CN1772876A - Formulation of aweto culture medium and oxygenating culture method of aweto - Google Patents
Formulation of aweto culture medium and oxygenating culture method of aweto Download PDFInfo
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Abstract
The present invention is formulation of aweto culture medium and oxygenating culture method of aweto. By means of high efficiency culture medium, deep culture of the liquid spawn and the oxygenating in the initial growth stage of sporophore, the present invention has high aweto sporophore producing efficiency and high yield. The culture medium for liquid spawn, which is obtained through spore or tissue separation and inoculated to culture medium solution, consists of glucose, peptone, yeast powder, potassium dihydrogen phosphate, magnesium sulfate, sodium chloride, vitamin B1 and water in certain weight proportion.
Description
Technical field
The bacterial classification and the sporophore that the present invention relates to a kind of Cordyceps militaris (L.) Link. are cultivated substrate formula, and adopt oxygenation to expand complicated strong method of promoting production with oxygenating in cultivating process.
Background technology
Along with the improvement of living condition, more and more to pay close attention to for quality of life and healthy raising, various nutritional supplementss also become the leading product in health care market.Cordyceps militaris (L.) Link. is a kind of precious Chinese medicine with very high medicinal and nourishing effects, and that wild Chinese caterpillar fungus distributes is rare, resource is extremely scarce.For meeting the need of market, existing with artificially breeding Chinese caterpillar fungus.And with the Chinese caterpillar fungus of artificial culture, its effective ingredient approaches natural, partial elements even be higher than more than several times or tens times of wild Chinese caterpillar fungus, and therefore the worm grass product of selling in the market is artificial culture mostly.
The strain cultivation of existing Cordyceps militaris (L.) Link. adopts the shaking table device usually, shakes by using shaking table will shake bottle, thereby impels the bacterial classification that shakes in the bottle to expand numerous growth.But this type of spawn culture mode can not be given regularly supplemental oxygen of liquid bacterial classification, and liquid spawn can only fill and shake bottle volumetrical below 30%, is spread otherwise will shake out leakage, so production efficiency is not high, takies meaninglessly and shakes a bottle volume.
When the sporophore of Cordyceps militaris (L.) Link. grows into 3-4 centimetre in culturing bottle, after promptly passing through primary growth, need to provide competent oxygen to grow fast in the culturing bottle for sporophore, also must effectively control the temperature and humidity in the culturing bottle simultaneously, otherwise sporophore the uneven of poor growth, length will appear and causes the underproduction.Existing way is that the bottleneck at culturing bottle is covered with film or the lid that one deck is pricked a plurality of ventilating pits, indoorly carries out repeatedly ventilation and improves oxygen supply amount in the culturing bottle by regularly carrying every day to train.But adopt this type of ventilation mode,, then do not have the effect of supplemental oxygen if the ventilating pit on film or the lid is too small and very few; If ventilating pit is excessive and too much, then sporophore is easy to dehydration and the influence growth; The excessive temperature differentials that humidity is excessive, oxygen-supplying amount is not enough or cause owing to ventilation all can cause sporophore rotten or be covered with mycelia and blackening is rotted, and then Cordyceps militaris (L.) Link. loses pharmaceutical use.
The bacterial classification of the difficult degeneration that obtains through separation and purification in addition, employed substratum is also very crucial in its cultivating process.Lack necessary promotor in the existing disclosed culture medium prescription, and nutrition has much room for improvement, the cultivation amount of surviving of liquid spawn is not high.
Prior art as above-mentioned content exists problems and deficiency, cultivating process for Cordyceps militaris (L.) Link., growing from the strain cultivation to the sporophore all exists and needs to improve and improve part, still do not have effective measure for regular replenishment oxygenation especially, thus Cordyceps militaris (L.) Link. production efficiency is limited, sporophore growth slowly, look uneven, nutrition and pharmaceutical use be not high.
Summary of the invention
Bacterial classification of a kind of Cordyceps militaris (L.) Link. of the present invention and sporophore are cultivated substrate formula, its purpose is to address the above problem with deficiency provides a kind of high-efficiency strain culture medium prescription, and realization improves the rejuvenation submerged culture of liquid spawn, and in the primary growth of the sporophore short rectangular method of oxygenation after the stage, thereby improve the production efficiency and the output of Cordyccps-militaris-(L.)-link. Sporophore.
For achieving the above object, the pure strain that obtains after by spore separation or separate tissue inserts in the culture medium solution, and for realizing that deep layer expands complicated strong cultivation, the culture medium prescription of described liquid spawn is,
By the part by weight of proportioning composition, glucose 2-3%, peptone 1.5-2%, yeast powder 1.5-2%, potassium primary phosphate 0.003-0.01%, sal epsom 0.002-0.005%, sodium-chlor 0.5-1%, vitamins B
10.002-0.005%, all the other are water.
Cordyceps militaris (L.) Link. oxygenation method of cultivation of the present invention, its main key points in design are to implement the short length technology of oxygenation at the critical stage of strain cultivation and sporophore growth.
Also can under the prerequisite that adopts above-mentioned substrate formula, implement the short long technology of oxygenation at the critical stage of strain cultivation and sporophore growth.
At the cultivation stage of liquid spawn, the shaking table device that changes available technology adopting shakes alr mode, expands numerous growth for impelling in the bottle liquid spawn, adopts the regularly way of supplemental oxygen of the liquid spawn of ventilate device in culturing bottle.Behind aerating oxygen, liquid spawn stirs with nutrient solution is upper and lower, thereby liquid spawn can deep layer evenly stir, and not only give supplemental oxygen in the culture environment, and bacterial classification has suitable vigor.
When the sporophore of Cordyceps militaris (L.) Link. grows into 3-4 centimetre through primary growth, adopt special ventilate device in bottle, to feed termly through the oxygen after filtering, under the prerequisite that does not influence humidity and temperature in the bottle, provide sporophore growth essential oxygen constantly, thus sporophore growth fast, long neat sporophore growth is full and have higher pharmaceutical use.
Described Cordyceps militaris (L.) Link. oxygenation method of cultivation, its realization flow is:
The first step, the preparation bacterial classification.
Gather the sporophore of wild cordyceps militaris and carry out the separation and purification cultivation, can adopt spore separation or tissue isolation that the bacterium pearl that obtains is seeded in and expand the system cultivation on the substratum, to obtain having stability and the pure strain that is difficult for degenerating.
Second step, the cultivation of liquid spawn.
Spore separation or tissue isolation will be cultivated in the pure strain access culture medium solution that obtain.The prescription of described culture medium solution can adopt above-mentioned prescription content, promptly by the part by weight of proportioning composition, and glucose 2-3%, peptone 1.5-2%, yeast powder 1.5-2%, potassium primary phosphate 0.003-0.01%, sal epsom 0.002-0.005%, sodium-chlor 0.5-1%, vitamins B
10.002-0.005%, all the other are water.
Select different model and volumetrical wide-necked bottle for use, hold culture medium solution, culture medium solution occupies whole bottle volumetrical about 75%.
Seal with tampon or soft rubber ball at bottleneck, the heat sterilization that under 110-130 ℃ of temperature condition culture medium solution was carried out about 25-30 minute is handled.
Treat culture medium solution cooling back access pure strain, in wide-necked bottle, feed termly through the oxygen after filtering by the ventilate device again, then in the culture medium solution behind the aerating oxygen, bacterial classification can obtain good grow aerobically environment, the carbonic acid gas that growth produced can in time be discharged, the degree of depth expands complicated strengthening under the bubble effect that bacterial classification produces after oxygen feeds, and can obtain liquid spawn One's name is legion, that have vigor.
During aerating oxygen, should continue to implement logical oxygen operation in culture medium solution, and temperature is controlled in the 18-23 ℃ of scope, logical oxygen time remaining is about 5 days.
As stated above in wide-necked bottle constantly aerating oxygen the time, still need leave venting hole at the bottle mouth position of culturing bottle, so that the carbonic acid gas that growth produced can in time be discharged.
In the 3rd step, liquid-spawn inoculation and primary growth are cultivated.
With the liquid spawn that above-mentioned cultivation obtains, be inoculated into and carry out the primary growth cultivation on the substratum in the culturing bottle.
The inoculation liquid spawn can be adopted rice medium or silkworm chrysalis vaccination ways.
The rice medium inoculation, be in culturing bottle, pack into rice and dried silkworm chrysalis meal, can be modulated into nutritive medium by following prescription, promptly by the part by weight of proportioning composition, glucose 1-2%, peptone 0.3-1%, potassium primary phosphate 0.05-0.1%, sal epsom 0.02-0.07%, triammonium citrate 0.03-0.08%, vitamins B
10.002-0.005%, all the other are water, and are modulated into the culture solution of pH value between 6.5-7.5, pour in the culturing bottle then.
The bottleneck of culturing bottle is tightened, and the heat sterilization that under 110-130 ℃ of temperature condition rice medium was carried out about 25-30 minute is handled.
After treating the rice medium cooling, adopt through spraying into liquid spawn in the rice medium of disinfectant spraying device in culturing bottle.
Behind the liquid-spawn inoculation, culturing bottle is moved to the lucifuge position, room temperature is controlled at about 20 ℃, through 2-3 days time, the surface of rice medium all covered with mycelia;
Through 4-5 days time, mycelia penetrated half degree of depth of substratum; At this moment, give culturing bottle naturally astigmatism irradiation and bottle mouth position prick get the part ventilation hole with controlling moisture about 85%;
Through about 10 days, increase strong illumination 4-8 hour of 500-1000 lux intensity every day again, between temperature is controlled 20-23 ℃.
Through 10-15 days primary growth breeding phase, the sporophore growth of Cordyceps militaris (L.) Link. was to the 3-4 cm height.
The silkworm chrysalis inoculation culture is earlier silkworm chrysalis, to be injected to liquid spawn in the silkworm chrysalis body with asepsis injector after 3-5 minute via 75% alcohol-pickled sterilization, and silkworm chrysalis is placed in the culture dish; Or silkworm chrysalis is placed in the culture dish, adopt through the disinfectant spraying device, liquid spawn is sprayed to the silkworm chrysalis whole body, then culture dish is sealed.
The silkworm chrysalis of inoculation liquid spawn is moved to the lucifuge position, room temperature is controlled at about 20 ℃, through 7-10 days time, pupal cell all covered with mycelia, and pupal cell ossifys.
The pupal cell that ossifys is moved in the culturing bottle, bottleneck is tightened processing.Through time of 5-7 days, pupal cell grew white or golden yellow mycelia, and bottleneck is left some ventilation holes, and astigmatism is shone naturally, indoor humidity is controlled at about 85% to give culturing bottle, and room temp is controlled at about 20 ℃.
Through about 5 days, increase strong illumination 4-8 hour of 500-1000 lux intensity every day again, between temperature is controlled 20-23 ℃.
Through 10-15 days primary growth breeding phase, the sporophore growth of Cordyceps militaris (L.) Link. was to the 3-4 cm height.
In the 4th step, the oxygenation growth of sporophore is cultivated.
Through the primary growth breeding phase, sporophore growth adopts ventilate device aerating oxygen in culturing bottle to the 3-4 cm height, by the venting hole of bottleneck the carbonic acid gas that sporophore growth produced is in time discharged simultaneously.
One of main points of Cordyceps militaris (L.) Link. oxygenation method of cultivation of the present invention are promptly to promote by oxygenation quick, the evenly growth of sporophore after sporophore is through the primary growth breeding phase.
Behind the primary growth breeding phase, sporophore enters the vigorous stage of growth, if the oxygen content in the culturing bottle is very few at this moment, then can cause sporophore growth slow and uneven, thereby the oxygen content that increases in its growing environment in the sporophore growth later stage can effectively promote the speed of growth and output.
During aerating oxygen, the temperature of aerating oxygen need be controlled at the room temperature of culture environment in culturing bottle, and the logical at every turn oxygen time is controlled in 10-15 scope second.
In the oxygenation growth nurturing period of sporophore, give the sporophore in the culturing bottle logical oxygen secondary at least, the timed interval was at 5-7 days.
When the sporophore oxygenation in culturing bottle promotes growth as stated above, need to leave the plurality of rows pore, so that the carbonic acid gas that sporophore growth produced can in time be discharged at the bottle mouth position of culturing bottle.
Cultivate through about 15 days oxygenation, sporophore enters the stage of maturity, and sporophore can grow to the bottle mouth position of culturing bottle uniformly, the Cordyceps militaris (L.) Link. of then can directly gathering.
As above content promptly is the general planning of Cordyceps militaris (L.) Link. oxygenation method of cultivation of the present invention.
In in strain cultivation solution and through the sporophore in primary growth stage, implementing the short long process of regular oxygenation, feed in the culturing bottle oxygen temperature and humidity should with culture environment in identical.
When the logical oxygen of the liquid spawn in culture medium solution, the ventilation mouth of pipe should be placed near the culturing bottle bottom, so that the oxygen that feeds enters culture medium solution near at the bottom of the bottle, then culture medium solution can fully stir to increase the oxygen content of growth environment.
When passing through the sporophore oxygenation of primary growth breeding phase, the ventpipe that feeds culturing bottle should be provided with a plurality of outlets, feed with multi-angle in the culturing bottle so that enter the oxygen of culturing bottle, guarantee that the oxygen content in the culturing bottle is even, evenly grow in order to sporophore.
Content to sum up, advantage and beneficial effect that described Cordyceps militaris (L.) Link. is cultivated substrate formula and Cordyceps militaris (L.) Link. oxygenation method of cultivation are:
1, adopt oxygenation to cultivate in the strain cultivation stage, can realize the bacterial classification rejuvenation submerged culture, bacterial classification has advantages such as disease-resistant bacterium, stalwartness and vigor foot; And do not adopt existing shaking table device, and can reduce equipment cost effectively, the bacterial classification solution capacity in the bottle improves, and also can improve integral production efficient.
2, when sporophore through primary growth after the stage, grow fast for sporophore by sufficient oxygen is provided, thereby sporophore growth is uniform, zoon, nutrition high the output height.
3, the critical stage of cultivating Cordyceps militaris (L.) Link. adopts the short rectangular method of oxygenation, the ventilate apparatus structure is simple, input cost is few and Cordyceps militaris (L.) Link. growth simultaneously fast, output is high, thereby aforesaid method is easy to promote the use of, economic return is obvious.
Description of drawings
Fig. 1 is described strain cultivation bottle and oxygen-increasing device synoptic diagram.
Fig. 2 is the oxygen-increasing device synoptic diagram of described sporophore oxygenation growth phase.
As depicted in figs. 1 and 2, nutrient solution 1, cotton plug 2, oxygen-supplying tube 3, air filter 4, connection rubber hose 5, oxygen increasing pump 6, wide-necked bottle 7.
Embodiment
Embodiment 1, and in conjunction with Fig. 1 and Fig. 2, described Chinese caterpillar fungus culture medium prescription is that the pure strain that obtains after by spore separation or separate tissue inserts in the culture medium solution, for realizing that deep layer expands the cultivation matrix that complicated strong cultivation provides.
Described culture medium prescription is, by the part by weight of proportioning composition, and glucose 2%, peptone 1.5%, yeast powder 1.5%, potassium primary phosphate 0.005%, sal epsom 0.002%, sodium-chlor 0.5%, VITMAIN B1 0.002-0.005%, all the other are water.
Cordyceps militaris (L.) Link. oxygenation method of cultivation of the present invention, its realization flow is:
The first step, the preparation bacterial classification.
Gather the sporophore of wild cordyceps militaris and carry out the separation and purification cultivation, adopt spore separation that the bacterium pearl that obtains is seeded in to expand on the substratum to make and cultivate, controlled temperature is about 20-23 ℃, and process promptly obtained to have stability and difficult pure strain of degenerating in 5-6 days.Pure strain is deposited in the environment of 4 ℃ in refrigerator and preserve.
Second step, the cultivation of liquid spawn.
The pure strain that spore separation is obtained inserts in the culture medium solution and cultivates.
Described culture medium prescription is, by the part by weight of proportioning composition, and glucose 2%, peptone 1.5%, yeast powder 1.5%, potassium primary phosphate 0.005%, sal epsom 0.002%, sodium-chlor 0.5%, VITMAIN B1 0.002-0.005%, all the other are water.
Select for use several wide-necked bottles of 3000 milliliters 7 to hold above-mentioned nutrient solution 1, nutrient solution 1 occupies whole bottle volumetrical about 75%.
Seal with alternative cotton plug 2 at bottleneck, the heat sterilization that carries out under 120 ℃ of temperature condition about 30 minutes is handled.
Treat to take out the alternative tampon 2 of bottleneck and insert pure strain after nutrient solution 1 cooling, adopt by oxygen-supplying tube 3, air filter 4, connect the oxygen the ventilate device that constitutes with rubber hose 5 and oxygen increasing pump 6 feeds the process filtration in wide-necked bottle 7 after.
When aerating oxygen in strain cultivation liquid 1, by oxygen increasing pump 6 suction oxygen, not influence the culture environment in the wide-necked bottle 7.
When leading to oxygen in nutrient solution 1, the mouth of pipe of oxygen-supplying tube 3 places near the bottom of contiguous wide-necked bottle 7, so that the oxygen that feeds enters nutrient solution 1 at the bottom of bottle, then nutrient solution 1 can fully stir.
In the nutrient solution 1 behind the aerating oxygen, bacterial classification can obtain good grow aerobically environment, the carbonic acid gas that growth produced can in time be discharged, and the degree of depth expands complicated strengthening under the bubble effect that bacterial classification produces after oxygen feeds, and can obtain liquid spawn One's name is legion, that have vigor.
During aerating oxygen, continue logical oxygen every day in nutrient solution 1, temperature is controlled at 23 ℃, and the logical oxygen time need continue 5 days.
In the 3rd step, liquid-spawn inoculation and primary growth are cultivated.
With the liquid spawn that above-mentioned cultivation obtains, be inoculated into and carry out the primary growth cultivation on the substratum in the culturing bottle.
The inoculation liquid spawn is adopted the rice medium vaccination ways.
Rice medium inoculation is pack in culturing bottle rice and dried silkworm chrysalis meal, is modulated into nutritive medium by following prescription, promptly by the part by weight of proportioning composition, glucose 1%, peptone 0.3%, potassium primary phosphate 0.05%, sal epsom 0.02%, triammonium citrate 0.03%, vitamins B
10.002%, all the other are water, and are modulated into the culture solution of pH value 6.5, pour in the culturing bottle then.
The bottleneck of culturing bottle is tightened, and the heat sterilization that under 120 ℃ of temperature condition rice medium was carried out about 30 minutes is handled.
After treating the rice medium cooling, adopt through spraying into liquid spawn in the rice medium of disinfectant spraying device in culturing bottle.
Behind the liquid-spawn inoculation, culturing bottle is moved to the lucifuge position, room temperature is controlled at about 20 ℃, through 3 day time, the surface of rice medium all covered with mycelia;
Through 5 day time, mycelia penetrated half degree of depth of substratum; At this moment, give culturing bottle naturally astigmatism irradiation and bottle mouth position prick get the part ventilation hole with controlling moisture about 85%;
Again through about 10 days, increase the strong illumination 4 hours of 1000 lux intensity every day, 23 ℃ of temperature controls.
Through 15 days primary growth breeding phase, the sporophore growth of Cordyceps militaris (L.) Link. was to the 3-4 cm height.
In the 4th step, the oxygenation growth of sporophore is cultivated.
Through the primary growth breeding phase, sporophore growth adopts by air cushioning pipe 11 jet needle tubing 12 to the 3-4 cm height, position fixing knob 13, ventilation handle 14, AFS 15, the ventilate device that rubber hose 16 and air compressor pump 17 constitutes, aerating oxygen constantly in culturing bottle.
When aerating oxygen in culturing bottle, by another spatial oxygen of air compressor pump 17 suction and culture environment uniform temp and humidity, not influence the sporophore growth environment in the culturing bottle.
Via the oxygen of air compressor pump 17 suction, enter in the air cushioning pipe 11 through rubber hose 16.Wherein, between rubber hose 16 and air cushioning pipe 11, be provided with one and be used to the ventilation handle 14 controlling and grip, on ventilation handle 14, an AFS 15 is set.
One location handle 13 is set on the top of air cushioning pipe 11.During operation, 6 jet needle tubings 12 are inserted in 6 culturing bottles simultaneously by gripping position fixing knob 13 may command.
Simultaneously, by the bottleneck venting hole of culturing bottle, the carbonic acid gas that sporophore growth produced in time can be discharged.
During aerating oxygen, the temperature of aerating oxygen need be controlled at the room temperature of culture environment in culturing bottle, and the logical at every turn oxygen time was controlled at 15 seconds.
In 15 days oxygenation cultivating process, give the sporophore in the culturing bottle logical oxygen secondary, the timed interval is 7 days.
Cultivate through about 15 days oxygenation, sporophore enters the stage of maturity, and sporophore can grow to the bottle mouth position of culturing bottle uniformly, the Cordyceps militaris (L.) Link. of then can directly gathering.
Claims (7)
1, a kind of Cordyceps militaris (L.) Link. is cultivated substrate formula, it is characterized in that: by the part by weight of proportioning composition, and glucose 2-3%, peptone 1.5-2%, yeast powder 1.5-2%, potassium primary phosphate 0.003-0.01%, sal epsom 0.002-0.005%, sodium-chlor 0.5-1%, vitamins B
10.002-0.005%, all the other are water.
2, a kind of Cordyceps militaris (L.) Link. oxygenation method of cultivation is characterized in that: in strain cultivation with through implementing the short long method of oxygenation in the sporophore cultivating process in primary growth stage;
At the cultivation stage of liquid spawn, adopt the ventilate device way of the liquid spawn supplemental oxygen in culturing bottle constantly, liquid spawn is stirred with nutrient solution is upper and lower;
When the sporophore of Cordyceps militaris (L.) Link. grows into 3-4 centimetre through primary growth, adopt ventilate device regular aerating oxygen in bottle, under the prerequisite that does not influence humidity and temperature in the bottle, provide sporophore growth essential oxygen.
3, Cordyceps militaris (L.) Link. oxygenation method of cultivation according to claim 2 is characterized in that: the realization flow of described method of cultivation is,
The first step, the preparation bacterial classification;
Gather the sporophore of wild cordyceps militaris and carry out the separation and purification cultivation, can adopt spore separation or tissue isolation that the bacterium pearl that obtains is seeded in and expand the system cultivation on the substratum, to obtain having stability and the pure strain that is difficult for degenerating;
Second step, the cultivation of liquid spawn;
With cultivating in the pure strain access culture medium solution that obtains, adopt wide-necked bottle to hold culture medium solution, culture medium solution occupies whole bottle volumetrical about 75%;
Seal with tampon or soft rubber ball at bottleneck, the heat sterilization that under 110-130 ℃ of temperature condition culture medium solution was carried out about 25-30 minute is handled;
Treat culture medium solution cooling back access pure strain, feed termly through the oxygen after filtering in wide-necked bottle by the ventilate device again that the carbonic acid gas that growth produced is in time discharged;
During aerating oxygen, logical constantly oxygen and temperature are controlled in the 18-23 ℃ of scope in culture medium solution, and the logical oxygen time need continue 5 days;
In the 3rd step, liquid-spawn inoculation and primary growth are cultivated;
With the liquid spawn that above-mentioned cultivation obtains, be inoculated into and carry out the primary growth cultivation on the substratum in the culturing bottle;
The inoculation liquid spawn is adopted rice medium or silkworm chrysalis vaccination ways;
Through 10-15 days primary growth breeding phase, the sporophore growth of Cordyceps militaris (L.) Link. was to the 3-4 cm height;
In the 4th step, the oxygenation growth of sporophore is cultivated;
Through the primary growth breeding phase, sporophore growth adopts the ventilate device to feed termly through the oxygen after filtering in culturing bottle to the 3-4 cm height, by the venting hole of bottleneck the carbonic acid gas that sporophore growth produced is in time discharged simultaneously;
During aerating oxygen, the temperature of aerating oxygen need be controlled at the room temperature of culture environment in culturing bottle, and the logical at every turn oxygen time is controlled in 10-15 scope second;
In the oxygenation growth nurturing period of sporophore, give the sporophore in the culturing bottle logical oxygen secondary at least, the timed interval was at 5-7 days.
When the sporophore oxygenation in culturing bottle promotes growth as stated above, need to leave the plurality of rows pore, so that the carbonic acid gas that sporophore growth produced can in time be discharged at the bottle mouth position of culturing bottle;
Cultivate through oxygenation, sporophore enters the stage of maturity, the Cordyceps militaris (L.) Link. of can directly gathering.
4, Cordyceps militaris (L.) Link. oxygenation method of cultivation according to claim 3 is characterized in that: when the liquid spawn in culture medium solution was led to oxygen, the mouth of pipe of will ventilating placed near the culturing bottle bottom, so that the oxygen that feeds upwards feeds culture medium solution at the bottom of bottle.
5, according to claim 3 or 4 described Cordyceps militaris (L.) Link. oxygenation method of cultivation, it is characterized in that: when passing through the sporophore oxygenation of primary growth breeding phase, the ventpipe that feeds culturing bottle should be provided with a plurality of air outlets, feeds in the culturing bottle with multi-angle so that enter the oxygen of culturing bottle.
6, Cordyceps militaris (L.) Link. oxygenation method of cultivation according to claim 3, it is characterized in that: described rice medium inoculation, be in culturing bottle, pack into rice and dried silkworm chrysalis meal, be modulated into nutritive medium, promptly by the part by weight of proportioning composition by following prescription, glucose 1-2%, peptone 0.3-1%, potassium primary phosphate 0.05-0.1%, sal epsom 0.02-0.07%, triammonium citrate 0.03-0.08%, vitamins B
10.002-0.005%, all the other are water, and are modulated into the culture solution of pH value between 6.5-7.5, pour in the culturing bottle then;
The bottleneck of culturing bottle is tightened, and the heat sterilization that under 110-130 ℃ of temperature condition rice medium was carried out about 25-30 minute is handled;
After treating the culture medium solution cooling, adopt through spraying into liquid spawn in the culture medium solution of disinfectant spraying device in culturing bottle;
Behind the liquid-spawn inoculation, culturing bottle is moved to the lucifuge position, room temperature is controlled at about 20 ℃, through 2-3 days time, the surface of rice medium all covered with mycelia;
Through 4-5 days time, mycelia penetrated half degree of depth of substratum; At this moment, give culturing bottle naturally astigmatism irradiation and bottle mouth position prick get the part ventilation hole with controlling moisture about 85%;
Through about 10 days, increase strong illumination 4-8 hour of 500-1000 lux intensity every day again, between temperature is controlled 20-23 ℃.
7, Cordyceps militaris (L.) Link. oxygenation method of cultivation according to claim 3, it is characterized in that: described silkworm chrysalis inoculation culture, be earlier silkworm chrysalis, to be injected to liquid spawn in the silkworm chrysalis body with asepsis injector after 3-5 minute via 75% alcohol-pickled sterilization, silkworm chrysalis is placed in the culture dish; Or silkworm chrysalis is placed in the culture dish, adopt through the disinfectant spraying device, liquid spawn is sprayed to the silkworm chrysalis whole body, then culture dish is sealed;
The silkworm chrysalis of inoculation liquid spawn is moved to the lucifuge position, room temperature is controlled at about 20 ℃, through 7-10 days time, pupal cell all covered with mycelia, and pupal cell ossifys;
The pupal cell that ossifys is moved in the culturing bottle, bottleneck is tightened processing.Through time of 5-10 days, pupal cell grew white or golden yellow mycelia, and bottleneck is left some ventilation holes, and astigmatism is shone naturally, indoor humidity is controlled at about 85% to give culturing bottle, and room temp is controlled at about 20 ℃;
Through about 10 days, increase strong illumination 4-8 hour of 500-1000 lux intensity every day again, between temperature is controlled 20-23 ℃.
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| CN103172424A (en) * | 2011-12-26 | 2013-06-26 | 江苏康迪生物科技有限公司 | Preparation method of culture solution for breeding organic Cordyceps militaris strain |
| CN103483037A (en) * | 2013-09-05 | 2014-01-01 | 东方中科生命科学有限责任公司 | Cordyceps liquid culture medium and application thereof |
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| CN1095104A (en) * | 1993-05-13 | 1994-11-16 | 刘文霞 | Cordyccps-militaris-(L.)-link. Sporophore and mycelium batch production production technology |
| CN1148623A (en) * | 1996-10-04 | 1997-04-30 | 海南兆隆实业有限公司 | North cordyceps mycelium fermentation technology |
| CN1152132C (en) * | 2000-06-09 | 2004-06-02 | 车振明 | Method for artificially cultivating cordyceps |
| CN1651568A (en) * | 2004-02-03 | 2005-08-10 | 李勇 | Edible fungus liquid culture submerged fermentation technology |
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