CN108901587A - A kind of solid culture method of cicada fungus - Google Patents
A kind of solid culture method of cicada fungus Download PDFInfo
- Publication number
- CN108901587A CN108901587A CN201710252724.9A CN201710252724A CN108901587A CN 108901587 A CN108901587 A CN 108901587A CN 201710252724 A CN201710252724 A CN 201710252724A CN 108901587 A CN108901587 A CN 108901587A
- Authority
- CN
- China
- Prior art keywords
- culture
- cultural method
- culture medium
- medium
- cicada fungus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 241000931705 Cicada Species 0.000 title claims abstract description 34
- 241000233866 Fungi Species 0.000 title claims abstract description 32
- 239000007787 solid Substances 0.000 title claims abstract description 28
- 238000012136 culture method Methods 0.000 title claims abstract description 7
- 230000003287 optical effect Effects 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 18
- 238000005286 illumination Methods 0.000 claims abstract description 13
- 241001625026 Cordyceps cicadae Species 0.000 claims abstract description 11
- 239000001963 growth medium Substances 0.000 claims description 23
- 239000002609 medium Substances 0.000 claims description 18
- 239000002054 inoculum Substances 0.000 claims description 7
- 235000010469 Glycine max Nutrition 0.000 claims description 5
- 240000007594 Oryza sativa Species 0.000 claims description 5
- 235000007164 Oryza sativa Nutrition 0.000 claims description 5
- 241000209140 Triticum Species 0.000 claims description 5
- 235000021307 Triticum Nutrition 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 235000009566 rice Nutrition 0.000 claims description 5
- 239000010902 straw Substances 0.000 claims description 5
- 241000894006 Bacteria Species 0.000 claims description 4
- 229920000742 Cotton Polymers 0.000 claims description 4
- 244000068988 Glycine max Species 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 235000007319 Avena orientalis Nutrition 0.000 claims description 2
- 241000209763 Avena sativa Species 0.000 claims description 2
- 235000007558 Avena sp Nutrition 0.000 claims description 2
- 240000008620 Fagopyrum esculentum Species 0.000 claims description 2
- 235000009419 Fagopyrum esculentum Nutrition 0.000 claims description 2
- 240000005979 Hordeum vulgare Species 0.000 claims description 2
- 235000007340 Hordeum vulgare Nutrition 0.000 claims description 2
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 2
- 244000000231 Sesamum indicum Species 0.000 claims description 2
- 235000003434 Sesamum indicum Nutrition 0.000 claims description 2
- 240000008042 Zea mays Species 0.000 claims description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 2
- 235000021329 brown rice Nutrition 0.000 claims description 2
- 235000005822 corn Nutrition 0.000 claims description 2
- 239000010985 leather Substances 0.000 claims description 2
- 235000015099 wheat brans Nutrition 0.000 claims description 2
- 235000013339 cereals Nutrition 0.000 claims 3
- 238000009313 farming Methods 0.000 claims 1
- 239000000463 material Substances 0.000 claims 1
- 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 10
- 238000011534 incubation Methods 0.000 description 9
- 239000000523 sample Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 238000001035 drying Methods 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 5
- 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
- 229930195725 Mannitol Natural products 0.000 description 5
- 229960005305 adenosine Drugs 0.000 description 5
- 239000000594 mannitol Substances 0.000 description 5
- 235000010355 mannitol Nutrition 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 239000013558 reference substance Substances 0.000 description 4
- 230000028070 sporulation Effects 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 239000000470 constituent Substances 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- 238000003306 harvesting Methods 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 238000009423 ventilation Methods 0.000 description 3
- 241001248590 Isaria Species 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 230000003698 anagen phase Effects 0.000 description 2
- 239000002518 antifoaming agent Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000000428 dust Substances 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000003020 moisturizing effect Effects 0.000 description 2
- 238000010899 nucleation Methods 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000004064 recycling Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000004904 shortening Methods 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 2
- 230000003519 ventilatory effect Effects 0.000 description 2
- NFMJXEMKAVCGPF-IOSLPCCCSA-N (2r,3r,4s,5r)-2-(6-amino-2-ethoxypurin-9-yl)-5-(hydroxymethyl)oxolane-3,4-diol Chemical compound C12=NC(OCC)=NC(N)=C2N=CN1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NFMJXEMKAVCGPF-IOSLPCCCSA-N 0.000 description 1
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 108010069514 Cyclic Peptides Proteins 0.000 description 1
- 102000001189 Cyclic Peptides Human genes 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000819999 Nymphes Species 0.000 description 1
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 1
- 241001465382 Physalis alkekengi Species 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 244000062793 Sorghum vulgare Species 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000002929 anti-fatigue Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 239000008236 heating water Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000000147 hypnotic effect Effects 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 235000019713 millet Nutrition 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- FJVZDOGVDJCCCR-UHFFFAOYSA-M potassium periodate Chemical compound [K+].[O-]I(=O)(=O)=O FJVZDOGVDJCCCR-UHFFFAOYSA-M 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 210000000697 sensory organ Anatomy 0.000 description 1
- 238000007493 shaping process Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
Abstract
The present invention provides a kind of solid culture method of cicada fungus, which is characterized in that the method be will be enlarged by culture Cordyceps cicadae strain be inoculated on solid medium, successively pass through dark culture and optical culture, and timely collecting fructification to get；Wherein, the optical culture stage uses the illumination in 16-20 hours/day.
Description
Technical field
The present invention relates to a kind of solid culture methods of cicada fungus, and in particular to light application time during a kind of fixed optical culture
Artificial culture cicada fungus method.
Background technique
Chinese medicine cicada fungus is cicada Isaria Isaria cicadae Miquel (Paecilomyces cicadae Paecilomyces cicadae
(Miquel) Samson) the bacterium worm complex formed on cicada nymph is colonized in, it is famous one of the Chinese medicine of China's tradition, medicine
With 800 years more early than cordyceps sinensis.Cicada fungus category dietotherapeutic fungi, nutritive value with higher and specific effect.Cicada fungus can
Generate a variety of physiological activators to play an important role in medical and health care, including ucleosides, cyclic peptide, polysaccharide, alcohols,
Sterols and organic acid etc., adjust it is immune, improve renal function, adjust lipid metabolism, antitumor, antifatigue, analgesia, hypnosis,
It is obvious to be depressured hypoglycemic etc. effect.
Natural cicada fungus resource is extremely limited, along with wild cicada fungus because the place of production is different, collecting time is different, easily by miscellaneous bacteria
Pollution mildew and the problems such as containing heavy metal, quality are under suspicion.The cicada fungus of artificial culture is free of heavy metal, pollution-free, and
Nutritional ingredient is stablized, and therefore, is increasingly valued by people.Currently, having been realized in the factory culture of cicada fungus.
Artificial culture cicada fungus is the liquid for obtaining cicada Isaria by inclined-plane seed, shaking flask culture and liquid deep layer fermenting
Body seed, which is inoculated on solid matrix, first carries out dark culture, and light-exposed culture later finally obtains the fructification of bifurcated racemosus, through adopting
Edible medicinal raw material can be made by receiving drying.The yield and active component content of raw material accept to pass the sense organ and quality of consumer
It is important, and different light application times has a certain impact for the content of fruiting body yield and effective active composition.
Currently, manually culture cicada fungus light application time used in the optical culture stage is mostly for 24 hours, to cause unnecessary energy
Source waste.
Summary of the invention
In order to solve the problems existing in the prior art, the present invention provides a kind of solid culture method of cicada fungus, this method be by
Cordyceps cicadae strain is inoculated on solid medium, successively pass through dark culture and optical culture, and timely collecting fructification to get；Wherein,
The optical culture stage uses the illumination in 16-20 hours/day.
Preferably, the Cordyceps cicadae strain is selected from the Cordyceps cicadae strain for expanding culture；Expansion culture include inclined-plane culture,
Any one or a few in the methods of shaking flask culture, seed tank culture fermentation tank culture.
Preferably, the optical culture stage uses the illumination of 18 hours/day.
In some embodiments, the solid medium is solid medium conventional in cicada fungus incubation, such as paddy
Object culture medium can be selected from wheat, corn, rice, millet, soybean powder, buckwheat, barley, oat, brown rice, any one in polished rice
Kind is several；Crop stalk culture medium, such as any one or a few in straw, cornstalk, cotton stalk, beans bar, sesame straw；Agriculture
Crop leather shell culture medium, such as wheat bran, soybean skin, cotton seed hulls.Preferably wheat culture medium.
The inoculum concentration is 5%~15%；Preferably inoculum concentration is 7%~12%；Further preferred inoculum concentration is 10%.
The condition of culture is：22-24 DEG C of cultivation temperature of the dark culture stage, relative humidity 70-85%；The optical culture stage
Cultivation temperature is 20-22 DEG C, relative humidity 70-85%.
The optical culture stage uses recombined white light, intensity of illumination 150-250Lx.
Beneficial effects of the present invention：The present invention uses LED white light in the cicada fungus optical culture stage and carries out irradiation in 18h/ days,
For irradiating compared with the existing technology using whole day illumination for 24 hours, following remarkable result is achieved：1. significantly reducing disappearing for the energy
Consumption extends lamps and lanterns and its controls the service life of equipment；2. shorten light application time do not influence fructification appearance character and effectively
Component content, yield are also increased slightly；3. greatly reducing sporulation quantity, dust recycling in incubation can be significantly reduced and brought
Trouble.
Specific embodiment
Cordyceps cicadae strain used is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms in following example 1
Center, (strain discloses in the patent of invention application No. is 201110120603.1 deposit number CGMCC No 3453, is
Known strain).The present invention is not research shows that Cordyceps cicadae strain type constitutes the factor for influencing effect of the present invention, present invention drying side
Method can be realized effect to different Cordyceps cicadae strains.
The expansion culture of 1 cicada fungus of embodiment
Inclined-plane culture：Strain is inoculated in slant tube, then the slant tube for being inoculated with strain is put into 22 DEG C of incubators,
Incubation time is 7 days, covers with test tube to mycelia；
Shaking flask culture：200m1 fluid nutrient medium, the high pressure sterilization 30 under 0.11Mpa pressure are packed into 500m1 triangular flask
Minute, it is cooled to room temperature, the strain of 1 slant tube is inoculated into 500m1 triangular flask culture medium, constant temperature oscillation is placed in
Incubator is cultivated under the conditions of 22 ± 1 DEG C of temperature, 150 revs/min, and incubation time is 3 days；
Seed tank culture：20L fluid nutrient medium is packed into 50L airlift fermentor, culture medium temperature is greater than 95 DEG C, adds
Entering edible defoaming agent, additional amount is the 0.03% of culture medium weight, under 0.11Mpa pressure, it is passed through steam and is heated to 121 DEG C,
And pressure maintaining 30-45 minutes；After sterilizing, when being cooled to 20 DEG C, 4 bottles of above-mentioned cultured 500m1 shaking flask strains are accessed into culture medium,
Cultivation temperature is 22 ± 1 DEG C, ventilatory capacity 1：0.5V/V min, pressure maintaining 4.903-7.0845 × 104Pa was cultivated through ventilation in 3 days,
After reaching logarithmic growth phase, whole volume of culture is 20L.
Fermentation tank culture：200L fluid nutrient medium is packed into 500L airlift fermentor, culture medium temperature is greater than 95 DEG C,
Edible defoaming agent is added, additional amount is the 0.03% of culture medium weight, under 0.11Mpa pressure, is passed through steam and is heated to 121
DEG C, and pressure maintaining 30-45 minutes；After sterilizing, when being cooled to 20 DEG C, by all access trainings of above-mentioned cultured 200L seeding tank seed
It supports, cultivation temperature is 22 ± 1 DEG C, ventilatory capacity 1：0.5V/V min, pressure maintaining 4.903-7.0845 × 104Pa was trained through ventilation in 3 days
It supports, after reaching logarithmic growth phase.Whole volume of culture is 200L.
Slant tube culture medium：200g potato liquor, 20g sucrose, 20g agar, remaining moisturizing to 1000ml, pH value 6.5；
Fluid nutrient medium in shaking flask, seeding tank and fermentor：30g yeast extract powder, 30g sucrose, 5g soy hydrolysate egg
White, surplus moisturizing is to 1000 milliliters, pH value 6.5.
The solid culture of 2 cicada fungus of embodiment
The preparation of solid medium：After wheat is cleaned control water, suitable water is added, weight ratio is wheat：Water is 1：
1.4, it is uniformly mixed；It is fitted into culture vessel after mixing thoroughly, culture medium thickness is controlled in 4cm.The culture vessel of culture medium will be installed
It places in autoclave, sterilize 30-50min at 121 DEG C；After sterilizing, in culture vessel dislocation surge chamber, naturally cool to 24 DEG C with
In lower dislocation transfer room.
Inoculation：Be inoculated with before culture vessel first in superclean bench or hundred-level laminar flow cover (under) use ultraviolet light 0.5h；
Each culture vessel is expanded the strain that culture obtains by 10% inoculum concentration inoculation embodiment 1, and the container after inoculation is put into culture
Room culture.
Condition of culture：Dark culture stage cultivation temperature is 22-24 DEG C, relative air humidity 70-85%；It is covered with to mycelium
Switch to optical culture when culture medium, light source uses LED white light, intensity of illumination 200Lx, and room temperature is cultivated in 18 hour/day of light application time
It is 20-22 DEG C, relative air humidity 70%-85%.Culturing room wants in due course ventilation, keeps bacterium germination room air fresh；To son
Entity is mature, not yet largely generates spore (about 23~26 days).
The solid culture of 3 cicada fungus of embodiment
Incubation is substantially the same manner as Example 2, and difference is that light application time is 16h.
The solid culture of 4 cicada fungus of embodiment
Incubation is substantially the same manner as Example 2, and difference is that light application time is 20h.
The solid culture of 5 cicada fungus of embodiment
Incubation is substantially the same manner as Example 2, and difference is that light application time is 22h.
The solid culture of 6 cicada fungus of embodiment
Incubation is substantially the same manner as Example 2, and difference is that light application time is for 24 hours.
The harvesting of embodiment 7 and drying
The fructification of any culture of embodiment 2~6 is harvested；Fructification after harvesting is directly entered drying equipment standard
Standby drying.60 DEG C of drying temperature, drying time 4-5h.
Influence of the light application time to fructification yield and quality
The fructification that Example 2-6 is cultivated is harvested and is dried by 7 method of embodiment, the production to fructification
Amount and quality are evaluated as follows.
1, influence of the light application time to cicada fungus fructification appearance
10 scoring people comment its appearance color the fructification that each embodiment harvests according to the standards of grading of table 1
Point, record average value is shown in Table 2.
1 appearance standards of grading of table
| Color | Score value |
| Light yellow-buff | 15-11 |
| Buff-dark gray | 10-6 |
| Dark gray-black | 5-0 |
Influence of 2 light application time of table to appearance color
| Embodiment | Optical culture stage daily light application time | Average |
| Embodiment 2 | 18h/ days | 13.8 |
| Embodiment 3 | 16h/ days | 13.0 |
| Embodiment 4 | 20h/ days | 10.4 |
| Embodiment 5 | 22h/ days | 12.7 |
| Embodiment 6 | For 24 hours/day | 13.4 |
The result shows that carrying out optical culture using 18h, fruit-body color is yellow, the light application time group phase several with other
Difference is not very big.As it can be seen that shortening light application time did not influenced the face shaping of cicada fungus fructification to 18h/ days.
2, influence of the light application time to cicada fungus yield
Every 1150g solid medium (dry weight) fruiting body yield of embodiment 2~6 is calculated, the results are shown in Table 3.
Influence result (n=3) of 3 light application time of table to yield
| Embodiment | Optical culture stage daily light application time | Yield g/1150g solid medium |
| Embodiment 2 | 18h/ days | 129.75±8.41 |
| Embodiment 3 | 16h/ days | 119.04±6.59 |
| Embodiment 4 | 20h/ days | 121.22±6.03 |
| Embodiment 5 | 22h/ days | 129.18±9.49 |
| Embodiment 6 | For 24 hours/day | 128.67±9.55 |
From table 3 it can be seen that giving daily illumination for 24 hours, fruiting body yield is 128.67g/1150g solid medium, with
The shortening of light application time, fruiting body yield be gradually reduced, when light application time be 20h/ days when, fruiting body yield is only
121.22g/1150g solid medium reduces 6%；But when giving daily 18h illumination, fruiting body yield improves instead,
It is suitable with for 24 hours/day illumination condition for 129.75g/1150g solid medium.As it can be seen that the light application time in optical culture stage is contracted
It is as short as 18h/ days, the reduction of fruiting body yield will not be caused.
In addition, the present invention, which is found surprisingly that be cultivated using illumination 18h/ days, also can significantly reduce sporulation quantity, the present invention
Research shows that cultivate its sporulation quantity in 1.2~1.4g/1150g solid medium using other several light application times, and adopt
It is cultivated with illumination 18h, sporulation quantity can significantly reduce in incubation in this way in 0.5~0.8g/1150g solid medium
Dust recycling bring trouble.
3, influence of the light application time to active constituent content
The cicada fungus fructification of Example 2-6 culture respectively, measures its effective component adenosine, HEA (i.e. N6- (2- ethoxy
Adenosine)), polyoses content.
3.1 detection method
3.1.1 the measurement of adenosine and HEA use high performance liquid chromatography：
Chromatographic condition and system suitability：Chromatographic column：Symmetry C18 (Waters, 4.6mm × 250mm, 5 μm,
L.N.0264313291)；Mobile phase：Acetonitrile -0.04mol/L potassium dihydrogen phosphate (5:95)；Flow velocity：1.0mL/min；Detect wave
It is long：260nm；Column temperature：35℃；Sample volume：10μL.Theoretical cam curve is calculated by adenosine peak should be not less than 3000.
The preparation of reference substance solution：Take adenosine and HEA reference substance appropriate, it is accurately weighed, add 50% methanol aqueous solution to be made
The solution of 25 μ g/mL to get.
The preparation of test solution：0.2g cicada fungus fructification powder is taken, it is accurately weighed, 20mL tool plug test tube is set, precision adds
Enter 5mL distilled water, ultrasonic (40KHz) is handled 30 minutes, is taken out, and it is accurate immediately that 5mL methanol is added, it mixes, crosses 0.22 μm of micropore
Filter membrane, take subsequent filtrate to get.
Measurement：It is accurate respectively to draw reference substance solution and each 10 μ L of test solution, inject high performance liquid chromatograph, sample introduction
Time is 20min, and measurement records reference substance and the corresponding peak area of test sample at this wavelength, according to concentration conversion result into
Row calculate to get.
3.1.2 the measurement of polysaccharide according to《Health food functional component detection method》The method of (Bai Hong chief editor).
3.1.3 mannitol is detected using UV-VIS spectrophotometry
The preparation of sample extracting solution：Precision weighs 0.1g cicada fungus fructification powder, and 70% ethanol water of 100mL is added,
85-90 DEG C of water-bath refluxing extraction, solution boil after 5min, take out, and filtering takes filtrate, liquid to be filtered is cooled to room temperature, with 70% second
Alcohol solution is settled to 100mL, shakes up, as testing sample solution.
The measurement of sample mannitol content：Precision pipettes sample extracting solution 1mL, sets in 15mL tool plug test tube, and 1mL is added in precision
Potassium metaperiodate solution mixes, is placed at room temperature for 10min, then plus 0.1%L- sandlwood sugar juice 2mL to remove excessive periodate,
Oscillation mixes, and finally plus the Nash reagent of 4mL Fresh, 53 DEG C of heating water bath 15min make its colour developing, are quickly cooled to room
Temperature.According to UV-VIS spectrophotometry (one annex VA of Pharmacopoeia of People's Republic of China), extinction is measured at 412nm wavelength
Value, brings standard curve into, finds out sample mannitol concentration (mg/mL), calculates sample mannitol content (mg/g) further according to formula.
Calculation formula：
3.2 measurement result
It the results are shown in Table 4.
Influence (n=3) of 4 light application time of table to active constituent content
| Solid culture method | Adenosine (%) | HEA (%) | Polysaccharide (%) | Mannitol (%) |
| Embodiment 2 | 0.107±0.004 | 0.088±0.007 | 7.27±0.462 | 5.14±0.387 |
| Embodiment 3 | 0.111±0.007 | 0.103±0.009 | 7.16±0.538 | 5.21±0.297 |
| Embodiment 4 | 0.100±0.005 | 0.094±0.007 | 7.21±0.665 | 5.17±0.384 |
| Embodiment 5 | 0.102±0.008 | 0.096±0.006 | 7.13±0.945 | 5.02±0.257 |
| Embodiment 6 | 0.097±0.004 | 0.093±0.021 | 7.58±0.970 | 5.18±0.265 |
The result shows that carrying out optical culture using 18h, fructification active constituent content is compared with other light application times without aobvious
Variation is write, shows not influencing product inherent quality using the irradiation of 18h light.
Claims (10)
1. a kind of solid culture method of cicada fungus, which is characterized in that the method is that Cordyceps cicadae strain is inoculated into solid medium
On, successively pass through dark culture and optical culture, and timely collecting fructification to get；Wherein, the optical culture stage using 16-20 hours/
It illumination.
2. cultural method as described in claim 1, which is characterized in that the Cordyceps cicadae strain is selected from the cicada fungus bacterium for expanding culture
Kind.
3. cultural method as claimed in claim 1 or 2, which is characterized in that the solid medium includes grain culture medium, agriculture
Crop material culture medium, crops bran mass.
4. cultural method as claimed in claim 3, which is characterized in that the grain culture medium is wheat, corn, rice, small
Rice, soybean powder, buckwheat, barley, oat, brown rice, any one or a few composition in polished rice culture medium；The crops straw
Bar culture medium is culture medium of straw, cornstalk, cotton stalk, beans bar, any one or a few composition in sesame straw；The farming
Object leather shell culture medium is culture medium of wheat bran, soybean skin, any one or a few composition in cotton seed hulls.
5. cultural method as claimed in claim 3, which is characterized in that institute's solid medium is grain culture medium.
6. cultural method as described in claim 1, which is characterized in that the inoculum concentration is 5%~15%.
7. cultural method as claimed in claim 6, which is characterized in that the inoculum concentration is 7%~12%.
8. cultural method as claimed in claim 7, which is characterized in that the inoculum concentration is 10%.
9. cultural method as described in claim 1, which is characterized in that the condition of culture is：Dark culture stage cultivation temperature
22-24 DEG C, relative humidity 70-85%；Optical culture stage cultivation temperature is 20-22 DEG C, relative humidity 70-85%.
10. cultural method as described in claim 1, which is characterized in that the optical culture stage uses recombined white light, and illumination is strong
Degree is 150-250Lx.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710252724.9A CN108901587B (en) | 2017-04-18 | 2017-04-18 | Solid culture method of cordyceps sobolifera |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710252724.9A CN108901587B (en) | 2017-04-18 | 2017-04-18 | Solid culture method of cordyceps sobolifera |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108901587A true CN108901587A (en) | 2018-11-30 |
| CN108901587B CN108901587B (en) | 2021-02-09 |
Family
ID=64402856
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710252724.9A Active CN108901587B (en) | 2017-04-18 | 2017-04-18 | Solid culture method of cordyceps sobolifera |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108901587B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112005814A (en) * | 2019-05-30 | 2020-12-01 | 大叶大学 | Cordyceps sobolifera, solid and liquid culture method thereof and application of extract thereof |
| CN114214199A (en) * | 2022-01-07 | 2022-03-22 | 沈阳农业大学 | Preservation method of isaria cicadae spore powder |
| CN114586606A (en) * | 2022-03-17 | 2022-06-07 | 连云港市农业科学院 | Culture method of cordyceps sobolifera |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1614003A (en) * | 2004-10-30 | 2005-05-11 | 浙江省农业科学院 | Culture of Chansi mould strains and extraction and use of anagentic compound therefrom |
| CN102763560A (en) * | 2012-06-19 | 2012-11-07 | 桐乡市中泰虫草专业合作社 | Cordyceps sobolifera fruiting body with cicada larva as host and culture method thereof |
| CN103283479A (en) * | 2012-02-24 | 2013-09-11 | 北京市弘科农场 | Cordyceps militaris strain and application thereof as well as method for producing cordyceps militaris |
| CN105543104A (en) * | 2016-01-05 | 2016-05-04 | 江苏农林职业技术学院 | Optimization method of solid culture medium for artificial acclimation and cultivation of wild Isaria cicadae Miquel |
| CN106034738A (en) * | 2016-06-06 | 2016-10-26 | 江苏科技大学 | Method for manually cultivating silkworm chrysalis cordyceps cicadae by using silkworm chrysalis of domestic silkworms as hosts |
| CN106190861A (en) * | 2016-07-19 | 2016-12-07 | 湖州新驰医药科技有限公司 | The bacterial screening of a kind of artificial isaria cicadae miq, preparation and productive culture method |
| CN106318875A (en) * | 2015-06-18 | 2017-01-11 | 浙江泛亚生物医药股份有限公司 | Cordyceps sobolifera two-way artificial culture method |
-
2017
- 2017-04-18 CN CN201710252724.9A patent/CN108901587B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1614003A (en) * | 2004-10-30 | 2005-05-11 | 浙江省农业科学院 | Culture of Chansi mould strains and extraction and use of anagentic compound therefrom |
| CN103283479A (en) * | 2012-02-24 | 2013-09-11 | 北京市弘科农场 | Cordyceps militaris strain and application thereof as well as method for producing cordyceps militaris |
| CN102763560A (en) * | 2012-06-19 | 2012-11-07 | 桐乡市中泰虫草专业合作社 | Cordyceps sobolifera fruiting body with cicada larva as host and culture method thereof |
| CN106318875A (en) * | 2015-06-18 | 2017-01-11 | 浙江泛亚生物医药股份有限公司 | Cordyceps sobolifera two-way artificial culture method |
| CN105543104A (en) * | 2016-01-05 | 2016-05-04 | 江苏农林职业技术学院 | Optimization method of solid culture medium for artificial acclimation and cultivation of wild Isaria cicadae Miquel |
| CN106034738A (en) * | 2016-06-06 | 2016-10-26 | 江苏科技大学 | Method for manually cultivating silkworm chrysalis cordyceps cicadae by using silkworm chrysalis of domestic silkworms as hosts |
| CN106190861A (en) * | 2016-07-19 | 2016-12-07 | 湖州新驰医药科技有限公司 | The bacterial screening of a kind of artificial isaria cicadae miq, preparation and productive culture method |
Non-Patent Citations (1)
| Title |
|---|
| 牟雪 等: "光照时数对蛹虫草生长发育的影响", 《河北农业科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112005814A (en) * | 2019-05-30 | 2020-12-01 | 大叶大学 | Cordyceps sobolifera, solid and liquid culture method thereof and application of extract thereof |
| CN114214199A (en) * | 2022-01-07 | 2022-03-22 | 沈阳农业大学 | Preservation method of isaria cicadae spore powder |
| CN114214199B (en) * | 2022-01-07 | 2023-08-22 | 沈阳农业大学 | Preservation method of Isaria cicadae spore powder |
| CN114586606A (en) * | 2022-03-17 | 2022-06-07 | 连云港市农业科学院 | Culture method of cordyceps sobolifera |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108901587B (en) | 2021-02-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107699499B (en) | One Aspergillus oryzae ZA127 and its application | |
| CN110476706A (en) | Improve the cultural method of ganoderma lucidum polysaccharide, Ganoderma total triterpenes acid content | |
| US10356988B2 (en) | Method for culturing Ophiocordyceps sinensis | |
| CN104388322A (en) | Solid state fermentation method of lipid-lowering Monascus purpureus Zhang-MP | |
| CN105981581B (en) | A kind of artificial culture method of cicada fungus | |
| CN105219655A (en) | Flavus biocontrol strain and the application thereof of aflatoxin do not produced to sterilant resistance | |
| CN105779360B (en) | A kind of bacillus subtilis and its application | |
| CN106318875A (en) | Cordyceps sobolifera two-way artificial culture method | |
| CN108901587A (en) | A kind of solid culture method of cicada fungus | |
| CN110157624A (en) | A kind of Paecilomyces lilacinus large-scale method for producing based on automation koji machine | |
| CN106212044A (en) | A kind of Antrodia Camphorata ware formula cultural method | |
| CN107586725B (en) | Cordyceps liquid culture medium and method for culturing cordyceps by using same | |
| CN110093283A (en) | Strain of Beauveria bassiana and its cultural method | |
| CN108175040B (en) | Preparation method of millet oat antrodia camphorata mycoplasm health food | |
| CN106922386A (en) | A kind of artificial culture method of cicada fungus | |
| CN106922387A (en) | A kind of artificial culture method of cicada fungus | |
| CN108624509A (en) | The circulation utilization method of solid medium in a kind of artificial incubation of cicada fungus | |
| CN106922389A (en) | A kind of artificial culture method of cicada fungus | |
| CN109762745A (en) | A kind of fermentation culture method of Pleurotus tuber-regium and the method that active constituent is extracted from culture | |
| CN108566982A (en) | A kind of drying means of cicada fungus fructification | |
| CN106258999B (en) | A kind of thickness pleat Aode mushroom strain, cultural method and application thereof | |
| CN108611381A (en) | It is a kind of to be conducive to the liquid fermentation culturing method that Flavonoid substances are enriched in Phellinus | |
| CN107048166A (en) | A kind of method of natural gamma Gamma-propalanine content in increase highland barley monascus | |
| JPH04304890A (en) | Production of ganoderic acids | |
| CN106922388A (en) | A kind of artificial culture method of cicada fungus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP03 | Change of name, title or address |
Address after: No.1938, xinqun Road, Pinghu Economic Development Zone, Jiaxing City, Zhejiang Province, 314200 Patentee after: Pan Asia Biopharmaceutical Co.,Ltd. Country or region after: China Address before: No.1938, xinqun Road, Pinghu Economic Development Zone, Jiaxing City, Zhejiang Province, 314200 Patentee before: ZHEJIANG BIOASIA PHARMACEUTICAL Co.,Ltd. Country or region before: China |