CN112931043A - A method for preparing strain of Lentinus Edodes, Hericium Erinaceus, Ganoderma, etc - Google Patents
A method for preparing strain of Lentinus Edodes, Hericium Erinaceus, Ganoderma, etc Download PDFInfo
- Publication number
- CN112931043A CN112931043A CN201911169420.1A CN201911169420A CN112931043A CN 112931043 A CN112931043 A CN 112931043A CN 201911169420 A CN201911169420 A CN 201911169420A CN 112931043 A CN112931043 A CN 112931043A
- Authority
- CN
- China
- Prior art keywords
- culture medium
- mushrooms
- seed
- water
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 240000000588 Hericium erinaceus Species 0.000 title claims abstract description 24
- 235000007328 Hericium erinaceus Nutrition 0.000 title claims abstract description 24
- 238000000034 method Methods 0.000 title claims abstract description 23
- 241000222336 Ganoderma Species 0.000 title claims abstract description 18
- 240000000599 Lentinula edodes Species 0.000 title claims abstract description 12
- 235000001715 Lentinula edodes Nutrition 0.000 title description 3
- 239000001963 growth medium Substances 0.000 claims abstract description 98
- 235000001674 Agaricus brunnescens Nutrition 0.000 claims abstract description 66
- 238000011081 inoculation Methods 0.000 claims abstract description 22
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 claims abstract description 18
- 229960000723 ampicillin Drugs 0.000 claims abstract description 18
- 238000002360 preparation method Methods 0.000 claims abstract description 14
- 239000000843 powder Substances 0.000 claims abstract description 13
- 229910052875 vesuvianite Inorganic materials 0.000 claims abstract description 12
- 230000000844 anti-bacterial effect Effects 0.000 claims abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 55
- 230000001954 sterilising effect Effects 0.000 claims description 47
- 238000004659 sterilization and disinfection Methods 0.000 claims description 35
- 238000012258 culturing Methods 0.000 claims description 24
- 235000015099 wheat brans Nutrition 0.000 claims description 21
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 20
- 229920001817 Agar Polymers 0.000 claims description 15
- 244000061456 Solanum tuberosum Species 0.000 claims description 15
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 15
- 239000008272 agar Substances 0.000 claims description 15
- 239000012153 distilled water Substances 0.000 claims description 15
- 239000012452 mother liquor Substances 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 15
- 208000015181 infectious disease Diseases 0.000 claims description 14
- 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 description 10
- 229930006000 Sucrose Natural products 0.000 claims description 10
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 10
- 238000009835 boiling Methods 0.000 claims description 10
- 238000001816 cooling Methods 0.000 claims description 10
- 239000000706 filtrate Substances 0.000 claims description 10
- 238000001914 filtration Methods 0.000 claims description 10
- 239000008103 glucose Substances 0.000 claims description 10
- 239000010440 gypsum Substances 0.000 claims description 10
- 229910052602 gypsum Inorganic materials 0.000 claims description 10
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 10
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 10
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 10
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 10
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 10
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 10
- 239000002994 raw material Substances 0.000 claims description 10
- 238000007789 sealing Methods 0.000 claims description 10
- 238000005507 spraying Methods 0.000 claims description 10
- 239000011435 rock Substances 0.000 claims description 9
- 239000002023 wood Substances 0.000 claims description 8
- 239000005720 sucrose Substances 0.000 claims description 7
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 claims description 6
- 229920000742 Cotton Polymers 0.000 claims description 5
- 229930003270 Vitamin B Natural products 0.000 claims description 5
- 229930003451 Vitamin B1 Natural products 0.000 claims description 5
- 239000003899 bactericide agent Substances 0.000 claims description 5
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 5
- VNFPBHJOKIVQEB-UHFFFAOYSA-N clotrimazole Chemical compound ClC1=CC=CC=C1C(N1C=NC=C1)(C=1C=CC=CC=1)C1=CC=CC=C1 VNFPBHJOKIVQEB-UHFFFAOYSA-N 0.000 claims description 5
- 229960004022 clotrimazole Drugs 0.000 claims description 5
- 238000005520 cutting process Methods 0.000 claims description 5
- 230000000249 desinfective effect Effects 0.000 claims description 5
- 235000001727 glucose Nutrition 0.000 claims description 5
- 238000000227 grinding Methods 0.000 claims description 5
- 229960002523 mercuric chloride Drugs 0.000 claims description 5
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 235000012015 potatoes Nutrition 0.000 claims description 5
- 238000002791 soaking Methods 0.000 claims description 5
- 239000007921 spray Substances 0.000 claims description 5
- 239000008223 sterile water Substances 0.000 claims description 5
- 239000006188 syrup Substances 0.000 claims description 5
- 235000020357 syrup Nutrition 0.000 claims description 5
- 229960003495 thiamine Drugs 0.000 claims description 5
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims description 5
- 235000019156 vitamin B Nutrition 0.000 claims description 5
- 239000011720 vitamin B Substances 0.000 claims description 5
- 239000011691 vitamin B1 Substances 0.000 claims description 5
- 235000010374 vitamin B1 Nutrition 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 239000000428 dust Substances 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 2
- 238000011177 media preparation Methods 0.000 claims description 2
- 240000008397 Ganoderma lucidum Species 0.000 claims 5
- 235000001637 Ganoderma lucidum Nutrition 0.000 claims 5
- 230000009286 beneficial effect Effects 0.000 abstract description 12
- 230000035699 permeability Effects 0.000 abstract description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 3
- 235000015097 nutrients Nutrition 0.000 abstract description 3
- 229910052760 oxygen Inorganic materials 0.000 abstract description 3
- 239000001301 oxygen Substances 0.000 abstract description 3
- 239000000126 substance Substances 0.000 abstract description 3
- 241000894006 Bacteria Species 0.000 description 17
- 229960004793 sucrose Drugs 0.000 description 8
- 230000002458 infectious effect Effects 0.000 description 6
- 241000233866 Fungi Species 0.000 description 4
- 238000011109 contamination Methods 0.000 description 3
- 206010011224 Cough Diseases 0.000 description 2
- 241000289669 Erinaceus europaeus Species 0.000 description 2
- 208000006673 asthma Diseases 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 241000959662 Hydnaceae Species 0.000 description 1
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 1
- 206010033557 Palpitations Diseases 0.000 description 1
- 241000222341 Polyporaceae Species 0.000 description 1
- 208000013738 Sleep Initiation and Maintenance disease Diseases 0.000 description 1
- 208000012886 Vertigo Diseases 0.000 description 1
- 206010003549 asthenia Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 206010022437 insomnia Diseases 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000002936 tranquilizing effect Effects 0.000 description 1
- 231100000889 vertigo Toxicity 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
- A01G18/30—Accessories for use before inoculation of spawn, e.g. sterilisers
-
- 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
- A01G18/50—Inoculation of spawn
-
- 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
- A01G18/60—Cultivation rooms; Equipment therefor
- A01G18/69—Arrangements for managing the environment, e.g. sprinklers
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Mushroom Cultivation (AREA)
Abstract
The embodiment of the invention discloses a method for preparing strains of mushrooms such as shiitake mushrooms, hericium erinaceus and lucid ganoderma, and particularly relates to the technical field of strain preparation, and the method comprises the following specific steps: s1, preparing a PDA culture medium; s2, preparing mother seeds; s3, preparing a strain culture medium; and S4, inoculation. The fine powder ground by adding the vesuvianite has the advantages of good air permeability of the prepared strain culture medium due to large specific surface area and high aperture ratio of the vesuvianite, being beneficial to strain contact adhesion and growth, effectively keeping more strains, simultaneously providing oxygen and nutrient substances required in the strain growth, simultaneously effectively improving the antibacterial capacity of the strains by adding the ampicillin in the strain culture medium, and further being beneficial to the culture of the strains.
Description
Technical Field
The embodiment of the invention relates to the technical field of strain production, in particular to a method for producing strains of mushrooms such as shiitake mushrooms, hericium erinaceus and lucid ganoderma.
Background
The mushroom is also called as dried mushroom, is edible fungus, the edible part is mushroom fruiting body, the fresh mushroom is dehydrated to form dried mushroom, it is convenient to transport and store, it is an important goods in south and north, the dried and fresh mushroom is widely used in Chinese dish. Hericium erinaceus is a fungus of the family Hydnaceae, has hairy fleshy spurs on the surface, is about 1-3 cm long, has round and thick fruiting body, is white when fresh, is dried from light yellow to light brown, has narrow base or slightly short handle, has expanded upper part and 3.5-10 cm diameter, is called Hericium erinaceus as the hedgehog fungus is far removed, and is called Hericium erinaceus as well as hedgehog fungus. Ganoderma is umbrella-shaped, pileus kidney-shaped, semicircular or nearly circular, is fruiting body of Ganoderma of Polyporaceae, has effects of invigorating qi, tranquilizing mind, relieving cough and asthma, and can be used for treating vertigo, insomnia, palpitation, short breath, and cough and asthma due to asthenia. Mushrooms such as shiitake mushrooms, hericium erinaceus and lucid ganoderma are usually artificially cultured, and the key to the success of the artificial cultivation of edible mushrooms is the strain, and only good strains can achieve high quality and high yield.
In the prior art, a culture medium is generally prepared to culture strains, but the existing culture medium has poor air permeability and is not beneficial to the attachment growth of the strains, so that the yield of strain culture is low.
Disclosure of Invention
Therefore, the embodiment of the invention provides a method for preparing strains of mushrooms such as mushrooms, hericium erinaceus, lucid ganoderma and the like, wherein the volcanic rock is added to be ground into fine powder, and the volcanic rock has large specific surface area and high aperture ratio, so that the prepared strain culture medium has good air permeability, is beneficial to strain contact adhesion and growth, effectively keeps more strains, can provide oxygen and nutrient substances required for the strain growth, and meanwhile, ampicillin added into the strain culture medium effectively improves the antibacterial capability of the strains, is further beneficial to strain culture, and solves the problem that the prior art is unfavorable for the strain adhesion growth and causes the strain culture quantity to be less due to poor air permeability of the culture medium.
In order to achieve the above object, the embodiments of the present invention provide the following technical solutions: a method for preparing strains of Lentinus Edodes, Hericium Erinaceus, Ganoderma, etc. comprises the following steps:
s1, preparing a PDA culture medium;
s2, mother seed preparation:
s2.1, selecting fresh mushrooms such as shiitake mushrooms, hericium erinaceus and lucid ganoderma and the like as seed mushrooms, selecting 8-9-minute mature fine varieties of the seed mushrooms, cleaning the seed mushrooms, cutting off two bases of the seed mushrooms, soaking the two bases of the seed mushrooms in 0.1% mercuric chloride water for 5-10min in an aseptic box, washing the seed mushrooms with sterile water and wiping the seed mushrooms dry;
s2.2, tearing off the seed mushrooms, picking a small tissue at the junction or the fold of the sprout cap and the stipe by using an inoculating needle, then transferring the tissue to a PDA culture medium, culturing for 3-5 days at the temperature of 25 ℃, generating white villous hyphae on the tissue, and rotating and expanding the tube to obtain a mother seed;
s3, preparation of a strain culture medium:
s3.1, preparing raw materials: broad-leaved tree wood dust 60-70%, wheat bran 10-20%, sucrose 1-1.5%, gypsum 1-1.5%, ampicillin 0.1-0.15%, and vesuvianite 10-20%;
s3.2, tedding the broad-leaved tree sawdust and the wheat bran for 3-5 days in the sun, and spraying lime water to the broad-leaved tree sawdust and the wheat bran by using a spray pot while tedding;
s3.3, grinding the vesuvianite into fine powder for later use by using a grinder, dissolving sucrose in a proper amount of water to prepare syrup for later use, adding distilled water with the concentration of 1000 times of that of ampicillin to prepare mother liquor, and then filtering and sterilizing the mother liquor for later use;
s3.4, uniformly stirring the wood chips, the wheat bran and the gypsum, adding the sugar water and the fine volcanic rock powder, and uniformly stirring the materials to prepare a culture medium, bottling the prepared culture medium, wherein the culture medium is required to be uniformly tightened up and down during bottling so as to facilitate the growth of hyphae;
s3.5, performing normal-pressure steam sterilization on the culture medium, and cooling the culture medium to 50-60 ℃ in a water bath after sterilization;
s3.6, adding one thousandth of ampicillin mother liquor of the volume of the cooled culture medium, and fully shaking up;
s4, inoculation:
s4.1, in an inoculation room, firstly, spraying and disinfecting a culture medium by using a clotrimazole bactericide, and inoculating when the temperature of the culture medium is reduced to room temperature;
s4.2, performing flame sterilization on the inoculation tool, then quickly inoculating the mother seeds into a culture medium by using the inoculation tool, then sealing, and then culturing in a culture chamber to obtain original seeds;
s4.3, inoculating the stock into a new culture medium, and then culturing in a culture room to obtain a strain.
Further, the PDA medium preparation step in step S1 is:
s1.1, preparing raw materials: 200g of peeled potato, 20g of glucose, 20g of agar, 2g of monopotassium phosphate, 0.5g of magnesium sulfate, 110 mg of vitamin B and 1000ml of water;
s1.2, slicing the potatoes, then placing the slices into a pot, adding a proper amount of distilled water, and boiling until the potato slices are soft but not rotten;
s1.3, filtering for 3-5 times by using gauze, and taking filtrate;
s1.4, adding agar into the filtrate, boiling until the agar is completely melted, then adding glucose, monopotassium phosphate, vitamin B1, magnesium sulfate and distilled water, and uniformly stirring;
s1.5, subpackaging the hot mixture in test tubes, and then plugging a cotton plug for sealing;
s1.6, sterilizing at high temperature for 40min, obliquely placing the test tube after sterilization, cooling, and placing the test tube in a culture room at the temperature of 30 ℃ for culture for 3-5 days.
Further, the water content of the culture medium prepared in step S3.4 is preferably 55-60%, and the judgment criteria are: the water overflows without dripping when the fingers are gripped by hands.
Further, in step S3.5, sterilization is performed by using a sterilization oven, wherein the sterilization temperature is 100 ℃ and the sterilization time is 12-14 h.
Further, the culture medium after inoculation in steps S4.2 and S4.3 was maintained at room temperature at 24-26 ℃ for the first 4 days of initial culture in the culture chamber, and was controlled at room temperature at 22-24 ℃ from day 5 and at 20-23 ℃ after day 16 until 45 days of culture.
Further, after culturing for 7 days in steps S4.2 and S4.3, the growth of hyphae and the contamination of other bacteria are checked, and if the contamination of other bacteria is found, the mixed bacteria is immediately cleaned and incinerated or buried for preventing infection.
The embodiment of the invention has the following advantages:
1. according to the invention, the volcanic rock is added and ground into fine powder, and the volcanic rock has large specific surface area and high aperture ratio, so that the prepared strain culture medium has good air permeability, is beneficial to strain contact adhesion and growth, effectively keeps more strains, can provide oxygen and nutrient substances required in the strain growth, and simultaneously effectively improves the antibacterial capacity of the strains by adding the ampicillin into the strain culture medium, thereby being further beneficial to the culture of the strains;
2. according to the invention, before the strain culture medium is prepared, the broadleaf tree sawdust and wheat bran are turned over and dried in the sun, and lime water is sprayed for disinfection, so that the quality of the prepared strain culture medium is improved.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. 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.
Example 1:
the invention provides a method for preparing strains of mushrooms, hericium erinaceus, lucid ganoderma and the like, which comprises the following steps:
s1, preparing a PDA culture medium, wherein the preparation steps are as follows:
s1.1, preparing raw materials: 200g of peeled potato, 20g of glucose, 20g of agar, 2g of monopotassium phosphate, 0.5g of magnesium sulfate, 110 mg of vitamin B and 1000ml of water;
s1.2, slicing the potatoes, then placing the slices into a pot, adding a proper amount of distilled water, and boiling until the potato slices are soft but not rotten;
s1.3, filtering for 3 times by using gauze, and taking filtrate;
s1.4, adding agar into the filtrate, boiling until the agar is completely melted, then adding glucose, monopotassium phosphate, vitamin B1, magnesium sulfate and distilled water, and uniformly stirring;
s1.5, subpackaging the hot mixture in test tubes, and then plugging a cotton plug for sealing;
s1.6, sterilizing at high temperature for 40min, obliquely placing a test tube after sterilizing, cooling, and placing in a culture room at 30 ℃ for 3 days;
s2, mother seed preparation:
s2.1, selecting fresh mushrooms such as shiitake mushrooms, hericium erinaceus and lucid ganoderma and the like as seed mushrooms, selecting 8-minute mature fine varieties of the seed mushrooms, cleaning the seed mushrooms, cutting off two bases of the seed mushrooms, soaking the seed mushrooms in 0.1% mercuric chloride water for 5min in an aseptic box, washing the seed mushrooms with sterile water and wiping the seed mushrooms dry;
s2.2, tearing off the seed mushrooms, picking a small tissue at the junction or the fold of the sprout cap and the stipe by using an inoculating needle, then transferring the tissue to a PDA culture medium, culturing for 3 days at the temperature of 25 ℃, generating white villous hyphae on the tissue, and rotating and expanding the tube to obtain a mother seed;
s3, preparation of a strain culture medium:
s3.1, preparing raw materials: broad-leaved tree wood dust 60%, wheat bran 10%, cane sugar 1%, gypsum 1%, ampicillin 0.1% and vesuvianite 10%;
s3.2, tedding the broad-leaved tree sawdust and the wheat bran for 3 days in the sun, and spraying lime water to the broad-leaved tree sawdust and the wheat bran by using a spray pot at the same time of tedding;
s3.3, grinding the vesuvianite into fine powder for later use by using a grinder, dissolving sucrose in a proper amount of water to prepare syrup for later use, adding distilled water with the concentration of 1000 times of that of ampicillin to prepare mother liquor, and then filtering and sterilizing the mother liquor for later use;
s3.4, uniformly stirring the wood chips, the wheat bran and the gypsum, adding the sugar water and the fine volcanic rock powder, and uniformly stirring to prepare a culture medium, wherein the prepared culture medium has a water content of 55 percent, and the judgment standard is as follows: the fingers are tightly held by hands, water overflows without dripping, the prepared culture medium is bottled, and the tightness of the culture medium is consistent up and down during bottling so as to be beneficial to the growth of hyphae;
s3.5, performing normal-pressure steam sterilization on the culture medium by using a sterilization stove, wherein the sterilization temperature is 100 ℃, the sterilization time is 12 hours, and cooling the culture medium to 50 ℃ in a water bath after sterilization;
s3.6, adding one thousandth of ampicillin mother liquor of the volume of the cooled culture medium, and fully shaking up;
s4, inoculation:
s4.1, in an inoculation room, firstly, spraying and disinfecting a culture medium by using a clotrimazole bactericide, and inoculating when the temperature of the culture medium is reduced to room temperature;
s4.2, performing flame sterilization on the inoculation tool, quickly inoculating the mother seeds into a culture medium by using the inoculation tool, sealing, culturing in a culture chamber, keeping the room temperature at 24 ℃ within 4 days before the inoculated culture medium is initially placed in the culture chamber, controlling the room temperature at 22 ℃ from 5 th day, culturing for 7 days, checking the growth condition of hyphae and whether to pollute infectious microbes, immediately clearing the infectious microbes for incineration or deep burying treatment to prevent infection if the infectious microbes are polluted, and controlling the room temperature at 20 ℃ after 16 th day until the original seeds are cultured for 45 days to obtain original seeds;
s4.3, inoculating the stock into a new culture medium, culturing in a culture room, keeping the room temperature at 24 ℃ within 4 days before the inoculated culture medium is initially placed in the culture room, controlling the room temperature at 22 ℃ from the 5 th day, culturing for 7 days, checking the growth condition of hyphae and whether the hyphae pollute the mixed bacteria, immediately clearing the mixed bacteria for incineration or deep burying treatment to prevent infection if the mixed bacteria are polluted, and controlling the room temperature at 20 ℃ after the 16 th day until the culture is cultured for 45 days to obtain the strain.
Example 2:
the invention provides a method for preparing strains of mushrooms, hericium erinaceus, lucid ganoderma and the like, which comprises the following steps:
s1, preparing a PDA culture medium, wherein the preparation steps are as follows:
s1.1, preparing raw materials: 200g of peeled potato, 20g of glucose, 20g of agar, 2g of monopotassium phosphate, 0.5g of magnesium sulfate, 110 mg of vitamin B and 1000ml of water;
s1.2, slicing the potatoes, then placing the slices into a pot, adding a proper amount of distilled water, and boiling until the potato slices are soft but not rotten;
s1.3, filtering for 4 times by using gauze, and taking filtrate;
s1.4, adding agar into the filtrate, boiling until the agar is completely melted, then adding glucose, monopotassium phosphate, vitamin B1, magnesium sulfate and distilled water, and uniformly stirring;
s1.5, subpackaging the hot mixture in test tubes, and then plugging a cotton plug for sealing;
s1.6, sterilizing at high temperature for 40min, obliquely placing a test tube after sterilizing, cooling, and placing in a culture room at 30 ℃ for culture for 4 days;
s2, mother seed preparation:
s2.1, selecting fresh mushrooms such as shiitake mushrooms, hericium erinaceus and lucid ganoderma and the like as seed mushrooms, selecting 8-minute mature fine varieties of the seed mushrooms, cleaning the seed mushrooms, cutting off two bases of the seed mushrooms, soaking the two bases of the seed mushrooms in 0.1% mercuric chloride water for 7.5min in an aseptic box, washing the seed mushrooms with sterile water and wiping the seed mushrooms dry;
s2.2, tearing off the seed mushrooms, picking a small tissue at the junction or the fold of the sprout cap and the stipe by using an inoculating needle, then transferring the tissue to a PDA culture medium, culturing for 4 days at the temperature of 25 ℃, generating white villous hyphae on the tissue, and rotating and expanding the tube to obtain a mother seed;
s3, preparation of a strain culture medium:
s3.1, preparing raw materials: 65% of broad-leaved tree sawdust, 15% of wheat bran, 1.25% of cane sugar, 1.25% of gypsum, 0.125% of ampicillin and 15% of vesuvianite;
s3.2, tedding the broad-leaved tree sawdust and the wheat bran for 4 days in the sun, and spraying lime water to the broad-leaved tree sawdust and the wheat bran by using a spray pot at the same time of tedding;
s3.3, grinding the vesuvianite into fine powder for later use by using a grinder, dissolving sucrose in a proper amount of water to prepare syrup for later use, adding distilled water with the concentration of 1000 times of that of ampicillin to prepare mother liquor, and then filtering and sterilizing the mother liquor for later use;
s3.4, uniformly stirring the wood chips, the wheat bran and the gypsum, adding the sugar water and the fine volcanic rock powder, and uniformly stirring to prepare a culture medium, wherein the prepared culture medium has a water content of 57.5 percent, and the judgment standard is as follows: the fingers are tightly held by hands, water overflows without dripping, the prepared culture medium is bottled, and the tightness of the culture medium is consistent up and down during bottling so as to be beneficial to the growth of hyphae;
s3.5, performing normal-pressure steam sterilization on the culture medium by using a sterilization stove, wherein the sterilization temperature is 100 ℃, the sterilization time is 13 hours, and cooling the culture medium to 55 ℃ in a water bath after sterilization;
s3.6, adding one thousandth of ampicillin mother liquor of the volume of the cooled culture medium, and fully shaking up;
s4, inoculation:
s4.1, in an inoculation room, firstly, spraying and disinfecting a culture medium by using a clotrimazole bactericide, and inoculating when the temperature of the culture medium is reduced to room temperature;
s4.2, performing flame sterilization on the inoculation tool, quickly inoculating the mother seeds into a culture medium by using the inoculation tool, sealing, culturing in a culture chamber, keeping the room temperature at 25 ℃ within 4 days before the inoculated culture medium is initially placed in the culture chamber, controlling the room temperature at 23 ℃ from 5 th day, culturing for 7 days, checking the growth condition of hyphae and whether to pollute infectious microbes, immediately clearing the infectious microbes for incineration or deep burying treatment to prevent infection if the infectious microbes are polluted, and controlling the room temperature at 21.5 ℃ after 16 th day until the original seeds are cultured for 45 days to obtain stock seeds;
s4.3, inoculating the stock into a new culture medium, culturing in a culture room, keeping the room temperature at 25 ℃ within 4 days before the inoculated culture medium is initially placed in the culture room, controlling the room temperature at 23 ℃ from the 5 th day, culturing for 7 days, checking the growth condition of hyphae and whether the hyphae pollute the mixed bacteria, immediately clearing the mixed bacteria for incineration or deep burying treatment to prevent infection if the mixed bacteria are polluted, and controlling the room temperature at 21.5 ℃ after the 16 th day until the culture is cultured for 45 days to obtain the strain.
Example 3:
the invention provides a method for preparing strains of mushrooms, hericium erinaceus, lucid ganoderma and the like, which comprises the following steps:
s1, preparing a PDA culture medium, wherein the preparation steps are as follows:
s1.1, preparing raw materials: 200g of peeled potato, 20g of glucose, 20g of agar, 2g of monopotassium phosphate, 0.5g of magnesium sulfate, 110 mg of vitamin B and 1000ml of water;
s1.2, slicing the potatoes, then placing the slices into a pot, adding a proper amount of distilled water, and boiling until the potato slices are soft but not rotten;
s1.3, filtering for 5 times by using gauze, and taking filtrate;
s1.4, adding agar into the filtrate, boiling until the agar is completely melted, then adding glucose, monopotassium phosphate, vitamin B1, magnesium sulfate and distilled water, and uniformly stirring;
s1.5, subpackaging the hot mixture in test tubes, and then plugging a cotton plug for sealing;
s1.6, sterilizing at high temperature for 40min, obliquely placing a test tube after sterilizing, cooling, and placing in a culture room at 30 ℃ for 5 days;
s2, mother seed preparation:
s2.1, selecting fresh mushrooms such as shiitake mushrooms, hericium erinaceus and lucid ganoderma as seed mushrooms, selecting 9-minute mature fine varieties of the seed mushrooms, cleaning the seed mushrooms, cutting off two bases of the seed mushrooms, soaking the seed mushrooms in 0.1% mercuric chloride water for 10min in an aseptic box, washing the seed mushrooms with sterile water, and wiping the seed mushrooms dry;
s2.2, tearing off the seed mushrooms, picking a small tissue at the junction or the fold of the sprout cap and the stipe by using an inoculating needle, then transferring the tissue to a PDA culture medium, culturing for 5 days at the temperature of 25 ℃, generating white villous hyphae on the tissue, and rotating and expanding the tube to obtain a mother seed;
s3, preparation of a strain culture medium:
s3.1, preparing raw materials: 70% of broad-leaved tree sawdust, 20% of wheat bran, 1.5% of cane sugar, 1.5% of gypsum, 0.15% of ampicillin and 20% of vesuvianite;
s3.2, tedding the broad-leaved tree sawdust and the wheat bran for 5 days in the sun, and spraying lime water to the broad-leaved tree sawdust and the wheat bran by using a spray pot while tedding;
s3.3, grinding the vesuvianite into fine powder for later use by using a grinder, dissolving sucrose in a proper amount of water to prepare syrup for later use, adding distilled water with the concentration of 1000 times of that of ampicillin to prepare mother liquor, and then filtering and sterilizing the mother liquor for later use;
s3.4, uniformly stirring the wood chips, the wheat bran and the gypsum, adding the sugar water and the fine volcanic rock powder, and uniformly stirring to prepare a culture medium, wherein the prepared culture medium has the water content of preferably 60 percent and has the judgment standard of: the fingers are tightly held by hands, water overflows without dripping, the prepared culture medium is bottled, and the tightness of the culture medium is consistent up and down during bottling so as to be beneficial to the growth of hyphae;
s3.5, performing normal-pressure steam sterilization on the culture medium by using a sterilization stove, wherein the sterilization temperature is 100 ℃, the sterilization time is 14 hours, and cooling the culture medium to 60 ℃ in a water bath after sterilization;
s3.6, adding one thousandth of ampicillin mother liquor of the volume of the cooled culture medium, and fully shaking up;
s4, inoculation:
s4.1, in an inoculation room, firstly, spraying and disinfecting a culture medium by using a clotrimazole bactericide, and inoculating when the temperature of the culture medium is reduced to room temperature;
s4.2, performing flame sterilization on the inoculation tool, quickly inoculating the mother seeds into a culture medium by using the inoculation tool, sealing, culturing in a culture chamber, keeping the room temperature of the inoculated culture medium at 26 ℃ within 4 days before the culture medium is initially placed in the culture chamber, controlling the room temperature at 24 ℃ from 5 th day, culturing for 7 days, checking the growth condition of hyphae and whether the hyphae pollute the mixed bacteria, immediately clearing the mixed bacteria to burn or deeply bury the mixed bacteria for infection prevention if the mixed bacteria are polluted, and controlling the room temperature at 23 ℃ after 16 th day until the mixed bacteria are cultured for 45 days to obtain original seeds;
s4.3, inoculating the stock into a new culture medium, culturing in a culture room, keeping the room temperature at 26 ℃ within 4 days before the inoculated culture medium is initially placed in the culture room, controlling the room temperature at 24 ℃ from the 5 th day, culturing for 7 days, checking the growth condition of hyphae and whether the hyphae pollute the mixed bacteria, immediately clearing the mixed bacteria for incineration or deep burying treatment to prevent infection if the mixed bacteria are polluted, and controlling the room temperature at 23 ℃ after the 16 th day until the culture is cultured for 45 days to obtain the strain.
Example 4:
the method of the above example 1-3 was used to prepare 90 strain culture media, and then the 90 strain culture media were inoculated with the prepared strain culture media, and the following data were obtained after culturing the strain:
| number of culture medium | Average degree of growth of strain number | Average degree of hypha contamination | The average degree of the strain quality improvement (compared with the common culture medium culture) | |
| Example 1 | 30 | 30％ | 32％ | 29％ |
| Example 2 | 30 | 65％ | 68％ | 58％ |
| Example 3 | 30 | 20％ | 24％ | 20％ |
As can be seen from the above table, the method in example 2 is suitable, and the prepared strain culture medium is beneficial to strain contact adhesion and growth, and simultaneously improves the antibacterial ability of the strain to the maximum extent, thereby being beneficial to strain culture.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (6)
1. A method for preparing strains of mushrooms such as shiitake mushrooms, hericium erinaceus and lucid ganoderma is characterized by comprising the following steps: the method comprises the following specific steps:
s1, preparing a PDA culture medium;
s2, mother seed preparation:
s2.1, selecting fresh mushrooms such as shiitake mushrooms, hericium erinaceus and lucid ganoderma and the like as seed mushrooms, selecting 8-9-minute mature fine varieties of the seed mushrooms, cleaning the seed mushrooms, cutting off two bases of the seed mushrooms, soaking the two bases of the seed mushrooms in 0.1% mercuric chloride water for 5-10min in an aseptic box, washing the seed mushrooms with sterile water and wiping the seed mushrooms dry;
s2.2, tearing off the seed mushrooms, picking a small tissue at the junction or the fold of the sprout cap and the stipe by using an inoculating needle, then transferring the tissue to a PDA culture medium, culturing for 3-5 days at the temperature of 25 ℃, generating white villous hyphae on the tissue, and rotating and expanding the tube to obtain a mother seed;
s3, preparation of a strain culture medium:
s3.1, preparing raw materials: broad-leaved tree wood dust 60-70%, wheat bran 10-20%, sucrose 1-1.5%, gypsum 1-1.5%, ampicillin 0.1-0.15%, and vesuvianite 10-20%;
s3.2, tedding the broad-leaved tree sawdust and the wheat bran for 3-5 days in the sun, and spraying lime water to the broad-leaved tree sawdust and the wheat bran by using a spray pot while tedding;
s3.3, grinding the vesuvianite into fine powder for later use by using a grinder, dissolving sucrose in a proper amount of water to prepare syrup for later use, adding distilled water with the concentration of 1000 times of that of ampicillin to prepare mother liquor, and then filtering and sterilizing the mother liquor for later use;
s3.4, uniformly stirring the wood chips, the wheat bran and the gypsum, adding the sugar water and the fine volcanic rock powder, and uniformly stirring the materials to prepare a culture medium, bottling the prepared culture medium, wherein the culture medium is required to be uniformly tightened up and down during bottling so as to facilitate the growth of hyphae;
s3.5, performing normal-pressure steam sterilization on the culture medium, and cooling the culture medium to 50-60 ℃ in a water bath after sterilization;
s3.6, adding one thousandth of ampicillin mother liquor of the volume of the cooled culture medium, and fully shaking up;
s4, inoculation:
s4.1, in an inoculation room, firstly, spraying and disinfecting a culture medium by using a clotrimazole bactericide, and inoculating when the temperature of the culture medium is reduced to room temperature;
s4.2, performing flame sterilization on the inoculation tool, then quickly inoculating the mother seeds into a culture medium by using the inoculation tool, then sealing, and then culturing in a culture chamber to obtain original seeds;
s4.3, inoculating the stock into a new culture medium, and then culturing in a culture room to obtain a strain.
2. The method for preparing strains of mushrooms, hericium erinaceus, ganoderma lucidum and the like according to claim 1, wherein the method comprises the following steps: the PDA medium preparation step in step S1 is:
s1.1, preparing raw materials: 200g of peeled potato, 20g of glucose, 20g of agar, 2g of monopotassium phosphate, 0.5g of magnesium sulfate, 110 mg of vitamin B and 1000ml of water;
s1.2, slicing the potatoes, then placing the slices into a pot, adding a proper amount of distilled water, and boiling until the potato slices are soft but not rotten;
s1.3, filtering for 3-5 times by using gauze, and taking filtrate;
s1.4, adding agar into the filtrate, boiling until the agar is completely melted, then adding glucose, monopotassium phosphate, vitamin B1, magnesium sulfate and distilled water, and uniformly stirring;
s1.5, subpackaging the hot mixture in test tubes, and then plugging a cotton plug for sealing;
s1.6, sterilizing at high temperature for 40min, obliquely placing the test tube after sterilization, cooling, and placing the test tube in a culture room at the temperature of 30 ℃ for culture for 3-5 days.
3. The method for preparing strains of mushrooms, hericium erinaceus, ganoderma lucidum and the like according to claim 1, wherein the method comprises the following steps: the water content of the culture medium prepared in step S3.4 is preferably 55-60%, and the judgment standard is as follows: the water overflows without dripping when the fingers are gripped by hands.
4. The method for preparing strains of mushrooms, hericium erinaceus, ganoderma lucidum and the like according to claim 1, wherein the method comprises the following steps: in step S3.5, a sterilization oven is used for sterilization, the sterilization temperature is 100 ℃, and the sterilization time is 12-14 h.
5. The method for preparing strains of mushrooms, hericium erinaceus, ganoderma lucidum and the like according to claim 1, wherein the method comprises the following steps: the culture medium inoculated in steps S4.2 and S4.3 was maintained at room temperature at 24-26 ℃ for the first 4 days of initial introduction into the culture chamber, and was controlled at 22-24 ℃ from day 5 and 20-23 ℃ after day 16 until 45 days of culture.
6. The method for preparing strains of mushrooms, hericium erinaceus, ganoderma lucidum and the like according to claim 1, wherein the method comprises the following steps: after culturing for 7 days in steps S4.2 and S4.3, checking the growth condition of hyphae and whether the hyphae are polluted, and immediately clearing and burning or deeply burying for preventing infection if the hyphae are polluted.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911169420.1A CN112931043A (en) | 2019-11-26 | 2019-11-26 | A method for preparing strain of Lentinus Edodes, Hericium Erinaceus, Ganoderma, etc |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911169420.1A CN112931043A (en) | 2019-11-26 | 2019-11-26 | A method for preparing strain of Lentinus Edodes, Hericium Erinaceus, Ganoderma, etc |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112931043A true CN112931043A (en) | 2021-06-11 |
Family
ID=76224861
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911169420.1A Pending CN112931043A (en) | 2019-11-26 | 2019-11-26 | A method for preparing strain of Lentinus Edodes, Hericium Erinaceus, Ganoderma, etc |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112931043A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114557233A (en) * | 2022-04-14 | 2022-05-31 | 福建省农业科学院食用菌研究所 | Industrialized production method of stropharia rugoso-annulata particle stock |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103404370A (en) * | 2013-07-31 | 2013-11-27 | 西南科技大学 | Method for cultivating edible mushroom strains by using lava |
| CN105248285A (en) * | 2015-09-24 | 2016-01-20 | 中国科学院昆明植物研究所 | Flammulina velutipes variety, and cultivation method thereof |
| CN107667776A (en) * | 2017-09-29 | 2018-02-09 | 山西禾嘉合信息科技有限公司 | A kind of Household edible mushroom planting method |
| CN109906872A (en) * | 2019-04-13 | 2019-06-21 | 中山市巨隆农业科技有限公司 | A kind of lucidum strain production method |
-
2019
- 2019-11-26 CN CN201911169420.1A patent/CN112931043A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103404370A (en) * | 2013-07-31 | 2013-11-27 | 西南科技大学 | Method for cultivating edible mushroom strains by using lava |
| CN105248285A (en) * | 2015-09-24 | 2016-01-20 | 中国科学院昆明植物研究所 | Flammulina velutipes variety, and cultivation method thereof |
| CN107667776A (en) * | 2017-09-29 | 2018-02-09 | 山西禾嘉合信息科技有限公司 | A kind of Household edible mushroom planting method |
| CN109906872A (en) * | 2019-04-13 | 2019-06-21 | 中山市巨隆农业科技有限公司 | A kind of lucidum strain production method |
Non-Patent Citations (1)
| Title |
|---|
| 刘敬等: "菇菌菌种制种技术", 现代农业科技(上半月刊) * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114557233A (en) * | 2022-04-14 | 2022-05-31 | 福建省农业科学院食用菌研究所 | Industrialized production method of stropharia rugoso-annulata particle stock |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102668878B (en) | Method for cultivating flammulina velutipes by using aquilaria-sinensis-chip edible mushroom culture medium | |
| WO2015180519A1 (en) | Method for cultivating high-cordyceps-polysaccharide cordyceps militaris | |
| CN102633552A (en) | Formula of culture medium for white beech mushroom and method for cultivating white beech mushroom | |
| CN106342515A (en) | High-quality tomato planting method | |
| CN102557792A (en) | Culture medium of beech mushrooms and culture method of beech mushrooms | |
| CN105900692A (en) | Method for cultivating hericium erinaceus by means of corncobs | |
| CN110604004B (en) | Method for separating strains by dried morchella sporocarp | |
| JP2016116487A (en) | Auricularia auricula-judae cultivation method | |
| KR100823541B1 (en) | Mushroom cultivation method | |
| CN105237139A (en) | Preparation and culturing method of tree branch bacterial strain of pleurotus eryngii and hypsizygus marmoreus | |
| CN111248026B (en) | Quercus matsutake culture medium and application thereof | |
| CN101461312B (en) | Thousandfold reproduction method for mother culture of Lyophyllum shimeji | |
| CN106358512A (en) | Low-temperature-resistant tomato seed treatment method | |
| CN107047068B (en) | Facility greenhouse mushroom yield-increasing cultivation method | |
| WO2021109291A1 (en) | Method for cultivating tremella strain | |
| CN109234171B (en) | Preparation method of selenium-rich ganoderma lucidum mycelium powder | |
| CN112931043A (en) | A method for preparing strain of Lentinus Edodes, Hericium Erinaceus, Ganoderma, etc | |
| KR101239711B1 (en) | Manufacturing process of lentinus edodes culture | |
| KR20110045866A (en) | Raw Material Medium Composition for Culturing Mushroom Using Cedar Sawdust and Mathod for Cultering Mushroom Using the Same | |
| CN103250564B (en) | Artificial cultivating method for chestnut mycorrhiza fungi | |
| CN109937975B (en) | Standardized earthworm breeding method | |
| CN103688744B (en) | A kind of Cordceps militaris breeding method and using method | |
| KR20200061556A (en) | A Method for Cultivating a White Shiitake Mushroom Containing a Selenium | |
| CN102612995B (en) | Method for preparing liquid strain of chanterelle | |
| CN106635826A (en) | Preparation method of edible fungi cultivation strain based on recycling of disposable bamboo chopsticks |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20210611 |