CN105779300A - Solid culture medium and cultivation method of cordyceps militaris fruiting bodies - Google Patents
Solid culture medium and cultivation method of cordyceps militaris fruiting bodies Download PDFInfo
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- CN105779300A CN105779300A CN201610096078.7A CN201610096078A CN105779300A CN 105779300 A CN105779300 A CN 105779300A CN 201610096078 A CN201610096078 A CN 201610096078A CN 105779300 A CN105779300 A CN 105779300A
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Abstract
The invention discloses a solid culture medium and cultivation method of cordyceps militaris fruiting bodies.The solid culture medium is prepared from 2 g-10 g of snail digestive gland dry powder, 10 g-18 g of rice and water which is 1.75 times the total mass of the snail digestive gland dry powder and the rice; the cultivation method of the cordyceps militaris fruiting bodies comprises the following steps of drying and smashing of snail digestive glands, preparation of liquid strains, sterilization, inoculation, darkness culture, color changing and growth and harvest of the fruiting bodies.By means of the solid culture medium and the cultivation method, waste in the snail production process can be effectively utilized, nutrients in the snail glands are utilized to the maximum extent, and the waste can be turned into wealth.Meanwhile, no pollution or waste exists in the production process, and the solid culture medium and cultivation method are beneficial for environmental protection.
Description
Technical field
The present invention relates to the cultural method of a kind of solid medium and Cordyceps militaris (L.) Link.sporophore for cultivating Cordyceps militaris (L.) Link.sporophore, specifically a kind of utilize Limax digestive gland to make film solid media the method for cultivating Cordyceps militaris (L.) Link.sporophore.
Background technology
White jade snail because of its head, skin, foot is as white as polished jade and gain the name, its nutritional labeling is extremely abundant.White jade snail whole body is precious, and comprehensive utilization value is high, and demand is big, and its meat fertilizer is tender, nutritious, is the senior nutrition that goes to zero of high protein, low fat, cholesterol.China's heliculture industry is to emerge from the eighties, and development speed is swift and violent, has now become China's characteristic agriculture development and the focus of foreign exchange earning project.Limax industry flourish, enriches the vegetable basket of the town and country masses dramatically, provides new high quality protein source for the popular common people.At present, snail product market at home and abroad is constantly in the situation that supply falls short of demand.But, while Limax industry is fast-developing, owing to Limax process of manufacture can produce substantial amounts of Limax digestive gland, and these Limax digestive glands are typically directly processed into animal feed or abandon as garbage as agricultural by-products, not only utilizing status is poor, industry added value is relatively low, and directly abandons and also can cause environmental pollution.
Summary of the invention
The technical problem to be solved is in that to propose a kind of garbage, the solid medium making the nutrient substance in Limax digestive gland obtain the Cordyceps militaris (L.) Link.sporophore utilized to the limit and cultural method that can effectively utilize in Limax production.
For solving above-mentioned technical problem, the solid medium of a kind of Cordyceps militaris (L.) Link.sporophore of the present invention, its composition is: Limax digestive gland dry powder 2～10g, rice 10～18g, the water of described Limax digestive gland dry powder and rice gross mass 1.75 times.
The solid medium of a kind of Cordyceps militaris (L.) Link.sporophore, its composition is: Limax digestive gland dry powder 4g, rice 16g and water 35mL.
The cultural method of a kind of Cordyceps militaris (L.) Link.sporophore, comprises the steps:
(1) drying of Limax digestive gland is pulverized: Limax digestive gland carries out drying and processing in an oven, dry, pulverize after Limax digestive gland dry powder；
(2) preparation of liquid spawn: prepare fluid medium by following compositing formula: described Limax digestive gland dry powder 20g/L that step 1 obtains, glucose 20g/L, KH2PO42g/L and MgSO4·7H2O1.0g/L；Cordyceps being inoculated on the described fluid medium prepared, under 25～27 DEG C of temperature conditions, shaking table is cultivated 5～7 days；
(3) sterilizing: prepare solid medium by following compositing formula: the water of described Limax digestive gland dry powder 2～10g, rice 10～18g and described Limax digestive gland dry powder and rice gross mass 1.75 times that step 1 obtains；Described solid medium is loaded the wide mouthed bottle of 200mL, then the bottleneck of described wide mouthed bottle is covered tightly or tightens, by taking out after bottle sterilizing, be cooled to room temperature；
(4) inoculation: aseptically every bottle graft enters described liquid spawn, and culture medium is stirred after terminating by inoculation with Glass rod, sealing；
(5) dark culturing: moved into by postvaccinal culture bottle under dark condition and cultivate, covers with to move on to after culture bottle until mycelia and cultivates under light；
(6) annesl: cultivating under light, first treat that mycelia is transferred to crocus by white, then treat that hyphal surface is covered with the projection of grain of rice size, annesl terminates；
(7) growth of sporophore: spray nutritional solution on the surface of described culture medium, treat sporophore growth；
(8) gather: when sporophore grows to 6～8 centimetres, sporophore is taken out from culture bottle, complete whole cultivation.
The cultural method of above-mentioned a kind of Cordyceps militaris (L.) Link.sporophore, after step (8), is placed on the sporophore gathered in 60 DEG C of baking ovens and dries, keep in Dark Place.
The cultural method of above-mentioned a kind of Cordyceps militaris (L.) Link.sporophore, in step (8), it also includes step (9) afterwards: after end of gathering, carry out ovendry power broken after described culture medium being taken out.
The cultural method of above-mentioned a kind of Cordyceps militaris (L.) Link.sporophore, the constituent of the described nutritional solution sprayed in step (7) is: triacontanol 0.5～1mg/L, vitaminB10 .08～0.12mg/L and potassium dihydrogen phosphate 1.0～1.5mg/L.
The cultural method of above-mentioned a kind of Cordyceps militaris (L.) Link.sporophore, described in step (4), the access amount of liquid spawn is 5 milliliters.
The cultural method of above-mentioned a kind of Cordyceps militaris (L.) Link.sporophore, described in step (1), the drying temperature conditions of Limax digestive gland is 60 DEG C.
The cultural method of above-mentioned a kind of Cordyceps militaris (L.) Link.sporophore, after wide mouthed bottle described in step (3) loads described solid medium, cover tightly with the lid with passage or tighten bottleneck with polyethylene plastic film, place the bottle in sterilizing 30 minutes under 120 DEG C of temperature conditions, then take out and be placed on superclean bench and be cooled to room temperature.
The cultural method of above-mentioned a kind of Cordyceps militaris (L.) Link.sporophore, described in step (5), the process of dark culturing includes: being moved into by postvaccinal culture bottle under dark condition and cultivate, incubation time 3～5 days, temperature is maintained at 16～18 DEG C；Then temperature is risen under 20～22 DEG C of conditions and be further cultured for 8～10 days, cover with to move on to after culture bottle until mycelia and cultivate under light.
The cultural method of above-mentioned a kind of Cordyceps militaris (L.) Link.sporophore, the process cultivated under light described in step (6) includes: keep indoor temperature 20～24 DEG C, humidity 60～70%, intensity of illumination 800 lux, light application time 12～14h every day, after treating that mycelia is transferred to crocus by white, nocturnal temperature is down to 10～15 DEG C, keep the temperature difference having 5～10 DEG C round the clock, when hyphal surface is covered with the projection of grain of rice size, stop day and night temperature and transfer 20～24 DEG C of constant temperature culture to.
The cultural method of above-mentioned a kind of Cordyceps militaris (L.) Link.sporophore, after described in step (7), nutritional solution is sprayed on the surface of culture medium, keep indoor temperature 20～24 DEG C, humidity 60～70%, intensity of illumination 600～800 lux, light application time 12～14h every day, brings up to 75～85% by indoor humidity after sporophore grows to 3～4 centimetres, and temperature and illumination condition remain unchanged.
Research finds, the crude protein content of Limax internal organs has reached 48.59%, possibly together with abundant trace element, and therefore can as good fermentable raw material.Due to the fact that and have employed technique scheme, the garbage produced in Limax process of manufacture and Limax digestive gland are used for planting Cordyceps militaris (L.) Link.sporophore by it, take full advantage of rich in protein and trace element in Limax digestive gland and produce required nutrition as Cordyceps militaris (L.) Link., substantially increase the biological transformation ratio of Limax digestive gland, the nutrient substance in Limax digestive gland is made to obtain maximum utilization, it is achieved that to turn waste into wealth.After Cordyceps is gathered, culture medium can pulverize after as feed additive for producing livestock feed, substantially increase the immunocompetence of poultry, domestic animal, it is achieved thereby that pollution-free in production process, without garbage, environmental conservation is significant.Therefore, the method have the advantages that (1) utilizes Limax digestive gland to produce required liquid spawn as culture matrix to prepare Cordyceps militaris (L.) Link., make full use of protein and the trace element of Limax digestive gland, Cordyceps growth is vigorous, and fermentation time is fast；(2) containing rich in protein and trace element in Limax digestive gland, as waste disposal, the wasting of resources is contaminated environment also, is added in Cordyceps militaris (L.) Link.sporophore culture medium by Limax digestive gland, turns waste into wealth, and makes full use of resource, improves the added value of Limax industry；(3) after liquid spawn, aseptically with Glass rod, culture medium is stirred in inoculation, can not only just culture medium mix homogeneously, and considerably increase the contact area of liquid spawn and culture medium, be conducive to the entrance of oxygen, accelerate and send out bacterium speed, decrease and send out the bacterium time.(4) after annesl terminates, spraying last layer nutritional solution in culture medium, the growth for Cordyceps sporophore provides sufficient nutrition, promotes the growth of Cordyceps sporophore.
Detailed description of the invention
Embodiment 1: the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore of the present invention comprises the steps:
(1) drying of Limax digestive gland is pulverized: Limax digestive gland is being carried out drying and processing in the baking oven of 60 DEG C, dry, pulverize after Limax digestive gland dry powder.
(2) preparation of liquid spawn: prepare fluid medium by following compositing formula: Limax digestive gland dry powder 20g/L that step 1 obtains, glucose 20g/L, KH2PO42g/L and MgSO4·7H2O1.0g/L；Cordyceps being inoculated on the fluid medium prepared, under 25～27 DEG C of temperature conditions, shaking table is cultivated 5～7 days；Wherein Chinese caterpillar fungus strain adopts the Cordyceps No. 1 of Ao Lang bio tech ltd, Shanghai.
(3) sterilizing: prepare solid medium of the present invention: Limax digestive gland 2g, rice 18g and the water 35mL that step 1 obtains by following compositing formula；Prepared solid medium is loaded the wide mouthed bottle of 200mL, cover tightly with the lid with passage or tighten bottleneck with polyethylene plastic film, place the bottle in sterilizing 30 minutes under 120 DEG C of conditions, taking-up be placed on superclean bench to be cooled to room temperature (adopt this temperature conditions and time conditions can not only sterilizing thorough, moreover it is possible to prevent the change of the culture medium nutrient substance that overlong time causes).
(4) inoculation: aseptically every bottle graft enters liquid spawn, and culture medium is stirred after terminating by inoculation with Glass rod, (finding in process of the test, after sterilizing, Limax digestive gland can arrive above culture medium, causes the uneven of culture medium in sealing.And adopt stirring means can solve this problem well)；The access amount of liquid spawn is 5 milliliters, can not only send out, for Cordyceps, the strain that bacterium provides enough, be unlikely to again to make culture medium moisture too high simultaneously.
(5) dark culturing: being moved into by postvaccinal culture bottle under dark condition and cultivate, starting temperature is maintained at 16～18 DEG C, 3～5 days, then temperature is risen to and is further cultured for 8～10 days under 20～22 DEG C of conditions, covers with to move on to after culture bottle until mycelia and cultivates under light；Wherein, temperature conditions and cultivated days can be adjusted according to actual needs in above-mentioned scope, adopt this temperature conditions and time conditions to be conducive to Cordyceps militaris (L.) Link. mycelial growth fast, and growing way is good, be conducive to controlling the growth of miscellaneous bacteria, be also beneficial to the success rate improving inoculation with sending out bacterium.
(6) annesl: keep indoor temperature 20～24 DEG C, humidity 60～70%, intensity of illumination 800 lux, light application time 12～14h every day, treats that mycelia is transferred to crocus by white, and nocturnal temperature is down to 10～15 DEG C afterwards, keep the temperature difference having 5～10 DEG C round the clock, when hyphal surface is covered with the projection of grain of rice size, stopping day and night temperature and transfer 20～24 DEG C of constant temperature culture to, annesl terminates；Wherein, temperature conditions, damp condition and light application time etc. can be adjusted according to actual needs in above-mentioned scope, and above-mentioned condition is conducive to the annesl of Cordyceps and the formation of former base.
(7) growth of sporophore: after annesl terminates, spraying nutritional solution on the surface of culture medium, consist of triacontanol 0.5mg/L, vitaminB10 .08mg/L and the potassium dihydrogen phosphate 1.0mg/L(of nutritional solution spray the nutritional solution of this component and can provide enough nutrition for Cordyceps growth)；Keep indoor temperature 20～24 DEG C (constant temperature), humidity 60～70%, intensity of illumination 600～800 lux, light application time 12～14h every day, after sporophore grows to 3～4 centimetres, indoor humidity is brought up to 75～85%, temperature and illumination invariant；Wherein, temperature conditions, damp condition, intensity of illumination and light application time etc. can be adjusted according to actual needs in above-mentioned scope, can accelerate the growth of Cordyceps sporophore under above-mentioned condition, improve the yield of Cordyceps.
(8) gather: when sporophore grows to 6～8 centimetres, sporophore is taken out from culture bottle, be placed in 60 DEG C of baking ovens and dry, keep in Dark Place.Under this condition, Cordyceps sporophore yield (dry weight) is 1.49g/ bottle；And under 60 DEG C of temperature conditions, dry the nutritional labeling that will not destroy in Cordyceps.
(9) gather after end, after culture medium is taken out, dry and pulverize, it is possible to as feed additive production livestock feed.
Embodiment 2: the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore of the present invention comprises the steps:
Other step is identical with embodiment 1, and difference is in that: in step (3), solid medium consists of Limax digestive gland 4g, rice 16g, water 35mL；Consisting of of step (7) Middle nutrition liquid: triacontanol 0.6mg/L, vitaminB10 .09mg/L, potassium dihydrogen phosphate 1.1mg/L.Under this condition, Cordyceps sporophore yield (dry weight) is 1.76g/ bottle.
Embodiment 3: the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore of the present invention comprises the steps:
Other step is identical with embodiment 1, and difference is in that: in step (3), solid medium consists of Limax digestive gland 5g, rice 15g, water 35mL；Consisting of of step 7 Middle nutrition liquid: triacontanol 0.7mg/L, vitaminB10 .10mg/L, potassium dihydrogen phosphate 1.2mg/L.Under this condition, Cordyceps sporophore yield (dry weight) is 1.64g/ bottle.
Embodiment 4: the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore of the present invention comprises the steps:
Other step is identical with embodiment 1, and difference is in that: in step (3), solid medium consists of Limax digestive gland 8g, rice 12g, water 35mL；Consisting of of step 7 Middle nutrition liquid: triacontanol 0.8mg/L, vitaminB10 .11mg/L, potassium dihydrogen phosphate 1.4mg/L.Under this condition, Cordyceps sporophore yield (dry weight) is 1.29g/ bottle.
Embodiment 5: the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore of the present invention comprises the steps:
Other step is identical with embodiment 1, and difference is in that: in step (3), solid medium consists of Limax digestive gland 10g, rice 10g, water 35mL；Consisting of of step 7 Middle nutrition liquid: triacontanol 1.0mg/L, vitaminB10 .12mg/L, potassium dihydrogen phosphate 1.5mg/L.Under this condition, Cordyceps sporophore yield (dry weight) is 1.18g/ bottle.
Claims (11)
1. the solid medium of a Cordyceps militaris (L.) Link.sporophore, it is characterised in that the composition of this solid medium is: Limax digestive gland dry powder 2～10g, rice 10～18g, the water of described Limax digestive gland dry powder and rice gross mass 1.75 times.
2. the solid medium of a kind of Cordyceps militaris (L.) Link.sporophore as claimed in claim 1, it is characterised in that the composition of this solid medium is: Limax digestive gland dry powder 4g, rice 16g and water 35mL.
3. the cultural method of a Cordyceps militaris (L.) Link.sporophore, it is characterised in that comprise the steps:
(1) drying of Limax digestive gland is pulverized: Limax digestive gland carries out drying and processing in an oven, dry, pulverize after Limax digestive gland dry powder；
(2) preparation of liquid spawn: prepare fluid medium by following compositing formula: described Limax digestive gland dry powder 20g/L that step 1 obtains, glucose 20g/L, KH2PO42g/L and MgSO4·7H2O1.0g/L；Cordyceps being inoculated on the described fluid medium prepared, under 25～27 DEG C of temperature conditions, shaking table is cultivated 5～7 days；
(3) sterilizing: prepare solid medium by following compositing formula: the water of described Limax digestive gland dry powder 2～10g, rice 10～18g and described Limax digestive gland dry powder and rice gross mass 1.75 times that step 1 obtains；Described solid medium is loaded the wide mouthed bottle of 200mL, then the bottleneck of described wide mouthed bottle is covered tightly or tightens, by taking out after bottle sterilizing, be cooled to room temperature；
(4) inoculation: aseptically every bottle graft enters described liquid spawn, and culture medium is stirred after terminating by inoculation with Glass rod, sealing；
(5) dark culturing: moved into by postvaccinal culture bottle under dark condition and cultivate, covers with to move on to after culture bottle until mycelia and cultivates under light；
(6) annesl: cultivating under light, first treat that mycelia is transferred to crocus by white, then treat that hyphal surface is covered with the projection of grain of rice size, annesl terminates；
(7) growth of sporophore: spray nutritional solution on the surface of described culture medium, treat sporophore growth；
(8) gather: when sporophore grows to 6～8 centimetres, sporophore is taken out from culture bottle, complete whole cultivation.
4. the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore as claimed in claim 3, it is characterised in that after step (8), is placed on the sporophore gathered in 60 DEG C of baking ovens and dries, keep in Dark Place.
5. the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore as described in claim 3 or 4, it is characterised in that also include step (9) after step (8): after end of gathering, carry out ovendry power after described culture medium being taken out broken.
6. the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore as described in claim 3 or 4, it is characterized in that, the constituent of the described nutritional solution sprayed in step (7) is: triacontanol 0.5～1mg/L, vitaminB10 .08～0.12mg/L and potassium dihydrogen phosphate 1.0～1.5mg/L.
7. the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore as described in claim 3 or 4, it is characterised in that described in step (4), the access amount of liquid spawn is 5 milliliters.
8. the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore as described in claim 3 or 4, it is characterized in that, after wide mouthed bottle described in step (3) loads described solid medium, cover tightly with the lid with passage or tighten bottleneck with polyethylene plastic film, place the bottle in sterilizing 30 minutes under 120 DEG C of temperature conditions, then take out and be placed on superclean bench and be cooled to room temperature.
9. the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore as described in claim 3 or 4, it is characterized in that, described in step (5), the process of dark culturing includes: being moved into by postvaccinal culture bottle under dark condition and cultivate, incubation time 3～5 days, temperature is maintained at 16～18 DEG C；Then temperature is risen under 20～22 DEG C of conditions and be further cultured for 8～10 days, cover with to move on to after culture bottle until mycelia and cultivate under light.
10. the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore as described in claim 3 or 4, it is characterized in that, the process cultivated under light described in step (6) includes: keep indoor temperature 20～24 DEG C, humidity 60～70%, intensity of illumination 800 lux, light application time 12～14h every day, after treating that mycelia is transferred to crocus by white, nocturnal temperature is down to 10～15 DEG C, keep the temperature difference having 5～10 DEG C round the clock, when hyphal surface is covered with the projection of grain of rice size, stop day and night temperature and transfer 20～24 DEG C of constant temperature culture to.
11. the cultural method of a kind of Cordyceps militaris (L.) Link.sporophore as described in claim 3 or 4, it is characterized in that, after described in step (7), nutritional solution is sprayed on the surface of culture medium, keep indoor temperature 20～24 DEG C, humidity 60～70%, intensity of illumination 600～800 lux, light application time 12～14h every day, after sporophore grows to 3～4 centimetres, indoor humidity being brought up to 75～85%, temperature and illumination condition remain unchanged.
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| 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 |
| WO2024159509A1 (en) * | 2023-02-03 | 2024-08-08 | 立万利创新股份有限公司 | Method for culturing cordyceps militaris and cordyceps militaris fruiting body extract |
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