CN112119838A - Pleurotus eryngii liquid stock culture solution and pleurotus eryngii stock preparation process - Google Patents
Pleurotus eryngii liquid stock culture solution and pleurotus eryngii stock preparation process Download PDFInfo
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- CN112119838A CN112119838A CN202010989623.1A CN202010989623A CN112119838A CN 112119838 A CN112119838 A CN 112119838A CN 202010989623 A CN202010989623 A CN 202010989623A CN 112119838 A CN112119838 A CN 112119838A
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- pleurotus eryngii
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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 invention relates to a mushroom culture solution, in particular to a pleurotus eryngii liquid stock culture solution which is prepared from the following raw materials in parts by weight: 17.9-18.1 parts of white sugar; 1.9-2.1 parts of soybean meal; 0.2-0.4 part of anhydrous magnesium sulfate; 0.2-0.4 part of monopotassium phosphate; 0.9-1.1 parts of a defoaming agent; 590-610 parts of water. The pleurotus eryngii liquid stock culture solution provided by the invention has the advantages of short culture period, high strain purity and stable strain activity.
Description
Technical Field
The invention relates to a culture solution of mushroom, in particular to a pleurotus eryngii liquid stock culture solution and a pleurotus eryngii stock preparation process.
Background
The pleurotus eryngii is cultured by adopting a solid culture medium mostly, the culture period of the solid culture medium is long, the purity of the strain is low, and the activity of the strain is unstable.
Disclosure of Invention
In order to solve the problems, the invention provides a pleurotus eryngii liquid stock culture solution with short culture period, high strain purity and stable strain activity, and the specific technical scheme is as follows:
the pleurotus eryngii liquid stock culture solution is prepared from the following raw materials in parts by weight: 17.9-18.1 parts of white sugar; 1.9-2.1 parts of soybean meal; 0.2-0.4 part of anhydrous magnesium sulfate; 0.2-0.4 part of monopotassium phosphate; 0.9-1.1 parts of a defoaming agent; 590-610 parts of water.
Because of adopting liquid culture, the outer surface of the strain is not easy to grow old fungus skin, the foreign fungus in the bag is easy to observe, the purity of the strain is high, the fungus growing speed is high, the foreign fungus is not easy to infect, and the fungus growing time is short.
Preferably, 18 parts of white sugar; 1.8 parts of soybean meal; 0.3 part of anhydrous magnesium sulfate; 0.3 part of monopotassium phosphate; 1 part of a defoaming agent; and 600 parts of water.
Further, the pH value of the pleurotus eryngii liquid stock culture solution before sterilization is 6.2 +/-0.2, and the pH value after sterilization is 5.8 +/-0.2.
The invention also aims to provide a pleurotus eryngii original seed production process, which comprises the following steps:
s10, preparing a pleurotus eryngii liquid stock culture solution, and pouring the culture solution into a triangular flask;
s20, sealing the bottle mouth of the triangular bottle;
s30, placing the sealed triangular flask into an autoclave for sterilization;
s40, cooling to 22-24 ℃ after sterilization;
s50, inoculating, namely inoculating the pleurotus eryngii mother seeds into a triangular flask;
s60, carrying out constant temperature shaking culture on the triangular flask inoculated with the mother strain of the pleurotus eryngii.
Further, after the bottle mouth is sealed in the step S20, a layer of kraft paper is firstly sleeved, and then a layer of plastic paper is sleeved.
Through adopting above-mentioned technical scheme, prevent that vapor water from drenching triangular flask cotton plug, lead to later stage pollution culture solution.
Further, in the step S30, the temperature is increased to 100-120 ℃, cold air is discharged, and the sterilization is carried out for not less than 25 minutes when the temperature is increased to 120-126 ℃.
Further, after the sterilization in the step S40 is finished, the autoclave is cooled, then the triangular flask is taken out, the plastic paper and the kraft paper are taken down, and the temperature is naturally cooled to 22-24 ℃.
Further, in step S50, a sterilization lamp is used to sterilize before inoculation, and a plurality of mother seeds with consistent specifications are loaded.
Furthermore, the triangular flask is 1000ml, the mother seeds are cubes with the side length of 3mm, and the number of the mother seeds is 11-13.
Further, the triangular flask in the step S60 is cultured in a constant temperature vibrator, the temperature is 22-23 ℃, and the oscillation speed is 145-155 rpm; and (5) observing the development condition of the hyphae after culturing for 6-7 days, and finishing culturing if fine particles are formed in the triangular flask.
By adopting the technical scheme, the pH value and the sugar content can be accurately detected to obtain the strain activity condition.
Compared with the prior art, the invention has the following beneficial effects:
the pleurotus eryngii liquid stock culture solution provided by the invention has the advantages of short culture period, high strain purity and stable strain activity.
Detailed Description
The present invention will now be further described with reference to examples.
The pleurotus eryngii liquid stock culture solution is prepared from the following raw materials in parts by weight: 17.9-18.1 parts of white sugar; 1.9-2.1 parts of soybean meal; 0.2-0.4 part of anhydrous magnesium sulfate; 0.2-0.4 part of monopotassium phosphate; 0.9-1.1 parts of a defoaming agent; 590-610 parts of water.
Wherein, when the parts of the solid raw materials are 'g', the parts of the antifoaming agent and the water are 'ml'.
The culture solution has advantages of hard old fungus skin generation on the outer surface, easy observation of the mixed fungus in the bag, high purity of the strain, fast spawn running speed, hard infection of the mixed fungus, short spawn running time, strong strain activity and short production period.
The above materials were put in a 1000ml Erlenmeyer flask and stirred.
The culture solution is sterilized after being prepared. The pH value of the pleurotus eryngii liquid stock culture solution before sterilization is 6.2 +/-0.2, and the pH value after sterilization is 5.8 +/-0.2. The growth environment of the pleurotus eryngii is optimal in the pH value range.
Example one
The pleurotus eryngii liquid stock culture solution is prepared from the following raw materials in parts by weight: 17.9 g of white sugar; 1.9 g of soybean meal; 0.2 g of anhydrous magnesium sulfate; potassium dihydrogen phosphate 0.2 g; 0.9ml of defoaming agent; 590ml of water.
Example two
The pleurotus eryngii liquid stock culture solution is prepared from the following raw materials in parts by weight: 18 g of white sugar; 2 g of soybean meal; 0.3 g of anhydrous magnesium sulfate; potassium dihydrogen phosphate 0.3 g; 1ml of defoaming agent; 600ml of water.
EXAMPLE III
The pleurotus eryngii liquid stock culture solution is prepared from the following raw materials in parts by weight: 18.1 g of white sugar; 2.1 g of soybean meal; anhydrous magnesium sulfate 0.4 g; potassium dihydrogen phosphate 0.4 g; 1.1ml of defoaming agent; 610ml of water.
The pH value of the pleurotus eryngii liquid stock culture solution in the embodiment before sterilization is 6.2 +/-0.2, and the pH value after sterilization is 6.0 +/-0.2.
The preparation process of the pleurotus eryngii stock comprises the following steps:
s10, preparing a pleurotus eryngii liquid stock culture solution, and pouring the culture solution into a triangular flask;
s20, sealing the bottle mouth of the triangular bottle;
s30, placing the sealed triangular flask into an autoclave for sterilization;
s40, cooling to 22-24 ℃ after sterilization;
s50, inoculating, namely inoculating the pleurotus eryngii mother seeds into a triangular flask;
s60, carrying out constant temperature shaking culture on the triangular flask inoculated with the mother strain of the pleurotus eryngii.
And step S20, after the bottle mouth is sealed, firstly sheathing a layer of kraft paper and then sheathing a layer of plastic paper. Prevent the steam water from entering the triangular flask to pollute the culture solution. Prevent the vapor from wetting the triangular flask cotton plug to cause the later pollution of the culture solution.
In step S30, the temperature is first raised to 100-120 deg.C, cold air is discharged, and sterilization is performed for not less than 25 minutes when the temperature is raised to 120-126 deg.C.
The operation was repeated twice while discharging the cold air.
The sterilization method can ensure that no other bacteria are propagated in the culture process.
In at least one embodiment, the temperature is raised to 110 + -1 deg.C during sterilization, and cold air is discharged. After the cold air is discharged twice, the temperature is raised to 123 +/-1 ℃ for sterilization for 30 minutes.
The sterilization temperature may be any temperature within the range of 120 to 126 ℃, or may be 120 ℃, 121 ℃, 122 ℃, 123 ℃, 124 ℃, 125 ℃, 126 ℃.
And (S40) cooling the autoclave after the sterilization is finished, taking out the triangular flask, taking down the plastic paper and the kraft paper, and naturally cooling to 22-24 ℃.
In step S50, a sterilization lamp is used for sterilization before inoculation, and a plurality of mother seeds with consistent specifications are loaded.
Wherein, the parent species have the same specification. The mother seeds require uniform hypha density, regular tip hypha and consistent color. The mother strain has the same particle size because the hyphae of the original strain in the triangular flask cultured by the small particle strain block are uniform, but the artificial inoculation is difficult to be infinitely small, so the size is standardized. After the mother seeds are consistent in size, the growth process of the hypha of the original seeds in the triangular flask at the later stage is easy to observe whether the hypha is abnormal or not, namely the hypha is easy to compare.
The triangular flask is 1000ml, the mother seeds are cubes with the side length of 3mm, and the number of the mother seeds is 11-13.
The sterilizing lamp is an ultraviolet lamp, and bacteria are prevented from entering the triangular flask.
The temperature of inoculation is within the range of 22-24 ℃, and can also be 22 ℃, 22.5 ℃, 23 ℃ and 23.5 ℃ and 24 ℃.
The culture solution is matched with the culture solution, so that the culture solution can be fully utilized, and the waste is reduced.
Culturing the triangular flask in the step S60 in a constant-temperature vibrator at the temperature of 22-23 ℃ and the oscillation speed of 145-155 rpm; and (5) observing the development condition of the hyphae after culturing for 6-7 days, and finishing culturing if fine particles are formed in the triangular flask.
The temperature may be any of 22 to 23 ℃ and may be 22 ℃, 22.5 ℃, 23 ℃, 23.5 ℃ and 24 ℃.
The oscillation speed may be any of 145 to 155rpm, and may be 145rpm, 146rpm, 147rpm, 148rpm, 149rpm, 150rpm, 151rpm, 152rpm, 153rpm, 154rpm, and 155 rpm.
After three days of culture, the development condition is observed, the culture is not good according to the condition, and the rest is continuously cultured.
The constant temperature culture improves the dissolved oxygen in the liquid culture medium, the strains are fully contacted with the culture solution, and the hyphae grow uniformly.
And (4) observing whether the culture is polluted after 2 days, observing the development condition after three days of culture, displaying fine particles in a triangular flask after 4-5 days of culture, and reserving for later use after 7 days of culture.
Compared with solid stock, the preparation method of the Pleurotus eryngii stock has the advantages of strong strain activity and short production period.
Claims (10)
1. The pleurotus eryngii liquid stock culture solution is characterized by being prepared from the following raw materials in parts by weight:
17.9-18.1 parts of white sugar;
1.9-2.1 parts of soybean meal;
0.2-0.4 part of anhydrous magnesium sulfate;
0.2-0.4 part of monopotassium phosphate;
0.9-1.1 parts of a defoaming agent;
590-610 parts of water.
2. The liquid stock culture solution of Pleurotus eryngii according to claim 1,
18 parts of white sugar;
1.8 parts of soybean meal;
0.3 part of anhydrous magnesium sulfate;
0.3 part of monopotassium phosphate;
1 part of a defoaming agent;
and 600 parts of water.
3. The liquid stock culture solution of Pleurotus eryngii according to claim 1,
the pH value of the pleurotus eryngii liquid stock culture solution before sterilization is 6.2 +/-0.2, and the pH value after sterilization is 5.8 +/-0.2.
4. The pleurotus eryngii original seed production process is characterized by comprising the following steps:
s10, preparing a pleurotus eryngii liquid stock culture solution, and pouring the culture solution into a triangular flask;
s20, sealing the bottle mouth of the triangular bottle;
s30, placing the sealed triangular flask into an autoclave for sterilization;
s40, cooling to 22-24 ℃ after sterilization;
s50, inoculating, namely inoculating the pleurotus eryngii mother seeds into a triangular flask;
s60, carrying out constant temperature shaking culture on the triangular flask inoculated with the mother strain of the pleurotus eryngii.
5. The Pleurotus eryngii stock culture preparation process according to claim 4,
and step S20, after the bottle mouth is sealed, firstly sheathing a layer of kraft paper and then sheathing a layer of plastic paper.
6. The Pleurotus eryngii stock culture preparation process according to claim 4,
in the step S30, the temperature is increased to 100-120 ℃, cold air is discharged, and the sterilization is carried out for not less than 25 minutes when the temperature is increased to 120-126 ℃.
7. The Pleurotus eryngii stock culture preparation process according to claim 4,
and after the sterilization in the step S40 is finished, cooling the autoclave, taking out the triangular flask, taking down the plastic paper and the kraft paper, and naturally cooling to 22-24 ℃.
8. The Pleurotus eryngii stock culture preparation process according to claim 4,
in step S50, a sterilization lamp is used to sterilize before inoculation, and a plurality of mother seeds with consistent specifications are loaded.
9. The Pleurotus eryngii stock culture preparation process according to claim 8,
the triangular flask is 1000ml, the mother seeds are cubes with the side length of 3mm, and the number of the mother seeds is 11-13.
10. The Pleurotus eryngii stock culture preparation process according to claim 4,
culturing the triangular flask in the step S60 in a constant-temperature vibrator at the temperature of 22-23 ℃ and the oscillation speed of 145-155 rpm; and (5) observing the development condition of the hyphae after culturing for 6-7 days, and finishing culturing if fine particles are formed in the triangular flask.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112125732A (en) * | 2020-09-19 | 2020-12-25 | 江苏丰收菇业有限公司 | Velvet antler mushroom liquid stock culture solution and velvet antler mushroom stock preparation process |
| CN115152532A (en) * | 2022-06-08 | 2022-10-11 | 湖南果秀食品有限公司 | Quantitative and visual pleurotus eryngii liquid shake flask stock seed production method |
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| CN102204474A (en) * | 2010-03-29 | 2011-10-05 | 上海市农业科学院 | Pleurotus eryngii fruiting method |
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2020
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112125732A (en) * | 2020-09-19 | 2020-12-25 | 江苏丰收菇业有限公司 | Velvet antler mushroom liquid stock culture solution and velvet antler mushroom stock preparation process |
| CN115152532A (en) * | 2022-06-08 | 2022-10-11 | 湖南果秀食品有限公司 | Quantitative and visual pleurotus eryngii liquid shake flask stock seed production method |
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