CN114456954A - Liquid germination bacterium and preparation method thereof - Google Patents
Liquid germination bacterium and preparation method thereof Download PDFInfo
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- CN114456954A CN114456954A CN202210180172.6A CN202210180172A CN114456954A CN 114456954 A CN114456954 A CN 114456954A CN 202210180172 A CN202210180172 A CN 202210180172A CN 114456954 A CN114456954 A CN 114456954A
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Abstract
The invention relates to a liquid germination bacterium and a preparation method thereof, wherein the liquid germination bacterium comprises the following raw materials in parts by mass: 3-12 parts of corn flour, 1-10 parts of soybean flour, 5-15 parts of glucose, 5-15 parts of white sugar, 1-8 parts of starch, 1-5 parts of peptone, 1-3 parts of potassium dihydrogen phosphate, 1-3 parts of magnesium sulfate, VB 11-2 parts and 0.3-0.5 part of defoaming agent. The invention has the advantages of low cost, simple production and processing, and can be used for industrial production and labor reduction.
Description
Technical Field
The invention relates to the technical field of industrial water treatment, in particular to a liquid germination bacterium and a preparation method thereof.
Background
The conventional sowing method is as follows: the method comprises the steps of firstly spreading a layer of wet leaves on a bed surface ready for planting, then spreading mixed germination fungus capsules on the leaves, pressing cultured armillaria mellea fungus sticks at intervals on the leaves, covering a culture material, covering a layer of leaves, carrying out moisture preservation and temperature control management, and harvesting after two or three years.
The germination rate of the gastrodia elata planting is directly determined by the quality of the germination bacteria, the vitality of the germination bacteria is strong, the gastrodia elata planting can be quickly inoculated due to long hyphae, the vitality of the germination bacteria is weak, the gastrodia elata planting has few chances to be inoculated due to short hyphae, nutrition cannot be obtained, the seeds continuously grow, self nutrition is consumed, and the gastrodia elata planting dies naturally after the nutrition is exhausted.
At present, in the process of cultivating sexual propagation germinating bacteria, a blank formed by mixing sawdust, white sugar, bran and the like is often adopted as a culture medium, the growth of the germinating bacteria is influenced by the powdery mixed culture medium generally due to the invasion of mixed bacteria and too much water, and the culture base material has complex components, complex process and higher manufacturing cost.
Disclosure of Invention
The invention aims to provide a liquid germinating bacterium and a preparation method thereof.
The above object of the present invention is achieved by the following technical solutions:
a liquid germination bacterium and a preparation method thereof are disclosed, which comprises the following raw materials by mass: 3-12 parts of corn flour, 1-10 parts of soybean flour, 5-15 parts of glucose, 5-15 parts of white sugar, 1-8 parts of starch, 1-5 parts of peptone, 1-3 parts of potassium dihydrogen phosphate, 1-3 parts of magnesium sulfate, VB 11-2 parts and 0.3-0.5 part of defoaming agent.
Preferably, the raw materials are calculated according to the mass parts: 3 parts of corn flour, 1 part of soybean flour, 5 parts of glucose, 5 parts of white sugar, 1 part of starch, 1 part of peptone, 1 part of monopotassium phosphate, 1 part of magnesium sulfate, VB11 parts and 0.3 part of defoaming agent.
Preferably, 12 parts of corn flour, 10 parts of soybean flour, 15 parts of glucose, 15 parts of white sugar, 8 parts of starch, 5 parts of peptone, 3 parts of monopotassium phosphate, 3 parts of magnesium sulfate, VB 12 parts and 0.5 part of defoaming agent.
Preferably, 8 parts of corn flour, 5 parts of soybean flour, 10 parts of glucose, 10 parts of white sugar, 4 parts of starch, 3 parts of peptone, 2 parts of monopotassium phosphate, 2 parts of magnesium sulfate, 11.5 parts of VB and 0.4 part of defoaming agent.
A preparation method of liquid germination bacteria comprises the following process steps:
(1) adding corn flour, soybean flour, glucose, white sugar, starch, peptone, potassium dihydrogen phosphate, magnesium sulfate, VB1 and a defoaming agent into a beaker according to the above components, then adding a proper amount of water, uniformly stirring by using a glass rod, then subpackaging the uniformly stirred solution into a triangular flask, and then plugging a cotton plug into the opening of the triangular flask for sealing;
(2) and (3) pot filling and sterilization: bundling the triangular bottle with the tampon by using double-layer newspaper, putting the triangular bottle into a sterilization pot, covering the sterilization pot with the newspaper, exhausting, starting timing when the temperature rises to 121 ℃, controlling the temperature to be 121-;
(3) and (3) drying the cotton plug: when the pressure is reduced to 0.05MPa, the cotton wool is pressed into a sterile room, when the pressure is about to be reduced to zero, the air is released, the pot cover is opened, the newspaper is taken out, the pot cover is embedded with a small seam, and the cotton wool is dried for 40-50 min.
(4) And (3) cooling: taking out the triangular flask, placing the triangular flask on a clean bench or an inoculation box, and cooling and inoculating;
(5) inoculation: the inoculation process of the mother strain A as the test tube strain comprises the following steps:
selecting 1 newly cultured test tube slant strain, pulling out a triangular flask tampon with the right hand, holding in the hand, picking 50% of the slant with an inoculation hook, cutting into 5-6 inoculation blocks of 4 × 4mm, and rapidly placing into the triangular flask; the test tube strains are quickly put back to the side of the alcohol lamp, the inoculation hook is also put back to the test tube, and the cotton plug is burned when being plugged again; bundling a bottle opening with double-layer newspaper, sticking a label on a triangular flask, noting the bacterial name, the culture medium formula and the inoculation date, and then putting the bottle on a shaking table for culturing;
inoculation process for taking B mother strain as plate strain
Selecting one newly cultured plate strain, marking on the plate strain to obtain an average inoculation block of 0.5 x 0.5cm, and temporarily placing an inoculation hook on a test tube rack after marking; pulling out the triangular flask tampon with the right hand, holding in the hand, picking 4-5 inoculation blocks with the inoculation hook, and rapidly placing in the triangular flask; quickly putting the flat strain back to the side of the alcohol lamp, putting the inoculation hook back to the flat dish, and burning when the cotton plug is plugged again; the bottle mouth is tied by double-layer newspaper, a label is stuck on a triangular flask, the bacterial name, the culture medium formula and the inoculation date are indicated, and then the bottle is placed on a shaking table for culture.
(6) Culturing liquid strains: placing the triangular flask on a shaker or a magnetic stirrer for culturing, controlling the temperature at 23-24 ℃, and gradually increasing the rotating speed or frequency; observing the color and the shape of the mycelium pellet at regular time, and identifying immediately if abnormality is found (ph detection, mycelium microscopic examination, sampling culture); the age of the fungus is generally controlled to be 6-8 days, a large number of fungus balls appear in the culture solution at the moment, and the culture solution is yellow-white and has a malic acid taste; and comprehensively detecting the cultured liquid seeds, then supplying the liquid seeds to a fermentation tank for inoculation, and simultaneously performing microscopic examination, empty culture and sampling on the liquid seeds after the fermentation tank is inoculated.
In summary, the beneficial technical effects of the invention are as follows:
in the traditional method, 15x30 bags of hypha are required to fully grow for 50-90 days by solid inoculation, the time for full growth is shortened by half by 30-40 days by liquid inoculation, 50-60 days are required for solid from mother seeds to secondary seeds, 16-20 days are required for liquid from the mother seeds to a fermentation tank, and the liquid preparation can be carried out in industrial production and also reduces labor.
Detailed Description
The present invention will be described in further detail with reference to examples.
Example 1:
a liquid germination bacterium and a preparation method thereof are disclosed, which comprises the following raw materials by mass: 3 parts of corn flour, 1 part of soybean flour, 5 parts of glucose, 5 parts of white sugar, 1 part of starch, 1 part of peptone, 1 part of monopotassium phosphate, 1 part of magnesium sulfate, VB11 parts and 0.3 part of defoaming agent.
Example 2
A liquid germination bacterium and a preparation method thereof are disclosed, which comprises the following raw materials by mass: 12 parts of corn flour, 10 parts of soybean flour, 15 parts of glucose, 15 parts of white sugar, 8 parts of starch, 5 parts of peptone, 3 parts of monopotassium phosphate, 3 parts of magnesium sulfate, VB 12 parts and 0.5 part of defoaming agent.
Example 3
A liquid germination bacterium and a preparation method thereof are disclosed, which are calculated by the following raw materials in parts by mass: 8 parts of corn flour, 5 parts of soybean flour, 10 parts of glucose, 10 parts of white sugar, 4 parts of starch, 3 parts of peptone, 2 parts of monopotassium phosphate, 2 parts of magnesium sulfate, 11.5 parts of VB and 0.4 part of defoaming agent.
A preparation method of liquid germination bacteria comprises the following process steps:
(1) adding corn flour, soybean flour, glucose, white sugar, starch, peptone, potassium dihydrogen phosphate, magnesium sulfate, VB1 and a defoaming agent into a beaker according to the above components, then adding a proper amount of water, uniformly stirring by using a glass rod, then subpackaging the uniformly stirred solution into a triangular flask, and then plugging a cotton plug into the opening of the triangular flask for sealing;
(2) and (3) pot filling and sterilization: bundling the triangular bottle with the tampon by using double-layer newspaper, putting the triangular bottle into a sterilization pot, covering the sterilization pot with the newspaper, exhausting, starting timing when the temperature rises to 121 ℃, controlling the temperature to be 121-;
(3) and (3) drying the cotton plug: when the pressure is reduced to 0.05MPa, the cotton plug enters the sterile room with pressure, when the pressure is about to be reduced to zero, the air is released, the pot cover is opened to take out the newspaper, and a small seam is embedded in the pot cover to dry the cotton plug for 40-50 min.
(4) And (3) cooling: taking out the triangular flask, placing the triangular flask on a clean bench or an inoculation box, and cooling and inoculating;
(5) inoculation: the inoculation process of the mother strain A as the test tube strain comprises the following steps:
selecting 1 newly cultured test tube slant strain, pulling out a triangular flask tampon with the right hand, holding in the hand, picking 50% of the slant with an inoculation hook, cutting into 5-6 inoculation blocks of 4 × 4mm, and rapidly placing into the triangular flask; the test tube strains are quickly put back to the side of the alcohol lamp, the inoculation hook is also put back to the test tube, and the cotton plug is burned when being plugged again; bundling a bottle opening with double-layer newspaper, sticking a label on a triangular flask, noting the bacterial name, the culture medium formula and the inoculation date, and then putting the bottle on a shaking table for culturing;
inoculation process for taking B mother strain as plate strain
Selecting one newly cultured plate strain, marking on the plate strain to obtain an average inoculation block of 0.5 x 0.5cm, and temporarily placing an inoculation hook on a test tube rack after marking; pulling out the triangular flask tampon with the right hand, holding in the hand, picking 4-5 inoculation blocks with the inoculation hook, and rapidly placing in the triangular flask; quickly putting the flat strain back to the side of the alcohol lamp, putting the inoculation hook back to the flat dish, and burning when the cotton plug is plugged again; the bottle mouth is tied by double-layer newspaper, a label is stuck on a triangular flask, the bacterial name, the culture medium formula and the inoculation date are indicated, and then the bottle is placed on a shaking table for culture.
(6) Culturing liquid strains: placing the triangular flask on a shaker or a magnetic stirrer for culturing, controlling the temperature at 23-24 ℃, and gradually increasing the rotating speed or frequency; observing the color and the shape of the mycelium pellet at regular time, and identifying immediately if abnormality is found (ph detection, mycelium microscopic examination, sampling culture); the age of the fungus is generally controlled to be 6-8 days, a large number of fungus balls appear in the culture solution at the moment, and the culture solution is yellow-white and has a malic acid taste; and comprehensively detecting the cultured liquid seeds, then supplying the liquid seeds to a fermentation tank for inoculation, and simultaneously performing microscopic examination, empty culture and sampling on the liquid seeds after the fermentation tank is inoculated.
In the traditional method, 15x30 bags of hypha are required to fully grow for 50-90 days by solid inoculation, the time for full growth is shortened by half by 30-40 days by liquid inoculation, 50-60 days are required for solid from mother seeds to secondary seeds, 16-20 days are required for liquid from the mother seeds to a fermentation tank, and the liquid preparation can be carried out in industrial production and also reduces labor.
The embodiments of the present invention are preferred embodiments of the present invention, and the scope of the present invention is not limited by these embodiments, so: all equivalent changes made according to the structure, shape and principle of the invention are covered by the protection scope of the invention.
Claims (5)
1. A liquid germination bacterium and a preparation method thereof are characterized in that: the material is prepared from the following raw materials in parts by weight: 3-12 parts of corn flour, 1-10 parts of soybean flour, 5-15 parts of glucose, 5-15 parts of white sugar, 1-8 parts of starch, 1-5 parts of peptone, 1-3 parts of potassium dihydrogen phosphate, 1-3 parts of magnesium sulfate, VB 11-2 parts and 0.3-0.5 part of defoaming agent.
2. The liquid germination bacteria of claim 1, wherein the raw materials comprise, by mass: 3 parts of corn flour, 1 part of soybean flour, 5 parts of glucose, 5 parts of white sugar, 1 part of starch, 1 part of peptone, 1 part of monopotassium phosphate, 1 part of magnesium sulfate, VB11 parts and 0.3 part of defoaming agent.
3. The liquid germination bacterium according to claim 1, wherein the raw materials are, by mass: 12 parts of corn flour, 10 parts of soybean flour, 15 parts of glucose, 15 parts of white sugar, 8 parts of starch, 5 parts of peptone, 3 parts of monopotassium phosphate, 3 parts of magnesium sulfate, VB 12 parts and 0.5 part of defoaming agent.
4. The liquid germination bacterium according to claim 1, wherein the raw materials are, by mass: 8 parts of corn flour, 5 parts of soybean flour, 10 parts of glucose, 10 parts of white sugar, 4 parts of starch, 3 parts of peptone, 2 parts of monopotassium phosphate, 2 parts of magnesium sulfate, 11.5 parts of VB and 0.4 part of defoaming agent.
5. The method for preparing liquid germinating bacteria according to claim 1, which comprises the following steps:
(1) adding corn flour, soybean flour, glucose, white sugar, starch, peptone, potassium dihydrogen phosphate, magnesium sulfate, VB1 and a defoaming agent into a beaker according to the above components, then adding a proper amount of water, uniformly stirring by using a glass rod, then subpackaging the uniformly stirred solution into a triangular flask, and then plugging a cotton plug into the opening of the triangular flask for sealing;
(2) and (3) pot filling and sterilization: bundling the triangular bottle with the tampon by using double-layer newspaper, putting the triangular bottle into a sterilization pot, covering the sterilization pot with the newspaper, exhausting, starting timing when the temperature rises to 121 ℃, controlling the temperature to be 121-;
(3) baking the cotton plug: when the pressure is reduced to 0.05MPa, the cotton plug enters the sterile room with pressure, when the pressure is about to be reduced to zero, the air is released, the pot cover is opened to take out the newspaper, and a small seam is embedded in the pot cover to dry the cotton plug for 40-50 min.
(4) And (3) cooling: taking out the triangular flask, placing the triangular flask on a clean bench or an inoculation box, and cooling and inoculating;
(5) inoculation: the inoculation process of the mother strain A as the test tube strain comprises the following steps:
selecting 1 newly cultured test tube slant strain, pulling out a triangular flask tampon with the right hand, holding in the hand, picking 50% of the slant with an inoculation hook, cutting into 5-6 inoculation blocks of 4 × 4mm, and rapidly placing into the triangular flask; the test tube strains are quickly put back to the side of the alcohol lamp, the inoculation hook is also put back to the test tube, and the cotton plug is burned when being plugged again; bundling a bottle opening with double-layer newspaper, sticking a label on a triangular flask, noting the bacterial name, the culture medium formula and the inoculation date, and then putting the bottle on a shaking table for culturing;
inoculation process for taking B mother strain as plate strain
Selecting one newly cultured plate strain, marking on the plate strain to obtain an average inoculation block of 0.5 x 0.5cm, and temporarily placing an inoculation hook on a test tube rack after marking; pulling out the triangular flask cotton plug by the right hand, holding in the hand, picking 4-5 inoculation blocks by an inoculation hook, and quickly putting into the triangular flask; quickly putting the flat strain back to the side of the alcohol lamp, putting the inoculation hook back to the flat dish, and burning when the cotton plug is plugged again; the bottle mouth is tied by double-layer newspaper, a label is stuck on a triangular flask, the bacterial name, the culture medium formula and the inoculation date are indicated, and then the bottle is placed on a shaking table for culture.
(6) Culturing liquid strains: placing the triangular flask on a shaker or a magnetic stirrer for culturing, controlling the temperature at 23-24 ℃, and gradually increasing the rotating speed or frequency; observing the color and the shape of the mycelium pellet at regular time, and identifying immediately if abnormality is found (ph detection, mycelium microscopic examination, sampling culture); the age of the fungus is generally controlled to be 6-8 days, a large number of fungus balls appear in the culture solution at the moment, and the culture solution is yellow-white and has a malic acid taste; and comprehensively detecting the cultured liquid seeds, then supplying the liquid seeds to a fermentation tank for inoculation, and simultaneously performing microscopic examination, empty culture and sampling on the liquid seeds after the fermentation tank is inoculated.
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| CN202210180172.6A CN114456954A (en) | 2022-02-26 | 2022-02-26 | Liquid germination bacterium and preparation method thereof |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20110044038A (en) * | 2009-10-22 | 2011-04-28 | 대한민국(관리부서 : 산림청 국립산림과학원장) | In vitro germination method of gastrodia elata seeds using symbiotic fungal species |
| CN103798056A (en) * | 2013-06-09 | 2014-05-21 | 程立君 | Production method of gastrodia elata seed germinating strains |
| CN107629967A (en) * | 2017-10-20 | 2018-01-26 | 贵州威门药业股份有限公司 | Fluid nutrient medium and rhizoma Gastrodiae implantation methods for the Germination Strain of rhizoma Gastrodiae plantation |
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- 2022-02-26 CN CN202210180172.6A patent/CN114456954A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20110044038A (en) * | 2009-10-22 | 2011-04-28 | 대한민국(관리부서 : 산림청 국립산림과학원장) | In vitro germination method of gastrodia elata seeds using symbiotic fungal species |
| CN103798056A (en) * | 2013-06-09 | 2014-05-21 | 程立君 | Production method of gastrodia elata seed germinating strains |
| CN107629967A (en) * | 2017-10-20 | 2018-01-26 | 贵州威门药业股份有限公司 | Fluid nutrient medium and rhizoma Gastrodiae implantation methods for the Germination Strain of rhizoma Gastrodiae plantation |
Non-Patent Citations (1)
| Title |
|---|
| 杨儒钦: "天麻蜜环菌、萌发菌母种生产技术", 特种经济动植物, no. 2, pages 28 * |
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