CN102428834A - Cultivation method of selenium-enriched Cordyceps militaris mother culture - Google Patents
Cultivation method of selenium-enriched Cordyceps militaris mother culture Download PDFInfo
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- CN102428834A CN102428834A CN2011102795211A CN201110279521A CN102428834A CN 102428834 A CN102428834 A CN 102428834A CN 2011102795211 A CN2011102795211 A CN 2011102795211A CN 201110279521 A CN201110279521 A CN 201110279521A CN 102428834 A CN102428834 A CN 102428834A
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Abstract
A cultivation method of selenium-enriched Cordyceps militaris mother fungi belongs to the technical field of the cultivation of Cordyceps militaris. The cultivation method comprises the following steps: 1) cultivating selenium-enriched strains on a test tube slant; 2) cultivating selenium-enriched Cordyceps militaris liquid strains; and 3) cultivating the selenium-enriched Cordyceps militaris. The cultivation method uses organic selenium yeast which has the lowest inhibitory effect on the growth of Cordyceps militaris, thus the selenium content of the culture medium is up to 50mg/L, the selenium content of the cultivated Cordyceps militaris is up to 35-50mg/kg which is triple of the selenium content of Cordyceps militaris with additional inorganic selenium, and the yield is increased by 20%.
Description
Technical field
The invention belongs to Cordyceps sinensis and cultivate technical field, relate in particular to the female cultivating method of planting of a kind of Se-enriched cordyceps militaris.
Background technology
Selenium is one of trace element of needed by human, and it has anti-cancer, anticancer, anti-oxidant, and the protection body is avoided the infringement of active oxygen radical, the protection heart, and treating liver necrosis improves immunity, and multiple pharmacological effect such as delay senility is described as " anticancer king ".Modern medicine study shows that the multiple disorders such as cancers of human body, cardiovascular and cerebrovascular disease, diabetes, hypertension etc. are all relevant with scarce selenium.The distributed pole of selenium in natural environment is inhomogeneous; The area of China 2/3rds is in and lacks the selenium state; In order to promote health, numerous scientific research institutions and biotechnology enterprise are converted into organic selenium with inorganic selenium through microorganism, plant, animal and produce safe, nontoxic, as to be convenient to absorption of human body utilization se-rich health food products.Along with people's is to selenium and immunologic function, the popularizing of cardiovascular and cerebrovascular diseases and cancer-resisting understanding, and the increasing demand of se-rich health food products increases, for the exploitation of se-rich health food products provides vast market.Se-enriched cordyceps militaris is in medium, to add trace elements of selenium, through to the absorption of mycelia in process of growth, transforms, and is enriched in the fruit body in conjunction with inorganic selenium being converted into organic selenium, produces Se-enriched cordyceps militaris.Prior art all is in medium, to add inorganic selenous acid or sodium selenite; The selenous acid of trace or sodium selenite have facilitation to the growth of northern Chinese caterpillar Fungus mycelia; But mycelial growth is suppressed when inorganic selenium reaches 20mg/L in medium; Surpassing the 30mg/L mycelia can not grow; Thereby limited the addition of inorganic selenium in medium, make cultivation the northern Chinese caterpillar Fungus selenium content one in 10-20mg/L, fail to demonstrate fully Se-enriched cordyceps militaris anti-cancer, anticancer, enhancing immunity, anti-ageing, prevent and treat the effect of Keshan disease, diabetes etc.
Summary of the invention
The purpose of this invention is to provide the female cultivating method of planting of a kind of Se-enriched cordyceps militaris; Be to adopt the organic selenium yeast; Avoid the murder by poisoning of inorganic selenous acid, sodium selenite, make the selenium content of medium reach 50mg/L, plant thereby the Se-enriched cordyceps militaris of cultivating is female to mycelial growth.
The present invention includes following steps:
One, the rich selenium culture of strains in test tube slant
1, medium composition: potato 200 grams, glucose 10-30 gram, peptone 10-20 gram, agar 15-30 gram, potassium dihydrogen phosphate are foretold 3 grams, the selenium yeast 10-50 gram of selenium content 2000mg/kg.
Compound method: take by weighing and clean peeling potato book sheet 200 grams, add 1000 milliliters and boiled 20 minutes, filter, get potato juice and put into 1000 ml beakers; Add glucose 20 grams, agar 20 grams, peptone 10 grams, potassium dihydrogen phosphate 05 gram; Selenium yeast 15 grams, water-bath is heated to agar and dissolves fully, adds water and supplies 1000 milliliters; Be divided in umbrella 20～200m.rn in vitro, 12 milliliters of every cuvette cartridges are used the tampon jam-pack; Keep sterilization 40 minutes for 120 ℃ at high-pressure sterilizing pot, while hot test tube is put into the inclined-plane that medium leaves tampon 5-6cm after taking the dish out of the pot, the cooling back bevel becomes solid, shaped subsequent use.
2, strain separating, connect slant strains: the wild cordyceps militaris fruit body that the sturdy circle of choosing growth, color and luster are orange or the cordyceps militaris fruit body of artificial culture, sterilized 1 minute with 75% alcohol, with aqua sterilisa flush away alcohol; In inoculating hood, fruit body being cut into 2mm book sheet inserts in the middle of the test tube medium; Otch is upwards cultivated to take out in 15 days in 20 ℃ of insulating boxs and is seen light, 3 days annesls of light scattering; It is smooth to select mycelial growth; Color and luster is orange, does not have female kind of Se-enriched cordyceps militaris of assorted bacterium, puts into 5 ℃ of insulating boxs and preserves subsequent use.
Two, the cultivation of Se-enriched cordyceps militaris liquid spawn
The prescription of nutrient solution: glucose 20 grams, peptone 10 grams, potassium dihydrogen phosphate 1 gram, magnesium sulfate 1 gram, selenium yeast 15 grams, 1000 milliliters in water.
Method of operating: take by weighing the listed raw material of prescription and put into 1000 ml waters and boiled 10 minutes, use the multilayer filtered through gauze after being cooled to room temperature, clear liquid is packed in the 500 mL of saline bottles; Every bottled 250 milliliters, use the tampon jam-pack, put into high-pressure sterilizing pot; 120 ℃ keep sterilization 30 minutes, are cooled to after the room temperature to plant together with mother to put into inoculating hood, cut with vaccinating lancet and femalely plant 0.5 square centimeter of access of an agreement that contracts a film or TV play to an actor or actress and float on the liquid level; Put into the darkroom; 20-22 ℃ of temperature of control leaves standstill cultivated 3 days, and female the kind grows white hypha, be placed in the shaking table 5 days the Se-enriched cordyceps militaris liquid spawn.
Three, the cultivation of Se-enriched cordyceps militaris
1, the preparation of solid culture medium nutrient solution: take by weighing glucose 20 grams, peptone 10 grams, potassium dihydrogen phosphate 1 gram, magnesium sulfate 1 gram, Cobastab, 100 milligrams, selenoprotein 35 grams are put into 1000 milliliters of 70-80 ℃ of hot water, are stirred to material dissolution.
2, the preparation of solid culture medium: in 600 milliliters big mouthful vials, every bottled rice 40 grams of going into, 60 milliliters of nutrient solutions; With polypropylene book film envelope bottleneck, put into the high pressure pot that goes out, under 120 ℃ of temperature, keep sterilization 60 minutes; Slowly take the dish out of the pot behind the blowdown presssure, cool off for use.
3, inoculation, cultivation: on super-clean bench, go into 3 milliliters of Se-enriched cordyceps militaris liquid spawns, move into 20-22 ℃ bacterium of darkroom control temperature, send out bacterium through 8 days lucifuges with the every bottle graft of inoculating gun of band nozzle; It is visible light that media surface all covers with white hypha, under light scattering 3-5 days mycelia transfer to orange to salmon pink, 20-22 ℃ of control temperature; Fruit body primordium appearred in 10 1 days in relative moisture 80-85%, again through 30 1 days growth; Fruit body is long to 4-8 centimetre; The fruit body maturation can be gathered when forming spore, in time is dried to moisture content and is no more than 5%, packs with plastic bag sealing.
The present invention adopts the organic selenium yeast minimum to the northern Chinese caterpillar Fungus growth inhibition, makes the selenium content of medium reach 50mg/L, and the Se-enriched cordyceps militaris selenium content of cultivating reaches 35-50mg/kg, and higher 2 times than the selenium content that adds inorganic selenium, output is high by 20%.
Further the present invention is explained below in conjunction with embodiment:
Embodiment
Embodiment: the first step, the rich selenium culture of strains in test tube slant: (1), medium composition: potato 200 grams, glucose 10-30 gram, peptone 10-20 gram, agar 15-30 gram, potassium dihydrogen phosphate are foretold 3 grams, the selenium yeast 10-50 gram of selenium content 2000mg/kg.Compound method: take by weighing and clean peeling potato book sheet 200 grams, add 1000 milliliters and boiled 20 minutes, filter, get potato juice and put into 1000 ml beakers; Add glucose 20 grams, agar 20 grams, peptone 10 grams, potassium dihydrogen phosphate 05 gram; Selenium yeast 15 grams, water-bath is heated to agar and dissolves fully, adds water and supplies 1000 milliliters; Be divided in umbrella 20～200m.rn in vitro, 12 milliliters of every cuvette cartridges are used the tampon jam-pack; Keep sterilization 40 minutes for 120 ℃ at high-pressure sterilizing pot, while hot test tube is put into the inclined-plane that medium leaves tampon 5-6cm after taking the dish out of the pot, the cooling back bevel becomes solid, shaped subsequent use.(2), strain separating, connect slant strains: the wild cordyceps militaris fruit body that the sturdy circle of choosing growth, color and luster are orange or the cordyceps militaris fruit body of artificial culture, sterilized 1 minute with 75% alcohol, with aqua sterilisa flush away alcohol; In inoculating hood, fruit body being cut into 2mm book sheet inserts in the middle of the test tube medium; Otch is upwards cultivated to take out in 15 days in 20 ℃ of insulating boxs and is seen light, 3 days annesls of light scattering; It is smooth to select mycelial growth; Color and luster is orange, does not have female kind of Se-enriched cordyceps militaris of assorted bacterium, puts into 5 ℃ of insulating boxs and preserves subsequent use.
The cultivation of second step, Se-enriched cordyceps militaris liquid spawn, the prescription of nutrient solution: glucose 20 grams, peptone 10 grams, potassium dihydrogen phosphate 1 gram, magnesium sulfate 1 gram, selenium yeast 15 grams, 1000 milliliters in water.Method of operating: take by weighing the listed raw material of prescription and put into 1000 ml waters and boiled 10 minutes, use the multilayer filtered through gauze after being cooled to room temperature, clear liquid is packed in the 500 mL of saline bottles; Every bottled 250 milliliters, use the tampon jam-pack, put into high-pressure sterilizing pot; 120 ℃ keep sterilization 30 minutes, are cooled to after the room temperature to plant together with mother to put into inoculating hood, cut with vaccinating lancet and femalely plant 0.5 square centimeter of access of an agreement that contracts a film or TV play to an actor or actress and float on the liquid level; Put into the darkroom; 20-22 ℃ of temperature of control leaves standstill cultivated 3 days, and female the kind grows white hypha, be placed in the shaking table 5 days the Se-enriched cordyceps militaris liquid spawn.
The cultivation of the 3rd step, Se-enriched cordyceps militaris: the preparation of (1), solid culture medium nutrient solution: take by weighing glucose 20 grams, peptone 10 grams, potassium dihydrogen phosphate 1 gram; Magnesium sulfate 1 gram, Cobastab, 100 milligrams; Selenoprotein 35 grams are put into 1000 milliliters of 70-80 ℃ of hot water, are stirred to material dissolution.(2), the preparation of solid culture medium: in 600 milliliters big mouthful vials, every bottled rice 40 grams of going into, 60 milliliters of nutrient solutions; With polypropylene book film envelope bottleneck, put into the high pressure pot that goes out, under 120 ℃ of temperature, keep sterilization 60 minutes; Slowly take the dish out of the pot behind the blowdown presssure, cool off for use.(3), inoculation, cultivation: on super-clean bench, go into 3 milliliters of Se-enriched cordyceps militaris liquid spawns, move into 20-22 ℃ bacterium of darkroom control temperature, send out bacterium through 8 days lucifuges with the every bottle graft of inoculating gun of band nozzle; It is visible light that media surface all covers with white hypha, under light scattering 3-5 days mycelia transfer to orange to salmon pink, 20-22 ℃ of control temperature; Fruit body primordium appearred in 10 1 days in relative moisture 80-85%, again through 30 1 days growth; Fruit body is long to 4-8 centimetre; The fruit body maturation can be gathered when forming spore, in time is dried to moisture content and is no more than 5%, packs with plastic bag sealing.
Claims (4)
1. the Se-enriched cordyceps militaris mother plants cultivating method, and characteristic is to comprise the steps: the first step, the rich selenium culture of strains in test tube slant; The cultivation of second step, Se-enriched cordyceps militaris liquid spawn; The cultivation of the 3rd step, Se-enriched cordyceps militaris.
2. the female cultivating method of planting of Se-enriched cordyceps militaris according to claim 1; It is characterized in that the rich selenium culture of strains in described test tube slant comprises: 1, medium composition: potato 200 grams; Glucose 10-30 gram, peptone 10-20 gram, agar 15-30 gram; Potassium dihydrogen phosphate is foretold 3 grams, the selenium yeast 10-50 gram of selenium content 2000mg/kg; Compound method: take by weighing and clean peeling potato book sheet 200 grams, add 1000 milliliters and boiled 20 minutes, filter, get potato juice and put into 1000 ml beakers; Add glucose 20 grams, agar 20 grams, peptone 10 grams, potassium dihydrogen phosphate 05 gram; Selenium yeast 15 grams, water-bath is heated to agar and dissolves fully, adds water and supplies 1000 milliliters; Be divided in umbrella 20～200m.rn in vitro, 12 milliliters of every cuvette cartridges are used the tampon jam-pack; Keep sterilization 40 minutes for 120 ℃ at high-pressure sterilizing pot, while hot test tube is put into the inclined-plane that medium leaves tampon 5-6cm after taking the dish out of the pot, the cooling back bevel becomes solid, shaped subsequent use; 2, strain separating, connect slant strains: the wild cordyceps militaris fruit body that the sturdy circle of choosing growth, color and luster are orange or the cordyceps militaris fruit body of artificial culture, sterilized 1 minute with 75% alcohol, with aqua sterilisa flush away alcohol; In inoculating hood, fruit body being cut into 2mm book sheet inserts in the middle of the test tube medium; Otch is upwards cultivated to take out in 15 days in 20 ℃ of insulating boxs and is seen light, 3 days annesls of light scattering; It is smooth to select mycelial growth; Color and luster is orange, does not have female kind of Se-enriched cordyceps militaris of assorted bacterium, puts into 5 ℃ of insulating boxs and preserves subsequent use.
3. the female cultivating method of planting of Se-enriched cordyceps militaris according to claim 1 is characterized in that the cultivation of Se-enriched cordyceps militaris liquid spawn comprises: the prescription of nutrient solution: glucose 20 grams, peptone 10 grams; Potassium dihydrogen phosphate 1 gram; Magnesium sulfate 1 gram, selenium yeast 15 grams, 1000 milliliters in water; Method of operating: take by weighing the listed raw material of prescription and put into 1000 ml waters and boiled 10 minutes, use the multilayer filtered through gauze after being cooled to room temperature, clear liquid is packed in the 500 mL of saline bottles; Every bottled 250 milliliters, use the tampon jam-pack, put into high-pressure sterilizing pot; 120 ℃ keep sterilization 30 minutes, are cooled to after the room temperature to plant together with mother to put into inoculating hood, cut with vaccinating lancet and femalely plant 0.5 square centimeter of access of an agreement that contracts a film or TV play to an actor or actress and float on the liquid level; Put into the darkroom; 20-22 ℃ of temperature of control leaves standstill cultivated 3 days, and female the kind grows white hypha, be placed in the shaking table 5 days the Se-enriched cordyceps militaris liquid spawn.
4. the female cultivating method of planting of Se-enriched cordyceps militaris according to claim 1 is characterized in that the cultivation of Se-enriched cordyceps militaris comprises: 1, the preparation of solid culture medium nutrient solution: take by weighing glucose 20 grams, peptone 10 grams; Potassium dihydrogen phosphate 1 gram, magnesium sulfate 1 gram, Cobastab; 100 milligrams; Selenoprotein 35 grams are put into 1000 milliliters of 70-80 ℃ of hot water, are stirred to material dissolution; 2, the preparation of solid culture medium: in 600 milliliters big mouthful vials, every bottled rice 40 grams of going into, 60 milliliters of nutrient solutions; With polypropylene book film envelope bottleneck, put into the high pressure pot that goes out, under 120 ℃ of temperature, keep sterilization 60 minutes; Slowly take the dish out of the pot behind the blowdown presssure, cool off for use; 3, inoculation, cultivation: on super-clean bench, go into 3 milliliters of Se-enriched cordyceps militaris liquid spawns, move into 20-22 ℃ bacterium of darkroom control temperature, send out bacterium through 8 days lucifuges with the every bottle graft of inoculating gun of band nozzle; It is visible light that media surface all covers with white hypha, under light scattering 3-5 days mycelia transfer to orange to salmon pink, 20-22 ℃ of control temperature; Fruit body primordium appearred in 10 1 days in relative moisture 80-85%, again through 30 1 days growth; Fruit body is long to 4-8 centimetre; The fruit body maturation can be gathered when forming spore, in time is dried to moisture content and is no more than 5%, packs with plastic bag sealing.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102771314A (en) * | 2012-08-15 | 2012-11-14 | 苏州硒谷科技有限公司 | Method for culturing selenium-enriched cordyceps militaris |
| CN102960184A (en) * | 2012-11-22 | 2013-03-13 | 兴隆县庆旺富硒食用菌农民专业合作社 | Edible fungus strain rich in organic selenium, preparation method therefore, edible fungus rich in organic selenium and culturing method thereof |
| CN103190291A (en) * | 2013-04-26 | 2013-07-10 | 上海亿佳亿生态科技有限公司 | Cultivation method of selenium-enriched cordyceps |
| CN104396571A (en) * | 2014-12-15 | 2015-03-11 | 重庆综艺制药有限公司 | High-cordycepin-content rich-selenium cordyceps sinensis cultivation method |
| CN104396572A (en) * | 2014-12-15 | 2015-03-11 | 重庆综艺制药有限公司 | Cordyceps sinensis strain spore separation, purification and rejuvenation method |
| CN105432319A (en) * | 2015-01-13 | 2016-03-30 | 罗大全 | Biological selenium-enriched fungi breeding method |
| CN106105780A (en) * | 2016-07-07 | 2016-11-16 | 孙备宽 | A kind of Se-rich pearl Cordyceps production method |
| CN109220536A (en) * | 2018-11-20 | 2019-01-18 | 常德炎帝牧源农业发展有限公司 | A kind of inoculation method of organic selenium-rich Cordyceps militaris |
| CN110215486A (en) * | 2019-07-08 | 2019-09-10 | 湖北圣峰药业有限公司 | A kind of selenium worm grass compound preparation and its preparation method and application |
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2011
- 2011-09-20 CN CN2011102795211A patent/CN102428834A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102771314A (en) * | 2012-08-15 | 2012-11-14 | 苏州硒谷科技有限公司 | Method for culturing selenium-enriched cordyceps militaris |
| CN102960184A (en) * | 2012-11-22 | 2013-03-13 | 兴隆县庆旺富硒食用菌农民专业合作社 | Edible fungus strain rich in organic selenium, preparation method therefore, edible fungus rich in organic selenium and culturing method thereof |
| CN102960184B (en) * | 2012-11-22 | 2015-02-11 | 兴隆县庆旺富硒食用菌农民专业合作社 | Preparation methodof edible fungus strain rich in organic selenium |
| CN103190291A (en) * | 2013-04-26 | 2013-07-10 | 上海亿佳亿生态科技有限公司 | Cultivation method of selenium-enriched cordyceps |
| CN104396571A (en) * | 2014-12-15 | 2015-03-11 | 重庆综艺制药有限公司 | High-cordycepin-content rich-selenium cordyceps sinensis cultivation method |
| CN104396572A (en) * | 2014-12-15 | 2015-03-11 | 重庆综艺制药有限公司 | Cordyceps sinensis strain spore separation, purification and rejuvenation method |
| CN105432319A (en) * | 2015-01-13 | 2016-03-30 | 罗大全 | Biological selenium-enriched fungi breeding method |
| CN106105780A (en) * | 2016-07-07 | 2016-11-16 | 孙备宽 | A kind of Se-rich pearl Cordyceps production method |
| CN109220536A (en) * | 2018-11-20 | 2019-01-18 | 常德炎帝牧源农业发展有限公司 | A kind of inoculation method of organic selenium-rich Cordyceps militaris |
| CN110215486A (en) * | 2019-07-08 | 2019-09-10 | 湖北圣峰药业有限公司 | A kind of selenium worm grass compound preparation and its preparation method and application |
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Application publication date: 20120502 |