CN102293122B - Cultivating method for cordyceps militaris by using manyprickle acathopanax root - Google Patents
Cultivating method for cordyceps militaris by using manyprickle acathopanax root Download PDFInfo
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- CN102293122B CN102293122B CN2011101734511A CN201110173451A CN102293122B CN 102293122 B CN102293122 B CN 102293122B CN 2011101734511 A CN2011101734511 A CN 2011101734511A CN 201110173451 A CN201110173451 A CN 201110173451A CN 102293122 B CN102293122 B CN 102293122B
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Abstract
The invention discloses a cultivating method for cordyceps militaris by using manyprickle acathopanax root and relates to a cultivating method for cordyceps militaris. The method is used for simplifying Chinese medicine compatibility dosage, increasing the utilization rate of cordyceps militaris and reducing the medicine consumption. The method comprises the following steps of: 1, activating strains; 2, preparing liquid strains; and 3, inoculating the liquid strains into a manyprickle acathopanax root solid culture medium for dark cultivation to obtain mycelia, then transferring to light and culturing until sporocarp grows up to 2 cm in height, after gas introduction, continuing culturing until the sporosarp grows up to 4-8 cm in height, yellow powdery substances appear on the surface of the sporosarp as well as small thorns appear on the top end, and harvesting. According to the method disclosed by the invention, active ingredients of the manyprickle acathopanax root are transferred into the sporocarp of cordyceps militaris, so that a beneficial curative effect of cordyceps militaris is indirectly enhanced, the method of compatibility of cordyceps militaris and manyprickle acathopanax root in traditional Chinese medical science is simplified, the utilization ratio, nutrition and medical value of cordyceps militaris are improved, the yield of cordyceps militaris is improved, the growth cycle of cordyceps militaris is shortened, and the quality of cordyceps militaris is improved; and moreover, implementing conditions are easily achieved.
Description
Technical field
The present invention relates to a kind of cultural method of Cordyceps militaris.
Background technology
Cordyceps militaris [Cordyceps militarise (L.) Link] is that the worm parasitism is had a liking for the low temperature fungi, has another name called Cordyceps militaris.Nourishing Northern cordyceps is abundant, contains cordycepin, adenosine, Cordyceps sinensis polysaccharide, cordycepic acid, protein, SOD, amino acid, vitamin and various trace elements to the human body beneficial; Have invigorate the lung and the kidney, the effect of hemostasis and phlegm, be used for that chronic cough void is breathed heavily, labor is coughed hemoptysis, impotence and seminal emission, soreness of waist and knee joint.Wilsonii has tonifying Qi benefit essence, wind-damp dispelling, strengthening the bones and muscles, invigorating qi for tranquilization; Being used for neurasthenia, hypertension, coronary heart disease, high fat of blood, diabetes, arteriosclerosis etc. has good curative effect, and compatibility is used in the prescriptions such as treatment miscarriage, impotence, soreness and weakness of waist and knees, neurasthenia, neuritis, vertigo, the rhinitis of being everlasting with Cordyceps militaris.
Often Cordyceps militaris and various traditional tonic medicine compatibility are used reaching the purpose of Synergistic on the tcm clinical practice, but the patient when compatibility uses Chinese medicine usually because dose is big, take the inconvenience interruption of serving as reasons and make medicament.Therefore simplify traditional Chinese medical science combination drug, the availability that improves Cordyceps militaris is the problem that must solve.
Summary of the invention
The invention provides a kind of cultural method of wilsonii Cordyceps militaris; Purpose is in order to simplify traditional Chinese medical science combination drug, to improve the availability of Cordyceps militaris, reducing dosage.
The cultural method of wilsonii Cordyceps militaris carries out: one, the Cordyceps militaris bacterial strain is inoculated on the PDA solid culture medium for beautiful rich No. two, in 28 ℃ of biochemical incubators, secretly cultivates 5d, obtain the bacterial classification of activation according to the following steps; Two, bacterial classification that activation is the good bacterium piece that is cut into 1cm * 1cm is inoculated in the liquid seed culture medium of 300ml; It is the dark 6d of cultivation under 23 ℃, the condition of 120r/min that static cultivation 24h is placed on temperature; Obtain bacteria suspension, add the 200ml sterile water then, get liquid spawn; Three, get the 5ml liquid-spawn inoculation in the blake bottle that the wilsonii solid culture medium is housed; Placing temperature is that 22 ℃, humidity are the dark 10～14d of cultivation under 55%～60% the condition; Get mycelium; Move to then under the light and be in temperature that 23 ℃, humidity are 55%～60%, intensity of illumination is 180～220lx, illumination every day 10～12h, day and night temperature after being cultured to mycelium under 10 ℃ the condition and all transferring yellow to by white, increase humidity to 85%～95%, it is long high to 2cm to continue to be cultured to fruit body; With the ventilation switch opens on the blake bottle; Continue to be cultured to that sporophore growth to 4～8cm is high, fruit body is surperficial yellow powder shape material occurs and spinule appears in the top, gathers, and promptly accomplishes the cultivation of wilsonii Cordyceps militaris;
Wherein the wilsonii solid culture medium is made up of the rice of 40g, the Manyprickle Acanthopanax Root of 0.1～0.6g and the nutrient solution of 60ml in the step 3, and the bottled amount of blake bottle is 1/4～1/3; The preparation of said Manyprickle Acanthopanax Root: wilsonii is pulverized the back cross 20 mesh sieves; The every L of said nutrient solution is by the peptone of 20g, the glucose of 20g, the KH of 1g
2PO
4, 0.5g MgSO
47H
2The potato liquid of the zinc sulphate of O, 0.5g, 200g, the Cobastab of 5mg
1Form with the water of surplus; The preparation of said potato liquid: 200g removes the peel potato, and 30min is boiled in stripping and slicing, complements to 1000ml after the filtered through gauze, pH nature, 121 ℃ of autoclaving 30min.
The cultural method of wilsonii Cordyceps militaris of the present invention; In the medium of Cordyceps militaris, add wilsonii, so that in the process of growth of Cordyceps militaris, change the active component of wilsonii over to, to reach the function of enhance immunity; Simplified instructions of taking; The wilsonii that played when using Cordyceps militaris compatibility has improved the availability of Cordyceps militaris, has strengthened the medical value of Cordyceps militaris.
The present invention adds the quality that wilsonii can improve the yield of Cordyceps militaris, the growth cycle that shortens Cordyceps militaris, improvement Cordyceps militaris in the medium of Cordyceps militaris; The Cordyceps militaris that adopts the inventive method to produce simultaneously can change the active component of wilsonii over to; Thereby play the help curative effect that strengthens Cordyceps militaris indirectly; Simplifying the purpose of traditional Chinese medical science compatibility, is a kind of technological invention that is worthy to be popularized.
The wilsonii Cordyceps militaris that the present invention cultivates can increase substantially Cordyceps militaris fruiting body cordycepin and adenosine content, and cordycepin can reach 3.4195 ± 0.0124mg/g, and adenosine can reach 0.9970 ± 0.0032mg/g, and commodity value is very high; And contain the Hyperoside of 0.0043 ± 0.0001mg/g and the Quercetin of 0.0008 ± 0.0001mg/g in the wilsonii Cordyceps militaris that the present invention cultivates; Thereby play wilsonii active ingredient is transferred to the purpose in the Cordyceps militaris fruiting body; Simplify the method for Cordyceps militaris and wilsonii compatibility in the traditional Chinese medicine, and improved nutrition and the medical value of Cordyceps militaris; In addition, implementation condition reaches easily, can promote in the whole industry.
Description of drawings
Fig. 1 is the mixing reference substance spectrogram that the HPLC method is measured adenosine and cordycepin content in the Cordyceps militaris in the embodiment four, curve 1 expression adenosine wherein, and curve 2 is represented cordycepins; Fig. 2 is a spectrogram of cultivating adenosine and cordycepin content in the gained wilsonii Cordyceps militaris fruiting body in the embodiment four, curve 1 expression adenosine wherein, and curve 2 is represented cordycepins; Fig. 3 is embodiment four empty control medium chromatograms; Fig. 4 is the chromatogram that adds wilsonii Cordyceps militaris fruiting body behind the reference substance in the embodiment four, curve 1 expression adenosine wherein, curve 2 expression cordycepins; Fig. 5 is the mixing reference substance spectrogram that the HPLC method is measured the active component content of wilsonii in the wilsonii Cordyceps militaris fruiting body in the embodiment five, and wherein curve 1 is a Hyperoside, and curve 2 is a Quercetin; Fig. 6 is a spectrogram of cultivating Hyperoside and quercetin content in the gained wilsonii Cordyceps militaris in the embodiment five, and wherein curve 1 is a Hyperoside, and curve 2 is a Quercetin; Fig. 7 is the chromatogram of embodiment five empty control group Cordyceps militaris.
Embodiment
Embodiment one: the cultural method of this embodiment wilsonii Cordyceps militaris, carry out: one, the Cordyceps militaris bacterial strain is inoculated on the PDA solid culture medium for beautiful rich No. two, in 28 ℃ of biochemical incubators, secretly cultivates 5d, obtain the bacterial classification of activation according to the following steps; Two, bacterial classification that activation is the good bacterium piece that is cut into 1cm * 1cm is inoculated in the liquid seed culture medium of 300ml; It is the dark 6d of cultivation under 23 ℃, the condition of 120r/min that static cultivation 24h is placed on temperature; Obtain bacteria suspension, add the 200ml sterile water then, get liquid spawn; Three, get the 5ml liquid-spawn inoculation in the blake bottle that the wilsonii solid culture medium is housed; Placing temperature is that 22 ℃, humidity are the dark 10～14d of cultivation under 55%～60% the condition; Get mycelium; Move to then under the light and be in temperature that 23 ℃, humidity are 55%～60%, intensity of illumination is 180～220lx, illumination every day 10～12h, day and night temperature after being cultured to mycelium under 10 ℃ the condition and all transferring yellow to by white, increase humidity to 85%～95%, it is long high to 2cm to continue to be cultured to fruit body; With the ventilation switch opens on the blake bottle; Continue to be cultured to that sporophore growth to 4～8cm is high, fruit body is surperficial yellow powder shape material occurs and spinule appears in the top, gathers, and promptly accomplishes the cultivation of wilsonii Cordyceps militaris;
Wherein the wilsonii solid culture medium is made up of the rice of 40g, the Manyprickle Acanthopanax Root of 0.1～0.6g and the nutrient solution of 60ml in the step 3, and the bottled amount of blake bottle is 1/4～1/3; The preparation of said Manyprickle Acanthopanax Root: wilsonii is pulverized the back cross 20 mesh sieves; The every L of said nutrient solution is by the peptone of 20g, the glucose of 20g, the KH of 1g
2PO
4, 0.5g MgSO
47H
2The potato liquid of the zinc sulphate of O, 0.5g, 200g, the Cobastab of 5mg
1Form with the water of surplus; The preparation of said potato liquid: 200g removes the peel potato, and 30min is boiled in stripping and slicing, complements to 1000ml after the filtered through gauze, pH nature, 121 ℃ of autoclaving 30min.
The step 1 of this embodiment is sterile working in superclean bench.
The Cordyceps militaris bacterial strain is beautiful rich No. two in this embodiment step 1, buys the Yu Feng bacterium industry Co., Ltd from Harbin.
1 bacterium piece of inoculation in every 300ml liquid seed culture medium in this embodiment step 2.
In the process of dark cultivation 10～14d, mycelium covers with the surface of whole medium and has thorough grasp medium in this embodiment step 3; And preceding 1 week of inoculation back is stuffy, later ventilation every day 1～2 time.
Move in this embodiment step 3 in the incubation under the light, to predominate, then adjust light source direction like fruit body, or every interval conversion in 3 days medium direction, so that daylighting is even, guarantee that sporophore growth to 4～8cm is high.
After gathering in this embodiment step 3, Cordyceps militaris placed under 45 ℃ dry to constant weight.
Embodiment two: what this embodiment and embodiment one were different is that the PDA solid culture medium is made up of the potato liquid of 200g, the glucose of 20g, the agar of 18g and the water of 1000ml in the step 1, the pH nature; The preparation of said potato liquid: 200g removes the peel potato, and 30min is boiled in stripping and slicing, complements to 1000ml after the filtered through gauze, pH nature, 121 ℃ of autoclaving 30min.Other step and parameter are identical with embodiment one.
Embodiment three: this embodiment and embodiment one are different is that the every L of liquid seed culture medium forms the pH nature by the water of magnesium sulfate, 1g potassium dihydrogen phosphate and the surplus of the glucose of the potato liquid of 200g, 20g, 1g in the step 2; The preparation of said potato liquid: 200g removes the peel potato, and 30min is boiled in stripping and slicing, complements to 1000ml after the filtered through gauze, pH nature, 121 ℃ of autoclaving 30min.Other step and parameter are identical with embodiment one.
Embodiment four: the cultural method of this embodiment wilsonii Cordyceps militaris, carry out: one, the Cordyceps militaris bacterial strain is inoculated on the PDA solid culture medium for beautiful rich No. two, in 28 ℃ of biochemical incubators, secretly cultivates 5d, obtain the bacterial classification of activation according to the following steps; Two, bacterial classification that activation is the good bacterium piece that is cut into 1cm * 1cm is inoculated in the liquid seed culture medium of 300ml; It is the dark 6d of cultivation under 23 ℃, the condition of 120r/min that static cultivation 24h is placed on temperature; Obtain bacteria suspension, add the 200ml sterile water then, get liquid spawn; Three, get the 5ml liquid-spawn inoculation in the blake bottle that the wilsonii solid culture medium is housed; Placing temperature is that 22 ℃, humidity are the dark 12d of cultivation under 60% the condition; Get mycelium; Move to then under the light and be in temperature that 23 ℃, humidity are 60%, intensity of illumination is 200lx, illumination every day 12h, day and night temperature after being cultured to mycelium under 10 ℃ the condition and all transferring yellow to by white, increase humidity to 90%, it is long high to 2cm to continue to be cultured to fruit body; With the ventilation switch opens on the blake bottle; Continue to be cultured to that sporophore growth is high to 7cm, fruit body is surperficial yellow powder shape material occurs and spinule appears in the top, gathers, and promptly accomplishes the cultivation of wilsonii Cordyceps militaris;
Wherein the wilsonii solid culture medium is made up of the rice of 40g, the Manyprickle Acanthopanax Root of 0.2g and the nutrient solution of 60ml in the step 3, and the bottled amount of blake bottle is 1/3; The preparation of said Manyprickle Acanthopanax Root: wilsonii is pulverized the back cross 20 mesh sieves; The every L of said nutrient solution is by the peptone of 20g, the glucose of 20g, the KH of 1g
2PO
4, 0.5g MgSO
47H
2The potato liquid of the zinc sulphate of O, 0.5g, 200g, the Cobastab of 5mg
1Form with the water of surplus; The preparation of said potato liquid: 200g removes the peel potato, and 30min is boiled in stripping and slicing, complements to 1000ml after the filtered through gauze, pH nature, 121 ℃ of autoclaving 30min.
The Cordyceps militaris bacterial strain is beautiful rich No. two in this embodiment step 1, buys the Yu Feng bacterium industry Co., Ltd from Harbin.
Wilsonii solid culture medium cultivation gained fruit body and blank medium (promptly not adding the medium of Manyprickle Acanthopanax Root) cultivation gained fruit body compare in this embodiment: with fruit body difference clean it up, blot surface moisture with filter paper, dry to constant weight in 45 ℃ of conditions; Cooling; Analytical balance weigh dry weight, and confirm to promote the optimum concentration of sporophore growth, the result is as shown in table 1; Compare with the blank group, the fruit body yield is the highest in the wilsonii solid culture medium.
Table 1
| Group | Fruit body yield (g) |
| Blank medium | 1.9330±0.0187 |
| The wilsonii solid culture medium | 2.7810±0.0143 |
Wilsonii is to the influence of fruit body growth cycle: calculate in this embodiment Cordyceps militaris from being seeded to the fate that maturation is gathered; And compare with blank group group; The result is as shown in table 2, and visible, wilsonii has the effect in tangible shortening sporophore growth cycle; Compare with the blank group, can shorten 5 days.
Table 2
| Group | Growth cycle (my god) |
| Blank medium | 55.25±1.29 |
| The wilsonii solid culture medium | 49.50±1.35 |
Cultivate gained wilsonii Cordyceps militaris in this embodiment, adenosine and content of cordycepin step are following in the employing HPLC method mensuration wilsonii Cordyceps militaris fruiting body:
(1) chromatographic condition
Chromatographic column: Venusil XBP-C
18Post (4.6mm * 250mm, 5 μ m, USA);
Flowing phase: methyl alcohol: water=85: 15;
Flow velocity: 1mL/min;
Column temperature: 25 ℃;
Detect wavelength: 260nm;
Sample size: 10 μ L.
(2) preparation of reference substance solution respectively precision take by weighing adenosine and the cordycepin reference substance is an amount of, process the reference substance storing solution that mass concentration is 1mg/mL with methyl alcohol; Accurately measure above-mentioned each 1mL of reference substance storing solution respectively, processing mass concentration is the mixing reference substance solution of adenosine 0.5mg/mL, cordycepin 0.5mg/mL.
(3) the preparation precision of fruit body need testing solution takes by weighing 2 parts of the Cordyceps militaris fruiting bodies that dry wilsonii solid culture medium cultivates, every part of 0.5g, accurate respectively 5mL 90% methyl alcohol that adds; Weigh; Ultrasonic 60min takes out, cooling; Supply the solvent that subtracts mistake with 90% methyl alcohol, filter the back as need testing solution with 0.45 μ m filter membrane.
(4) sample determination is under chromatographic condition, get reference substance solution and need testing solution with 0.45 μ m membrane filtration after, the accurate respectively 10 μ L sample introductions of drawing, the record peak area calculates content by external standard method.
(5), result:
1. the separating of adenosine and cordycepin under the chromatographic condition
Under chromatographic condition, adenosine, cordycepin are all separated preferably in the test sample, and not only the retention time of its chromatographic peak is consistent with reference substance, and each chromatographic peak has also all obtained confirming that preferably the result sees Fig. 1, Fig. 2, Fig. 3 and Fig. 4 through standard addition method.
2. sample determination result
Adenosine in the Cordyceps militaris fruiting body that the wilsonii solid culture medium is turned out and content of cordycepin result see table 3, and adenosine and cordycepin content are the highest in the Cordyceps militaris fruiting body that the wilsonii solid culture medium is turned out, and compare significant difference with the blank group.
Table 3
| Group | Adenosine (mg/g) | Cordycepin (mg/g) |
| Blank medium | 0.2012±0.0018 | 1.8241±0.0045 |
| The wilsonii solid culture medium | 0.9970±0.0032 | 3.4195±0.0124 |
Embodiment five: the cultural method of this embodiment wilsonii Cordyceps militaris, carry out: one, the Cordyceps militaris bacterial strain is inoculated on the PDA solid culture medium for beautiful rich No. two, in 28 ℃ of biochemical incubators, secretly cultivates 5d, obtain the bacterial classification of activation according to the following steps; Two, bacterial classification that activation is the good bacterium piece that is cut into 1cm * 1cm is inoculated in the liquid seed culture medium of 300ml; It is the dark 6d of cultivation under 23 ℃, the condition of 120r/min that static cultivation 24h is placed on temperature; Obtain bacteria suspension, add the 200ml sterile water then, get liquid spawn; Three, get the 5ml liquid-spawn inoculation in the blake bottle that the wilsonii solid culture medium is housed; Placing temperature is that 22 ℃, humidity are the dark 12d of cultivation under 55% the condition; Get mycelium; Move to then under the light and be in temperature that 23 ℃, humidity are 60%, intensity of illumination is 200lx, illumination every day 12h, day and night temperature after being cultured to mycelium under 10 ℃ the condition and all transferring yellow to by white, increase humidity to 95%, it is long high to 2cm to continue to be cultured to fruit body; With the ventilation switch opens on the blake bottle; Continue to be cultured to that sporophore growth is high to 8cm, fruit body is surperficial yellow powder shape material occurs and spinule appears in the top, gathers, and promptly accomplishes the cultivation of wilsonii Cordyceps militaris;
Wherein the wilsonii solid culture medium is made up of the rice of 40g, the Manyprickle Acanthopanax Root of 0.1～0.6g and the nutrient solution of 60ml in the step 3, and the bottled amount of blake bottle is 1/3; The preparation of said Manyprickle Acanthopanax Root: wilsonii is pulverized the back cross 20 mesh sieves; The every L of said nutrient solution is by the peptone of 20g, the glucose of 20g, the KH of 1g
2PO
4, 0.5g MgSO
47H
2The potato liquid of the zinc sulphate of O, 0.5g, 200g, the Cobastab of 5mg
1Form with the water of surplus; The preparation of said potato liquid: 200g removes the peel potato, and 30min is boiled in stripping and slicing, complements to 1000ml after the filtered through gauze, pH nature, 121 ℃ of autoclaving 30min.
The step 1 of this embodiment is sterile working in superclean bench.
The Cordyceps militaris bacterial strain is beautiful rich No. two in this embodiment step 1, buys the Yu Feng bacterium industry Co., Ltd from Harbin.
The wilsonii solid culture medium of different wilsonii content is cultivated gained wilsonii Cordyceps militaris in this embodiment, adopts the HPLC method to measure the active component of wilsonii in the fruit body, and step is following:
(1) chromatographic condition
Chromatographic column: Venusil XBP-C
18Post (4.6mm * 250mm, 5 μ m, USA);
Flowing phase: methyl alcohol-0.2% phosphoric acid (55: 45);
Flow velocity: 1mL/min;
Column temperature: 25 ℃;
Detect wavelength: 350nm;
Sample size: 10 μ L.
(2) precision takes by weighing Hyperoside respectively, the Quercetin reference substance is an amount of in the preparation of reference substance solution, processes the reference substance storing solution that mass concentration is 1mg/mL respectively with methyl alcohol.Accurately measure above-mentioned each 1mL of reference substance storing solution respectively, process Hyperoside, the Quercetin that mass concentration is 0.5mg/mL and mix reference substance solution.
(3) the preparation precision of need testing solution takes by weighing each 2 parts of the cordyceps militaris fruit bodies that the wilsonii solid culture medium of dry different wilsonii content cultivates, every part of 2g; The accurate respectively 5mL methyl alcohol that adds is weighed, and ultrasonic 60min takes out, and cooling is supplied the solvent that subtracts mistake with methyl alcohol, filters the back as need testing solution with 0.45 μ m filter membrane.
(4) sample determination is under chromatographic condition, get reference substance solution and need testing solution with 0.45 μ m membrane filtration after, the accurate respectively 10 μ L sample introductions of drawing, the record peak area calculates content by external standard method.
(5) result:
1. the separation under the chromatographic condition
Under chromatographic condition; Wilsonii active component Hyperoside, Quercetin are all separated preferably in the test sample; Not only the retention time of its chromatographic peak is consistent with reference substance, and each chromatographic peak has also all obtained confirming that preferably the result sees Fig. 5, Fig. 6 and Fig. 7 through standard addition method.
2. sample determination result
The Hyperoside in the Cordyceps militaris fruiting body that the wilsonii solid culture medium is turned out and the content results of Quercetin are seen table 4; It is thus clear that; In cordyceps culturing medium, add wilsonii, the active component that has wilsonii accordingly changes in the Cordyceps militaris fruiting body, and does not detect the active component of wilsonii in the blank group Cordyceps militaris fruiting body; Explain through adding wilsonii and cultivate Cordyceps militaris; Can produce the Cordyceps militaris that is rich in the wilsonii active component,, simplify the traditional Chinese medicine compatibility and lay the foundation for strengthening the effect of northern Chinese caterpillar Fungus.
Table 4
Annotate: compare * P＜0.05 with the blank group, * * P＜0.01
Claims (3)
1. the cultural method of a wilsonii Cordyceps militaris; The cultural method that it is characterized in that the wilsonii Cordyceps militaris; Carry out according to the following steps: one, the Cordyceps militaris bacterial strain is inoculated on the PDA solid culture medium for beautiful rich No. two, in 28 ℃ of biochemical incubators, secretly cultivates 5d, obtain the bacterial classification of activation; Two, bacterial classification that activation is the good bacterium piece that is cut into 1cm * 1cm is inoculated in the liquid seed culture medium of 300ml; It is the dark 6d of cultivation under 23 ℃, the condition of 120r/min that static cultivation 24h is placed on temperature; Obtain bacteria suspension, add the 200ml sterile water then, get liquid spawn; Three, get the 5ml liquid-spawn inoculation in the blake bottle that the wilsonii solid culture medium is housed; Placing temperature is that 22 ℃, humidity are the dark 10～14d of cultivation under 55%～60% the condition; Get mycelium; Move to then under the light and be in temperature that 23 ℃, humidity are 55%～60%, intensity of illumination is 180～220lx, illumination every day 10～12h, day and night temperature after being cultured to mycelium under 10 ℃ the condition and all transferring yellow to by white, increase humidity to 85%～95%, it is long high to 2cm to continue to be cultured to fruit body; With the ventilation switch opens on the blake bottle; Continue to be cultured to that sporophore growth to 4～8cm is high, fruit body is surperficial yellow powder shape material occurs and spinule appears in the top, gathers, and promptly accomplishes the cultivation of wilsonii Cordyceps militaris;
Wherein the wilsonii solid culture medium is made up of the rice of 40g, the Manyprickle Acanthopanax Root of 0.1～0.6g and the nutrient solution of 60ml in the step 3, and the bottled amount of blake bottle is 1/4～1/3; The preparation of said Manyprickle Acanthopanax Root: wilsonii is pulverized the back cross 20 mesh sieves; The every L of said nutrient solution is by the peptone of 20g, the glucose of 20g, the KH of 1g
2PO
4, 0.5g MgSO
47H
2The potato liquid of the zinc sulphate of O, 0.5g, 200g, the Cobastab of 5mg
1Form with the water of surplus; The preparation of said potato liquid: 200g removes the peel potato, and 30min is boiled in stripping and slicing, complements to 1000ml after the filtered through gauze, pH nature, 121 ℃ of autoclaving 30min.
2. the cultural method of a kind of wilsonii Cordyceps militaris according to claim 1 is characterized in that the PDA solid culture medium is made up of the potato liquid of 200g, the glucose of 20g, the agar of 18g and the water of 1000ml in the step 1, the pH nature; The preparation of said potato liquid: 200g removes the peel potato, and 30min is boiled in stripping and slicing, complements to 1000ml after the filtered through gauze, pH nature, 121 ℃ of autoclaving 30min.
3. the cultural method of a kind of wilsonii Cordyceps militaris according to claim 1 is characterized in that the every L of liquid seed culture medium in the step 2 forms the pH nature by the water of magnesium sulfate, 1g potassium dihydrogen phosphate and the surplus of the glucose of the potato liquid of 200g, 20g, 1g; The preparation of said potato liquid: 200g removes the peel potato, and 30min is boiled in stripping and slicing, complements to 1000ml after the filtered through gauze, pH nature, 121 ℃ of autoclaving 30min.
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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 |
| CN103688748B (en) * | 2013-12-13 | 2015-06-03 | 福建农林大学 | Epimedium cordyceps militaris cultivation and product processing method |
| CN104823699A (en) * | 2015-04-21 | 2015-08-12 | 吴中区胥口精益生物医药研究所 | Culture method of cordyceps militaris |
| CN108676825B (en) * | 2018-05-22 | 2021-10-22 | 福建农林大学 | Fermentation culture medium for promoting cordyceps militaris to produce extracellular substances |
| CN109429896A (en) * | 2018-12-14 | 2019-03-08 | 大连森源菌业有限公司 | A kind of method of Artificial Cultivation of Cordyceps militaris Link |
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|---|---|---|---|---|
| KR100876820B1 (en) * | 2008-10-23 | 2009-01-07 | 박재근 | A cultivating method for cordyceps sp |
| CN101513161A (en) * | 2009-03-18 | 2009-08-26 | 朱忠贵 | Formula for culture medium of cordyceps militaris liquid strains and method for culturing same |
| CN101791329A (en) * | 2010-03-30 | 2010-08-04 | 天津科技大学 | Astragalus waste cordyceps sinensis fermentation technique and application thereof |
| CN102091100A (en) * | 2009-12-15 | 2011-06-15 | 云南云百草生物技术有限公司 | Method for improving main medicinal ingredients through solid fermentation of Indian buead and cordyceps militaris |
-
2011
- 2011-06-24 CN CN2011101734511A patent/CN102293122B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100876820B1 (en) * | 2008-10-23 | 2009-01-07 | 박재근 | A cultivating method for cordyceps sp |
| CN101513161A (en) * | 2009-03-18 | 2009-08-26 | 朱忠贵 | Formula for culture medium of cordyceps militaris liquid strains and method for culturing same |
| CN102091100A (en) * | 2009-12-15 | 2011-06-15 | 云南云百草生物技术有限公司 | Method for improving main medicinal ingredients through solid fermentation of Indian buead and cordyceps militaris |
| CN101791329A (en) * | 2010-03-30 | 2010-08-04 | 天津科技大学 | Astragalus waste cordyceps sinensis fermentation technique and application thereof |
Non-Patent Citations (2)
| Title |
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| 张金艳等.培养瓶透气性对蛹虫草生长的影响.《广东农业科学》.2010,(第4期), * |
| 张雪超.北虫草标准化栽培新技术.《吉林农业》.2009,(第3期), * |
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