CN110305795B - Hirsutella sinensis and application thereof - Google Patents
Hirsutella sinensis and application thereof Download PDFInfo
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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
- A01G18/20—Culture media, e.g. compost
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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
- A01G18/40—Cultivation of spawn
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H15/00—Fungi; Lichens
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N3/00—Spore forming or isolating processes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
Abstract
The invention discloses hirsutella sinensis and application thereof. Hirsutella sinensis (Ophiocerdyceps sinensis) KD11-2 with the deposit number GDMCC No. 60594. The hirsutella sinensis (Ophiocerociceps sinensis) KD11-2 can grow fruiting bodies on an artificial culture medium and form ascospores (ascospores) and has the outstanding capability of forming ascospores, so that the hirsutella sinensis KD11-2 can be used for producing and collecting the hirsutella sinensis ascospores under the artificial culture condition. The invention promotes the artificial cultivation of hirsutella sinensis sporocarp and provides a new method for infecting the larva of hepialus armoricanus which is a host of cordyceps sinensis by using the obtained ascospores.
Description
The technical field is as follows:
the invention belongs to the field of artificial cultivation of cordyceps sinensis sporocarp, and particularly relates to hirsutella sinensis (Ophiococcus sinensis) KD11-2 capable of growing sporocarp on an artificial culture medium and forming ascocarp (ascospore) and application thereof.
Background art:
the cordyceps sinensis is the most distinctive biological resource in China, belongs to Ascomycota (Ascomycota), Chaetomium (Sordariomycetes), Hypocrea (Hypocrea), nematoda (Ophiomorphic picaceae) and Cordyceps (Ophiomorphic ceps), and is mainly produced in high and cold snow mountain areas with the altitude of more than 3000 meters, such as Tibet, Qinghai, Yunnan, Sichuan and Gansu, in China.
The cordyceps sinensis is a host insect infected by hirsutella sinensis fungus, namely a hepialus larva, is rigidified, is infected with stiff worm to grow under a proper condition, and forms a structure with a complex shape of worm (stiff worm) grass (fungus fruiting body).
Cordyceps sinensis has many effects of tonifying lung, strengthening kidney, benefiting essence and qi, treating deficiency and impairment, and the like. Modern medicine proves that the cordyceps sinensis has the functions of resisting bacteria, viruses, tumors, radiation, immunity and the like, and is widely applied to the aspects of medicines, foods, modern biotechnology and the like. In the traditional tonic market of our country, cordyceps sinensis has been advocated and relied on by the national people and is hot sold in international markets such as japan, korea, south east asian countries and the usa. Exhaustion of resources, flourishing of demand, and protection of policies lead to a rapid increase in market price. Wild Cordyceps sinensis has been classified as a national secondary protective plant. In order to protect plateau ecology and cordyceps sinensis resources and enable cordyceps sinensis to serve human health better, the only choice is artificial cultivation and substitute. At present, fermentation culture of hirsutella sinensis or related fungi and products thereof, which are anamorphs of Cordyceps sinensis, are put into the market (Zhou et al, 2013, Informata Heathcare, DOI: 10.3109/07388551.791245; Yue et al, 2013, International Journal of Medicinal Mushooms, 15: 425-. From the market point of view, the value of the Chinese caterpillar fungus mycelium is far lower than that of the Chinese caterpillar fungus fruiting body. The report of Zhuxinyan et al (2013) discloses that 80% of nutrients and active ingredients in a complete cordyceps sinensis are concentrated in stroma (sporocarp), and only 20% of nutrients and active ingredients are concentrated in polypide (mycelium), so that the cordyceps sinensis sporocarp is the essence of cordyceps sinensis. At present, only a few units in China publicize that the cordyceps sinensis fruiting body is successfully cultured on an artificial culture medium, such as Xiehan et al (2016) (construction of cDNA library before and after sexual development of hirsutella sinensis in Zhejiang agriculture, 2016, 28(11):1895-1900), Caoli et al (2014) (CN201410289703.0) and Zhuxinyan et al (2013) (CN 201310432723.4). However, the patents and publications disclosed so far do not mention whether the cultured fruit bodies can finally develop ascocarp.
The invention content is as follows:
the invention aims to provide hirsutella sinensis (Ophiocerdyceps sinensis) KD11-2 capable of growing fruiting bodies and forming ascospores on artificial culture media, which is preserved in Guangdong province microbial culture collection center (GDMCC) in 2019, 02 and 28 days, and has the address: no. 59 building 5 of No. 100 Dazhong Jie of Pieli, Guangzhou city, with the collection number GDMCC No. 60594. The invention promotes the artificial cultivation of hirsutella sinensis sporocarp and provides a new method for infecting the larva of hepialus armoricanus which is a host of cordyceps sinensis by using the obtained ascospores.
The method comprises the steps of inoculating hirsutella sinensis (Ophiococcus sinensis) KD11-2 into a sterile culture medium, culturing at 9-13 ℃ in the dark, inducing at 2-8 ℃ in the dark or under the light until mycelia grow over the culture medium until sporophore primordium grows out, culturing at 11-16 ℃ until sporophore grows out, and continuously culturing to form granular ascocarp at the tail end of the sporophore.
Among hundreds of cordyceps fungi isolated and preserved in the laboratory, only one hirsutella sinensis (Ophiocordyceps sinensis) KD11-2, which can grow fruiting bodies and form ascospores on artificial media, was obtained. That is, the present strain has an outstanding ability to form ascospores as compared with other strains.
Therefore, the second purpose of the invention is to provide the application of the hirsutella sinensis (Ophiocerdyceps sinensis) KD11-2 in the production of hirsutella sinensis ascospores on artificial culture medium.
The third purpose of the invention is to provide the application of hirsutella sinensis (Ophiocerdyceps sinensis) KD11-2 in obtaining cordyceps sinensis by infecting hepialus armoricanus larvae.
The hirsutella sinensis (Ophiocerociceps sinensis) KD11-2 can grow fruiting bodies on an artificial culture medium and form ascospores (ascospores) and has the outstanding capability of forming ascospores, so that the hirsutella sinensis KD11-2 can be used for producing and collecting the hirsutella sinensis ascospores under the artificial culture condition. The invention promotes the artificial cultivation of hirsutella sinensis sporocarp and provides a new method for infecting the larva of hepialus armoricanus which is a host of cordyceps sinensis by using the obtained ascospores.
Hirsutella sinensis (ophiocordyces sinensis) KD11-2 is preserved in Guangdong province microbial culture collection center (GDMCC) in 2019 at 28 th 02, and the address is as follows: no. 59 building 5 of No. 100 Dazhong Jie of Pieli, Guangzhou city, with the collection number GDMCC No. 60594.
Description of the drawings:
FIG. 1 is a photograph of a colony of hirsutella sinensis cultured in the dark at KD 11-212 ℃ for 75 days.
FIG. 2 shows an artificially cultivated fruiting body of Cordyceps sinensis with Saccharum sinensis Roxb.
The specific implementation mode is as follows:
the following examples are further illustrative of the present invention and are not intended to be limiting thereof.
The media in the following examples:
the formula of the solid PPDA culture medium is as follows: contains glucose 20g, potato 200g, peptone 10g, KH per liter2PO43g、MgSO4·7H2O 1.5g、VB10.02g of agar and 15g of agar, and the balance of H2O, natural pH, and the preparation method comprises the following steps: cleaning potato, peeling, weighing 200g peeled potato, shredding, decocting in water, filtering with gauze, adding glucose 20g, peptone 10g, KH2PO4 3g、MgSO4·7H2O 1.5g、VB120mg of agar and 15g of agar, adding water to a volume of 1L, and sterilizing at 121 ℃ for 30 minutes for later use. The liquid PPDA medium refers to a medium obtained by removing agar from a solid PPDA medium, and is prepared in the same manner as above except that agar is not added.
The dishes used to prepare the solid medium plates in the following examples were identical and had a diameter of 9.0 cm.
Example 1: isolation and characterization of hirsutella sinensis (Ophiocerdyceps sinensis) KD11-2
1. Acquisition of KD11-2 Strain: fresh cordyceps sinensis is collected from Yalaxiang of a furnace town of Kangding, Ganzui, Sichuan province in a cordyceps production area and is transported to a laboratory through low-temperature preservation. Removing soil and mycoderm around Cordyceps in sterile operating room or clean bench, and sterilizing 75% alcohol body surface; removing epidermis of sclerotium and stroma with sterile scalpel, cutting sclerotium and stroma into small pieces (avoiding intestinal tract of insect), respectively planting in PPDA solid plate culture medium, and culturing at 9-16 deg.C; and (3) carrying out molecular identification on the separated and obtained pollution-free typical colony of the cordyceps sinensis, confirming that the typical colony is cordyceps sinensis, obtaining the strain, and numbering the strain as KD 11-2.
2. Characteristics of the KD11-2 strain:
morphological characteristics: on PPDA solid medium, the colony is earthy yellow, the surface of the colony is convex, and a layer of fine white loose hairy mycelium and wrinkles are formed (see figure 1). The mycelium was observed by fluorescent staining and had septa, branches and a width of 2.2 to 4.9. mu.m. The kidney-type spore is of double kidney type.
Molecular biological characteristics: the molecular identification is hirsutella sinensis, named hirsutella sinensis (Ophiocerdyceps sinensis) KD11-2, which is preserved in Guangdong province microorganism culture Collection (GDMCC) in 2019 at 28 days 02 and 9, and the address: no. 59 building 5 of No. 100 Dazhong Jie of Pieli, Guangzhou city, with the collection number GDMCC No. 60594.
The culture characteristics are as follows: the most important feature of strain No. KD11-2 was that mature ascocarps (ascospores) were obtained under the following culture conditions. The method comprises the following steps: inoculating hirsutella sinensis (Ophiococcus sinensis) KD11-2 into a sterile culture medium, culturing at 9-13 deg.C in dark for 40-60 days, inducing at 2-8 deg.C in dark or under illumination (various light sources including white light, blue light, red light, etc. or various color light sources mixed) for 55-65 days after mycelia overgrow the culture medium, culturing at 11-16 deg.C (dark or light illumination) for 20-45 days until fruiting body length is 4-8cm, and culturing for 25-45 days to obtain fruiting body with ascocarp, wherein the obtained fruiting body has rod shape, no branch, and grey brown color, and the granular ascocarp is distributed at the tail end of the fruiting body, and has similar shape to wild collected Cordyceps sinensis fruiting body.
The culture medium is prepared by mixing rice and wheat singly or after mixing the rice and the wheat according to the weight ratio of 1:1 and nutrient solution according to the weight ratio of 1: 1.2-1.5; the nutrient solution comprises 2 percent of glucose and KH according to 100 percent of the total mass fraction2PO40.2％，MgSO40.1 percent, 0.1 percent of ammonium citrate, 0.5 percent of peptone, 0.2 percent of silkworm chrysalis meal and the balance of water, and the pH value is 6.0-6.5.
The other hirsutella sinensis strains are numbered as KD1223, are separated from Cordyceps sinensis in Yaraxiang of furnace town of Kangding, Cumingzi, Sichuan province of Cordyceps sinensis production area, and are identified as hirsutella sinensis by molecule.
The invention can achieve the aim of the invention through repeated operation for many times, namely, the cordyceps sinensis fruiting body can be successfully cultivated in a large scale and the ascocarp (ascospore) is formed. The establishment of the selected strains and the culture conditions of the fruiting bodies are the key to the realization of the invention.
Example 2:
the experimental site: the following cultures were all at conventional oxygen concentrations at the institute for the application of biological resources in Guangdong province, Guangzhou City.
Respectively inoculating hirsutella sinensis KD11-2 strain and other hirsutella sinensis (KD1223) strains into a sterile solid PPDA culture medium, performing dark culture at 9 deg.C for 60 days, selecting colonies, inoculating into sterile liquid PPDA culture medium, performing shaking culture (dark culture) at 9 deg.C and 100rpm for 65 days, and selecting light brown fermentation culture with uniform mycelium pellet size and diameter of 2-3 mm as liquid strain for culturing and producing fruiting body of Cordyceps sinensis.
In a sterile room or a clean bench, a liquid seed culture diluted 5 times with sterile water (30 mL of the diluted liquid seed culture per 100g of the culture medium) is inoculated into a sterile culture medium. Placing the inoculated culture bottle at 9 ℃ for dark culture for 60 days, inducing at 2 ℃ and dark for 60 days after hypha grows over the culture medium, then growing fruiting body primordium, turning to 11 ℃ for dark culture for 45 days, harvesting fruiting bodies with the length of 4-8cm, continuing culturing for 45 days, and forming granular ascocarp at the tail end of the fruiting body of hirsutella sinensis KD11-2, wherein the artificially cultured cordyceps sinensis fruiting body with ascocarp is shown in figure 2, the harvested fruiting bodies are rod-shaped, not branched and grey brown, the granular ascocarp is distributed at the tail end of the fruiting body, the shape of the harvested fruiting body is similar to that of the wild collected cordyceps sinensis fruiting body, and the fruiting bodies of other strains do not obtain ascocarp.
The preparation method of the culture medium comprises the following steps: mixing glucose 20g and KH2PO4 2g，MgSO41g of ammonium citrate, 5g of peptone and 2g of silkworm chrysalis meal, dissolving in a small amount of water, and adding water to a constant volume of 1L to obtain a nutrient solution, wherein the pH value is 6.0; mixing rice and nutrient solution at a weight ratio of 1:1.2, stirring to obtain culture medium, placing into culture bottle, and sterilizing at 121 deg.C for 60 min.
Example 3:
the experimental site: the following cultures were all at conventional oxygen concentrations at the institute for the application of biological resources in Guangdong province, Guangzhou City.
Respectively inoculating hirsutella sinensis KD11-2 strain and other hirsutella sinensis (KD1223) strains into a sterile solid PPDA culture medium, performing dark culture at 16 deg.C for 45 days, inoculating the selected colonies into a sterile liquid PPDA culture medium, performing shaking culture (dark culture) at 100rpm at 16 deg.C for 40 days, and selecting fermentation culture with uniform mycelium pellet size and diameter of 2-3 mm as liquid strain for cultivating and producing Cordyceps fruiting body.
In a sterile room or a clean bench, a liquid seed culture diluted 10 times with sterile water (30 mL of the diluted liquid seed culture per 100g of the culture medium) is inoculated into a sterile culture medium. Placing the inoculated culture bottle at 13 ℃ for dark culture for 40 days, after hypha grows over the culture medium, inducing at 4 ℃ for 55 days by white light (illumination intensity 300lux and illumination 10h/d) every day to grow fruiting body primordium, turning to 16 ℃ for 20 days by illumination culture (illumination intensity 300lux and illumination 10h/d) to harvest fruiting bodies with the length of 4-6cm, continuing to culture for 25 days, forming granular ascocarp at the tail end of the fruiting body by hirsutella sinensis KD11-2, and distributing the harvested fruiting bodies at the tail end of the fruiting bodies in a rod shape, without branching and grey brown, wherein the granular ascocarp is similar to the wild collected fruiting bodies of cordyceps sinensis. Fruiting bodies of other strains did not obtain ascocarp.
The preparation method of the culture medium comprises the following steps: mixing glucose 20g and KH2PO4 2g，MgSO41g of ammonium citrate, 5g of peptone and 2g of silkworm chrysalis meal, dissolving in a small amount of water, adjusting the pH value to 6.5, and adding water to a constant volume of 1L to obtain a nutrient solution; mixing rice and wheat in equal weight, mixing with nutrient solution at a weight ratio of 1:1.5, stirring to obtain culture medium, placing into culture bottle, and sterilizing at 121 deg.C for 60 min.
Example 4:
the experimental site: the following cultures were all at conventional oxygen concentrations at the institute for the application of biological resources in Guangdong province, Guangzhou City.
The hirsutella sinensis strain KD11-2 and other hirsutella sinensis strains (KD1223) are respectively inoculated into a sterile solid PPDA culture medium, after dark culture at 11 ℃ for 53 days, colonies are selected to be inoculated into a sterile liquid PPDA culture medium, shaking culture (dark culture) is carried out at 100rpm at 11 ℃ for 50 days, and a fermentation culture with uniform mycelium pellet size and 2-3 mm diameter is selected as a liquid strain for cultivation and production of cordyceps sinensis.
In a sterile room or a clean bench, a liquid seed culture diluted 7 times with sterile water (30 mL of the diluted liquid seed culture per 100g of the culture medium) is inoculated into a sterile culture medium. Placing the inoculated culture bottle at 11 ℃ for dark culture for 50 days, inducing the culture bottle at 8 ℃ for 65 days by blue light every day (illumination intensity is 200lux, illumination time is 10h/d) after hypha grows over a culture medium, then harvesting sporocarp primordium with the length of 4-7cm after dark culture for 35 days at 13 ℃, continuing culturing for 30 days, forming granular ascocarp at the tail end of the sporocarp by hirsutella sinensis KD11-2, wherein the harvested sporocarp is rod-shaped, not branched and grey brown, and the granular ascocarp is distributed at the tail end of the sporocarp and is similar to the shape of the cordyceps sinensis sporocarp collected in the field. Fruiting bodies of other strains did not obtain ascocarp.
The preparation method of the culture medium comprises the following steps: mixing glucose 20g and KH2PO4 2g，MgSO41g of ammonium citrate, 5g of peptone and 2g of silkworm chrysalis meal, dissolving the components in a small amount of water, adjusting the pH value to 6.5, adding water to a constant volume of 1L to obtain the nutrient solutionCultivating liquid; mixing and stirring wheat and the nutrient solution according to the weight ratio of 1:1.5 to obtain a culture medium, filling the culture medium into a culture bottle, and sterilizing for 60 minutes at 121 ℃ for later use.
Claims (1)
1. Hirsutella sinensis (A) and (B)Ophiocordyceps sinensis) Application of KD11-2 in producing hirsutella sinensis ascospore on artificial culture medium (hirsutella sinensis) (A)Ophiocordyceps sinensis) The KD11-2 has a preservation number of GDMC C No. 60594, and the application specifically comprises the following steps: hirsutella sinensis (A) and (B)Ophiocordyceps sinensis) Inoculating KD11-2 into sterile culture medium, culturing at 13 deg.C in dark, inducing fruiting body primordium to grow at 4 deg.C, illumination intensity of 300lux per day and illumination time of 10h/d after mycelia overgrow the culture medium, and culturing at 16 deg.C to obtain fruiting body with ascocarp.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN85101971A (en) * | 1985-04-01 | 1987-03-04 | 青海省畜牧兽医科学院 | China Cordyceps sinensis fungi and artificial culture method thereof |
| CN1560228A (en) * | 2004-02-27 | 2005-01-05 | 俞永信 | Method of whole manual culture for chinese caterpillar fungus |
| JP2007135422A (en) * | 2005-11-15 | 2007-06-07 | Yukimori Kondo | Method for producing carpophore of insect parasite fungus |
| CN101353628A (en) * | 2008-09-12 | 2009-01-28 | 江苏省农业科学院 | Rapid identification method of Chinese caterpillar fungus fertile bacterial strain |
| CN102283022A (en) * | 2011-07-22 | 2011-12-21 | 重庆市中药研究院 | Method for isolating Cordyceps sinensis Sacc. asexual-stage spawn |
| CN104472221A (en) * | 2014-12-22 | 2015-04-01 | 四川藏宝虫草生物科技有限公司 | Method for fermenting mycelia from natural ophiocordyceps sinensis fruiting bodies |
| CN106538902A (en) * | 2016-10-25 | 2017-03-29 | 广东省生物资源应用研究所 | A kind of man-made feeds of Vespertilio moth larvae and preparation method thereof |
| CN108739050A (en) * | 2018-05-25 | 2018-11-06 | 广东省生物资源应用研究所 | A kind of aweto fluid nutrient medium and the efficient method for obtaining host of Cordyceps sinensis insect infection blastopore |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106010978A (en) * | 2016-05-25 | 2016-10-12 | 浙江农林大学 | Culture method of cordyceps sinensis anamorphic hirsutella sinensis pure strain |
-
2019
- 2019-03-15 CN CN201910198974.8A patent/CN110305795B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN85101971A (en) * | 1985-04-01 | 1987-03-04 | 青海省畜牧兽医科学院 | China Cordyceps sinensis fungi and artificial culture method thereof |
| CN1560228A (en) * | 2004-02-27 | 2005-01-05 | 俞永信 | Method of whole manual culture for chinese caterpillar fungus |
| JP2007135422A (en) * | 2005-11-15 | 2007-06-07 | Yukimori Kondo | Method for producing carpophore of insect parasite fungus |
| CN101353628A (en) * | 2008-09-12 | 2009-01-28 | 江苏省农业科学院 | Rapid identification method of Chinese caterpillar fungus fertile bacterial strain |
| CN102283022A (en) * | 2011-07-22 | 2011-12-21 | 重庆市中药研究院 | Method for isolating Cordyceps sinensis Sacc. asexual-stage spawn |
| CN104472221A (en) * | 2014-12-22 | 2015-04-01 | 四川藏宝虫草生物科技有限公司 | Method for fermenting mycelia from natural ophiocordyceps sinensis fruiting bodies |
| CN106538902A (en) * | 2016-10-25 | 2017-03-29 | 广东省生物资源应用研究所 | A kind of man-made feeds of Vespertilio moth larvae and preparation method thereof |
| CN108739050A (en) * | 2018-05-25 | 2018-11-06 | 广东省生物资源应用研究所 | A kind of aweto fluid nutrient medium and the efficient method for obtaining host of Cordyceps sinensis insect infection blastopore |
Non-Patent Citations (4)
| Title |
|---|
| CHARACTERIZATION OF AN ENTOMOPHAGOUS MEDICINAL FUNGUS CORDYCEPS SINENSIS (BERK.) SACC. OF UTTARAKHAND，INDIA;RUPESH KUMAR ARORA et al.;《The bioscan》;20131231;第8卷(第1期);第195-200页 * |
| Fruiting Body Production of the Medicinal ChineseCaterpillar Mushroom, Ophiocordyceps sinensis(Ascomycetes), in Artificial Medium;Li Cao et al.;《International Journal of Medicinal Mushrooms》;20151231;第17卷(第11期);第1109-1111页 * |
| 冬虫夏草的研究进展、现存问题与研究展望;丘雪红等;《环境昆虫学报》;20160131;第38卷(第1期);第1-23页 * |
| 冬虫夏草菌子实体发育起始阶段cDNA文库的构建及ESTs分析;李建宏;《中国优秀硕士学位论文全文数据库 农业科技辑》;20140215(第2期);第D047-531页 * |
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