CN114793755A - Method for cultivating silkworm cordyceps militaris with high rigidification rate - Google Patents
Method for cultivating silkworm cordyceps militaris with high rigidification rate Download PDFInfo
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- CN114793755A CN114793755A CN202210406740.XA CN202210406740A CN114793755A CN 114793755 A CN114793755 A CN 114793755A CN 202210406740 A CN202210406740 A CN 202210406740A CN 114793755 A CN114793755 A CN 114793755A
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- 241000255789 Bombyx mori Species 0.000 title claims abstract description 78
- 241001264174 Cordyceps militaris Species 0.000 title claims abstract description 50
- 238000000034 method Methods 0.000 title claims abstract description 16
- 241000382353 Pupa Species 0.000 claims abstract description 23
- 238000011081 inoculation Methods 0.000 claims abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 15
- 238000006243 chemical reaction Methods 0.000 claims abstract description 13
- 230000003020 moisturizing effect Effects 0.000 claims abstract description 9
- 241000894006 Bacteria Species 0.000 claims abstract description 8
- 244000025254 Cannabis sativa Species 0.000 claims abstract description 8
- 238000007599 discharging Methods 0.000 claims abstract description 8
- 238000003306 harvesting Methods 0.000 claims abstract description 8
- 239000007921 spray Substances 0.000 claims abstract 2
- 238000005286 illumination Methods 0.000 claims description 29
- 239000000243 solution Substances 0.000 claims description 9
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 5
- 238000002347 injection Methods 0.000 claims description 4
- 239000007924 injection Substances 0.000 claims description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 244000061456 Solanum tuberosum Species 0.000 claims description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 3
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- JZRWCGZRTZMZEH-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 2
- 240000000249 Morus alba Species 0.000 abstract 1
- 235000008708 Morus alba Nutrition 0.000 abstract 1
- 238000004321 preservation Methods 0.000 description 8
- 239000001963 growth medium Substances 0.000 description 6
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 5
- 239000011362 coarse particle Substances 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 241000190633 Cordyceps Species 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 239000000306 component Substances 0.000 description 2
- 238000012136 culture method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000004886 process control Methods 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000012364 cultivation method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000003716 rejuvenation Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/50—Inoculation of spawn
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- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Mushroom Cultivation (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for cultivating silkworm cordyceps militaris with high rigidification rate, which comprises the following steps: (1) preparing mulberry silkworm chrysalis: selecting healthy living mulberry silkworm pupas which are just cut from silkworm cocoons; (2) preparing cordyceps militaris bacterium liquid: inoculating Cordyceps militaris strain into culture solution, standing at 25 deg.C for 48 hr, and performing shake culture at 25 deg.C and 108rpm for 7 days; (3) inoculation: injecting cordyceps militaris bacterium liquid into the head inlet of the living silkworm pupae by using a spray continuous injector, quantitatively injecting 0.15mL of each silkworm pupae, and inoculating until the tail part is straight; (4) rigidizing; (5) moisturizing and cultivating; (6) color conversion; (7) grass discharging; (8) harvesting; the method has the advantages of high rigidification rate, short growth period, easy control of conditions and living body cultivation of the silkworm cordyceps militaris, and the prepared mulberry cordyceps militaris stroma has high content of effective components.
Description
Technical Field
The invention relates to a method for cultivating high-rigidity silkworm cordyceps militaris, which is used for cultivating silkworm chrysalis cordyceps militaris and belongs to the technical field of edible fungus cultivation.
Background
The natural resources of the cordyceps militaris are limited, but the artificial culture is successful, and the following three culture methods are mainly adopted at present: firstly, natural animal culture medium is adopted for cultivation, and silkworm chrysalis is mainly adopted as a culture medium. Injecting cordyceps sinensis strain into host silkworm pupa through strict strain breeding and silkworm pupa selection to provide proper growth conditions for cordyceps militaris fruiting bodies, and finally obtaining mature cordyceps militaris; and secondly, artificial solid culture medium cultivation, wherein rice and wheat are mostly selected as culture mediums. Selecting wild or artificially cultured Cordyceps strains, culturing to obtain mother strains by separating, breeding, rejuvenating, etc., transferring to prepared inactivated solid culture medium after propagation, adding various necessary nutrients, and culturing to obtain mature Cordyceps militaris; thirdly, artificial liquid fermentation cultivation, which mostly adopts a liquid fermentation tank for cultivation. Putting the culture medium into a test tube, sterilizing at high temperature, placing into a slope, inoculating slant test tube strains into a culture solution, placing on a shaking table for shaking culture, then performing seed tank amplification culture, performing liquid fermentation tank deep fermentation on high-concentration bacteria solution, and finally performing centrifugal separation to obtain mature Cordyceps militaris. Liquid culture medium components suitable for cordyceps militaris growth can be selected according to different production index requirements.
The defects of the prior art mainly comprise:
(1) the inoculation pollution rate is high, the cost of the silkworm chrysalis is high, and the income is low.
(2) Because the inoculation is carried out by using the injector, the inoculation amount is controlled by using hands, on one hand, the inoculation is not accurate enough, on the other hand, the force is small, the pupa body cannot be filled with the bacterial liquid, and the hypha germination time is long.
(3) The inoculation part is on the soft back, the inoculation force is not easy to master, and the liquid is easy to seep after the syringe is punctured by inoculation, thereby causing pollution.
(4) The rigidness time is short, the rigidness time is 4-6 days after the new inoculation method, and the rigidness time for the syringe inoculation is 10-15 days.
For example, in the invention patent 'an efficient cultivation method of tussah silkworm chrysalis cordyceps' with application number 201811034151.3, the Duxing model and the like, the invention mentions that the disinfected tussah silkworm living chrysalis is inoculated with 0.3-0.7ml of cordyceps militaris liquid strain from the 3 rd-5 th body node of the back of the tussah silkworm living chrysalis, placed in a plastic tray, and cultivated for 45-50 days under the conditions of proper temperature, humidity, illumination and the like to obtain the mature cordyceps militaris. However, the published literature does not provide specific technical process control parameters and production effects, and it can be understood that the inventor only provides a concept of utilizing the fruiting body of the living tussah cordyceps militaris, and does not have a specific production technology and method. YanghuangGen, etc. provide specific technological process control parameters and production effects in "a culture method of Cordyceps militaris" (application No. 202011161622.4), but there is no specific improvement on the most critical inoculation technique, and inoculation is carried out by using an injector.
Disclosure of Invention
The invention aims to provide a method for cultivating high-rigidity cordyceps militaris, which is characterized in that the cordyceps militaris is cultivated through living bodies, and the content of active ingredients of stroma of the prepared cordyceps militaris is high.
In order to achieve the aim, the invention provides a method for cultivating cordyceps militaris with high rigidification rate, which comprises the following steps:
(1) preparing mulberry silkworm chrysalis: selecting healthy living silkworm pupa (the head and tail parts are shaken) which are just cut from silkworm cocoons.
(2) Preparing cordyceps militaris bacterium liquid: inoculating Cordyceps militaris (Cordyceps militaris) strain into the culture solution, standing at 25 deg.C for 48 hr, and performing shake culture at 25 deg.C and 108rpm for 7 days. Specifically, the components of the culture solution are as follows: 220 g of potato, 20 g of glucose, 1 g of monopotassium phosphate, 1 g of magnesium sulfate and vitamin B 1 20 mg, sodium chloride 1.8 g, peptone 4 g, 1000ml water, pH 6.5.
(3) Inoculation: the cordyceps militaris bacterium liquid is injected to the head inlet (the mouth part is the softest) of the living silkworm pupae by using a spraying continuous injector, 0.15mL of cordyceps militaris bacterium liquid is quantitatively injected into each silkworm pupae, and the inoculation is carried out until the tail part is stretched. After injection, if the pupa Bombycis is still shaken, another injection is performed.
(4) Rigidification: after the cordyceps militaris strain is inoculated into the silkworm pupa, feeding the silkworm pupa into a cultivation workshop with the temperature of 15-18 ℃ and the relative humidity of 40-60% for cultivation; after 6 days, the inoculated silkworm pupae gradually hardened. The pupae can be picked out in sequence and transferred to a new culture tray for culture after being completely hardened.
(5) And (5) moisturizing and cultivating: the humidity is 70-90%, and the moisture-preserving culture is carried out for 3-5 d in the environment with the temperature of 18-22 ℃.
(6) Color conversion: after preserving moisture and culturing for 3-5 days, auxiliary illumination is carried out, the illumination intensity is 1000 Lx-3000 Lx, the humidity is 70-90%, the temperature is 18-22 ℃, and after continuous illumination culture for 4-6 days, the surface hypha of the silkworm chrysalis is changed from white to orange.
(7) Grass discharging: after color-changing culture for 7-12 days, fruiting bodies begin to grow on the surface of the silkworm chrysalis; when the length of the sporocarp is 1cm, the illumination intensity is reduced to 200 Lx-1000 Lx, so that the sporocarp is prevented from being matured prematurely.
(8) Harvesting: when the fruiting body grows to 4-7 cm, the head of the fruiting body is expanded, and rough particles appear for collection.
The method can realize continuous inoculation, improve the efficiency, shorten the production period, have high rigidification rate, reduce the rigidification time from the original 10-15 days to 4-6 days, improve the rigidification rate from about 50 percent to more than 95 percent, have easily controlled conditions, and have high content of the active ingredients of the stroma of the prepared cordyceps militaris.
Detailed Description
Inoculating Cordyceps militaris (Cordyceps militaris) strain to living silkworm pupa, and culturing, wherein ACCC50562 is selected as the strain.
The culture solution is: 220 g of potato, 20 g of glucose, 1 g of monopotassium phosphate, 1 g of magnesium sulfate and vitamin B 1 20 mg, 1.8 g of sodium chloride, 4 g of peptone, naturally about pH6.5, 300ml in a 500ml shake flask; 1000ml of water.
Example 1
(1) Preparing mulberry silkworm chrysalis: selecting healthy living silkworm pupa (the head and tail parts are shaken) which are just cut from silkworm cocoons.
(2) Preparing bacterial liquid: inoculating Cordyceps militaris strain into culture solution, standing at 25 deg.C for 48 hr, and performing shake culture at 25 deg.C and 108rpm for 7 days.
(3) Inoculation: inoculating the prepared cordyceps militaris bacterial liquid to living silkworm chrysalis in an inoculation workshop. The strain is directly injected into living silkworm pupae by a self-made continuous inoculator to achieve the purpose of infection, the strain is injected into the head inlet (the softest part of the mouth) of each silkworm pupae, 0.15mL of strain is quantitatively injected into each silkworm pupae, the strain is inoculated until the tail part is straight, and the strain is injected once again by shaking.
(4) Rigidification: after the cordyceps militaris strain is inoculated in the silkworm pupa, the silkworm pupa is fed into a cultivation workshop with the temperature of 15 ℃ and the relative humidity of 60% for cultivation. After 6 days, the inoculated silkworm pupae gradually hardened. The pupae can be picked out in sequence and transferred to a new culture tray for culture after being completely hardened.
(5) And (5) moisturizing and cultivating: and performing moisture preservation culture for 3d in an environment with the humidity of 80% and the temperature of 20 ℃.
(6) Color conversion: and (3) after the silkworm chrysalis is subjected to moisture preservation culture for 5 days, auxiliary illumination is carried out, the illumination intensity is preferably 2000Lx, the humidity is 90%, the temperature is 20 ℃, and after the illumination culture is continuously carried out for 5 days, the surface hypha of the silkworm chrysalis is changed from white to orange.
(7) Grass discharging: after color conversion culture for 10 days, fruiting bodies begin to grow on the surface of the silkworm chrysalis. When the sporocarp grows to about 1cm in length, the illumination intensity can be adjusted to 200Lx, so that the sporocarp is prevented from being matured prematurely.
(8) Harvesting: when the fruiting body grows to 5cm, the head of the fruiting body is expanded and coarse particles appear, and the fruiting body is mature and can be harvested.
Example 2
The same as in example 1, except that:
(4) rigidification: after the cordyceps militaris strain is inoculated into the silkworm pupa, the silkworm pupa is fed into a cultivation workshop with the temperature of 16 ℃ and the relative humidity of 60% for cultivation. After 6 days, the inoculated silkworm pupae gradually hardened. The pupae can be picked out in sequence and transferred to a new culture tray for culture after being completely hardened.
(5) And (5) moisturizing and cultivating: and carrying out moisture preservation culture for 4d in an environment with the humidity of 90% and the temperature of 22 ℃.
(6) Color conversion: and (3) after the silkworm chrysalis is subjected to moisture preservation culture for 5 days, auxiliary illumination is carried out, the illumination intensity is preferably 2000Lx, the humidity is 90%, the temperature is 18 ℃, and after the illumination culture is continuously carried out for 4 days, the surface hypha of the silkworm chrysalis is changed from white to orange.
(7) Grass discharging: after color conversion culture for 8 days, fruiting bodies begin to grow on the surface of the silkworm chrysalis. When the sporocarp grows to about 1cm in length, the illumination intensity can be adjusted to 1000Lx, and the sporocarp is prevented from being matured prematurely.
(8) Harvesting: when the fruiting body grows to 6cm, the head of the fruiting body is expanded and coarse particles appear, and the fruiting body is mature and can be harvested.
Example 3
The same as in example 1, except that:
(4) rigidification: after the cordyceps militaris strain is inoculated in the silkworm pupa, the silkworm pupa is sent into a cultivation workshop with the temperature of 18 ℃ and the relative humidity of 50% for cultivation. After 6 days, the inoculated silkworm pupae gradually hardened. The pupae can be picked out in sequence and transferred to a new culture tray for culture after being completely hardened.
(5) And (5) moisturizing and cultivating: and performing moisture preservation culture for 5d in an environment with the humidity of 90% and the temperature of 18 ℃.
(6) Color conversion: and after the silkworm chrysalis is subjected to moisture preservation culture for 3 days, auxiliary illumination is performed, the illumination intensity is 3000Lx, the humidity is 70%, the temperature is 20 ℃, and after the illumination culture is continued for 6 days, the surface hyphae of the silkworm chrysalis are changed from white to orange.
(7) Grass discharging: after color conversion culture for 10 days, fruiting bodies begin to grow on the surface of the silkworm chrysalis. When the sporocarp grows to about 1cm in length, the illumination intensity can be adjusted to 800Lx, and the sporocarp is prevented from being matured prematurely.
(8) Harvesting: when the fruiting body grows to 4cm, the head of the fruiting body is expanded and coarse particles appear, and the fruiting body is mature and can be harvested.
Example 4
The same as in example 1, except that:
(4) rigidification: after the cordyceps militaris strain is inoculated in the silkworm pupa, the silkworm pupa is fed into a cultivation workshop with the temperature of 18 ℃ and the relative humidity of 40% for cultivation. After 6 days, the inoculated silkworm pupae gradually hardened. The pupae can be picked out in sequence and transferred to a new culture tray for culture after being completely hardened.
(5) And (5) moisturizing and cultivating: and carrying out moisture-keeping culture for 4d in an environment with the humidity of 70% and the temperature of 22 ℃.
(6) Color conversion: and (4) after the silkworm chrysalis is subjected to moisture preservation culture for 4 days, auxiliary illumination is carried out, the illumination intensity is preferably 1000Lx, the humidity is 70%, the temperature is 22 ℃, and after the illumination culture is continued for 4 days, the surface hyphae of the silkworm chrysalis are changed from white to orange.
(7) Grass discharging: after 12 days of color conversion culture, fruiting bodies begin to grow on the surface of the silkworm chrysalis. When the sporocarp grows to about 1cm in length, the illumination intensity can be adjusted to 600Lx, and the sporocarp is prevented from being matured too early.
(8) Harvesting: when the fruiting body grows to 7cm, the head of the fruiting body is expanded and coarse particles appear, and the fruiting body is mature and can be harvested.
Example 5
The same as in example 1, except that:
(4) rigidification: after the cordyceps militaris strain is inoculated in the silkworm pupa, the silkworm pupa is fed into a cultivation workshop with the temperature of 15 ℃ and the relative humidity of 40% for cultivation. After 6 days, the inoculated silkworm pupae gradually hardened. The pupae can be picked out in sequence and transferred to a new culture tray for culture after being completely hardened.
(5) And (5) moisturizing and cultivating: and performing moisture-keeping culture at 20 deg.C with humidity of 90% for 5 d.
(6) Color conversion: and (3) after the silkworm chrysalis is subjected to moisture preservation culture for 5 days, auxiliary illumination is carried out, the illumination intensity is 3000Lx, the humidity is 80%, the temperature is 22 ℃, and after the illumination culture is continued for 5 days, the surface hyphae of the silkworm chrysalis are changed from white to orange.
(7) Grass discharging: after color conversion culture for 7d, fruiting bodies begin to grow on the surface of the silkworm chrysalis. When the sporocarp grows to about 1cm in length, the illumination intensity can be adjusted to 400Lx, so that the sporocarp is prevented from being matured prematurely.
(8) Harvesting: when the fruiting body grows to 7cm, the head of the fruiting body is expanded and coarse particles appear, and the fruiting body is mature and can be harvested.
The detection basis is agricultural industry standard NY/T2116-2012 (high performance liquid chromatography for measuring cordycepic acid and adenosine in cordyceps products) of the people's republic of China, and the results are as follows:
| sample (I) | Cordycepic acid (mg/g) |
| Example 1 | 1.57×10 2 |
| Example 2 | 1.64×10 2 |
| Example 3 | 1.63×10 2 |
| Example 4 | 1.53×10 2 |
| Example 5 | 1.62×10 2 |
The cordycepic acid content of common Cordyceps militaris (Cordyceps militaris) is generally 8.12 × 10 1 ～8.72×10 1 The cordycepic acid content of Cordyceps militaris (rice stroma) is 6.13 × 10 1 And the content of the cordycepic acid in the embodiment is obviously improved.
The above embodiments do not limit the present invention in any way, and all technical solutions obtained by means of equivalent substitution or equivalent transformation fall within the protection scope of the present invention.
Claims (3)
1. The method for cultivating the silkworm cordyceps militaris with high rigidification rate is characterized by comprising the following steps:
(1) preparing mulberry silkworm chrysalis: selecting healthy living mulberry silkworm pupas which are just cut from silkworm cocoons;
(2) preparing cordyceps militaris bacterium liquid: inoculating Cordyceps militaris strain into culture solution, standing at 25 deg.C for 48 hr, and performing shake culture at 25 deg.C and 108rpm for 7 days;
(3) inoculation: injecting cordyceps militaris bacterium liquid into the head inlet of the living silkworm pupae by using a spray continuous injector, quantitatively injecting 0.15mL of each silkworm pupae, and inoculating until the tail part is straight;
(4) rigidification: after the cordyceps militaris strain is inoculated in the silkworm pupa, the silkworm pupa is sent into a cultivation workshop with the temperature of 15-18 ℃ and the relative humidity of 40-60% for cultivation; after 6 days, the inoculated silkworm pupae gradually hardened. The pupae can be picked out in sequence and transferred into a new culture tray for culture after being completely hardened;
(5) and (5) moisturizing and cultivating: carrying out moisturizing culture for 3-5 d in an environment with the humidity of 70-90% and the temperature of 18-22 ℃;
(6) color conversion: after preserving moisture and culturing for 3-5 days, carrying out auxiliary illumination, wherein the illumination intensity is 1000 Lx-3000 Lx, the humidity is 70-90%, the temperature is 18-22 ℃, and after continuously carrying out illumination culture for 4-6 days, the surface hypha of the silkworm chrysalis is changed from white to orange;
(7) grass discharging: after color-changing culture for 7-12 days, fruiting bodies begin to grow on the surface of the silkworm chrysalis; when the sporocarp grows to 1cm in length, the illumination intensity is reduced to 200 Lx-1000 Lx, so that the sporocarp is prevented from being matured prematurely;
(8) harvesting: when the fruiting body grows to 4-7 cm, the head of the fruiting body is expanded, and rough particles appear for collection.
2. The method for cultivating cordyceps militaris with high rigidification rate as claimed in claim 1, wherein the method comprises the following steps: in the step (2), the components of the culture solution are as follows: 220 g of potato, 20 g of glucose, 1 g of monopotassium phosphate, 1 g of magnesium sulfate and vitamin B 1 20 mg, sodium chloride 1.8 g, peptone 4 g, 1000ml water, pH 6.5.
3. The method for cultivating cordyceps militaris with high rigidification rate as claimed in claim 1, wherein the method comprises the following steps: in the step (3), after injection, if the silkworm pupae are still shaken, the injection is performed once again.
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|---|---|---|---|---|
| CN102440149A (en) * | 2011-10-10 | 2012-05-09 | 江苏科技大学 | Method for breeding cordyceps militaris by utilizing silkworm larvae |
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