture solution is obtained after being sufficiently mixed with 200ml distilled water.
Above-mentioned culture solution 500ml is taken, 500g uncooked rices is weighed and adds in culture solution, under the conditions of 20 DEG C, is impregnated 10 hours
After be put into high-pressure sterilizing pot, heating 40 minutes temperature made in high-pressure sterilizing pot reach 102 DEG C；Then air valve decompression 10 is opened
Minute, make high-pressure sterilizing pot external and internal pressure consistent；Closing air valve continues 20 minutes temperature made in high-pressure sterilizing pot of heating and reaches
128 DEG C, pressure reach 0.148mpa, kept for 20 minutes；Then it opens air valve to depressurize 10 minutes, makes high-pressure sterilizing pot external and internal pressure
It is consistent.
Inoculation Cordyceps militaris kind bacterium after culture medium cooling, is sent into darkroom at 20 DEG C, stands and cultivate 6 days；Then by strain
Culture medium is put into middle between 20 DEG C, cultivation of the intensity of illumination for 24 hours full spectral illuminations of 50 luxs cultivate 6 days；Treat mycelia by
When white is changed into golden yellow, by manually delineating cut in Spawn incubation primary surface, the spacing of cut is maintained at 0.5cm；Then
Be transferred to 20 DEG C, the full spectrum of 90 lux of intensity of illumination cultivate between 5 days；Daytime period to light, is protected from light at night；Treat that vaccine is grown into
During 0.3cm, by vaccine be transferred to 20 DEG C, the full spectrum of 200 lux of intensity of illumination cultivate between 26 days；Daytime period is to light, evening
It is protected from light, need to divulge information daily twice, 30 minutes every time, ventilation for the first time between 5-6 points, was divulged information 9 points at night for the second time in the morning
Left and right.
The Cordyceps militaris cultivated to the cultivation period of embodiment 1-4, survival rate, the cost of 1kg culture mediums, 1kg culture mediums
Weight and high-quality Cordyceps militaris account for the ratio of total amount and counted, compared with the prior art, as a result such as the following table 1 institute
Show：
1 cultivation period comparing result of table
By upper table 1 it is found that the culture medium of the application offer and the method that Cordyceps militaris is cultivated using the culture medium, cultivation period
It greatly shortens, survival rate greatly improves, at low cost.In terms of weight in wet base, the culture medium and breeding method of the application, Cordyceps militaris yield
Height, in terms of dry weight, the quality of Cordyceps militaris is high.High quality Cordyceps militaris refers to that length is more than or equal to 9cm, foundation diameter exists in table
The Cordyceps militaris of more than 0.5cm.Quality is higher, condition is better, and economic benefit is better.
Comparative example 1
8g potassium nitrate, 15g ammonium nitrate, 18g potassium dihydrogen phosphates, 2g magnesium sulfate and 20g calcium nitrate are dissolved in 1000ml distillations
In water, fully dissolving obtains a great number of elements aqueous solution；15g boric acid, 21g zinc sulfate, 2g manganese sulfates and 8g copper sulphate are dissolved in
In 1000ml distilled water, fully dissolving obtains trace element water-soluble liquid；By 0.51g vitamin B1s, 0.20g vitamin B6s, 20g cigarettes
Acid and 18g glycine are dissolved in 1000ml distilled water, and fully dissolving obtains organic aqueous solution；By 18g iron sulfites and 29g second
Diacid tetraacethyl disodium is dissolved in 1000ml distilled water, and fully dissolving obtains the aqueous solution of salt；500g edible glucoses are dissolved in
In 1000ml distilled water, fully dissolving obtains glucose solution.
Measure the water of a great number of elements aqueous solution 150ml, trace element water-soluble liquid 150ml, organic aqueous solution 150ml, salt
Solution 150ml and glucose solution 200ml, culture solution is obtained after being sufficiently mixed with 200ml distilled water.
Above-mentioned culture solution 500ml is taken, 500g uncooked rices is weighed and adds in culture solution, under the conditions of 20 DEG C, is impregnated 10 hours
After be put into high-pressure sterilizing pot, heating 45 minutes temperature made in high-pressure sterilizing pot reach 105 DEG C；Then air valve decompression 12 is opened
Minute, make high-pressure sterilizing pot external and internal pressure consistent；Closing air valve continues 22 minutes temperature made in high-pressure sterilizing pot of heating and reaches
130 DEG C, pressure reach 0.150mpa, kept for 22 minutes；Then it opens air valve to depressurize 12 minutes, makes high-pressure sterilizing pot external and internal pressure
It is consistent.
Inoculation Cordyceps militaris kind bacterium after culture medium cooling, is sent into darkroom at 20 DEG C, stands and cultivate 7 days；Then by strain
Culture medium is put into middle between 20 DEG C, cultivation of the intensity of illumination for 24 hours full spectral illuminations of 50 luxs cultivate 5 days；Treat mycelia by
When white is changed into golden yellow, be transferred to 20 DEG C, the full spectrum of 90 lux of intensity of illumination cultivate between 4 days；Daytime period is to light, evening
On be protected from light；When vaccine grows into 0.3cm, by vaccine be transferred to 20 DEG C, the full spectrum of 200 lux of intensity of illumination cultivate between 25
My god；Daytime period to light, is protected from light, need to divulge information daily twice at night, 30 minutes every time, and ventilation for the first time is in the morning between 5-6 points,
9 points or so at night of second of ventilation.
Comparative example 1 and embodiment 3 the difference is that, when mycelia is changed into golden yellow by white, without artificial cut.
50 groups of Cordyceps militaris is cultivated according to embodiment 3 and the method for comparative example 1, statistics high-quality Cordyceps militaris accounts for the ratio of total amount
Example, result are as shown in table 2 below：
2 high-quality Cordyceps militaris of table accounts for the ratio comparison of total amount
| Label | Embodiment 3 | Comparative example 1 |
| Ratio/% | 88 | 71 |
By upper table 2 it is found that artificial cut can guide Cordyceps militaris ordering growth, can improve finished product Cordyceps militaris quality and
Condition.
Comparative example 2
8g potassium nitrate, 15g ammonium nitrate, 18g potassium dihydrogen phosphates, 2g magnesium sulfate and 20g calcium nitrate are dissolved in 1000ml distillations
In water, fully dissolving obtains a great number of elements aqueous solution；15g boric acid, 21g zinc sulfate, 2g manganese sulfates and 8g copper sulphate are dissolved in
In 1000ml distilled water, fully dissolving obtains trace element water-soluble liquid；By 0.51g vitamin B1s, 0.20g vitamin B6s, 20g cigarettes
Acid and 18g glycine are dissolved in 1000ml distilled water, and fully dissolving obtains organic aqueous solution；By 18g iron sulfites and 29g second
Diacid tetraacethyl disodium is dissolved in 1000ml distilled water, and fully dissolving obtains the aqueous solution of salt；500g edible glucoses are dissolved in
In 1000ml distilled water, fully dissolving obtains glucose solution.
Measure the water of a great number of elements aqueous solution 150ml, trace element water-soluble liquid 150ml, organic aqueous solution 150ml, salt
Solution 150ml and glucose solution 200ml, culture solution is obtained after being sufficiently mixed with 200ml distilled water.
Above-mentioned culture solution 500ml is taken, 500g uncooked rices is weighed and adds in culture solution, under the conditions of 20 DEG C, is impregnated 10 hours
After be put into high-pressure sterilizing pot, heating 45 minutes temperature made in high-pressure sterilizing pot reach 105 DEG C；Then air valve decompression 5 is opened
Minute, make high-pressure sterilizing pot external and internal pressure consistent；Closing air valve continues 22 minutes temperature made in high-pressure sterilizing pot of heating and reaches
130 DEG C, pressure reach 0.150mpa, kept for 22 minutes；Then it opens air valve to depressurize 5 minutes, makes high-pressure sterilizing pot external and internal pressure
It is consistent.
Inoculation Cordyceps militaris kind bacterium after culture medium cooling, is sent into darkroom at 20 DEG C, stands and cultivate 7 days；Then by strain
Culture medium is put into middle between 20 DEG C, cultivation of the intensity of illumination for 24 hours full spectral illuminations of 50 luxs cultivate 5 days；Treat mycelia by
When white is changed into golden yellow, be transferred to 20 DEG C, the full spectrum of 90 lux of intensity of illumination cultivate between 4 days；Daytime period is to light, evening
On be protected from light；When vaccine grows into 0.3cm, by vaccine be transferred to 20 DEG C, the full spectrum of 200 lux of intensity of illumination cultivate between 25
My god；Daytime period to light, is protected from light, need to divulge information daily twice at night, 30 minutes every time, and ventilation for the first time is in the morning between 5-6 points,
9 points or so at night of second of ventilation.
Comparative example 2 and embodiment 3 the difference is that, shorten the time depressurized in disinfecting action.
50 groups of Cordyceps militaris is cultivated according to embodiment 3 and the method for comparative example 2, statistics high-quality Cordyceps militaris accounts for the ratio of total amount
Example, result are as shown in table 3 below：
3 high-quality Cordyceps militaris of table accounts for the ratio comparison of total amount
| Label | Embodiment 3 | Comparative example 2 |
| Ratio/% | 88 | 60 |
By upper table 3 it is found that too fast decompression, can reduce the quality of culture medium, and then reduce the quality of finished product Cordyceps militaris.
The culture medium that the application provides is full of nutrition, balanced；It makes simple, at low cost；Culture medium enrich balanced nutrition,
Appropriate cultural method can effectively shorten cultivation period, improve antibacterial disease resistance, increase yield；The Cordyceps militaris cultivated
Condition is good, quality is high, good in economic efficiency.
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from the present invention's
Many other change and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (8)
1. a kind of production method of culture medium, which is characterized in that the method is：
Under the conditions of 18-22 DEG C, impregnate substrate 8-12 hours using culture solution, then sterilize, obtain the culture medium；
The culture solution with volume percent, including：A great number of elements aqueous solution 10-20%, trace element water-soluble liquid 10-
20%th, organic aqueous solution 10-20%, the aqueous solution 10-20% of salt and glucose solution 15-25%, surplus are distilled water；
In a great number of elements aqueous solution, 7-9g potassium nitrate, 13-17g ammonium nitrate, 15-20g phosphoric acid are corresponded to per 1000ml distilled water
Potassium dihydrogen, 1-3g magnesium sulfate and 18-22g calcium nitrate；
In the trace element water-soluble liquid, 12-18g boric acid, 17-25g zinc sulfate, 1-3g manganese sulfates are corresponded to per 1000ml distilled water
With 5-10g copper sulphate；
In the organic aqueous solution, 0.49-0.52g vitamin B1s, 0.18-0.22g vitamins are corresponded to per 1000ml distilled water
B6,18-22g niacin and 15-20g glycine；
In the aqueous solution of the salt, 15-20g iron sulfites and 27-31g ethanedioic acids tetraacethyl two are corresponded to per 1000ml distilled water
Sodium；
In the glucose solution, 450-550g glucose is corresponded to per 1000ml distilled water；
The substrate is uncooked rice；
The dosage of the culture solution and the substrate is that culture solution correspondence uses substrate described in 0.8-1.2g described in per 1ml；
Wherein, the sterilization steps are specially：
Culture medium before sterilizing is put into high-pressure sterilizing pot, the 35-45 minutes temperature made in the high-pressure sterilizing pot of heating reach
To 100-105 DEG C；
Then it opens air valve to depressurize 8-12 minutes, makes the high-pressure sterilizing pot external and internal pressure consistent；
It closes the air valve and continues to heat that the 18-22 minutes temperature made in the high-pressure sterilizing pot reach 125-130 DEG C, pressure reaches
To 0.145-0.150MPa, kept for 18-22 minutes；
Then it opens the air valve to depressurize 8-12 minutes, makes the high-pressure sterilizing pot external and internal pressure consistent.
2. the production method of culture medium according to claim 1, which is characterized in that the culture solution is with percentage by volume
Meter, including：A great number of elements aqueous solution 15%, trace element water-soluble liquid 15%, organic aqueous solution 15%, salt aqueous solution 15%
With glucose solution 20%, surplus is distilled water.
3. the production method of culture medium according to claim 2, which is characterized in that in a great number of elements aqueous solution, often
1000ml distilled water corresponds to 8g potassium nitrate, 15g ammonium nitrate, 18g potassium dihydrogen phosphates, 2g magnesium sulfate and 20g calcium nitrate；
In the trace element water-soluble liquid, 15g boric acid, 21g zinc sulfate, 2g manganese sulfates and 8g sulfuric acid are corresponded to per 1000ml distilled water
Copper；
In the organic aqueous solution, 0.51g vitamin B1s, 0.20g vitamin B6s, 20g niacin are corresponded to per 1000ml distilled water
With 18g glycine；
In the aqueous solution of the salt, 18g iron sulfites and 29g ethanedioic acid tetraacethyl disodiums are corresponded to per 1000ml distilled water；
In the glucose solution, 500g glucose is corresponded to per 1000ml distilled water.
A kind of 4. method for cultivating Cordyceps militaris, which is characterized in that the described method comprises the following steps：
A. culture medium is made according to the method described in any one of claim 1-3；
Cordyceps militaris kind bacterium is inoculated with by the medium sterilization and after cooling down, is sent into stand in darkroom and cultivate；Then strain is trained
Foster base is put into middle cultivation between the cultivation of 24 hours full spectral illuminations；
B. when mycelia is changed into golden yellow by white, by manually delineating cut in Spawn incubation primary surface；
C. by the bacterium culture medium after scratching is transferred to 18-22 DEG C, the full spectrum of intensity of illumination 80-100 luxs trains
It educates 3-5 days；Daytime period to light, is protected from light at night；
D. when vaccine grows into 0.2-0.4cm, vaccine is transferred to 18-22 DEG C, the full spectrum of intensity of illumination 190-210 luxs
23-27 days between cultivation；Daytime period to light, is protected from light at night.
5. according to the method described in claim 4, it is characterized in that, in the step a, the cultivation temperature in the darkroom is 18-
22 DEG C, it is 6-8 days to cultivate the time；Cultivation temperature between the cultivation is 18-22 DEG C, and it is 4-7 days to cultivate the time, and intensity of illumination is
45-55 luxs.
6. according to the method described in claim 5, it is characterized in that, in the step b, the spacing of the adjacent cut is kept
In 0.2-0.8cm.
7. according to the method described in claim 6, it is characterized in that, in the step d, need to divulge information daily twice, often between cultivation
It is 25-35 minutes secondary.
8. the method according to the description of claim 7 is characterized in that 5-6 points in the morning of divulging information for the first time, second of ventilation is in evening
Upper 8-10 points.
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| CN108485983B (en) * | 2018-06-04 | 2021-07-30 | 海南源泉生物科技有限公司 | Culture medium for large-scale high-density cultivation of green algae and culture method thereof |
| CN110547141A (en) * | 2019-09-30 | 2019-12-10 | 江苏康能生物工程股份有限公司 | Cultivation method for increasing polysaccharide content in cordyceps militaris sporocarp |
| CN112825728A (en) * | 2019-11-25 | 2021-05-25 | 湖南金芙农业科技有限公司 | Method for cultivating cordyceps militaris with high cordycepin content |
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| CN102626036A (en) * | 2012-04-24 | 2012-08-08 | 珠海市先康生物科技有限公司 | Large-scale cultivation method and quality detection method of cordyceps militaris fruit bodies |
| CN103609329A (en) * | 2013-11-05 | 2014-03-05 | 昆山市康乐虫草专业合作社 | Cordyceps militaris culturing method capable of improving cordycepin content |
| CN104163684A (en) * | 2014-06-30 | 2014-11-26 | 桂林丰茂源农业技术开发有限公司 | A cordyceps militaris sporophore culture medium comprising shaddock peel |
| CN104177182A (en) * | 2014-08-19 | 2014-12-03 | 中华全国供销合作总社南京野生植物综合利用研究所 | Cultural method for producing cordyceps militaris fruiting body by use of corn cobs |
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| CN102626036A (en) * | 2012-04-24 | 2012-08-08 | 珠海市先康生物科技有限公司 | Large-scale cultivation method and quality detection method of cordyceps militaris fruit bodies |
| CN103609329A (en) * | 2013-11-05 | 2014-03-05 | 昆山市康乐虫草专业合作社 | Cordyceps militaris culturing method capable of improving cordycepin content |
| CN104163684A (en) * | 2014-06-30 | 2014-11-26 | 桂林丰茂源农业技术开发有限公司 | A cordyceps militaris sporophore culture medium comprising shaddock peel |
| CN104177182A (en) * | 2014-08-19 | 2014-12-03 | 中华全国供销合作总社南京野生植物综合利用研究所 | Cultural method for producing cordyceps militaris fruiting body by use of corn cobs |
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