CN111345199A - Black skin termitomyces liquid strain culture solution and preparation method thereof - Google Patents
Black skin termitomyces liquid strain culture solution and preparation method thereof Download PDFInfo
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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
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Abstract
The invention discloses a black skin termitomyces liquid strain culture solution and a preparation method thereof, wherein each liter of the culture solution comprises the following components: 1.5-2.5 g of peptone, 2.0-3.0 g of flour, 3.2-4.2 g of corn flour, 6.8-7.8 g of soybean flour, 4.0-6.0 g of glucose, 9.2-10.2 g of brown sugar, 0.5-1.5 g of magnesium sulfate, 1.5-2.5 g of monopotassium phosphate, 1.5-2.5 g of calcium carbonate, 0.3-1.0 mL of defoaming agent and the balance of water; the culture solution can be simultaneously used for the culture of the black skin termitomyces albuminosus and the production process of products, can ensure that the black skin termitomyces albuminosus and the hypha quickly germinate, is convenient to inoculate, effectively improves the production efficiency, and greatly reduces the production cost. The culture solution has the advantages of cheap and easily obtained components, simple preparation method, obvious effect and wide practical production and application value.
Description
Technical Field
The invention relates to the field of artificial cultivation of termitomyces albuminosus, in particular to a black termitomyces albuminosus liquid strain culture solution and a preparation method thereof.
Background
The black skin termitomyces albuminosus is a nutritious edible fungus, is rich in various amino acids, and is one of edible fungi and medicinal fungi. The fresh, tender and mellow meat, delicate and white meat, unique taste, edible raw and cooked, dual-purpose of food and medicine and rich nutrition are popular among people.
At present, people adopt advanced scientific technology to introduce the black-skin termitomyces albuminosus into a greenhouse for cultivation, so that the artificial cultivation of the black-skin termitomyces albuminosus is realized. At present, the artificial cultivation of the black skin termitomyces albuminosus mainly adopts a solid strain culture medium, has a plurality of defects and deficiencies, and is mainly represented as follows: the growth period of the strains is long, the management is inconvenient, the production cost of the strains is high, the fungus ages are inconsistent, the fruiting is irregular, and the like.
Disclosure of Invention
The invention mainly solves the technical problem of providing the black termitomyces albuminosus liquid strain culture solution and the preparation method thereof, and the black termitomyces albuminosus can be quickly and efficiently cultured.
In order to solve the technical problems, the invention adopts a technical scheme that:
the black skin termitomyces liquid strain culture solution is characterized by comprising the following components: each liter of culture medium comprises: 1.5-2.5 g of peptone, 2.0-3.0 g of flour, 3.2-4.2 g of corn flour, 6.8-7.8 g of soybean flour, 4.0-6.0 g of glucose, 9.2-10.2 g of brown sugar, 0.5-1.5 g of magnesium sulfate, 1.5-2.5 g of monopotassium phosphate, 1.5-2.5 g of calcium carbonate and 0.3-1.0 mL of defoaming agent;
further, the composition comprises the following components: each liter of culture medium comprises: 1.8-2.2 g of peptone, 2.2-2.8 g of flour, 3.5-4.0 g of corn flour, 7.0-7.5 g of soybean flour, 4.5-5.5 g of glucose, 9.5-10 g of brown sugar, 0.8-1.2 g of magnesium sulfate, 1.8-2.2 g of monopotassium phosphate, 1.8-2.2 g of calcium carbonate and 0.4-0.8 mL of defoaming agent.
Further, the composition comprises the following components: each liter of culture medium comprises: 2.0g of peptone, 2.5g of flour, 3.7g of corn flour, 7.3g of soybean flour, 5.0g of glucose, 9.7g of brown sugar, 1.0g of magnesium sulfate, 2.0g of monopotassium phosphate, 2.0g of calcium carbonate and 0.6mL of defoaming agent.
In a particular embodiment of the invention, the balance is water; further, the water is distilled water.
Further, the defoaming agent is a polyether defoaming agent; further, the polyether defoaming agent is selected from one of GP type polyether defoaming agent, GPE type polyether defoaming agent and PPE type polyether defoaming agent.
Further, the peptone is selected from fresh bone peptone and/or yeast peptone.
In a particular embodiment of the invention, the peptone comprises: 14.5 percent of nitrogen, more than or equal to 2 percent of amino acid, 0.8 percent of tryptophan, less than or equal to 10 percent of ash, less than or equal to 5 percent of drying weight loss, less than 3 percent of sodium chloride, no alkaline precipitate and no phosphate precipitate, wherein the percentages refer to the weight ratio.
Further, the flour is selected from one or more of strong flour, medium flour, low flour and non-strong flour; furthermore, the fineness of the flour is more than or equal to 60 meshes, the fineness of the corn flour is more than or equal to 60 meshes, and the fineness of the soybean flour is more than or equal to 60 meshes.
The corn meal is a powder of corn particles.
The invention also provides a preparation method of the black skin termitomyces liquid strain culture solution, which comprises the following steps:
(1) mixing peptone, flour, corn flour, soybean flour, glucose, brown sugar, magnesium sulfate, potassium dihydrogen phosphate, calcium carbonate, defoaming agent and water, and stirring;
(2) filtering the stirred mixture to remove particles to obtain the product;
further, stirring all the raw materials for 10-40 min at 1500-4000 revolutions per minute by a stirrer; further, stirring is carried out for 20-30 min at 2000-3000 r/min.
Further, the culture solution is sterilized before use; further, the sterilization is steam sterilization; further, the sterilization temperature is 120-125 ℃, and the sterilization time is 1-3 h.
The invention also provides a culture method of the black skin termitomyces albuminosus liquid strain, which is used for culturing the black skin termitomyces albuminosus liquid strain culture solution.
Further, inoculating a black skin termitomyces albuminosus mother strain into the black skin termitomyces albuminosus liquid strain culture solution, and performing shake culture for 2-4 days at 22-25 ℃ by using a constant temperature oscillator to obtain the black skin termitomyces albuminosus liquid strain culture solution.
Further, the cultivation method of the black skin termitomyces albuminosus liquid strain further comprises an expanding propagation step.
Further, the expanding propagation step is to mix the prepared liquid strain and the culture solution of the black skin termitomyces albuminosus liquid strain according to the volume ratio of 1:100, and continuously stir and culture for 4-6 days to obtain the black skin termitomyces albuminosus liquid strain.
Further, the incubation process is performed in a sterile environment.
The invention has the beneficial effects that:
(1) the culture solution can be used for preparing the liquid strains of the black skin termitomyces albuminosus by high-efficiency artificial culture, the cost is lower than that of a solid culture medium, each bag of the strains only needs a few minutes of money, and the cost is about one fifth of that of the solid strains, so that the production cost is greatly reduced; meanwhile, liquid strain inoculation is adopted, the working efficiency can be improved by 4-5 times compared with solid strain inoculation, the production efficiency is effectively improved, and more economic benefits are brought.
(2) The liquid strain of the black skin termitomyces albuminosus cultured by the culture solution has the advantages of simple process, high strain germination speed and high strain purity, and can ensure the subsequent robust fruiting.
(3) The germination speed of the black skin termitomyces albuminosus in the culture solution of the invention is far faster than the breeding speed of mixed fungi, and the mixed fungi hardly have the opportunity of breeding, thereby overcoming the technical problem that the black skin termitomyces albuminosus liquid culture is easy to be polluted by the mixed fungi, and ensuring the quality of the culture.
(4) The culture solution can also be used for producing black termitomyces albuminosus bags, the black termitomyces albuminosus strains have strong permeability in the culture solution, germinate at multiple points after inoculation, grow inside and outside together, the hyphae can germinate after 6-12 hours, can grow 7-10 days earlier than a solid culture medium and fill the fungus bags, can collect mushrooms 10 days earlier, has the advantages of vigorous hypha growth and short fungus age, and has higher quality and yield than the solid culture medium production mode, thereby winning the market and gaining benefits.
(5) The culture solution can be used for cultivating the black skin termitomyces liquid strain, can also be used for the expanded propagation cultivation of the black skin termitomyces liquid strain, can also be used as a culture solution for production to directly cultivate the black skin termitomyces, does not need to prepare various different culture solutions in the production process, is further convenient for production, and improves the production efficiency.
Detailed Description
The technical solutions of the present invention are described clearly and completely below, and it is obvious that the described embodiments are some, not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In this embodiment:
the peptone and the polyether antifoaming agent are all conventional products sold in the market, wherein in the peptone: 14.5 percent of nitrogen, more than or equal to 2 percent of amino acid, 0.8 percent of tryptophan, less than or equal to 10 percent of ash, less than or equal to 5 percent of drying weight loss, less than 3 percent of sodium chloride, no alkaline precipitate and no phosphate precipitate.
The wheat flour, the corn flour and the soybean flour are all products with the fineness larger than or equal to 60 meshes sold in the market;
the glucose is pure glucose without sodium, protein and fat, and the carbohydrate is more than or equal to 90 percent;
the stainless steel culture tank is a special stainless steel culture tank for edible fungi sold in the market, and has stirring and cooling functions.
Example 1
(1) Uniformly mixing 2g of peptone, 2.5g of wheat flour, 3.7g of corn flour, 7.3g of soybean flour, 5g of glucose, 9.7g of brown sugar, 1g of magnesium sulfate, 2g of monopotassium phosphate, 2g of calcium carbonate and 0.6mL of GP type polyether defoamer, adding 1L of distilled water, stirring at a high speed of a stirrer at the speed of 2000 rpm for 30 minutes, filtering with a fine net until no particles exist, adding the obtained liquid without particles into a steam sterilization pot, sterilizing at 125 ℃ with steam for 2 hours, and naturally cooling to 20-30 ℃ to obtain the black skin termitomyces liquid strain culture solution.
(2) Adding the black skin termitomyces liquid strain culture solution into a triangular flask, inoculating the black skin termitomyces mother strain into the high culture solution in the triangular flask, placing into a constant temperature oscillator, culturing at 23 ℃ for 3 days by continuous oscillation, performing microscope examination on the black skin termitomyces strain, observing that the strain balls germinate fully, and indicating that high-quality liquid strains are obtained, and further performing expanded propagation on the liquid strains.
(3) Preparing 100 liters of culture solution according to the formula of the raw materials according to the proportion in the step (1), and sterilizing the culture solution by using steam in a steam sterilization pot at the temperature of 120-125 ℃ for 2 hours. Introducing the prepared culture solution into a stainless steel culture tank, naturally cooling to 20-30 ℃, and preparing liquid strains according to the following steps: pouring the culture solution in a volume ratio of 1:100, continuously stirring and culturing for 5 days, carrying out microscopy on the black termitomyces albuminosus strain, and observing that the fungus balls germinate fully.
Example 2
(1) Uniformly mixing 1.8g of peptone, 2.2g of wheat flour, 4.0g of corn flour, 7.5g of soybean flour, 5.5g of glucose, 9.5g of brown sugar, 0.8g of magnesium sulfate, 1.8g of monopotassium phosphate, 1.8g of calcium carbonate and 0.4mL of GPE type polyether defoamer, adding 1L of distilled water, stirring at a high speed of 3000 rpm for 20 minutes, filtering with a fine net until no particles exist, adding the obtained particle-free liquid into a steam sterilization pot, sterilizing with steam at a temperature of 120 ℃ for 3 hours, and naturally cooling to 20-30 ℃ to obtain the termitomyces nigripes liquid strain culture solution.
(2) Adding the black skin termitomyces liquid strain culture solution into a triangular flask, inoculating the black skin termitomyces mother strain into the high culture solution in the triangular flask, placing into a constant temperature oscillator, continuously oscillating and culturing at 25 ℃ for 2 days, carrying out microscope examination on the black skin termitomyces strain, observing that a fungus ball is fully germinated, and indicating that high-quality liquid strains are obtained, and further carrying out expanded propagation on the liquid strains.
(3) Preparing 100 liters of culture solution according to the formula of the raw materials according to the proportion in the step (1), and sterilizing the culture solution by using steam in a steam sterilization pot at the temperature of 120 ℃ for 3 hours. Introducing the prepared culture solution into a stainless steel culture tank, naturally cooling to 20-30 ℃, and preparing liquid strains according to the following steps: pouring the culture solution in a volume ratio of 1:100, continuously stirring and culturing for 6 days, carrying out microscopy on the black termitomyces albuminosus strain, and observing that the fungus balls germinate fully.
Example 3
(1) 2.2g of peptone, 2.8g of wheat flour, 3.5g of corn flour, 7.0g of soybean flour, 4.5g of glucose, 10.0g of brown sugar, 1.2g of magnesium sulfate, 2.2g of monopotassium phosphate, 2.2g of calcium carbonate and 0.8mL of PPE type polyether defoamer are uniformly mixed, 1L of distilled water is added for high-speed stirring, the speed of a stirrer is 2000 rpm, the stirring time is 30 minutes, the mixture is filtered by a fine mesh until no particles exist, the obtained particle-free liquid is added into a steam sterilization pot, the steam sterilization temperature is 125 ℃, the time is 1 hour, and the liquid is naturally cooled to 20-30 ℃ to obtain the black skin termitomyces liquid strain culture solution.
(2) Adding the black skin termitomyces liquid strain culture solution into a triangular flask, inoculating the black skin termitomyces mother strain into the high culture solution in the triangular flask, placing into a constant temperature oscillator, culturing at 22 ℃ for 4 days by continuous oscillation, performing microscope examination on the black skin termitomyces strain, observing that the strain balls germinate fully, and indicating that high-quality liquid strains are obtained, and further performing expanded propagation on the liquid strains.
(3) Preparing 100 liters of culture solution according to the formula of the raw materials in proportion according to the step (1), and sterilizing the culture solution by using steam in a steam sterilization pot at the temperature of 125 ℃ for 1 hour. Introducing the prepared culture solution into a stainless steel culture tank, naturally cooling to 20-30 ℃, and preparing liquid strains according to the following steps: pouring the culture solution in a volume ratio of 1:100, continuously stirring and culturing for 5 days, carrying out microscopy on the black termitomyces albuminosus strain, and observing that the fungus balls germinate fully.
Example 4
(1) Uniformly mixing 2.2g of peptone, 2.3g of wheat flour, 3.7g of corn flour, 7.3g of soybean flour, 5.5g of glucose, 9.7g of brown sugar, 1g of magnesium sulfate, 2g of monopotassium phosphate, 2g of calcium carbonate and 0.6mL of GP type polyether defoamer, adding 1L of distilled water, stirring at a high speed of a stirrer of 2000 rpm for 30 minutes, filtering with a fine mesh until no particles exist, adding the obtained liquid without particles into a steam sterilization pot, sterilizing with steam at a temperature of 125 ℃ for 2 hours, and naturally cooling to 20-30 ℃ to obtain the liquid culture solution of the termitomyces nigripes.
(2) Adding the black skin termitomyces liquid strain culture solution into a triangular flask, inoculating the black skin termitomyces mother strain into the high culture solution in the triangular flask, placing into a constant temperature oscillator, culturing at 23 ℃ for 3 days by continuous oscillation, performing microscope examination on the black skin termitomyces strain, observing that the strain balls germinate fully, and indicating that high-quality liquid strains are obtained, and further performing expanded propagation on the liquid strains.
(3) Preparing 100 liters of culture solution according to the formula of the raw materials according to the proportion in the step (1), and sterilizing the culture solution by using steam in a steam sterilization pot at the temperature of 125 ℃ for 2 hours. Introducing the prepared culture solution into a stainless steel culture tank, naturally cooling to 20-30 ℃, and preparing liquid strains according to the following steps: pouring the culture solution in a volume ratio of 1:100, continuously stirring and culturing for 5 days, carrying out microscopy on the black termitomyces albuminosus strain, and observing that the fungus balls germinate fully.
Example 5
The liquid strains prepared in the embodiments 1 to 4 are used as a culture solution for production, inoculation is carried out according to a conventional method to prepare a strain bag and culture, after inoculation, multiple points germinate, the inner side and the outer side grow up and down together, hyphae can germinate within 6 to 12 hours, the hyphae can grow over a strain bag 7 to 10 days earlier than a solid culture medium, the fruiting is robust, and the fruiting can be picked about 10 days earlier.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. The black skin termitomyces liquid strain culture solution is characterized by comprising the following components: each liter of culture medium comprises: 1.5-2.5 g of peptone, 2.0-3.0 g of flour, 3.2-4.2 g of corn flour, 6.8-7.8 g of soybean flour, 4.0-6.0 g of glucose, 9.2-10.2 g of brown sugar, 0.5-1.5 g of magnesium sulfate, 1.5-2.5 g of monopotassium phosphate, 1.5-2.5 g of calcium carbonate and 0.3-1.0 mL of defoaming agent;
further, the composition comprises the following components: each liter of culture medium comprises: 1.8-2.2 g of peptone, 2.2-2.8 g of flour, 3.5-4.0 g of corn flour, 7.0-7.5 g of soybean flour, 4.5-5.5 g of glucose, 9.5-10 g of brown sugar, 0.8-1.2 g of magnesium sulfate, 1.8-2.2 g of monopotassium phosphate, 1.8-2.2 g of calcium carbonate and 0.4-0.8 mL of defoaming agent.
Further, the composition comprises the following components: each liter of culture medium comprises: 2.0g of peptone, 2.5g of flour, 3.7g of corn flour, 7.3g of soybean flour, 5.0g of glucose, 9.7g of brown sugar, 1.0g of magnesium sulfate, 2.0g of monopotassium phosphate, 2.0g of calcium carbonate and 0.6mL of defoaming agent.
2. The liquid culture medium for termitomyces nigricans according to claim 1, wherein the balance is water; further, the water is distilled water.
3. The liquid termitomyces albuminosus strain culture solution according to claim 1 or 2, wherein the antifoaming agent is a polyether antifoaming agent; further, the polyether defoaming agent is selected from one of GP type polyether defoaming agent, GPE type polyether defoaming agent and PPE type polyether defoaming agent.
4. The liquid culture medium for the termitomyces melanocorticus according to claim 1 or 2, wherein the peptone is selected from the group consisting of fresh bone peptone and/or yeast peptone; further, in the peptone: 14.5 percent of nitrogen, more than or equal to 2 percent of amino acid, 0.8 percent of tryptophan, less than or equal to 10 percent of ash, less than or equal to 5 percent of drying weight loss, less than 3 percent of sodium chloride, no alkaline precipitate and no phosphate precipitate, wherein the percentages refer to the weight ratio.
5. The liquid culture medium for termitomyces albuminosus according to claim 1 or 2, wherein the flour is selected from one or more of strong flour, medium flour, low flour and weak flour; furthermore, the fineness of the flour is more than or equal to 60 meshes, the fineness of the corn flour is more than or equal to 60 meshes, and the fineness of the soybean flour is more than or equal to 60 meshes.
6. The method for preparing a culture solution of Collybia nigricans liquid strain according to any one of claims 1 to 5, comprising the following steps:
(1) mixing peptone, flour, corn flour, soybean flour, glucose, brown sugar, magnesium sulfate, potassium dihydrogen phosphate, calcium carbonate, defoaming agent and water, and stirring;
(2) filtering the stirred mixture to remove particles to obtain the product;
further, stirring all the raw materials for 10-40 min at 1500-4000 revolutions per minute by a stirrer; further, stirring is carried out for 20-30 min at 2000-3000 r/min.
7. The method for preparing a culture solution of liquid spawn of termitomyces nigricans according to claim 6, wherein the culture solution is sterilized before use; further, the sterilization is steam sterilization; further, the sterilization temperature is 120-125 ℃, and the sterilization time is 1-3 h.
8. A method for culturing a liquid culture of Collybia melanocortis, which is characterized in that the culture solution of the liquid culture of Collybia melanocortis according to claim 1 or 2 is used for culturing; further, inoculating a black skin termitomyces albuminosus mother strain into the black skin termitomyces albuminosus liquid strain culture solution, and performing shake culture for 2-4 days at 22-25 ℃ by using a constant temperature oscillator to obtain the black skin termitomyces albuminosus liquid strain culture solution.
9. The method for cultivating a liquid spawn of Collybia nigricans according to claim 8, further comprising the step of expanding propagation; further, the expanding propagation step is to mix the prepared liquid spawn and the black skin termitomyces albuminosus liquid spawn culture solution according to the volume ratio of 1:100, and continuously stir and culture for 4-6 days, thus obtaining the black skin termitomyces albuminosus liquid spawn culture solution.
10. The method for cultivating a liquid spawn of Collybia nigricans according to claim 8, wherein the cultivation process is performed in an aseptic environment.
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