CN109220559A - A kind of production method and growth condition of Stropharia rugoso-annulata reduction strain - Google Patents
A kind of production method and growth condition of Stropharia rugoso-annulata reduction strain Download PDFInfo
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- CN109220559A CN109220559A CN201811160855.5A CN201811160855A CN109220559A CN 109220559 A CN109220559 A CN 109220559A CN 201811160855 A CN201811160855 A CN 201811160855A CN 109220559 A CN109220559 A CN 109220559A
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- annulata
- stropharia rugoso
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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
-
- 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
Abstract
A kind of production method and growth condition of Stropharia rugoso-annulata reduction strain, this method breaks the mode of conventional solid strain, it is made of liquid spawn, strain and culture substrate separation, pure culture transport, the new model of destination liquid feeding body reduction strain, cooperate the liquid formulations of our reduction strain simultaneously: yeast powder 0.01-0.05% being added in the distilled water of 1L, K2HPO4 0.03-0.08%, KH2PO4 0.03-0.08%, MgSO4 0.01-0.05%, Theravite 0.1%(Theravite be 0.1% thiamine hydrochloride and 0.01% riboflavin mixing after plus water match and be made), it can carry out crop field or hayashishita sprinkling inoculation.One side liquid cultivation seed seeding process is simple, low manufacture cost and bacterium germination are fast, improves the efficiency of cultivation Stropharia rugoso-annulata significantly；On the other hand pure culture is small in size after separation, and convenient transportation, strain is separated with culture medium, and calorific value is few, effectively prevents the generation that bacterium is burnt in fever in strain transportational process.
Description
Technical field
The present invention relates to the production methods and growth condition of a kind of Stropharia rugoso-annulata reduction strain, belong to edible mashroom cultivating, plant
Train technical field.
Background technique
Stropharia rugoso-annulata (Stropharia rugoso-annulata), belong to Agaricales, Strophariaceae, Stropharia is connection
Ten big kinds on the new mushroom species that He Guo food and agricultural organization (FAO) is recommended to developing country, and international mushroom class trade market it
One.Its fructification amino acid and vitamin needed by human containing there are many, and there are the diseases such as prevention coronary heart disease, antitumor
Medical value is welcomed by consumers.At present in production, inoculated and cultured usually is carried out using solid state cultivation kind, is needed
Solid state cultivation kind amount is big and cell age difference is big, sows low efficiency.More distinct issues are, stropharia rugosoannulata strain vigor compared with
Vigorous, for solid spawn since mycelium and solid matrix mix, mycelium can generate a large amount of heat, cause burning bacterium existing
The generation of elephant, it is difficult to meet the needs of stropharia rugosoannulata strain quality and long-distance transport.
Summary of the invention
It is an object of that present invention to provide the production methods and growth condition of a kind of Stropharia rugoso-annulata reduction strain.
To achieve the above object, present invention provide the technical scheme that a kind of Stropharia rugoso-annulata reduction strain production method,
Include the following steps:
It is activated on the plate of PDA culture medium step 1: Stropharia rugoso-annulata parent species are transferred to one and are accommodated, keeps Stropharia rugoso-annulata female
The mycelia vitality restoration of kind, then segmentation obtains Stropharia rugoso-annulata original seed PDA fungus block；
Step 2: the Stropharia rugoso-annulata original seed PDA fungus block is inoculated into the PDB fluid nutrient medium after sterilizing, in 22-26 DEG C of condition
Under, it is cultivated 7-8 days with shaking table culture case in dark place, obtains Stropharia rugoso-annulata level-one kind；
Step 3: the Stropharia rugoso-annulata level-one kind to be inoculated into the fermentor for accommodating Stropharia rugoso-annulata liquid cultivation seed culture medium
In, then by the fermentor after inoculation with 22-26 DEG C culture 7-8 days；The preparation of the Stropharia rugoso-annulata liquid cultivation seed culture medium
Method are as follows: be added the sucrose of its quality 1.0-2.0% to 1 liter of distilled water, the ferment of the albumen powder of 0.10-0.50%, 0.20-0.30%
Female powder, the bean cake powder of 0.6-0.7%, the K of 0.05-0.10%2HPO4, the KH of 0.05-0.20%2PO4With the MgSO of 0.05-0.10%4,
It is made after adjusting pH value to 6.3-6.8, then sterilized, cooling step after mixing；
Step 4: being concentrated the Stropharia rugoso-annulata liquid spawn that third step culture obtains to get big ball using vacuum filtration machine
The pure cultivation strain of lid mushroom；
Step 5: the sterile valve bag of Stropharia rugoso-annulata pure culture is dispensed, preservation and transport are then carried out under the conditions of 4-8 DEG C；
Step 6: transporting to after destination, by the Stropharia rugoso-annulata pure culture with strain reducing solution according to the volume of 1:0.9-1.1
Than pulverizer is added, stirring adds 8-10 times of volume of Stropharia rugoso-annulata pure culture of strain reducing solution, obtains big spherical cap after crushing
Mushroom pure culture.
Preferred technical solution are as follows: the strain reducing solution the preparation method comprises the following steps: its quality is added in the distilled water of 1L
The yeast powder of 0.01-0.05%, the K of 0.03-0.08%2HPO4, the KH of 0.03-0.08%2PO4, the MgSO of 0.01-0.05%4, 0.8-
1.2% Theravite.
Preferred technical solution are as follows: the Theravite is dissolved in water by 1g thiamine hydrochloride and 0.1g riboflavin
In, it after being settled to 1L, sterilizes 30 minutes in 104 DEG C, keeps the temperature 2 hours then at 70 DEG C, be made after cooling.
To achieve the above object, present invention provide the technical scheme that a kind of Stropharia rugoso-annulata restores the growth condition of strain,
By Stropharia rugoso-annulata pure culture made from the above method, crop field to be seeded or hayashishita are sprayed with spray appliance.
Preferred technical solution are as follows: the spray appliance is water pump.
Since above-mentioned technical proposal is used, the present invention has the advantage, that compared with prior art
The present invention is directed to traditional Stropharia rugoso-annulata solid spawn, solid spawn freight is high, to occupy volume big, complicated for operation at
The disadvantages of this height and cell age difference are big obtains cultivar using Stropharia rugoso, then carries out pure culture and Liquid Culture
Base separation, low-temperature preservation transport arrive at the destination addition special liquid matrix reduction mycelium, carry out the sprinkling pure bacterium of Stropharia rugoso-annulata
The cultural method of kind obtains a kind of production method that Stropharia rugoso-annulata restores strain and its application method in cultivation production.
Specific embodiment
Embodiments of the present invention are illustrated by particular specific embodiment below, those skilled in the art can be by this explanation
Content disclosed by book is understood other advantages and efficacy of the present invention easily.
Embodiment one: a kind of production method and growth condition of Stropharia rugoso-annulata reduction strain
The application of Stropharia rugoso-annulata liquid cultivation seed
(1) preparation of Stropharia rugoso-annulata original seed
Stropharia rugoso-annulata parent species in test tube are transferred on PDA plate and are activated, continue switching activation 3 times in this way, to parent species
Mycelia vitality restoration.
(2) preparation of Stropharia rugoso-annulata liquid level-one kind
100mLPDB fluid nutrient medium is added in the triangular flask of 250mL, is 121 DEG C in temperature, goes out under conditions of pressure 0.15Mpa
Bacterium 15min.
(3) it by after sterilization and cooling, access diameter is 6 pieces of fungus block of Stropharia rugoso-annulata original seed PDA after the activation of 1cm, sets
It is cultivated 7 days in the speed dark place of 23 DEG C of shaking table culture case 150rpm.
(4) culture medium prescription of Stropharia rugoso-annulata liquid cultivation seed is: the formula of fluid nutrient medium is: in the distilled water of 1L
The sucrose of distillation water quality 2.0%, 0.5% albumen powder, 0.3% yeast powder, 0.7% bean cake powder, 0.1% K is added2HPO4,
0.2% KH2PO4With 0.10% MgSO4, pH 6.8 is being 121 DEG C in temperature, sterilizing under conditions of pressure 0.15Mpa
22min, it is cooling.
(5) by the level liquid kind of step (2) cultured Stropharia rugoso-annulata be inoculated into sterilizing cool down be equipped with step (4)
Stropharia rugoso-annulata liquid cultivation seed fermentor in.Then, the fermentor after inoculation is cultivated 7 days with 25 DEG C.
(6) after to the liquid cultivation seed culture of Stropharia rugoso-annulata, using vacuum filtration machine to Stropharia rugoso-annulata liquid spawn
It is concentrated, the stropharia rugosoannulata strain after concentration is placed in 4 DEG C of incubator, sowing place is transported, by the big spherical cap
Mushroom pure culture and strain reducing solution add Stropharia rugoso-annulata pure culture after stirring crushing according to the volume ratio addition pulverizer of 1:1
9 times of volume of strain reducing solution, obtains Stropharia rugoso-annulata pure culture.
The strain reducing solution the preparation method comprises the following steps: the yeast powder of its quality 0.03% is added in the distilled water of 1L,
0.055% K2HPO4, 0.055% KH2PO4, 0.03% MgSO4, 1% Theravite.
The Theravite is soluble in water by 1g thiamine hydrochloride and 0.1g riboflavin, after being settled to 1L, in 104
DEG C sterilizing 30 minutes, 2 hours are kept the temperature then at 70 DEG C, is made after cooling.
By Stropharia rugoso-annulata pure culture made from the above method, crop field to be seeded is sprayed with spray appliance.
The spray appliance is water pump.
Embodiment two: a kind of production method and growth condition of Stropharia rugoso-annulata reduction strain
A kind of production method of Stropharia rugoso-annulata reduction strain, including the following steps:
It is activated on the plate of PDA culture medium step 1: Stropharia rugoso-annulata parent species are transferred to one and are accommodated, keeps Stropharia rugoso-annulata female
The mycelia vitality restoration of kind, then segmentation obtains Stropharia rugoso-annulata original seed PDA fungus block；
Step 2: the Stropharia rugoso-annulata original seed PDA fungus block is inoculated into the PDB fluid nutrient medium after sterilizing, under the conditions of 22 DEG C,
It is cultivated 7 days with shaking table culture case in dark place, obtains Stropharia rugoso-annulata level-one kind；
Step 3: the Stropharia rugoso-annulata level-one kind to be inoculated into the fermentor for accommodating Stropharia rugoso-annulata liquid cultivation seed culture medium
In, then the fermentor after inoculation is cultivated 7 days with 22 DEG C；The preparation method of the Stropharia rugoso-annulata liquid cultivation seed culture medium
Are as follows: be added the sucrose of its quality 1% to 1 liter of distilled water, 0.1% albumen powder, 0.2% yeast powder, 0.6% bean cake powder, 0.05%
K2HPO4, 0.05% KH2PO4With 0.05% MgSO4, pH value is adjusted after mixing to 6.3, then sterilized, cooling step
After be made；
Step 4: being concentrated the Stropharia rugoso-annulata liquid spawn that third step culture obtains to get big ball using vacuum filtration machine
The pure cultivation strain of lid mushroom；
Step 5: the sterile valve bag of Stropharia rugoso-annulata pure culture is dispensed, preservation and transport are then carried out under the conditions of 4 DEG C；
Step 6: transporting to after destination, the Stropharia rugoso-annulata pure culture is added with strain reducing solution according to the volume ratio of 1:0.9
Enter pulverizer, stirring adds the strain reducing solution of 8 times of Stropharia rugoso-annulata pure culture volume, obtains the pure bacterium of Stropharia rugoso-annulata after crushing
Kind.
Preferred technical solution are as follows: the strain reducing solution the preparation method comprises the following steps: its quality is added in the distilled water of 1L
0.01% yeast powder, 0.03% K2HPO4, 0.03% KH2PO4, 0.01% MgSO4, 0.8% Theravite.
Preferred technical solution are as follows: the Theravite is dissolved in water by 1g thiamine hydrochloride and 0.1g riboflavin
In, it after being settled to 1L, sterilizes 30 minutes in 104 DEG C, keeps the temperature 2 hours then at 70 DEG C, be made after cooling.
To achieve the above object, present invention provide the technical scheme that a kind of Stropharia rugoso-annulata restores the growth condition of strain,
By Stropharia rugoso-annulata pure culture made from the above method, crop field to be seeded or hayashishita are sprayed with spray appliance.
Preferred technical solution are as follows: the spray appliance is water pump.
Embodiment three: a kind of production method and growth condition of Stropharia rugoso-annulata reduction strain
A kind of production method of Stropharia rugoso-annulata reduction strain, including the following steps:
It is activated on the plate of PDA culture medium step 1: Stropharia rugoso-annulata parent species are transferred to one and are accommodated, keeps Stropharia rugoso-annulata female
The mycelia vitality restoration of kind, then segmentation obtains Stropharia rugoso-annulata original seed PDA fungus block；
Step 2: the Stropharia rugoso-annulata original seed PDA fungus block is inoculated into the PDB fluid nutrient medium after sterilizing, under the conditions of 26 DEG C,
It is cultivated 8 days with shaking table culture case in dark place, obtains Stropharia rugoso-annulata level-one kind；
Step 3: the Stropharia rugoso-annulata level-one kind to be inoculated into the fermentor for accommodating Stropharia rugoso-annulata liquid cultivation seed culture medium
In, then the fermentor after inoculation is cultivated 8 days with 26 DEG C；The preparation method of the Stropharia rugoso-annulata liquid cultivation seed culture medium
Are as follows: be added the sucrose of its quality 2.0% to 1 liter of distilled water, 0.50% albumen powder, 0.30% yeast powder, 0.7% bean cake powder,
0.10% K2HPO4, 0.20% KH2PO4With 0.10% MgSO4, pH value is adjusted after mixing to 6.8, then sterilized, cooling
It is made after step；
Step 4: being concentrated the Stropharia rugoso-annulata liquid spawn that third step culture obtains to get big ball using vacuum filtration machine
The pure cultivation strain of lid mushroom；
Step 5: the sterile valve bag of Stropharia rugoso-annulata pure culture is dispensed, preservation and transport are then carried out under the conditions of 8 DEG C；
Step 6: transporting to after destination, by the Stropharia rugoso-annulata pure culture with strain reducing solution according to the volume ratio of 1:1.1
Pulverizer is added, stirring adds the strain reducing solution of 10 times of Stropharia rugoso-annulata pure culture volume, it is pure to obtain Stropharia rugoso-annulata after crushing
Strain.
Preferred technical solution are as follows: the strain reducing solution the preparation method comprises the following steps: its quality is added in the distilled water of 1L
0.05% yeast powder, 0.08% K2HPO4, 0.08% KH2PO4, 0.05% MgSO4, 1.2% Theravite.
Preferred technical solution are as follows: the Theravite is dissolved in water by 1g thiamine hydrochloride and 0.1g riboflavin
In, it after being settled to 1L, sterilizes 30 minutes in 104 DEG C, keeps the temperature 2 hours then at 70 DEG C, be made after cooling.
A kind of growth condition of Stropharia rugoso-annulata reduction strain, by Stropharia rugoso-annulata pure culture made from the above method, with sprinkling
Equipment sprays crop field to be seeded or hayashishita.
Preferred technical solution are as follows: the spray appliance is water pump.
The above-described embodiments merely illustrate the principles and effects of the present invention, and is not intended to limit the present invention.It is any ripe
The personage for knowing this technology all without departing from the spirit and scope of the present invention, carries out modifications and changes to above-described embodiment.Cause
This, institute is complete without departing from the spirit and technical ideas disclosed in the present invention by those of ordinary skill in the art such as
At all equivalent modifications or change, should be covered by the claims of the present invention.
Claims (5)
1. a kind of production method of Stropharia rugoso-annulata reduction strain, it is characterised in that: include the following steps:
It is activated on the plate of PDA culture medium step 1: Stropharia rugoso-annulata parent species are transferred to one and are accommodated, keeps Stropharia rugoso-annulata female
The mycelia vitality restoration of kind, then segmentation obtains Stropharia rugoso-annulata original seed PDA fungus block；
Step 2: the Stropharia rugoso-annulata original seed PDA fungus block is inoculated into the PDB fluid nutrient medium after sterilizing, in 22-26 DEG C of condition
Under, it is cultivated 7-8 days with shaking table culture case in dark place, obtains Stropharia rugoso-annulata level-one kind；
Step 3: the Stropharia rugoso-annulata level-one kind to be inoculated into the fermentor for accommodating Stropharia rugoso-annulata liquid cultivation seed culture medium
In, then by the fermentor after inoculation with 22-26 DEG C culture 7-8 days；The preparation of the Stropharia rugoso-annulata liquid cultivation seed culture medium
Method are as follows: be added the sucrose of its quality 1.0-2.0% to 1 liter of distilled water, the ferment of the albumen powder of 0.10-0.50%, 0.20-0.30%
Female powder, the bean cake powder of 0.6-0.7%, the K of 0.05-0.10%2HPO4, the KH of 0.05-0.20%2PO4With the MgSO of 0.05-0.10%4,
It is made after adjusting pH value to 6.3-6.8, then sterilized, cooling step after mixing；
Step 4: being concentrated the Stropharia rugoso-annulata liquid spawn that third step culture obtains to get big ball using vacuum filtration machine
The pure cultivation strain of lid mushroom；
Step 5: the sterile valve bag of Stropharia rugoso-annulata pure culture is dispensed, preservation and transport are then carried out under the conditions of 4-8 DEG C；
Step 6: transporting to after destination, by the Stropharia rugoso-annulata pure culture with strain reducing solution according to the volume of 1:0.9-1.1
Than pulverizer is added, stirring adds 8-10 times of volume of Stropharia rugoso-annulata pure culture of strain reducing solution, obtains big spherical cap after crushing
Mushroom pure culture.
2. the production method of Stropharia rugoso-annulata reduction strain according to claim 1, it is characterised in that: the strain reducing solution
The preparation method comprises the following steps: the yeast powder of its quality 0.01-0.05% is added in the distilled water of 1L, the K of 0.03-0.08%2HPO4,
The KH of 0.03-0.08%2PO4, the MgSO of 0.01-0.05%4, the Theravite of 0.8-1.2%.
3. the production method of Stropharia rugoso-annulata reduction strain according to claim 2, it is characterised in that: the multi-vitamins
Solution is soluble in water by 1g thiamine hydrochloride and 0.1g riboflavin, after being settled to 1L, sterilizes 30 minutes in 104 DEG C, then at 70 DEG C
Heat preservation 2 hours is made after cooling.
4. a kind of growth condition of Stropharia rugoso-annulata reduction strain, it is characterised in that: claim 1-3 any claim to be made
Stropharia rugoso-annulata pure culture, crop field to be seeded or hayashishita are sprayed with spray appliance.
5. the growth condition of Stropharia rugoso-annulata according to claim 4 reduction strain, it is characterised in that: the spray appliance is
Water pump.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110574633A (en) * | 2019-08-22 | 2019-12-17 | 江苏东越生物技术发展股份有限公司 | liquid culture medium for industrialized production of stropharia rugoso-annulata and preparation method thereof |
| CN111615996A (en) * | 2020-05-19 | 2020-09-04 | 江西省农业科学院农业应用微生物研究所（江西省农村能源研究中心） | Stropharia rugosoannulata strain breeding method |
| CN113348968A (en) * | 2021-07-13 | 2021-09-07 | 石忠斌 | Preparation of stropharia rugoso-annulata comprehensive culture medium and stropharia rugoso-annulata cultivation method |
| CN114342734A (en) * | 2021-11-10 | 2022-04-15 | 中国科学院微生物研究所 | Preparation of liquid reduction strain of stropharia rugoso-annulata and cultivation method of fruiting body |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110574633A (en) * | 2019-08-22 | 2019-12-17 | 江苏东越生物技术发展股份有限公司 | liquid culture medium for industrialized production of stropharia rugoso-annulata and preparation method thereof |
| CN111615996A (en) * | 2020-05-19 | 2020-09-04 | 江西省农业科学院农业应用微生物研究所（江西省农村能源研究中心） | Stropharia rugosoannulata strain breeding method |
| CN113348968A (en) * | 2021-07-13 | 2021-09-07 | 石忠斌 | Preparation of stropharia rugoso-annulata comprehensive culture medium and stropharia rugoso-annulata cultivation method |
| CN114342734A (en) * | 2021-11-10 | 2022-04-15 | 中国科学院微生物研究所 | Preparation of liquid reduction strain of stropharia rugoso-annulata and cultivation method of fruiting body |
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