CN112779165A - Mushroom foot powder preservation culture medium for hypsizigus marmoreus strains and preparation method of mushroom foot powder preservation culture medium - Google Patents
Mushroom foot powder preservation culture medium for hypsizigus marmoreus strains and preparation method of mushroom foot powder preservation culture medium Download PDFInfo
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Abstract
The invention discloses a mushroom foot powder preservation culture medium for hypsizigus marmoreus strains and a preparation method thereof, and belongs to the field of strain preservation. The mushroom foot powder preservation culture medium comprises potato leach liquor, glucose (20g/L), agar (20g/L), mushroom foot powder and the like, wherein the potato leach liquor is prepared by putting 200g of diced potatoes into 800ml of water, heating and boiling the diced potatoes until the diced potatoes can be squeezed by a glass rod, filtering the diced potatoes by 8 layers of gauze to obtain filtrate, and preparing 1L of culture medium by using the filtrate; the mushroom foot powder is one of edible mushroom foot powders such as shiitake mushroom, crab-flavor mushroom, white beech mushroom, pleurotus eryngii, seafood mushroom and the like. The preservation culture medium can ensure that the preserved hypsizigus marmoreus strain hyphae has high growth speed, high activity and difficult degeneration, and the culture medium has low price, simple and convenient preparation method, and the obtained strain has high purity, can be widely applied in industrial production, and has obvious market prospect.
Description
Technical Field
The invention belongs to the field of strain preservation, and particularly relates to a culture medium for preserving mushroom foot powder of a hypsizigus marmoreus strain and a preparation method thereof.
Background
The hypsizigus marmoreus is one of important edible fungi which are industrially produced at present, enjoys the reputation of smelling tricholoma matsutake and eating Hypsizigus marmoreus, is a rare edible fungus which is rich in nutrition and can be used as both medicine and food, contains eight essential amino acids required by a human body, has various effects of improving the immunity of the human body, preventing aging, prolonging the service life and the like, particularly has higher content of lysine and arginine than that of common mushrooms, is beneficial to improving the intelligence of teenagers and increasing the height, and has the effects of resisting cancer and reducing cholesterol.
The strain is an important biological resource and production data, and is an indispensable important material in the production link of edible fungi. The preservation of the strain generally requires that the excellent characteristics of the edible fungi are kept unchanged, no variation occurs within a certain period of time, the living ability is kept, and the metabolism of the edible fungi is in a state of being least active or relatively static. These requirements determine that the seed culture preservation medium should provide all the nutrients necessary for seed growth and should be supplied slowly. At present, in the factory production, hypsizigus marmoreus strains are generally subjected to slant preservation in a test tube by using a PDA (Potato dextrose agar) culture medium, and subculture is performed every 90 days, so that the problems of high preservation cost, poor quality, high pollution rate and the like exist.
A large amount of mushroom feet can be produced in the production process of mushrooms, the nutritional value of the mushroom feet is similar to that of fruiting bodies, the mushroom feet contain active ingredients such as polysaccharide, crude protein, crude fat, amino acid, small peptide, flavone, saponin, vitamin and the like, and the mushroom feet are usually thrown away as waste, so that the environment is polluted, and resources are wasted. The invention provides a culture medium for preserving mushroom foot powder of hypsizigus marmoreus strains, which has the characteristics of low price, good purity of preserved strains, high activity, simple preparation method, easy popularization and the like.
Disclosure of Invention
The invention aims to provide a culture medium for preserving mushroom foot powder of hypsizigus marmoreus strains and a preparation method thereof, wherein the culture medium is low in price, the preserved strains are high in purity and activity and difficult to degenerate, the preparation method is simple and easy to popularize, and the variety and range of the culture medium for preserving the hypsizigus marmoreus strains can be effectively widened.
The invention comprises the following two aspects:
a culture medium for preserving mushroom foot powder of hypsizigus marmoreus strain is characterized by comprising potato extract, glucose, agar, mushroom foot powder and the like.
Preferably, the potato leaching liquor is a filtrate obtained by putting 200g of potato dices in 800ml of water, heating and boiling until the potato dices can be squeezed by a glass rod, and filtering by 8 layers of gauze, wherein the filtrate can be used for preparing 1L of culture medium.
Preferably, the glucose is analytically pure and has a concentration of 20-30 g/L.
Preferably, the agar is analytically pure and has a concentration of 20-30 g/L.
Preferably, the mushroom foot powder is one of edible mushroom foot powders such as shiitake mushroom, hypsizygus marmoreus, pleurotus eryngii, hypsizygus marmoreus and the like.
More preferably, the mushroom foot powder is mushroom foot powder.
Preferably, the concentration of the mushroom foot powder is 1-20 g/L.
More preferably, the concentration of the shiitake mushroom foot powder is 5 g/L.
The invention also aims to provide a preparation method of the mushroom foot powder preservation medium, which comprises the following specific steps:
(1) preparing a potato extract, and specifically preparing: putting 200g of potato dices in 800ml of water, heating to boiling with strong fire, boiling with slow fire until the potato dices can be squeezed by a glass rod, and filtering to obtain filtrate, namely potato leaching liquor.
(2) And (2) transferring the potato leaching liquor obtained in the step (1) into a 2L big beaker, adding glucose, stirring until the glucose is dissolved, adding agar powder, and heating to boil while stirring.
(3) Adding the mushroom foot powder while the mixture is hot under stirring, continuously heating and stirring, and fixing the volume to 1L, and cooling.
(4) When the temperature is reduced to about 90 ℃, subpackaging the obtained culture medium, and sterilizing at 121 ℃.
Preferably, in the step (1), the screening is performed by using gauze, and the number of layers of the gauze is 8. The gauze after filtration was also squeezed out and the squeezed liquid was combined into the filtrate.
Preferably, in the step (3), the mushroom foot powder is obtained by tearing fresh mushroom feet into filaments, naturally drying or baking, grinding into powder, and sieving with a 100-mesh sieve.
Preferably, in the step (3), the heating and stirring time is 20 minutes.
Compared with the prior art, the invention has the beneficial effects that:
the mushroom foot preservation culture medium provided by the invention provides a large amount of nutrient elements and trace elements beneficial to the growth of hypsizigus marmoreus hyphae, can meet the basic requirements of hypsizigus marmoreus strain growth, not only keeps the original activity of the strain, but also ensures that the hyphae in the culture medium has high growth speed and active growth metabolism. The culture medium for preserving the mushroom foot of the hypsizigus marmoreus strain provided by the invention has a preservation effect superior to that of the existing slant preservation method, the activity of the preserved hyphae is the same as that of a first-generation mother strain, powerful guarantee is provided for simple preservation and large-scale cultivation of the hypsizigus marmoreus strain, and the culture medium has high application value, economic significance and social significance.
The specific implementation mode is as follows:
the invention is described below by means of specific embodiments. Unless otherwise specified, the technical means used in the present invention are well known to those skilled in the art. In addition, the embodiments should be considered illustrative, and not restrictive, of the scope of the invention, which is defined solely by the claims. It will be apparent to those skilled in the art that various changes or modifications in the components and amounts of the materials used in these embodiments can be made without departing from the spirit and scope of the invention. The present invention is further illustrated by the following specific examples.
Example 1: air-dried shiitake mushroom foot powder preservation culture medium and preparation method thereof
(1) Putting 200g of potato dices in 800ml of water, heating to boiling with strong fire, boiling with slow fire until the potato dices can be squeezed by a glass rod, filtering for 8 layers, and obtaining filtrate which is potato leaching liquor.
(2) And (2) transferring the potato leaching liquor obtained in the step (1) into a 2L big beaker, adding 20g of glucose, stirring and dissolving, adding 20g of agar powder, and heating to boil while stirring.
(3) Adding air-dried shiitake mushroom foot powder 5g sieved by a 100-mesh sieve while stirring, continuously heating and stirring for 20 minutes, metering the volume to 1L, and cooling.
(4) When the temperature is reduced to about 90 ℃, subpackaging the obtained culture medium, and sterilizing at 121 ℃.
Example 2: dried mushroom foot powder preservation culture medium and preparation method thereof
(1) Putting 200g of potato pieces into 800ml of water, heating to boil with strong fire, boiling with slow fire until the potato pieces can be squeezed by a glass rod, filtering for 9 layers, and obtaining filtrate which is potato leaching liquor.
(2) And (2) transferring the potato leaching liquor obtained in the step (1) into a 2L big beaker, adding 20g of glucose, stirring and dissolving, adding 20g of agar powder, and heating to boil while stirring.
(3) Adding 10g of dried mushroom residue powder which is sieved by a 100-mesh sieve into the hot mushroom residue powder while stirring, continuously heating and stirring the mixture for 20 minutes, and carrying out constant volume treatment to 1L and cooling the mixture.
(4) When the temperature is reduced to about 90 ℃, subpackaging the obtained culture medium, and sterilizing at 121 ℃.
Example 3: air-dried white beech mushroom foot powder preservation culture medium and preparation method thereof
(1) Putting 200g of potato dices in 800ml of water, heating to boiling with strong fire, boiling with slow fire until the potato dices can be squeezed by a glass rod, filtering for 8 layers, and obtaining filtrate which is potato leaching liquor.
(2) And (2) transferring the potato leaching liquor obtained in the step (1) into a 2L big beaker, adding 20g of glucose, stirring and dissolving, adding 20g of agar powder, and heating to boil while stirring.
(3) Adding 15g of white beech mushroom foot powder which is air-dried and sieved by a 100-mesh sieve while the mixture is hot under stirring, continuously heating and stirring for 20 minutes, metering the volume to 1L, and cooling.
(4) When the temperature is reduced to about 90 ℃, subpackaging the obtained culture medium, and sterilizing at 121 ℃.
Example 4: preservation culture medium for dried hypsizygus marmoreus foot powder and preparation method thereof
(1) Putting 200g of potato dices in 800ml of water, heating to boiling with strong fire, boiling with slow fire until the potato dices can be squeezed by a glass rod, filtering for 8 layers, and obtaining filtrate which is potato leaching liquor.
(2) And (2) transferring the potato leaching liquor obtained in the step (1) into a 2L big beaker, adding 20g of glucose, stirring and dissolving, adding 20g of agar powder, and heating to boil while stirring.
(3) Adding 20g of white beech mushroom foot powder which is dried and sieved by a 100-mesh sieve while stirring, continuously heating and stirring for 20 minutes, metering the volume to 1L, and cooling.
(4) When the temperature is reduced to about 90 ℃, subpackaging the obtained culture medium, and sterilizing at 121 ℃.
Comparative example: PDA slant preservation culture medium
Weighing 25g of potato starch and glucose mixture (PD powder) and 20g of agar, placing in a beaker, adding a proper amount of water, heating to dissolve, and adding water to a constant volume of 1L. Subpackaging, and sterilizing at 121 deg.C for 20 min. And (5) after sterilization, swinging an inclined plane.
Determination of Strain preservation Properties
Transferring the hypsizigus marmoreus slant mother strain to a PDA plate, culturing in an incubator at 25 deg.C until the plate is full of mycelia.
A9 mm-diameter punch was used to punch a plate with hyphae to obtain 9mm clumps, and one clump was picked up and inoculated on the test tube slant of the medium obtained in the different examples and the control and cultured in an incubator at 25 ℃. After 15 days of culture, the growth morphology of the colonies and other characteristics were observed to examine whether or not they were contaminated with infectious microbes. After checking and confirming that no error exists, the well grown inclined test tube is placed in a refrigerator at 4 ℃ for preservation, and 3 test tube inclined planes are preserved in parallel. Hypha growth rate test was performed after 3 months of storage.
And (3) detecting the strain preservation effect: hyphal growth rate V
Resuscitating and culturing a slant test tube preserved for 3 months at 25 ℃ for 3 days, inoculating a strain block with the diameter of 9mm to a PDA plate, measuring the diameter of a bacterial colony after hyphae germinates to be 1, and measuring the diameter of the bacterial colony after continuously culturing for 48 hours at 25 ℃ to be 2, and calculating by the following formula:
hypha growth rate V (cm/d) ═ colony diameter 2-colony diameter 1/time (d)
The effect of strain preservation is shown in Table 1.
TABLE 1 preservation Effect of culture Medium for preserving Agaricus blazei Murill
As can be seen from Table 1, when the culture medium for preserving the mushroom foot powder provided by the invention preserves hypsizigus marmoreus strains, the hypha growth rate V of the rest culture mediums for preserving the mushroom foot powder is obviously higher than that of a control group except that the hypha growth rate of the 5mg/L air-dried hypsizigus marmoreus foot powder culture medium is slightly lower than that of the control group, particularly the hypha growth rate of the 5mg/L dried culture medium for preserving the mushroom foot powder reaches 3.07mm/d and is 34% higher than that of the control group, and the results show that the culture medium for preserving the mushroom foot powder not only can effectively preserve strains, but also can effectively improve the hypha growth rate of the preserved strains.
Claims (10)
1. A mushroom foot powder culture medium for preserving hypsizigus marmoreus strains is characterized in that: comprises potato extract, glucose, agar and mushroom foot powder.
2. The culture medium for preserving mushroom foot powder for hypsizigus marmoreus strain according to claim 1, wherein: the potato leaching liquor is prepared by putting 200g of diced potatoes in 800ml of water, heating and boiling until the diced potatoes can be squeezed by a glass rod, filtering by 8 layers of gauze to obtain filtrate, and preparing 1L of culture medium by using the filtrate.
3. The culture medium for preserving mushroom foot powder for hypsizigus marmoreus strain according to claim 1, wherein: the glucose is analytically pure, and the concentration of the glucose is 20-30 g/L.
4. The culture medium for preserving mushroom foot powder for hypsizigus marmoreus strain according to claim 1, wherein: the agar is analytically pure, and the concentration of the agar is 20-30 g/L.
5. The culture medium for preserving mushroom foot powder for hypsizigus marmoreus strain according to claim 1, wherein: the mushroom foot powder is one of edible mushroom foot powders such as shiitake mushroom, crab-flavor mushroom, white jade mushroom, pleurotus eryngii, seafood mushroom and the like.
6. The culture medium for preserving mushroom foot powder for hypsizigus marmoreus strain according to claim 5, wherein: the concentration of the mushroom foot powder is 5-20 g/L.
7. The method for preparing a culture medium for preserving mushroom foot powder according to any one of claims 1 to 6, wherein: the method comprises the following specific steps:
(1) preparing a potato extract, and specifically preparing: putting 200g of potato dices in 800ml of water, heating to boiling with strong fire, boiling with slow fire until the potato dices can be squeezed by a glass rod, and filtering to obtain filtrate, namely potato leaching liquor.
(2) And (2) transferring the potato leaching liquor obtained in the step (1) into a 2L big beaker, adding glucose, stirring until the glucose is dissolved, adding agar powder, and heating to boil while stirring.
(3) Adding the mushroom foot powder while the mixture is hot under stirring, continuously heating and stirring, and fixing the volume to 1L, and cooling.
(4) When the temperature is reduced to about 90 ℃, subpackaging the obtained culture medium and sterilizing.
8. The method for preparing a culture medium for preserving mushroom foot powder according to claim 7, wherein: in the step (1), gauze is used for sieving, and the number of layers of the gauze is 8. The gauze after filtration was also squeezed out and the squeezed liquid was combined into the filtrate.
9. The method for preparing a culture medium for preserving mushroom foot powder according to claim 7, wherein: in the step (3), the mushroom foot powder is obtained by tearing fresh mushroom feet into filaments, naturally drying or baking, grinding into powder and sieving with a 100-mesh sieve.
10. The method for preparing a culture medium for preserving mushroom foot powder according to claim 7, wherein: in the step (3), the heating and stirring time is 20 minutes.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113528351A (en) * | 2020-04-20 | 2021-10-22 | 天津农学院 | Sugar-free crab-flavor mushroom mother culture medium containing mushroom foot powder and preparation method thereof |
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| CN103155785A (en) * | 2011-12-09 | 2013-06-19 | 天津市金三农农业科技开发有限公司 | Establishment method for brown mushroom three-level strain propagation system |
| CN105493897A (en) * | 2016-01-08 | 2016-04-20 | 周琪 | Industrialized cultivation method of beech mushrooms |
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- 2019-11-08 CN CN201911080642.6A patent/CN112779165B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103155785A (en) * | 2011-12-09 | 2013-06-19 | 天津市金三农农业科技开发有限公司 | Establishment method for brown mushroom three-level strain propagation system |
| CN105493897A (en) * | 2016-01-08 | 2016-04-20 | 周琪 | Industrialized cultivation method of beech mushrooms |
Non-Patent Citations (3)
| Title |
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| MIN LIU ET AL.: "The characterization,renoprotection and antioxidation of enzymatic and acidic exopolysaccharides from Hypsizigus marmoreus", 《SCI REP》 * |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113528351A (en) * | 2020-04-20 | 2021-10-22 | 天津农学院 | Sugar-free crab-flavor mushroom mother culture medium containing mushroom foot powder and preparation method thereof |
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