CN113383677A - Hypsizigus marmoreus tissue isolation medium and preparation method thereof - Google Patents
Hypsizigus marmoreus tissue isolation medium and preparation method thereof Download PDFInfo
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- 238000002955 isolation Methods 0.000 title claims abstract description 37
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 239000001963 growth medium Substances 0.000 claims abstract description 83
- 238000012360 testing method Methods 0.000 claims abstract description 82
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 36
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 32
- 240000008042 Zea mays Species 0.000 claims abstract description 28
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 28
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 28
- 235000005822 corn Nutrition 0.000 claims abstract description 28
- 239000002994 raw material Substances 0.000 claims abstract description 24
- 239000002609 medium Substances 0.000 claims abstract description 23
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims abstract description 22
- 230000001954 sterilising effect Effects 0.000 claims abstract description 20
- 239000000843 powder Substances 0.000 claims abstract description 19
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 18
- 229920001817 Agar Polymers 0.000 claims abstract description 17
- 239000008272 agar Substances 0.000 claims abstract description 17
- 239000001888 Peptone Substances 0.000 claims abstract description 16
- 108010080698 Peptones Proteins 0.000 claims abstract description 16
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims abstract description 16
- 235000019341 magnesium sulphate Nutrition 0.000 claims abstract description 16
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 16
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 16
- 235000019319 peptone Nutrition 0.000 claims abstract description 16
- 238000005303 weighing Methods 0.000 claims abstract description 16
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 15
- 239000008103 glucose Substances 0.000 claims abstract description 15
- 229910052742 iron Inorganic materials 0.000 claims abstract description 11
- 239000008213 purified water Substances 0.000 claims abstract description 11
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims abstract description 10
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 9
- 238000010411 cooking Methods 0.000 claims abstract description 6
- 239000003814 drug Substances 0.000 claims abstract description 6
- 238000001914 filtration Methods 0.000 claims abstract description 6
- 238000010438 heat treatment Methods 0.000 claims abstract description 6
- 238000003756 stirring Methods 0.000 claims abstract description 6
- 239000000706 filtrate Substances 0.000 claims description 21
- 238000000034 method Methods 0.000 claims description 16
- 238000001816 cooling Methods 0.000 claims description 10
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 6
- 239000012774 insulation material Substances 0.000 claims description 5
- 238000007789 sealing Methods 0.000 claims description 5
- 239000013589 supplement Substances 0.000 claims description 5
- 238000011049 filling Methods 0.000 claims description 2
- 239000002245 particle Substances 0.000 abstract description 7
- 240000007594 Oryza sativa Species 0.000 abstract 1
- 235000007164 Oryza sativa Nutrition 0.000 abstract 1
- 235000009566 rice Nutrition 0.000 abstract 1
- 230000012010 growth Effects 0.000 description 13
- 230000000052 comparative effect Effects 0.000 description 7
- 230000008901 benefit Effects 0.000 description 5
- 230000035784 germination Effects 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 239000001965 potato dextrose agar Substances 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 description 1
- 241001534815 Hypsizygus marmoreus Species 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241001558929 Sclerotium <basidiomycota> Species 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000028644 hyphal growth Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000000582 semen Anatomy 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 239000011716 vitamin B2 Substances 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
Classifications
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a hypsizigus marmoreus tissue isolation medium and a preparation method thereof, belonging to the technical field of biological strain cultivation and comprising the following components in parts by weight: 20 parts of corn; 20 parts of glucose; 20 parts of agar powder; 1 part of peptone; 1 part of magnesium sulfate; 1 part of monopotassium phosphate; 1000 parts of water. The preparation method comprises the following steps: step 1, weighing the medicine; step 2, pouring 1000mL of purified water into an iron pan, adding weighed corn particles, cooking, filtering the cooked liquid by using four layers of thick gauze, pouring the liquid into a beaker with the raw materials of the culture medium, stirring the raw materials uniformly, and putting the raw materials into the electric rice pan again for water bath heating until the raw materials are completely dissolved; step 3, subpackaging the test tubes and putting the test tubes into a sterilization pot for sterilization; step 4, after the temperature is reduced to a certain temperature, swinging an inclined plane; and 5, finishing the preparation of the culture medium and then starting to use.
Description
Technical Field
The invention relates to the technical field of biological strain cultivation, and designs a tissue isolation culture medium for hypsizigus marmoreus and a preparation method thereof.
Background
A method of placing a small piece of tissue such as fruit body, sclerotium, and cordis on a slant culture medium to germinate it into pure hyphae is called tissue isolation. The tissue separation has the advantages of simplicity, convenience, easy success, capability of keeping the original strain character and the like, but is easy to degenerate after being repeated for many times, and the tissue separation and the spore separation are carried out alternately. The technical field of strain cultivation in the prior art provides a formula and a preparation process of a hypsizigus marmoreus tissue isolation strain test tube culture medium, wherein the formula of the tissue isolation test tube culture medium is used in the prior art, and the formula in the prior art generally comprises 39g of PDA (potato dextrose agar) dry powder, 10g of agar powder, 3g of peptone, 1.5g of magnesium sulfate, 3g of monopotassium phosphate, 1g of yeast extract, vitamin B16 tablets and 1L of water.
The hypsizigus marmoreus prepared by the formula in the prior art has slow hypha growth, general strain quality, long strain growth period and high cost of the used culture medium, and cannot obtain better economic benefit.
Disclosure of Invention
The invention aims to provide a hypsizigus marmoreus tissue isolation culture medium, which improves the strain quality, shortens the hypha growth period, improves the hypha growth uniformity and reduces the ingredient cost of a culture medium.
A hypsizigus marmoreus tissue isolation culture medium comprises the following components in parts by weight:
15-25 parts of corn; 15-25 parts of glucose; 15-25 parts of agar powder; 0.5-1.5 parts of peptone; 0.5-1.5 parts of magnesium sulfate; 0.5-1.5 parts of monopotassium phosphate; 800 portions of water and 1200 portions of water.
Further, the hypsizigus marmoreus tissue isolation medium comprises the following components in parts by weight: 20 parts of corn; 20 parts of glucose; 20 parts of agar powder; 1 part of peptone; 1 part of magnesium sulfate; 1 part of monopotassium phosphate; 1000 parts of water.
The invention also provides a preparation method of the hypsizigus marmoreus tissue isolation medium.
A preparation method of hypsizigus marmoreus tissue isolation medium comprises the following steps:
step 1, weighing the medicine
Accurately weighing glucose, agar powder, peptone, magnesium sulfate and potassium dihydrogen phosphate according to a standard formula, and placing into a 1000ml beaker;
step 2 preparation of the Medium
Pouring 1000mL of purified water into an iron pan, adding the crushed corn into the iron pan, cooking, filtering the cooked liquid with four layers of thick gauze to obtain a filtrate, and pouring the filtrate into the beaker containing the culture medium raw material in the step 1; if the filtered filtrate is less than 1000mL, adding purified water to supplement 1000m L, stirring the raw materials and the filtrate uniformly, and heating in water bath again until the raw materials are completely dissolved to obtain a culture medium;
step 3, subpackaging test tubes
Subpackaging the culture medium obtained in the step 2 in test tubes, pouring the test tubes into clean test tubes when the test tubes are subpackaged before the culture medium is not solidified, filling 18ml of culture medium liquid into each used test tube, sealing after subpackaging is finished, and sterilizing;
step 4, cooling
Placing the test tube on a clean, spacious and flat table top after the culture medium in the test tube is cooled, placing and positioning the test tube at one time, wherein the length of the culture medium at the bottom end of the test tube is required to be 1cm, and covering a layer of heat insulation material on the surface of the test tube filled with the culture medium after placing is finished, so that the temperature of the culture medium is slowly reduced to be cooled, and the generation of test tube condensate water is reduced;
and 5, cooling the culture medium, and performing tissue isolation culture on the culture medium.
Preferably, in the step 1, the weighing precision is controlled within the range of +/-0.1.
Preferably, the sterilization condition in the step 3 is sterilization at 121 ℃ for 20min, and the whole process needs 50 min.
Preferably, the temperature before the bevel is swung in the step 4 is reduced to 55-60 ℃.
The hypsizigus marmoreus tissue isolation medium and the preparation method have the beneficial effects that:
(1) the invention adopts the formula of the cornAdjusting, and decocting semen Maydis in water to obtain filtrate containing abundant sugar, amino acids such as lysine, tryptophan and methionine, and trace elements such as vitamin A and vitamin B1Vitamin B2The added amino acid can be directly used by tissues, so that the strain quality is improved, the hypha growth period is shortened, the hypha growth uniformity is improved, and better economic benefit is obtained.
(2) The strain hyphae of the invention grow fast, the whole tissue separation experiment process is shortened, more labor cost is saved, and the aging degree of the hyphae is slowed down. And the strain using the corn formula culture medium has obvious and fast growth vigor, thick and white hypha state and good hypha quality.
(3) The method has the advantages that the used raw materials are low in cost, the yield and the quality of the hyphae can be improved, the cost of the culture medium can be reduced at the same time through the special matching among the corn kernels, the glucose and the agar powder, and the method has strong practicability and popularization.
Detailed Description
Example 1
A hypsizigus marmoreus tissue isolation culture medium comprises the following components in parts by weight:
15g of corn; 15g of glucose; 15g of agar powder; peptone 0.5 g; magnesium sulfate 0.5 g; potassium dihydrogen phosphate 0.5 g; 800mL of water.
Example 2
A hypsizigus marmoreus tissue isolation culture medium comprises the following components in parts by weight:
25g of corn; 25g of glucose; 25g of agar powder; peptone 1.5 g; magnesium sulfate 1.5 g; 1.5g of monopotassium phosphate; 1200mL of water.
Example 3
A hypsizigus marmoreus tissue isolation culture medium comprises the following components in parts by weight:
25g of corn; 15g of glucose; 15g of agar powder; peptone 1 g; 1g of magnesium sulfate; 1g of monopotassium phosphate; 1000mL of water.
Example 4
A hypsizigus marmoreus tissue isolation culture medium comprises the following components in parts by weight:
20g of corn; 20g of glucose; 20g of agar powder; peptone 1 g; 1g of magnesium sulfate; 1g of monopotassium phosphate; 1000mL of water.
Comparative example 1
The tissue isolation test tube culture medium in the prior art is enriched with 39g of PDA dry powder, 10g of agar powder, 3g of peptone, 1.5g of magnesium sulfate, 3g of monopotassium phosphate, 1g of yeast extract, vitamin B16 tablets and 1000mL of water.
A cost comparison of example 4 with comparative example 1 gave the results shown in tables 1 and 2;
TABLE 1 cost of enrichment formulation for comparative example 1 tissue isolation tube culture media
TABLE 2 EXAMPLE 4 cost of tissue isolation tube media
As can be seen from tables 1 and 2, the corn-based tissue isolation tube medium used in the present invention is less costly and provides greater economic benefits.
Example 5
A preparation method of hypsizigus marmoreus tissue isolation medium comprises the following steps:
step 1, weighing the medicine
Accurately weighing 20 parts of glucose, 20 parts of agar powder, 1 part of peptone, 1 part of magnesium sulfate and 1 part of potassium dihydrogen phosphate according to the raw material components of the culture medium in the embodiment 4, and putting the raw materials into a 1000ml beaker for later use;
specifically, the weighing precision is controlled within the range of +/-0.1;
step 2 preparation of the Medium
Pouring 1000mL of purified water into an iron pan, crushing corn in parts by mass into corn particles, putting the corn particles into the iron pan, cooking, and filtering the cooked liquid by using gauze with a thickness of four layers to obtain a filtrate; pouring the filtrate into the beaker containing the culture medium raw materials in the step 1, heating in a water bath and stirring to ensure that the culture medium raw materials are fully and completely dissolved in the filtrate;
in the actual operation process, if the filtrate of the corn is less than 1000mL, purified water can be added to supplement 1000 mL;
step 3, subpackaging test tubes
And (4) subpackaging the culture medium dissolved in the step (2) in a test tube. When the test tubes are subpackaged, before the culture medium is not solidified, subpackaging the test tubes by using a funnel and pouring the test tubes into prepared clean test tubes, wherein each used test tube contains 18ml of the culture medium, sealing by plugging a silica gel plug after subpackaging is finished, placing the test tubes containing the culture medium on a test tube rack, wrapping the whole test tube rack by using newspaper, and placing the test tubes into a sterilization pot for batch sterilization; wherein the sterilization condition is sterilization at 121 deg.C for 20min, and the whole test tube subpackaging process is controlled within 50 min.
Step 4, cooling
Placing the test tube on a clean, spacious and flat table top after the temperature is reduced to 60 ℃, placing and positioning the test tube at one time, wherein the length of a culture medium at the bottom end of the test tube is required to be 1cm, and covering a layer of heat insulation material on the test tube filled with the culture medium after placing is finished, so that the temperature of the culture medium is slowly reduced and cooled to room temperature, and the generation of test tube condensate water can be reduced;
and 5, cooling the culture medium, and performing tissue isolation culture on the culture medium.
Example 6
A preparation method of hypsizigus marmoreus tissue isolation medium comprises the following steps:
step 1, weighing the medicine
Accurately weighing 20 parts of glucose, 20 parts of agar powder, 1 part of peptone, 1 part of magnesium sulfate and 1 part of potassium dihydrogen phosphate according to the raw material components of the culture medium in the embodiment 4, and putting the raw materials into a 1000ml beaker for later use;
specifically, the weighing precision is controlled within the range of +/-0.1;
step 2 preparation of the Medium
Pouring 1000mL of purified water into an iron pan, crushing corn in parts by mass into corn particles, putting the corn particles into the iron pan, cooking, and filtering the cooked liquid by using gauze with a thickness of four layers to obtain a filtrate; pouring the filtrate into the beaker containing the culture medium raw materials in the step 1, heating in a water bath and stirring to ensure that the culture medium raw materials are fully and completely dissolved in the filtrate;
in the actual operation process, if the filtrate of the corn is less than 1000mL, purified water can be added to supplement 1000 mL;
step 3, subpackaging test tubes
And (4) subpackaging the culture medium dissolved in the step (2) in a test tube. When the test tubes are subpackaged, before the culture medium is not solidified, subpackaging the test tubes by using a funnel and pouring the test tubes into prepared clean test tubes, wherein each used test tube contains 18ml of the culture medium, sealing by plugging a silica gel plug after subpackaging is finished, placing the test tubes containing the culture medium on a test tube rack, wrapping the whole test tube rack by using newspaper, and placing the test tubes into a sterilization pot for batch sterilization; wherein the sterilization condition is sterilization at 121 deg.C for 20min, and the whole test tube subpackaging process is controlled within 50 min.
Step 4, cooling
Placing the test tube on a clean, spacious and flat table top after the temperature is reduced to 56 ℃, placing and positioning the test tube at one time, wherein the length of a culture medium at the bottom end of the test tube is required to be 1cm, and covering a layer of heat insulation material on the test tube filled with the culture medium after placing is finished, so that the temperature of the culture medium is slowly reduced and cooled to room temperature, and the generation of test tube condensate water can be reduced;
and 5, cooling the culture medium, and performing tissue isolation culture on the culture medium.
Example 7
A preparation method of hypsizigus marmoreus tissue isolation medium comprises the following steps:
step 1, weighing the medicine
Accurately weighing 20 parts of glucose, 20 parts of agar powder, 1 part of peptone, 1 part of magnesium sulfate and 1 part of potassium dihydrogen phosphate according to the raw material components of the culture medium in the embodiment 4, and putting the raw materials into a 1000ml beaker for later use;
specifically, the weighing precision is controlled within the range of +/-0.1;
step 2 preparation of the Medium
Pouring 1000mL of purified water into an iron pan, crushing corn in parts by mass into corn particles, putting the corn particles into the iron pan, cooking, and filtering the cooked liquid by using gauze with a thickness of four layers to obtain a filtrate; pouring the filtrate into the beaker containing the culture medium raw materials in the step 1, heating in a water bath and stirring to ensure that the culture medium raw materials are fully and completely dissolved in the filtrate;
in the actual operation process, if the filtrate of the corn is less than 1000mL, purified water can be added to supplement 1000 mL;
step 3, subpackaging test tubes
And (4) subpackaging the culture medium dissolved in the step (2) in a test tube. When the test tubes are subpackaged, before the culture medium is not solidified, subpackaging the test tubes by using a funnel and pouring the test tubes into prepared clean test tubes, wherein each used test tube contains 18ml of the culture medium, sealing by plugging a silica gel plug after subpackaging is finished, placing the test tubes containing the culture medium on a test tube rack, wrapping the whole test tube rack by using newspaper, and placing the test tubes into a sterilization pot for batch sterilization; wherein the sterilization condition is sterilization at 121 deg.C for 20min, and the whole test tube subpackaging process is controlled within 50 min.
Step 4, cooling
Placing the test tube on a clean, spacious and flat table top after the temperature is reduced to 55 ℃, placing and positioning the test tube at one time, wherein the length of a culture medium at the bottom end of the test tube is required to be 1cm, and covering a layer of heat insulation material on the test tube filled with the culture medium after placing is finished, so that the temperature of the culture medium is slowly reduced and cooled to room temperature, and the generation of test tube condensate water can be reduced;
and 5, cooling the culture medium, and performing tissue isolation culture on the culture medium.
Test group
The small pieces of tissue in the stem of hypsizygus marmoreus were inoculated into the medium of example 6 and the tissue isolation tube medium of comparative example 1, each tube containing 18mL of the medium of comparative example 1, which was the same as the medium of example 6, and all the media were covered and placed in an incubator for growth, and the experimental data obtained are shown in table 3:
TABLE 3 comparison of hyphal growth of tissue pieces in the media of example 6 and comparative example 1
| Incubation time | Example 6 | Comparative example 1 |
| Day 3 | Germination of tissue mass | Germination of tissue mass |
| Day 8 | Hypha growth of 25mm | The tissue mass begins to eat |
| Day 12 | 50mm | 22-25mm |
| Color of hypha | Dense white | White colour (Bai) |
| Growth of hypha | Dense (very high density of hypha) | Denser (higher density of hypha) |
| Hyphal morphology | Rough and strong | Slender |
As can be seen from Table 3, the present invention shows good properties of hyphal germination, growth and growth compared to the existing enriched medium.
It can be seen from the combination of tables 1, 2 and 3 that the corn-based tissue isolation test tube culture medium of the present invention can provide good germination and growth of hyphae, and can greatly reduce the cost of culture medium raw materials, and has unexpected effects.
The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. A hypsizigus marmoreus tissue isolation medium is characterized in that: the culture medium comprises the following components in parts by weight:
15-25 parts of corn;
15-25 parts of glucose;
15-25 parts of agar powder;
0.5-1.5 parts of peptone;
0.5-1.5 parts of magnesium sulfate;
0.5-1.5 parts of monopotassium phosphate;
800 portions of water and 1200 portions of water.
2. The hypsizigus marmoreus tissue isolation medium according to claim 1, wherein: the culture medium comprises the following components in parts by weight:
20 parts of corn;
20 parts of glucose;
20 parts of agar powder;
1 part of peptone;
1 part of magnesium sulfate;
1 part of monopotassium phosphate;
1000 parts of water.
3. A method for preparing the hypsizigus marmoreus tissue isolation medium according to claim 1 or 2, wherein: the method comprises the following steps:
step 1, weighing the medicine
Accurately weighing glucose, agar powder, peptone, magnesium sulfate and potassium dihydrogen phosphate according to a standard formula, and placing into a 1000ml beaker;
step 2 preparation of the Medium
Pouring 1000mL of purified water into an iron pan, adding the crushed corn into the iron pan, cooking, filtering the cooked liquid with four layers of thick gauze to obtain a filtrate, and pouring the filtrate into the beaker containing the culture medium raw material in the step 1; if the filtered filtrate is less than 1000mL, adding purified water to supplement 1000m L, stirring the raw materials and the filtrate uniformly, and heating in water bath again until the raw materials are completely dissolved to obtain a culture medium;
step 3, subpackaging test tubes
Subpackaging the culture medium obtained in the step 2 in test tubes, pouring the test tubes into clean test tubes when the test tubes are subpackaged before the culture medium is not solidified, filling 18ml of culture medium liquid into each used test tube, sealing after subpackaging is finished, and sterilizing;
step 4, cooling
Placing the test tube on a clean, spacious and flat table top after the culture medium in the test tube is cooled, placing and positioning the test tube at one time, wherein the length of the culture medium at the bottom end of the test tube is required to be 1cm, and covering a layer of heat insulation material on the surface of the test tube filled with the culture medium after placing is finished, so that the temperature of the culture medium is slowly reduced to be cooled, and the generation of test tube condensate water is reduced;
and 5, cooling the culture medium, and performing tissue isolation culture on the culture medium.
4. The method for preparing a culture medium for tissue isolation of hypsizigus marmoreus according to claim 3, wherein: in the step 1, the weighing precision is controlled within the range of +/-0.1.
5. The method for preparing a culture medium for tissue isolation of hypsizigus marmoreus according to claim 3, wherein: the sterilization condition in the step 3 is sterilization at 121 ℃ for 20 min.
6. The method for preparing a culture medium for tissue isolation of hypsizigus marmoreus according to claim 3, wherein: and (3) reducing the temperature before swinging the inclined plane in the step (4) to 55-60 ℃.
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| CN202110827328.0A CN113383677A (en) | 2021-07-21 | 2021-07-21 | Hypsizigus marmoreus tissue isolation medium and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110827328.0A CN113383677A (en) | 2021-07-21 | 2021-07-21 | Hypsizigus marmoreus tissue isolation medium and preparation method thereof |
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| CN104541975A (en) * | 2014-09-25 | 2015-04-29 | 山东晨阳菌业有限公司 | Rapid preparation method and use method of hypsizigus marmoreus liquid strains |
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| CN107488599A (en) * | 2017-09-14 | 2017-12-19 | 西充星河生物科技有限公司 | A kind of fine and soft mushroom strains manufacture craft of frost |
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| CN105837324A (en) * | 2016-03-28 | 2016-08-10 | 东莞市合心生物科技有限公司 | Hypsizigus marmoreus liquid strain medium and planting method |
| CN107488599A (en) * | 2017-09-14 | 2017-12-19 | 西充星河生物科技有限公司 | A kind of fine and soft mushroom strains manufacture craft of frost |
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Application publication date: 20210914 |