CN108401794A - A kind of armillaria mellea accreting with Rhizoma Gastrodiae liquid spawn production method and cultigen special culture media - Google Patents
A kind of armillaria mellea accreting with Rhizoma Gastrodiae liquid spawn production method and cultigen special culture media Download PDFInfo
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- CN108401794A CN108401794A CN201810144260.4A CN201810144260A CN108401794A CN 108401794 A CN108401794 A CN 108401794A CN 201810144260 A CN201810144260 A CN 201810144260A CN 108401794 A CN108401794 A CN 108401794A
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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
- 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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Abstract
The present invention for halimasch Armillaria gallica (A.gallica) and Armillaria ostoyae (A.ostoyae) provide dedicated armillaria mellea accreting with Rhizoma Gastrodiae liquid strain cultivation kind special combination culture medium, including first group of fluid nutrient medium, second group of fluid nutrient medium and solid medium composition.The present invention utilizes halimasch liquid bacterial strains, is inoculated in dedicated solid medium, ensure that higher yield rate, shorten cultivation cycle, improve strain quality.
Description
Technical field
The present invention relates to a kind of associations to give birth to strain production method, more specifically to a kind of armillaria mellea accreting with Rhizoma Gastrodiae liquid spawn
Production method and cultigen special culture media.
Technical background
It is a kind of summer and autumn that halimasch (Armillaria spp.), which belongs to Basidiomycotina Agaricales Tricholomataceae Armillaria,
Generation energy facultative parasitism is to be found for the first time by Vahl for 1790 in a variety of woody herbal famous dietotherapeutic bacterium halimasch
And identify so far in the world altogether find 36 biological species, wherein Europe be distributed with 5 kinds, North America be distributed 7 kinds, Australia it is big
5 kinds of Leah distribution, 9 kinds of Japan's distribution, 2 kinds of Africa distribution, China are distributed with 9 kinds, mainly divide in China halimasch
Cloth is in the provinces and regions such as Yunnan Inner Mongol Tibet, Hebei Shanxi, Zhejiang Fujian, Heilungkiang Jilin and Taiwan.Suitable for Rhizoma Gastrodiae bacterium halimasch kind
There are 6 kinds or so.
Rhizoma Gastrodiae is rare medicinal herbs, and demand increases year by year, by being completed with the mode planted with halimasch culture bacterium material in production,
The quality of halimasch culture determines the yield and quality of Rhizoma Gastrodiae.Armillaria mellea culture denominator kind, original seed and cultigen, are more preferable
Ground utilize local resource, cost-effective, halimasch cultigen branch makees raw material, block mixed with other composts, training of bottling
Bacterium branch cultigen is raised, mode has two kinds of liquid and solid, claims halimasch liquid bacterium branch cultigen and solid bacterium branch cultigen.
Between halimasch and Rhizoma Gastrodiae there is association relationship studies have shown that Rhizoma Gastrodiae in growth course with the relationship of halimasch
Main experience refuses bacterium, control bacterium, open three phases, and " bacterium eats fiber crops " and " fiber crops eat bacterium " two kinds of phenomenons are presented.Halimasch is facultative posts
The important concomitance bacterium of raw fungi and gastrodin cultivation, not only can complete independently growth and development, colleague also infect Rhizoma Gastrodiae, be Rhizoma Gastrodiae
Growth, development, breeding provide abundant nutrition, realize the artificial cultivation of Rhizoma Gastrodiae.
In the liquid spawn for the fermentation tank culture that the end of the nineties in last century, Japan are studied with South Korea, it is used for needle mushroom
Factory culture.The country, Shen Min in 2010 researches and develops the liquid spawn of Pleurotus eryngii, and is successfully applied to batch production, and realizes
High yield, high-quality, efficient target.2013, Li Zengzhi et al. successfully realized that the batch production of Cordyceps militaris is planted using liquid spawn
Training.Prepared by edible mushroom traditional bacterial classification, the seed bottle bottleneck used is small, and charging is difficult, and gas permeability is looked into, and sealing tampon is easy pollution of making moist
The cultivation cycle of miscellaneous bacteria, strain is long, and strain quality is irregular, and cost is higher.Patent CN106010975 discloses liquid production honey
Ring bacterium strain method, hence it is evident that shorten cultivation cycle, improve yield rate.
Invention content
The significant difference of influence of the different halimasch to Rhizoma Gastrodiae biological yield and gastrodin content, China high altitude localities
The significant difference of influence of the different type halimasch to Rhizoma Gastrodiae biological yield and gastrodin content.Halimasch different strains are to day
Numb yield tool has a certain impact, and the present invention is directed to halimasch Armillaria gallica (A.gallica) and Armillaria ostoyae
(A.ostoyae) dedicated armillaria mellea accreting with Rhizoma Gastrodiae liquid strain cultivation kind special combination culture medium, including first group of liquid are provided
Culture medium, second group of fluid nutrient medium and solid medium composition：
First group of fluid nutrient medium formulation weight is than composition：0.5-3 parts of glucose, 5-30 parts of potato liquid, potassium dihydrogen phosphate
0-0.5 parts, 0.05 part of magnesium sulfate, vitamin B1 additive amount be 0-1mg/100ml；
Second group of fluid nutrient medium formulation weight is than composition：0-3 parts of glucose, 0-5 parts of cornstarch, beancake powder 0.5-5
Part, 0.5-5 parts of groundnut meal, 0-0.5 parts of potassium dihydrogen phosphate, 0.5 part of magnesium sulfate, vitamin B1 additive amount are 0-1mg/100ml；
Solid medium formulation weight is than composition：10-50 parts of the broad-leaf forest branch section of length 5-10cm, broad-leaf forest trees bits
20-50 parts, 0-20 parts of corn flour, 5-20 parts of wheat bran, 0-10 parts of beancake powder, 0.5-2 parts of sucrose, 0.5-2 parts of land plaster.Water content
60-70 parts.
A kind of armillaria mellea accreting with Rhizoma Gastrodiae liquid spawn production method, for Armillaria gallica or the Austria with Rhizoma Gastrodiae with association relationship
For family name halimasch as halimasch kind, specific production method is as follows：
1) Armillaria mellea triangular flask Liquid Culture is used into 200ml/500ml triangular flasks, it is 5-7 to be cultivated on shaking table
It, 200rpm, 22-28 DEG C of temperature is seeded to liquid spawn is homogeneous before fermentation tank, and liquid bacteria is obtained through culture；
2) aforesaid liquid strain is inoculated into the fermentation tank for being equipped with fluid nutrient medium, incubation time is 5-7 days, culture temperature
22-28 DEG C of degree, culture pressure are 0.02-0.04MP, and rotating speed 200rpm, ventilatory capacity 0.5vvm-0.8vvm are obtained by culture sweet ring
Bacterium strain；
3) by above-mentioned Armillaria mellea it is homogeneous after be inoculated into special culture solid medium, through culture obtain halimasch solid
Cultigen.
The method have the benefit that：Using halimasch liquid bacterial strains, it is inoculated in dedicated solid medium, is ensured
Higher yield rate, shortens cultivation cycle, improves strain quality.
Specific implementation mode
The preparation of 1 armillaria mellea accreting with Rhizoma Gastrodiae liquid strain cultivation kind special combination culture medium of embodiment
1, raw material is weighed according to weight ratio formula：2 parts of glucose, 20 parts of potato liquid, 0.3 part of potassium dihydrogen phosphate, magnesium sulfate
0.15 part, addition vitamin B1 to 1mg/100ml, obtain first group of fluid nutrient medium.
2, raw material is weighed according to weight ratio formula：2 parts of glucose, 3 parts of cornstarch, 1 part of beancake powder, groundnut meal 0.5
Part, 0.3 part of potassium dihydrogen phosphate, 0.15 part of magnesium sulfate, addition vitamin B1 obtain second group of fluid nutrient medium to 1mg/100ml.
3, raw material is weighed according to weight ratio formula：40 parts of broad-leaf forest branch section, 45 parts of broad-leaf forest trees bits, 6 parts of corn flour,
6 parts of wheat bran, 1 part of beancake powder, 1 part of sucrose, 1 part of land plaster.Add and adds water to water content 65%.
4, Armillaria mellea triangular flask Liquid Culture is used into 200ml/500ml triangular flasks, adds first group of liquid training
Support base, culture is 5-7 days, 200rpm on shaking table, 22-28 DEG C of temperature, be seeded to it is liquid spawn is homogeneous before fermentation tank, through cultivating
Obtain liquid bacteria；
5, aforesaid liquid strain is inoculated into the fermentation tank for being equipped with fluid nutrient medium, adds second group of fluid nutrient medium,
Incubation time is 5-7 days, 22-28 DEG C of cultivation temperature, and culture pressure is 0.02-0.04MP, rotating speed 150-200rpm, ventilatory capacity
0.5vvm-0.8vvm is obtained by culture Armillaria mellea；
6, by above-mentioned Armillaria mellea it is homogeneous after be inoculated into solid medium, access solid after fluid nutrient medium strain homogeneous
In culture medium, inoculum concentration 20ml cultivates, keeps flat and be protected from light culture 33 days, cover with bottle, obtains halimasch solid state cultivation for 25 days
Kind.
Special solid culture medium described in above-mentioned 6th step is placed in wide mouth glass bottle, and every bottle of branch section is evenly distributed on
At radius 2/3, length 5-10cm is around filled up with special culture media, and punching is inserted into centre with the branch of diameter 5mm, and surrounding is used
Special culture media fills up, and is sealed using breathable sealing film.Compared with traditional carboy, charging is suitable, with tampon
It compares, breathable sealing film of the present invention increases the gas permeability of cultigen culture, substantially reduces and reduces because tampon makes moist and brings
Pollution.
Include before culture：121 sterilizings 2 hours, natural cooling uses preceding ozonization.It is small using high-temperature sterilization 2
When, the microorganism in solid ingredient can be effectively killed, pollution rate is effectively reduced.Ozonization can penetrate the gap between bottle, have
Effect sterilization, compared to the ultraviolet various microorganisms that can only kill surface, germicidal efficiency improves, in addition, ozone is safe environment protection type, it can
It is decomposed into oxygen.Ozonization 4h.
Using the present invention provide special combination culture medium cultivate making and traditional seeding technique relatively compared with, specific such as table
Shown in 1.
The production of hybrid seeds effect expedition of 1 special combination culture medium of table
Embodiment 2 cultivates comparative test result
Using for Armillaria gallica (A.gallica), Armillaria ostoyae (A.ostoyae), mustard Huang halimasch
(A.sinapina), yellow lid halimasch (A.luteopileata) is trained using the liquid of special combination culture medium provided by the invention
It supports the production of hybrid seeds and cultivates comparative test result (50m with traditional seeding technique2Area), it is specific as shown in table 2.
Influence of 2 different culture media of table to different Rhizoma Gastrodiae growths
Remarks：Wei meter Ma under 10g, 10g or more 50g or less are hemp, and 50g or more is commodity fiber crops.
The test data of table 2 proves, using Liquid Culture producing method for seed provided by the invention and special culture media for day
The yield and quality of fiber crops all have significant raising, while fluid present invention culture producing method for seed and special culture media are directed to Gaul
Halimasch and Armillaria ostoyae have significant especially effect with Rhizoma Gastrodiae association plantation, this shows different bacterial strains to breeding strain
Culture nutriment demand it is different, this needs targetedly to research and develop dedicated culture medium prescription.
Claims (2)
1. armillaria mellea accreting with Rhizoma Gastrodiae liquid strain cultivation kind special combination culture medium, it is characterised in that：Including first group of fluid nutrient medium,
Second group of fluid nutrient medium and solid medium composition：
First group of fluid nutrient medium formulation weight is than composition：0.5-3 parts of glucose, 5-30 parts of potato liquid, potassium dihydrogen phosphate 0-0.5
Part, 0.05 part of magnesium sulfate, vitamin B1 additive amount are 0.1-1mg/100ml；
Second group of fluid nutrient medium formulation weight is than composition：0-3 parts of glucose, 0-5 parts of cornstarch, 0.5-5 parts of beancake powder, flower
0.5-5 parts of raw cake powder, 0-0.5 parts of potassium dihydrogen phosphate, 0.1-0.5 parts of magnesium sulfate, vitamin B1 additive amount are 0.1-1mg/
100ml；
Solid medium formulation weight is than composition：10-50 parts of the broad-leaf forest branch section of length 5-10cm, broad-leaf forest trees consider 20- to be worth doing
50 parts, 0-20 parts of corn flour, 5-20 parts of wheat bran, 0-10 parts of beancake powder, 0.5-2 parts of sucrose, 0.5-2 parts of land plaster.Water content 60-
70 parts.
2. a kind of armillaria mellea accreting with Rhizoma Gastrodiae liquid spawn production method, it is characterised in that：For the Gaul with Rhizoma Gastrodiae with association relationship
As halimasch kind, specific production method is as follows for halimasch or Armillaria ostoyae：
1) Armillaria mellea triangular flask Liquid Culture is used into 200ml/500ml triangular flasks, addition described in claim 1 the
One group of fluid nutrient medium, culture is 5-7 days, 150-200rpm on shaking table, 22-28 DEG C of temperature, is seeded to liquid before fermentation tank
Strain is homogeneous, and liquid bacteria is obtained through culture；
2) aforesaid liquid strain is inoculated into the fermentation tank for being equipped with fluid nutrient medium, second group described in claim 1 of addition
Fluid nutrient medium, incubation time are 5-7 days, and 22-28 DEG C of cultivation temperature, culture pressure is 0.02-0.04MP, rotating speed 150-
200rmp, ventilatory capacity 0.5vvm-0.8vvm, is obtained by culture Armillaria mellea；
3) by above-mentioned Armillaria mellea it is homogeneous after be inoculated into solid medium described in claim 1, through culture obtain halimasch
Solid state cultivation kind.
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Cited By (8)
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| CN109207380A (en) * | 2018-10-22 | 2019-01-15 | 长春圣金诺生物制药有限公司 | A kind of deep liquid fermentation process of high yield halimasch |
| CN109370916A (en) * | 2018-12-04 | 2019-02-22 | 昆明理工大学 | A kind of process for rapid activation of halimasch aging strain |
| CN110184376A (en) * | 2019-05-20 | 2019-08-30 | 中国中医科学院中药研究所 | The PCR-RFLP discrimination method of Armillaria gallica |
| CN111019835A (en) * | 2019-11-01 | 2020-04-17 | 汉中市汉台区汉麓生物科技有限公司 | Method for separating Armillaria mellea from ambary endophytic fungi |
| CN113403202A (en) * | 2021-05-28 | 2021-09-17 | 重庆市中药研究院 | Method for rapidly breeding gastrodia elata armillaria mellea |
| CN113475316A (en) * | 2021-06-04 | 2021-10-08 | 李俊 | Preparation method of stable-yield high-yield liquid armillaria mellea strain |
| CN115336503A (en) * | 2022-08-30 | 2022-11-15 | 西南林业大学 | Preparation method of substitute bacterial material for gastrodia elata cultivation |
| CN115399190A (en) * | 2022-03-31 | 2022-11-29 | 吉林农业大学 | Armillaria mellea strain culture medium and preparation method thereof |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109207380A (en) * | 2018-10-22 | 2019-01-15 | 长春圣金诺生物制药有限公司 | A kind of deep liquid fermentation process of high yield halimasch |
| CN109370916A (en) * | 2018-12-04 | 2019-02-22 | 昆明理工大学 | A kind of process for rapid activation of halimasch aging strain |
| CN110184376A (en) * | 2019-05-20 | 2019-08-30 | 中国中医科学院中药研究所 | The PCR-RFLP discrimination method of Armillaria gallica |
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| CN111019835A (en) * | 2019-11-01 | 2020-04-17 | 汉中市汉台区汉麓生物科技有限公司 | Method for separating Armillaria mellea from ambary endophytic fungi |
| CN113403202A (en) * | 2021-05-28 | 2021-09-17 | 重庆市中药研究院 | Method for rapidly breeding gastrodia elata armillaria mellea |
| CN113403202B (en) * | 2021-05-28 | 2023-04-07 | 重庆市中药研究院 | Method for rapidly breeding gastrodia elata armillaria mellea |
| CN113475316A (en) * | 2021-06-04 | 2021-10-08 | 李俊 | Preparation method of stable-yield high-yield liquid armillaria mellea strain |
| CN115399190A (en) * | 2022-03-31 | 2022-11-29 | 吉林农业大学 | Armillaria mellea strain culture medium and preparation method thereof |
| CN115336503A (en) * | 2022-08-30 | 2022-11-15 | 西南林业大学 | Preparation method of substitute bacterial material for gastrodia elata cultivation |
| CN115336503B (en) * | 2022-08-30 | 2024-01-16 | 西南林业大学 | Preparation method of substitute bacteria for gastrodia elata cultivation |
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Application publication date: 20180817 |