WO2015196734A1 - 一种冬虫夏草子实体人工栽培方法 - Google Patents

一种冬虫夏草子实体人工栽培方法 Download PDF

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Publication number
WO2015196734A1
WO2015196734A1 PCT/CN2014/092747 CN2014092747W WO2015196734A1 WO 2015196734 A1 WO2015196734 A1 WO 2015196734A1 CN 2014092747 W CN2014092747 W CN 2014092747W WO 2015196734 A1 WO2015196734 A1 WO 2015196734A1
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WIPO (PCT)
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medium
days
cordyceps sinensis
sterile
cultivation
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PCT/CN2014/092747
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English (en)
French (fr)
Chinese (zh)
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曹莉
韩日畴
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广东省昆虫研究所
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Priority to JP2016557177A priority Critical patent/JP6143245B2/ja
Priority to US15/122,763 priority patent/US10400209B2/en
Publication of WO2015196734A1 publication Critical patent/WO2015196734A1/zh

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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
    • 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
    • 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

Definitions

  • Cordyceps sinensis is a homologous product of Chinese medicine and food. It has many functions such as “reinforcing the lungs, strengthening the kidneys, benefiting the essence, and alleviating the damage”. Modern medicine regards Cordyceps sinensis as a natural immune regulator, and is a "natural compound" for caring for human health. Cordyceps sinensis can produce a variety of physiologically active substances with antibacterial, antiviral, antitumor, anti-radiation and immune regulation functions. It has a wide range of applications in medicine, food and modern biotechnology, especially in the traditional tonic market in China. A very important position has always been respected and trusted by the nationals and is popular in international markets such as Japan, South Korea, Southeast Asian countries and the United States.
  • Patent application No. 201310432723.4 the invention name is: a Cordyceps sinensis fruit body and its cultivation method, although it can cultivate the Cordyceps sinensis fruit body, it has the following three disadvantages: 1. Its induced fruiting body to fruiting body growth needs to be At low oxygen concentrations (10 to 15% oxygen concentration), if the low oxygen concentration is maintained for 5-6 months at low altitudes, the cost is considerable. 2. It takes a period of 5-6 months for the eliciting fruiting body to the fruiting body to grow for a long time. 3, the cost of its medium is relatively high, not suitable for commercial large-scale cultivation.
  • Ophiocordyceps sinensis was inoculated into a sterile culture medium and cultured at 9-13 ° C for 40-60 days. After the mycelium was overgrown with the medium, the temperature was induced at 1-8 ° C. - The primordium of the fruiting body can be grown in -80 days, and the fruiting body with a length of 4-12 cm can be harvested by culturing for 30-40 days at 11-16 ° C. The harvested fruiting bodies are rod-shaped, unbranched, taupe, and wild. The collected fruit bodies of Cordyceps sinensis are similar in morphology;
  • the Chinese bacterium is inoculated into a sterile culture medium, and the Chinese bacterium is prepared by the following method:
  • the solid PPDA medium of the present invention is a medium commonly used in the prior art, and its formulation is: glucose 20g, potato 200g, peptone 10g, KH 2 PO 4 3g, MgSO 4 ⁇ 7H 2 O 1.5g, VB1 0.02g, agar 15g, H 2 O 1000mL, natural pH, prepared by washing and peeling the potato, adding water to boil, filtering with gauze, adding glucose, peptone, KH 2 PO 4 , MgSO 4 ⁇ 7H 2 O to the filtrate. , VB1, agar, dilute to 1L with water, sterilize at 121 ° C for 30 minutes.
  • the liquid PPDA medium refers to a medium obtained by removing the agar in the solid PPDA medium, and the preparation method is the same as above, except that no agar is added.
  • the invention has been repeatedly repeated to achieve the object of the present invention, that is, the fruiting body of the Cordyceps sinensis can be successfully grown on a large scale.
  • the establishment of the culture conditions of the fruiting bodies is the key to the realization of the present invention.
  • Table 1 Analysis results of the main components of the artificially cultivated Cordyceps sinensis fruit body and the wild Cordyceps sinensis fruit body of the present invention
  • the artificial cultivation method of the Cordyceps sinensis fruit body of the present invention is cultured under normal oxygen concentration in a low altitude region, and does not require a low oxygen concentration of 10 to 15%, thereby significantly reducing the cultivation cost and enabling the growth from the fruiting body to the growth of the fruiting body.
  • the harvested fruit body takes only 90-120 days (ie, 3-4 months), shortens the culture time, and the rice medium used in the present invention is low in cost and is suitable for commercial large-scale cultivation of Cordyceps sinensis.
  • Figure 1 is a view of a Cordyceps sinensis fruiting body cultured in accordance with the present invention.
  • the Chinese genus Ophiocordyceps sinensis was inserted into the sterile solid PPDA medium, and after 60 days of dark culture at 9 °C, the typical colony of Cordyceps sinensis was selected as the parent species;
  • the parental fungus was dropped into a sterile liquid PPDA medium, and cultured at 100 ° C for 60 days under shaking at 9 ° C.
  • the hyphae were uniformly sized and the diameter was 2-3 mm as a liquid strain for the cultivation of Cordyceps sinensis. produce.
  • the fruit body of the Cordyceps sinensis is similar in morphology to the fruit body of the wild Cordyceps sinensis, and the main component content is not lower than that of the wild Cordyceps sinensis fruit body, and can be used as a food application.
  • the preparation method of the cultivation medium comprises: 20 g of glucose, 2 g of KH 2 PO 4 , 1 g of MgSO 4 , 1 g of ammonium citrate, 5 g of peptone, 2 g of silkworm cocoon powder, dissolved in a small amount of water, pH 6.0-6.5, and then adjusted to volume To 1L, a nutrient solution is obtained.
  • the rice and the nutrient solution were mixed and stirred at a weight ratio of 1:1, and then placed in a culture flask and sterilized at 121 ° C for 60 minutes for use.
  • the Chinese genus Ophiocordyceps sinensis was inserted into the aseptic solid PPDA medium, and after 45 days of dark culture at 16 °C, the typical colony of Cordyceps sinensis was selected as the parent species;
  • the parental fungus was dropped into a sterile liquid PPDA medium, and cultured at 16 ° C for 10 days with shaking at 100 rpm.
  • the hyphae ball was uniformly sized and the diameter was 2-3 mm as a liquid strain for the cultivation of Cordyceps sinensis. produce.
  • the liquid strain diluted 10 times with sterile water is placed in a sterile culture medium in a sterile room or a clean bench.
  • the inoculated culture flask was cultured at 13 ° C for 40 days.
  • the fruiting body primordia could be grown by inducing the temperature at 8 ° C for 80 days, and the length was harvested by transferring to 16 ° C for 30 days.
  • the fruiting body reaching 4-6 cm, that is, the time from the low temperature induction of the fruit body to the growth of the fruit body capable of harvesting is 110 days.
  • the artificially cultivated Cordyceps sinensis fruit body is shown in Figure 1.
  • the fruit body of the Cordyceps sinensis is similar in morphology to the fruit body of the wild Cordyceps sinensis, and the main component content is not lower than that of the wild Cordyceps sinensis fruit body, and can be used as a food application.
  • the preparation method of the cultivation medium comprises: 20 g of glucose, 2 g of KH 2 PO 4 , 1 g of MgSO 4 , 1 g of ammonium citrate, 5 g of peptone, 2 g of silkworm cocoon powder, dissolved in a small amount of water, pH 6.0-6.5, and then adjusted to volume To 1L, a nutrient solution is obtained.
  • the rice and the nutrient solution were mixed and stirred at a weight ratio of 1:1.5, and then placed in a culture flask and sterilized at 121 ° C for 60 minutes for use.
  • the Chinese genus Ophiocordyceps sinensis was inserted into the sterile solid PPDA medium, and after 53 days of dark culture at 11 °C, the typical colony of Cordyceps sinensis was selected as the parent species;
  • the parental fungus was dropped into a sterile liquid PPDA medium, and cultured at 100 ° C for 50 days at 11 ° C.
  • the hyphae were uniformly sized and the diameter was 2-3 mm as a liquid strain for the cultivation of Cordyceps sinensis. produce.
  • the preparation method of the cultivation medium comprises: 20 g of glucose, 2 g of KH 2 PO 4 , 1 g of MgSO 4 , 1 g of ammonium citrate, 5 g of peptone, 2 g of silkworm cocoon powder, dissolved in a small amount of water, pH 6.0-6.5, and then adjusted to volume To 1L, a nutrient solution is obtained.
  • the rice and the nutrient solution were mixed and stirred at a weight ratio of 1:1.3, and then placed in a culture flask and sterilized for use.
PCT/CN2014/092747 2014-06-25 2014-12-02 一种冬虫夏草子实体人工栽培方法 WO2015196734A1 (zh)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2016557177A JP6143245B2 (ja) 2014-06-25 2014-12-02 冬虫夏草の子実体の人工培養方法
US15/122,763 US10400209B2 (en) 2014-06-25 2014-12-02 Method for artificial cultivation of ophiocordyceps sinensis fruiting bodies

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CN201410289703.0 2014-06-25
CN201410289703.0A CN104082034B (zh) 2014-06-25 2014-06-25 一种冬虫夏草子实体人工栽培方法

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CN112322507A (zh) * 2020-12-09 2021-02-05 盛世荣恩生物科技有限公司 一种类变形被孢霉的菌种培养基及其培养方法
CN114868599A (zh) * 2022-06-09 2022-08-09 河南省农业科学院植物营养与资源环境研究所 一种利用麦芽糖加快诱导平菇原基形成的方法
CN115299291A (zh) * 2022-09-16 2022-11-08 沧州职业技术学院 一种茅窝菌仿野生栽培方法

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CN104082034B (zh) * 2014-06-25 2015-05-20 广东省昆虫研究所 一种冬虫夏草子实体人工栽培方法
CN104473978B (zh) * 2014-11-26 2017-12-15 南京中科药业有限公司 一种富含活性成分的虫草菌丝体加工方法
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CN108739044A (zh) * 2017-12-29 2018-11-06 谢远泰 一种家蚕幼虫感染母种菌株及北冬虫夏草人工栽培方法
CN108739050B (zh) * 2018-05-25 2020-12-01 广东省科学院动物研究所 一种冬虫夏草菌液体培养基及高效获得冬虫夏草寄主昆虫感染用芽生孢子的方法
CN108739068B (zh) * 2018-06-08 2020-05-19 山西万海澳生物科技有限责任公司 一种冬虫夏草菌丝体和子实体的生产工艺
CN109937792B (zh) * 2019-03-21 2021-11-09 广东省微生物研究所（广东省微生物分析检测中心） 一种野生柱状环伞子实体的人工栽培方法
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CN111436330B (zh) * 2020-04-09 2021-08-10 广东省科学院动物研究所 一种利用弗氏假丝酵母菌促进冬虫夏草子实体生长的人工栽培方法
CN111713336A (zh) * 2020-07-17 2020-09-29 杭州明煦生物科技有限公司 一种改善人工冬虫夏草成品品相的方法
CN112029669B (zh) * 2020-09-15 2022-03-18 柯邦生物科技(上海)有限公司 一种线虫草菌、线虫草发酵液及方法、抗氧化剂、应用
CN112243798B (zh) * 2020-10-22 2022-02-22 山东同康农业开发有限公司 一种节能高效银耳菌株的选育及种植方法
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CN112322507A (zh) * 2020-12-09 2021-02-05 盛世荣恩生物科技有限公司 一种类变形被孢霉的菌种培养基及其培养方法
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CN115299291B (zh) * 2022-09-16 2023-08-15 沧州职业技术学院 一种茅窝菌仿野生栽培方法

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JP2017500059A (ja) 2017-01-05
JP6143245B2 (ja) 2017-06-07
CN104082034A (zh) 2014-10-08
US20170067011A1 (en) 2017-03-09
US10400209B2 (en) 2019-09-03
CN104082034B (zh) 2015-05-20

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