CN108476861A - A kind of nunchakus spore Chinese caterpillar fungus strain and its cultural method - Google Patents

A kind of nunchakus spore Chinese caterpillar fungus strain and its cultural method Download PDF

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CN108476861A
CN108476861A CN201810176973.9A CN201810176973A CN108476861A CN 108476861 A CN108476861 A CN 108476861A CN 201810176973 A CN201810176973 A CN 201810176973A CN 108476861 A CN108476861 A CN 108476861A
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nunchakus
spore
culture
caterpillar fungus
chinese caterpillar
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胡殿明
张俊波
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Jiangxi Agricultural University
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G18/00Cultivation of mushrooms
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G18/00Cultivation of mushrooms
    • A01G18/20Culture media, e.g. compost

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  • Life Sciences & Earth Sciences (AREA)
  • Mycology (AREA)
  • Environmental Sciences (AREA)
  • Mushroom Cultivation (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention discloses a kind of nunchakus spore Chinese caterpillar fungus strain and its cultural method, the nunchakus spore Chinese caterpillar fungus strains(Cordyceps ninchukispora(C.H. Su & H.H. Wang) G.H. Sung,J.M. Sung,Hywel‑Jones & Spatafora)It has been stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), deposit number:CGMCC No. 15089.Domesticating cultivation method includes fermented liquid bacterium, cultivation matrix preparation, Mycelium culture base flower bud, fruiting, and the present invention has tamed one plant of wild nunchakus spore Chinese caterpillar fungus strain, provides a new strains and an artificial domesticating cultivation method.

Description

A kind of nunchakus spore Chinese caterpillar fungus strain and its cultural method
Technical field
The present invention relates to a kind of domesticating and cultivating new methods of nunchakus spore cordyceps sinensis, more particularly to one plant of nunchakus spore Cordyceps Militaris Strain and cultivation nunchakus spore cordyceps sinensis method.
Background technology
Nunchakus spore cordyceps sinensis(Cordyceps ninchukispora (C.H. Su & H.H. Wang) G.H. Sung, J.M. Sung, Hywel-Jones & Spatafora)It is under the jurisdiction of mycota, Ascomycota, Ascomycotina, excrement shell Gammaproteobacteria, Meat seat bacterium subclass, Hypocreales, cordyceps sinensis section, Cordyceps.Cordyceps sinensis fungus multiparasitization is in the segmental appendages such as insect larvae or adult, spider The Partial Species of animal and Elaphomyces, and stroma is generated on host.The nunchakus spore cordyceps sinensis that this patent is related to parasitizes On agglomerate capsule category fungi.
Cordyceps sinensis is important medicinal fungi, and China cordyceps sinensis, which is studied and utilizes, long history, such as widely known Cordyceps sinensis, buddhist flower, Cordyceps militaris etc., refer to cordyceps sinensis, and numerous people both know about it with higher medical value, can be due to it Can not artificial cultivation to cause wild resource to destroy serious and expensive etc..Cordyceps sinensis is the living resources of preciousness, some of which Type has higher nutritive value, health care and medical functions.The cordyceps sinensis industry in China relates to cordyceps sinensis and its phase at present The Related products such as fungi, Cordyceps militaris, cicada fungus are closed, have formd a huge industry, and realize higher economic value And social benefit.Have become industry active in world's biological economy epoch by the entire fungi industry of representative of cordyceps sinensis industry One of.
Nunchakus spore cordyceps sinensis promotes wound tissue to secrete growth factor some researches show that it has, and is that wound climate reaches balance, So that wound is quickly spent the inflammation phase, reaches wound healing into the hyperplasia reformation phase;And has to Lung Tumor and inhibit Effect etc..For nunchakus spore cordyceps sinensis not yet by artificial cultivation, the present invention is to complete Planting to nunchakus spore cordyceps sinensis for the first time at present Training.Its value could be utilized preferably after only reaching artificial cultivation, by for more people, play its maximum value.
Invention content
To solve the above problems, the purpose of the present invention is to provide one plant of nunchakus spore Chinese caterpillar fungus strain and its domesticating and cultivating sides Method realizes artificial cultivation nunchakus spore cordyceps sinensis.
In order to achieve the above objectives, the present invention is achieved through the following technical solutions:
A kind of nunchakus spore Chinese caterpillar fungus strain (Cordyceps ninchukispora (C.H.Su&H.H.Wang) G.H.Sung, J.M. Sung, Hywel-Jones&Spatafora), which has been stored in China Committee for Culture Collection of Microorganisms Common micro-organisms center (CGMCC), preservation address:Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, preservation date:2017 12 The moon 27, deposit number:CGMCC No. 15089.
A kind of preparation method of the liquid spawn of nunchakus spore cordyceps sinensis, includes the following steps:
Step 1,300 g of peeled potatoes cut fritter, add 500ml boilings rotten;
Step 2, filtering potato, take filtrate, and glucose 30g, peptone 2g, potassium dihydrogen phosphate 0.5g, magnesium sulfate 1g, dimension life is added Plain B1-3 pieces;
Step 3, Jia Shui are settled to 1000 ml;
Step 4, stirring and dissolving, in packing to triangular flask;
Step 5, liquid triangular flask are as 121 DEG C of sterilizing 20min;
It is taken out after step 6, bacterium of having gone out and is put into super bacterium workbench cooling, addition nunchakus spore cordyceps species inoculation after cooling;
Triangular flask is positioned over 25 ~ 28 DEG C of shaking table cultures by step 7 after connecting kind, and fermentation can be used after 4 ~ 6 days.
A kind of nunchakus spore cordyceps sinensis domesticating cultivation method, using the liquid spawn of above-mentioned nunchakus spore cordyceps sinensis, including it is following Step:
Step 1, culture material are mixed by any one to two kinds in polished rice, wheat, three kinds of niblet;
Step 2, using 240ml tissue culture bottles as cultivating container, every bottle of culture material is that 16g feeds;
Step 3 is mixed with nutrient solution by the ratio of following mass percent:Magnesium sulfate 0.2%, potassium dihydrogen phosphate 0.3%, dimension life Plain B1-2 pieces/L;
Step 4, every bottle culture material and nutrient solution are added according to quantity and is made cultivation matrix, the ratio of culture material and nutrient solution is 1:1.3~ 1.5;
Step 5, sterilizing, using 121 DEG C of high pressure steam sterilization 40-60min of temperature;
Cooling is taken out after the completion of step 6, sterilizing;
Step 7, cooling, inoculation, carry out liquid inoculation when by cultivation matrix cooling down to 25 DEG C or so, use nunchakus spore worm The liquid spawn of grass, inoculum concentration are the 5%-8% that strain accounts for compost quality;
Step 8, cultural hypha, the light culture mycelia 10 days to 15 days under 20 DEG C -30 DEG C, 30%-50% air humidity conditions;
Step 9, Cui Lei, the temperature difference, light culture 2-5 days under the conditions of 15 DEG C -25 DEG C, air humidity 50%-60%;
Step 10 goes out careless culture, 18 DEG C -25 DEG C, air humidity 50%-60%, natural light can be irradiated under the conditions of cultivate 25-40 It;
Step 11, nunchakus spore cordyceps militaris sporocarp head turn yellow, coarse rear grass is ripe, picking.
The present invention has the following advantages:
(1)It can cultivate to obtain nunchakus spore cordyceps militaris sporocarp by artificial condition.
(2)Domesticating cultivation method is simple, easily grasps.
(3)Less investment, at low cost, matrix preparation manipulation is simple and practicable.
(4)Culture material is essentially coarse food grain, can meet needed for extensive cultivation.
(5)Yield is high, and the period is short.
Specific implementation mode
The present invention is in order to which nunchakus spore cordyceps sinensis is reached artificial domesticating cultivation, recently as people to the continuous of cordyceps sinensis Research, finds its higher medical value, but wild resource much can not meet people's needs, so cordyceps sinensis is artificial Domesticating and cultivating is inevitable.The present invention is by nunchakus spore cordyceps sinensis success domesticating and cultivating, and the advantages that method is simple, raw material is sufficient, Further to study used and playing its bigger value.It is multigroup it is demonstrated experimentally that nunchakus spore Chinese caterpillar fungus strain of the invention can lead to It crosses cultural method in the present invention and completes domestication.Different ratios of raw materials domesticating and cultivating nunchakus spore cordyceps sinensis effect is shown in Table 1.
1. different ratios of raw materials domesticating and cultivating nunchakus spore cordyceps sinensis Cordyceps ninchukispora effects of table
Raw material 1 Raw material 2 Grass output amount Grass is long
Proportioning 1 Polished rice 50% Niblet 50% Largely, average 150 or so/bottle It is 7 centimetres average
Proportioning 2 Niblet 50% Wheat 50% Largely, average 180 or so/bottle It is 8 centimetres average
Proportioning 3 Polished rice 50% Wheat 50% Largely, average 100 or so/bottle It is 5 centimetres average
Proportioning 4 Wheat 100% 0% It is medium, average 80 or so/bottle It is 5 centimetres average
Proportioning 5 Polished rice 100% 0% Largely, average 150 or so/bottle It is 5 centimetres average
Case study on implementation 1:Using proportioning be polished rice 50%, niblet 50% as culture material, domesticating and cultivating nunchakus spore cordyceps sinensis include with Lower step:
(1)Use formula:Potato 300g, glucose 30g, peptone 2g, potassium dihydrogen phosphate 0.5g, magnesium sulfate 1g, vitamin B1- 3.By 28 DEG C of shaking tables, 4 days liquid fermentations, liquid spawn is made.
(2)It is matched using polished rice 8g, niblet 8g, 240ml tissue culture bottles are container, per bottled material 16g.
(3)Nutrient solution is mixed with by the ratio of following mass percent:Magnesium sulfate 0.2%, potassium dihydrogen phosphate 0.3%, dimension Raw element B1-2 pieces/L.
(4)Every bottle is added nutrient solution 24ml.
(5)The tissue culture bottle sterilizing of culture material will be installed, using 121 DEG C of high pressure steam sterilization 60min of temperature.
(6)Cooling is taken out after the completion of sterilizing.
(7)It is the 5%-8% that strain accounts for compost quality that liquid inoculation, inoculum concentration are used when being cooled to 25 DEG C or so.
(9)Cultural hypha, is positioned over 24 DEG C of light culture casees, culture mycelia 7 days, and surface is yellow to golden yellow.
(10)Cui Lei, 22 DEG C of light culture case are cultivated 6 hours, and 25 DEG C are cultivated 18 hours, and it is white that Cui Lei a large amount of rice-shapeds occurs in 4 days The small protrusion of color.
(11)Go out careless culture, culture bottle is converted to 25 DEG C to be irradiated under the conditions of natural light and is cultivated 28 days.
(12)Head turns yellow, coarse rear grass is ripe, and picking is average careless 7 centimetres or so long, average every bottle 150 grass left and right.
Case study on implementation 2:It is niblet 50%, wheat 50% as culture material, domesticating and cultivating nunchakus spore cordyceps sinensis packet using proportioning Include following steps:
(1)Use formula:Potato 300g, glucose 30g, peptone 2g, potassium dihydrogen phosphate 0.5g, magnesium sulfate 1g, vitamin B1- 3.By 28 DEG C of shaking tables, 4 days liquid fermentations, liquid spawn is made.
(2)It is matched using niblet 8g, wheat 8g, 240ml tissue culture bottles are container, per bottled material 16g.
(3)Nutrient solution is mixed with by the ratio of following mass percent:Magnesium sulfate 0.2%, potassium dihydrogen phosphate 0.3%, dimension Raw element B1-2 pieces/L.
(4)Every bottle is added nutrient solution 24ml.
(5)The tissue culture bottle sterilizing of culture material will be installed, using 121 DEG C of high pressure steam sterilization 60min of temperature.
(6)Cooling is taken out after the completion of sterilizing.
(7)It is the 5%-8% that strain accounts for compost quality that liquid inoculation, inoculum concentration are used when being cooled to 25 DEG C or so.
(9)Cultural hypha, is positioned over 24 DEG C of light culture casees, culture mycelia 7 days, and surface is yellow to golden yellow.
(10)Cui Lei, 22 DEG C of light culture case are cultivated 6 hours, and 25 DEG C are cultivated 18 hours, and it is white that Cui Lei a large amount of rice-shapeds occurs in 4 days The small protrusion of color.
(11)Go out careless culture, culture bottle is converted to 25 DEG C to be irradiated under the conditions of natural light and is cultivated 28 days.
(12)Head turns yellow, coarse rear grass is ripe, and picking is average careless 8 centimetres or so long, average every bottle 180 grass left and right.
Case study on implementation 3:To match as polished rice 50%, wheat 50% as culture material, domesticating and cultivating nunchakus spore cordyceps sinensis includes Following steps:
(1)Use formula:Potato 300g, glucose 30g, peptone 2g, potassium dihydrogen phosphate 0.5g, magnesium sulfate 1g, vitamin B1- 3.By 28 DEG C of shaking tables, 4 days liquid fermentations, liquid spawn is made.
(2)It is matched using polished rice 8g, wheat 8g, 240ml tissue culture bottles are container, per bottled material 16g.
(3)Nutrient solution is mixed with by the ratio of following mass percent:Magnesium sulfate 0.2%, potassium dihydrogen phosphate 0.3%, dimension Raw element B1-2 pieces/L.
(4)Every bottle is added nutrient solution 24ml.
(5)The tissue culture bottle sterilizing of culture material will be installed, using 121 DEG C of high pressure steam sterilization 60min of temperature.
(6)Cooling is taken out after the completion of sterilizing.
(7)It is the 5%-8% that strain accounts for compost quality that liquid inoculation, inoculum concentration are used when being cooled to 25 DEG C or so.
(9)Cultural hypha, is positioned over 24 DEG C of light culture casees, culture mycelia 7 days, and surface is yellow to golden yellow.
(10)Cui Lei, 22 DEG C of light culture case are cultivated 6 hours, and 25 DEG C are cultivated 18 hours, and it is white that Cui Lei a large amount of rice-shapeds occurs in 4 days The small protrusion of color.
(11)Go out careless culture, culture bottle is converted to 25 DEG C to be irradiated under the conditions of natural light and is cultivated 28 days.
(12)Head turns yellow, coarse rear grass is ripe, and picking is average careless 5 centimetres or so long, average every bottle 100 grass left and right.
Case study on implementation 4:To match as wheat 100% as culture material, domesticating and cultivating nunchakus spore cordyceps sinensis includes following step Suddenly:
(1)Use formula:Potato 300g, glucose 30g, peptone 2g, potassium dihydrogen phosphate 0.5g, magnesium sulfate 1g, vitamin B1- 3.By 28 DEG C of shaking tables, 4 days liquid fermentations, liquid spawn is made.
(2)Using wheat 16g, 240ml tissue culture bottles are container, per bottled material 16g.
(3)Nutrient solution is mixed with by the ratio of following mass percent:Magnesium sulfate 0.2%, potassium dihydrogen phosphate 0.3%, dimension Raw element B1-2 pieces/L.
(4)Every bottle is added nutrient solution 24ml.
(5)The tissue culture bottle sterilizing of culture material will be installed, using 121 DEG C of high pressure steam sterilization 60min of temperature.
(6)Cooling is taken out after the completion of sterilizing.
(7)It is the 5%-8% that strain accounts for compost quality that liquid inoculation, inoculum concentration are used when being cooled to 25 DEG C or so.
(9)Cultural hypha, is positioned over 24 DEG C of light culture casees, culture mycelia 7 days, and surface is yellow to golden yellow.
(10)Cui Lei, 22 DEG C of light culture case are cultivated 6 hours, and 25 DEG C are cultivated 18 hours, and it is white that Cui Lei a large amount of rice-shapeds occurs in 4 days The small protrusion of color.
(11)Go out careless culture, culture bottle is converted to 25 DEG C to be irradiated under the conditions of natural light and is cultivated 28 days.
(12)Head turns yellow, coarse rear grass is ripe, and picking is average careless 5 centimetres or so long, average every bottle 80 grass left and right.
Case study on implementation 5:To match as polished rice 100% as culture material, domesticating and cultivating nunchakus spore cordyceps sinensis includes the following steps:
(1)Use formula:Potato 300g, glucose 30g, peptone 2g, potassium dihydrogen phosphate 0.5g, magnesium sulfate 1g, vitamin B1- 3.By 28 DEG C of shaking tables, 4 days liquid fermentations, liquid spawn is made.
(2)It is matched using polished rice 16g, 240ml tissue culture bottles are container, per bottled material 16g.
(3)Nutrient solution is mixed with by the ratio of following mass percent:Magnesium sulfate 0.2%, potassium dihydrogen phosphate 0.3%, dimension Raw element B1-2 pieces/L.
(4)Every bottle is added nutrient solution 24ml.
(5)The tissue culture bottle sterilizing of culture material will be installed, using 121 DEG C of high pressure steam sterilization 60min of temperature.
(6)Cooling is taken out after the completion of sterilizing.
(7)It is the 5%-8% that strain accounts for compost quality that liquid inoculation, inoculum concentration are used when being cooled to 25 DEG C or so.
(9)Cultural hypha, is positioned over 24 DEG C of light culture casees, culture mycelia 7 days, and surface is yellow to golden yellow.
(10)Cui Lei, 22 DEG C of light culture case are cultivated 6 hours, and 25 DEG C are cultivated 18 hours, and it is white that Cui Lei a large amount of rice-shapeds occurs in 4 days The small protrusion of color.
(11)Go out careless culture, culture bottle is converted to 25 DEG C to be irradiated under the conditions of natural light and is cultivated 28 days.
(12)Head turns yellow, coarse rear grass is ripe, and picking is average careless 5 centimetres or so long, average every bottle 150 grass left and right.

Claims (3)

1. a kind of nunchakus spore Chinese caterpillar fungus strain(Cordyceps ninchukispora (C.H. Su & H.H. Wang) G.H. Sung, J.M. Sung, Hywel-Jones & Spatafora), which has been stored in Chinese microorganism strain preservation Administration committee's common micro-organisms center (CGMCC), deposit number:CGMCC No. 15089.
2. a kind of preparation method of the liquid spawn of nunchakus spore Chinese caterpillar fungus strain according to claim 1, including following step Suddenly:
Step 1,300 g of peeled potatoes cut fritter, add 500 ml boilings rotten;
Step 2, filtering potato, take filtrate, 30 g of glucose, 2 g of peptone, 0.5 g of potassium dihydrogen phosphate, 1 g of magnesium sulfate are added, Vitamin B 1-3 pieces;
Step 3, Jia Shui are settled to 1000 ml;
Step 4, stirring and dissolving, in packing to triangular flask;
Step 5, liquid triangular flask are as 121 DEG C of 20 min of sterilizing;
It is taken out after step 6, bacterium of having gone out and is put into superclean bench cooling, inoculation nunchakus spore Cordyceps Militaris after being cooled to 30 DEG C or less About 1 g of filament;
Triangular flask is positioned over 25 ~ 28 DEG C of shaking table cultures, 160r/min by step 7 after connecting kind, and fermentation is fabricated to after 4 ~ 6 days Liquid spawn.
3. a kind of nunchakus spore cordyceps sinensis domesticating cultivation method uses the liquid spawn of nunchakus spore cordyceps sinensis described in claim 2, packet Include the following steps:
Step 1, culture material are mixed by any one or two kinds in polished rice, wheat, three kinds of niblet;
Step 2, using 240 ml tissue culture bottles as cultivating container, every bottle of culture material is that 16 g feed;
Step 3 is mixed with nutrient solution by the ratio of following mass percent:Magnesium sulfate 0.2%, potassium dihydrogen phosphate 0.3%, dimension life Plain B 1-2 pieces/L;
Step 4, every bottle culture material and nutrient solution are added according to quantity and is made cultivation matrix, the ratio of culture material and nutrient solution is 1:1.3~ 1.5;
Step 5, sterilizing, using 121 DEG C of temperature high pressure steam sterilization 40-60 minutes;
Cooling is taken out after the completion of step 6, sterilizing;
Step 7, cooling, inoculation, carry out liquid inoculation when by cultivation matrix cooling down to 25 DEG C or so, use nunchakus spore worm The liquid spawn of grass, inoculum concentration are the 5-8% that strain accounts for compost quality;
Step 8, cultural hypha, are protected from light culture mycelia 10 to 15 days under the conditions of 20-30 DEG C, air humidity 30-50%;
Step 9, Cui Lei, in 10 DEG C of day and night temperature, 15 DEG C -25 DEG C, light culture 2-5 days under the conditions of air humidity 50%-60%;
Step 10 goes out careless culture, is cultivated 25-40 days under the conditions of 18-25 DEG C, air humidity 50-60%, natural light;
Step 11, nunchakus spore cordyceps militaris sporocarp head turn yellow, are coarse rear i.e. ripe, picking.
CN201810176973.9A 2018-03-04 2018-03-04 A kind of nunchakus spore Chinese caterpillar fungus strain and its cultural method Pending CN108476861A (en)

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Citations (5)

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CN102630481A (en) * 2012-04-11 2012-08-15 广东省微生物研究所 Cultivation method for oospore oudemansiella mucida
CN102771314A (en) * 2012-08-15 2012-11-14 苏州硒谷科技有限公司 Method for culturing selenium-enriched cordyceps militaris
CN103843583A (en) * 2014-03-06 2014-06-11 刘彦君 Industrialized production method of green cordyceps militaris
CN104609943A (en) * 2014-12-28 2015-05-13 江西农业大学 Novel medium for cultivating pleurotus ostreatus and method for cultivating pleurotus ostreatus by using novel medium
CN106754427A (en) * 2017-01-20 2017-05-31 山西省农业科学院食用菌研究所 Thermophilic blue spore pore fungi new strains and its acclimation method

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中国工程院环境保护部编: "《中国环境宏观战略研究-环境要素保护战略卷(下)》", 31 January 2011, 中国环境科学出版社 *
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