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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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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medium
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cordyceps sinensis
sterile
cultivation
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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 US15/122,763 priority Critical patent/US10400209B2/en
Priority to JP2016557177A priority patent/JP6143245B2/ja
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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

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  • 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.

Abstract

一种冬虫夏草子实体人工栽培方法，是将中国被毛孢菌（Ophiocordyceps sinensis）接种到无菌的大米培养基中，置于9-13℃下培养40-60天，待菌丝长满培养基后，1-8℃下低温诱导60-80天即可长出子实体原基，转至11-16℃下培养至收获子实体。所述冬虫夏草子实体人工栽培方法无需低氧环境，可降低培养成本，从诱导到收获子实体只需3-4个月，使用的大米培养基成本低廉，适合于冬虫夏草子实体商业化栽培。

Description

一种冬虫夏草子实体人工栽培方法 技术领域：
本发明属于微生物技术领域，具体涉及一种冬虫夏草子实体人工栽培方法。
背景技术：
冬虫夏草(Ophiocordyceps sinensis,异名Cordyceps sinensis)是我国最具特色的生物资源，隶属子囊菌门(Ascomycota)、粪壳菌纲(Sordariomycetes)、肉座菌目(Hypocreales)、线虫草科(Ophiocordycipitaceae)、虫草属(Ophiocordyceps)，国内主要产于西藏、青海、云南、四川、甘肃等海拔3000米以上的雪山高寒山区。正宗的药用冬虫夏草是中国被毛孢(Ophiocordyceps sinensis)真菌感染寄主昆虫-蝙蝠蛾幼虫，并使其僵化，在适宜条件下感菌僵虫生长，形成具有虫(僵虫)草(真菌子实体)复合形态的结构。
冬虫夏草是我国药食同源佳品，具有“补肺、强肾、益精气、理诸虚百损”等众多功效。现代医学则把冬虫夏草视作天然免疫调节剂，是呵护人体健康的“天然大复方”。冬虫夏草可产生多种具有抗菌、抗病毒、抗肿瘤、抗辐射及免疫调节等功能的生理活性物质，在医药、食品以及现代生物技术等方面具有广泛的应用，尤其在我国传统滋补品市场中占有非常重要的地位，一直被国民推崇和信赖，并在日本、韩国、东南亚国家和美国等国际市场热销。
资源的枯竭、需求的旺盛和政策的保护导致其市场价格飞涨。野生冬虫夏草已被列为国家二级保护物种。为了保护青藏高原生态、虫草资源，让冬虫夏草更好地为人类健康服务，唯一选择是人工培育。但是，冬虫夏草的生长条件苛刻，目前尚无冬虫夏草人工规模化培养技术问世，尽管冬虫夏草无性型-中国被毛孢或相关真菌的发酵培养及其产品业已投入市场(Zhou et al.,2013，Informa Heathcare,DOI:10.3109/07388551.791245；Yue et al.,2013，International Journal of Medicinal Mushrooms,15:425–434.)。
目前，世界上能够成功人工培育的虫草子实体种类不多。除了利用含米饭的培养基栽培蛹虫草子实体外，还通过以昆虫为寄主来培养蛹虫草。潘中华等(2002)(江苏蚕业，24:21-24)和汪西强等(2002)(安徽农业科学，30：965-968) 分别利用家蚕蛹(Bombyx mori L.)和蚱蝉蛹(Cryptotympana pustulata F.)培植蛹虫草。李如春等(2005)(菌物学报，24：349-355)以台湾虫草(C.formosana)菌感染黄粉虫(Tenebrio molitor L.)形成菌索。李泰辉等(2005)(国家发明专利，ZL200510101348.0)利用人工培养的黄粉虫蛹成功培植出蛹虫草子座。一些专利也描述了利用蚕蛹培植蛹虫草方法(徐道田等，2000，ZL00130381.3；李健，2001，ZL01141386.7；陈瑞英等，1997，ZL97113190.2)。韩日畴等(2006)(国家发明专利，ZL200610123355.5)利用大蜡螟(Galleria mellonella)幼虫成功培植出蛹虫草子实体。
专利申请号为201310432723.4，发明名称为：一种冬虫夏草子实体及其培植方法，虽然其能够培养出冬虫夏草子实体，但是其具有以下三个缺点：1、其诱导子实体至子实体长出需要在低氧浓度(10～15％氧气浓度)下，如果在低海拔地区，维持低氧浓度长达5-6个月的话，其成本是非常可观的。2、其诱导子实体至子实体长出需要长达5-6个月的时间，时间比较长。3、其培养基的成本比较高，不适合商业化大规模培养。
发明内容：
本发明的目的是提供一种无需低氧环境，可在低海拔地区的正常氧浓度下商业化人工培养冬虫夏草子实体的方法。
本发明的冬虫夏草子实体人工栽培方法，其特征在于，包括以下步骤：
将中国被毛孢菌(Ophiocordyceps sinensis)接种到无菌的栽培培养基中，置于9-13℃下培养40-60天，待菌丝长满培养基后，1-8℃下低温诱导60-80天即可长出子实体原基，转至11-16℃下培养30-40天可收获长度为4-12cm的子实体，收获的子实体棒状、不分枝、灰褐色，与野外采集的冬虫夏草的子实体形态相似；
所述的栽培培养基由大米和营养液按重量比1:1～1.5混合而成，所述的营养液，按总质量分数100％计，包括葡萄糖2％，KH2PO40.2％，MgSO40.1％，柠檬酸铵0.1％，蛋白胨0.5％,蚕蛹粉0.2％，余量为水，pH6.0-6.5。
作为优选，所述的将中国被毛孢菌接种到无菌的栽培培养基中，其中国被毛孢菌是通过以下方法制备：
(1)母种制备：将中国被毛孢菌接入固体PPDA培养基中，9-16℃下培养 45-60天后，选取冬虫夏草典型菌落作为母种；
(2)液体菌种制备：将母种菌落接至液体PPDA培养基中，9-16℃下，振荡培养40-60天，选取菌丝球大小均匀一致、直径为2-3毫米的作为液体菌种；
在无菌环境下，用无菌水稀释液体菌种5-10倍，再接入到无菌的栽培培养基上。
本发明的固体PPDA培养基是现有技术中常用的培养基，其配方为：葡萄糖20g，马铃薯200g，蛋白胨10g，KH2PO4 3g，MgSO4·7H2O 1.5g，VB1 0.02g，琼脂15g，H2O 1000mL，自然pH，其配制方法为：将马铃薯洗净去皮，加水煮烂，再用纱布过滤，在滤液中加入葡萄糖、蛋白胨，KH2PO4，MgSO4·7H2O，VB1，琼脂，用水定容至1L，121℃灭菌30分钟备用。所述的液体PPDA培养基是指将固体PPDA培养基中的琼脂去掉后得到的培养基，其配制方法同上，只是不加琼脂。
本发明经过多次重复，均能达到本发明的目的，即能成功规模化栽培出冬虫夏草子实体。子实体培养条件建立是本发明得以实现的关键。
按照本发明的栽培方法栽培获得的冬虫夏草子实体在形态上与野生冬虫夏草的子实体相似，主要成分含量未低于野生冬虫夏草子实体，可作为食品应用。以下是野生冬虫夏草和按照本发明的人工栽培方法获得的冬虫夏草子实体主要成分的分析结果，具体如表1所示。
表1：本发明的人工栽培的冬虫夏草子实体和野生冬虫夏草子实体主要成分的分析结果
分析项目 人工栽培的冬虫夏草子实体 野生冬虫夏草子实体
多糖(g/100g子实体) 3.37 1.07
麦角甾醇(mg/kg子实体) 3500 813
虫草酸(g/100g子实体) 11 14
本发明的冬虫夏草子实体人工栽培方法，其是在低海拔地区的正常氧浓度下培养，无需10～15％的低氧浓度，因此显著地降低了培养成本，并且从子实体诱导到长出能够收获的子实体只需90-120天(即3-4个月)即可，缩短了培养时间，并且本发明所使用的大米培养基成本低廉，适合于冬虫夏草的商业化大规模栽培。
附图说明：
图1是本发明培养的冬虫夏草子实体的视图。
具体实施方式：
以下实施例是对本发明的进一步说明，而不是对本发明的限制。
实施例1：
实验地点：位于广州市的广东省昆虫研究所，下述培养都是在空气的常规氧浓度下。
将中国被毛孢(Ophiocordyceps sinensis)菌种接入无菌固体PPDA培养基中，在9℃暗培养60天后，选取冬虫夏草典型菌落作为母种；
将母种菌落接至无菌液体PPDA培养基中，在9℃下，100rpm振荡培养60天，选取菌丝球大小均匀一致、直径为2-3毫米的作为液体菌种，用于冬虫夏草的栽培生产。
于无菌室或超净工作台中，将经无菌水稀释5倍的液体菌种接入无菌的栽培培养基中。将接种后的培养瓶置于9℃下培养60天，待菌丝长满培养基后，于1℃下低温诱导60天即可长出子实体原基，转至11℃培养40天便可收获长度达到4-8cm的子实体，即从子实体低温诱导到长出能够收获的子实体的时间为100天。人工栽培的冬虫夏草子实体如图1所示。该冬虫夏草子实体在形态上与野生冬虫夏草的子实体相似，主要成分含量未低于野生冬虫夏草子实体的，可作为食品应用。
所述的栽培培养基的配制方法：将葡萄糖20g，KH2PO4 2g，MgSO4 1g，柠檬酸铵1g，蛋白胨5g，蚕蛹粉2g，溶于少量水中，pH值6.0-6.5，再定容至1L，得到营养液。将大米和营养液按重量比1:1混合搅拌后，装于培养瓶中，121℃灭菌60分钟备用。
实施例2：
实验地点：位于广州市的广东省昆虫研究所，下述培养都是在空气的常规氧浓度下。
将中国被毛孢(Ophiocordyceps sinensis)菌种接入无菌固体PPDA培养基中，在16℃暗培养45天后，选取冬虫夏草典型菌落作为母种；
将母种菌落接至无菌液体PPDA培养基中，在16℃下，100rpm振荡培养40天，选取菌丝球大小均匀一致、直径为2-3毫米的作为液体菌种，用于冬虫夏草的栽培生产。
于无菌室或超净工作台中，将经无菌水稀释10倍的液体菌种接入无菌的栽培培养基中。将接种后的培养瓶置于13℃下培养40天，待菌丝长满培养基后，8℃低温诱导80天即可长出子实体原基，转至16℃培养30天便可收获长度达到4-6cm的子实体，即从子实体低温诱导到长出能够收获的子实体的时间为110天。人工栽培的冬虫夏草子实体如图1所示。该冬虫夏草子实体在形态上与野生冬虫夏草的子实体相似，主要成分含量未低于野生冬虫夏草子实体的，可作为食品应用。
所述的栽培培养基的配制方法：将葡萄糖20g，KH2PO4 2g，MgSO4 1g，柠檬酸铵1g，蛋白胨5g，蚕蛹粉2g，溶于少量水中，pH值6.0-6.5，再定容至1L，得到营养液。将大米和营养液按重量比1:1.5混合搅拌后，装于培养瓶中，121℃灭菌60分钟备用。
实施例3：
实验地点：位于广州市的广东省昆虫研究所，下述培养都是在空气的常规氧浓度下。
将中国被毛孢(Ophiocordyceps sinensis)菌种接入无菌固体PPDA培养基中，在11℃暗培养53天后，选取冬虫夏草典型菌落作为母种；
将母种菌落接至无菌液体PPDA培养基中，在11℃下，100rpm振荡培养50天，选取菌丝球大小均匀一致、直径为2-3毫米的作为液体菌种，用于冬虫夏草的栽培生产。
于无菌室或超净工作台中，将经无菌水稀释7倍的液体菌种接入无菌的栽培培养基中。将接种后的培养瓶置于11℃下培养50天，待菌丝长满培养基后，5℃低温诱导65天即可长出子实体原基，再经13℃培养35天便可收获长度达到4-12cm的子实体，即从子实体低温诱导到长出能够收获的子实体的时间为100天。人工栽培的冬虫夏草子实体如图1所示，该冬虫夏草子实体在形态上与野生冬虫夏草的子实体相似，主要成分含量未低于野生冬虫夏草子实体的，可作为食品应用。
所述的栽培培养基的配制方法：将葡萄糖20g，KH2PO4 2g，MgSO4 1g，柠檬酸铵1g，蛋白胨5g，蚕蛹粉2g,溶于少量水中，pH值6.0-6.5，再定容至1L，得到营养液。将大米和营养液按重量比1:1.3混合搅拌后，装于培养瓶中，灭菌 备用。

Claims (2)

  1. 一种冬虫夏草子实体人工栽培方法，其特征在于，包括以下步骤：
    将中国被毛孢菌(Ophiocordyceps sinensis)接种到无菌的栽培培养基中，置于9-13℃下培养40-60天，待菌丝长满培养基后，1-8℃下低温诱导60-80天即可长出子实体原基，转至11-16℃下培养至收获子实体；
    所述的栽培培养基由大米和营养液按重量比1:1～1.5混合而成，所述的营养液，按总质量分数100％计，包括葡萄糖2％，KH2PO40.2％，MgSO40.1％，柠檬酸铵0.1％，蛋白胨0.5％，蚕蛹粉0.2％，余量为水，pH6.0-6.5。
  2. 根据权利要求1所述的冬虫夏草子实体人工栽培方法，其特征在于，所述的将中国被毛孢菌接种到无菌的栽培培养基中，其中国被毛孢菌是通过以下方法制备：
    (1)母种制备：将中国被毛孢菌接入固体PPDA培养基中，9-16℃下，暗培养45-60天后，选取冬虫夏草典型菌落作为母种；
    (2)液体菌种制备：将母种菌落接至液体PPDA培养基中，9-16℃下，振荡培养40-60天，选取菌丝球大小均匀一致、直径为2-3毫米的作为液体菌种；
    然后在无菌环境下，用无菌水稀释液体菌种5-10倍，再接入到无菌的栽培培养基上；
    所述的固体PPDA培养基，每升含有葡萄糖20g，马铃薯200g，蛋白胨10g，KH2PO43g，MgSO4·7H2O 1.5g，VB1 0.02g，琼脂15g，余量为水，自然pH；
    所述的液体PPDA培养基，每升含有葡萄糖20g，马铃薯200g，蛋白胨10g，KH2PO43g，MgSO4·7H2O 1.5g，VB1 0.02g，余量为水，自然pH。
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Families Citing this family (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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CN115067152B (zh) * 2022-06-28 2023-07-25 广东省科学院动物研究所 一种人工培养冬虫夏草子实体的培养基和培养方法
CN117099612B (zh) * 2023-09-04 2024-03-26 西藏月王药诊生态藏药科技有限公司 一种蛹虫草培育和提纯方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101773052A (zh) * 2010-03-16 2010-07-14 孟庆学 冬虫夏草菌种制备与工厂化栽培
CN102138437A (zh) * 2011-01-13 2011-08-03 安徽农业大学 一种人工培养台湾虫草子实体的方法
CN102187786A (zh) * 2011-03-25 2011-09-21 杜保华 野生冬虫夏草菌种无性繁殖方法
CN102613007A (zh) * 2012-04-25 2012-08-01 常熟市新港农产品产销有限公司 一种北冬虫夏草无公害栽培方法
CN103650910A (zh) * 2012-09-20 2014-03-26 乳源南岭好山好水冬虫夏草有限公司 一种冬虫夏草子实体及其培植方法
CN104082034A (zh) * 2014-06-25 2014-10-08 广东省昆虫研究所 一种冬虫夏草子实体人工栽培方法

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH01296991A (ja) * 1988-05-23 1989-11-30 Takeo Tabata 冬虫夏草類からの抗菌性物質の製造法及び用途
JP2937967B2 (ja) * 1997-10-07 1999-08-23 有限会社吉井菌学研究所 冬虫夏草菌の培養方法
JP4054584B2 (ja) * 2002-02-15 2008-02-27 スン、ジャ−モ 冬虫夏草子実体の大量生産のための液体種菌培養装置
CN1478886A (zh) * 2003-06-14 2004-03-03 安徽林苑虫草研究所 一种中国被毛孢液体菌种发酵工艺方法
CN1704469A (zh) * 2004-05-31 2005-12-07 沈南英 一种中国冬虫夏草真菌的生产方法
CN104322451B (zh) * 2014-08-20 2015-08-19 广东省昆虫研究所 一种冬虫夏草寄主昆虫蝙蝠蛾的人工低海拔饲养方法
CA2980173C (en) * 2015-03-31 2021-07-27 Paul Edward Stamets Antiviral activity from medicinal mushrooms containing phenyl carboxylate/acrylate compounds
TWI580780B (zh) * 2015-05-29 2017-05-01 東海大學 The cultivation method of Cordyceps sinensis

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101773052A (zh) * 2010-03-16 2010-07-14 孟庆学 冬虫夏草菌种制备与工厂化栽培
CN102138437A (zh) * 2011-01-13 2011-08-03 安徽农业大学 一种人工培养台湾虫草子实体的方法
CN102187786A (zh) * 2011-03-25 2011-09-21 杜保华 野生冬虫夏草菌种无性繁殖方法
CN102613007A (zh) * 2012-04-25 2012-08-01 常熟市新港农产品产销有限公司 一种北冬虫夏草无公害栽培方法
CN103650910A (zh) * 2012-09-20 2014-03-26 乳源南岭好山好水冬虫夏草有限公司 一种冬虫夏草子实体及其培植方法
CN104082034A (zh) * 2014-06-25 2014-10-08 广东省昆虫研究所 一种冬虫夏草子实体人工栽培方法

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
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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