CN112772298A - Isolation culture medium for boletus brevicaulis strain - Google Patents
Isolation culture medium for boletus brevicaulis strain Download PDFInfo
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- CN112772298A CN112772298A CN202110328895.1A CN202110328895A CN112772298A CN 112772298 A CN112772298 A CN 112772298A CN 202110328895 A CN202110328895 A CN 202110328895A CN 112772298 A CN112772298 A CN 112772298A
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- sulfate
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- potassium
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- 239000001963 growth medium Substances 0.000 title claims abstract description 30
- 241000222455 Boletus Species 0.000 title claims abstract description 21
- 238000002955 isolation Methods 0.000 title abstract description 8
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 claims abstract description 36
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims abstract description 24
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims abstract description 24
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims abstract description 24
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 22
- 229920001817 Agar Polymers 0.000 claims abstract description 13
- 239000000843 powder Substances 0.000 claims abstract description 13
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 claims abstract description 12
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 12
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000004471 Glycine Substances 0.000 claims abstract description 12
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000008272 agar Substances 0.000 claims abstract description 12
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000004327 boric acid Substances 0.000 claims abstract description 12
- 239000001110 calcium chloride Substances 0.000 claims abstract description 12
- 229910001628 calcium chloride Inorganic materials 0.000 claims abstract description 12
- 235000011148 calcium chloride Nutrition 0.000 claims abstract description 12
- GFHNAMRJFCEERV-UHFFFAOYSA-L cobalt chloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].[Cl-].[Co+2] GFHNAMRJFCEERV-UHFFFAOYSA-L 0.000 claims abstract description 12
- JZCCFEFSEZPSOG-UHFFFAOYSA-L copper(II) sulfate pentahydrate Chemical compound O.O.O.O.O.[Cu+2].[O-]S([O-])(=O)=O JZCCFEFSEZPSOG-UHFFFAOYSA-L 0.000 claims abstract description 12
- 239000011790 ferrous sulphate Substances 0.000 claims abstract description 12
- 235000003891 ferrous sulphate Nutrition 0.000 claims abstract description 12
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 claims abstract description 12
- 229960000367 inositol Drugs 0.000 claims abstract description 12
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims abstract description 12
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 12
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 claims abstract description 12
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 12
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 12
- 235000001968 nicotinic acid Nutrition 0.000 claims abstract description 12
- 229960003512 nicotinic acid Drugs 0.000 claims abstract description 12
- 239000011664 nicotinic acid Substances 0.000 claims abstract description 12
- 239000004323 potassium nitrate Substances 0.000 claims abstract description 12
- 235000010333 potassium nitrate Nutrition 0.000 claims abstract description 12
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 claims abstract description 12
- 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 abstract description 12
- 229960000344 thiamine hydrochloride Drugs 0.000 claims abstract description 12
- 235000019190 thiamine hydrochloride Nutrition 0.000 claims abstract description 12
- 239000011747 thiamine hydrochloride Substances 0.000 claims abstract description 12
- RZLVQBNCHSJZPX-UHFFFAOYSA-L zinc sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Zn+2].[O-]S([O-])(=O)=O RZLVQBNCHSJZPX-UHFFFAOYSA-L 0.000 claims abstract description 12
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims abstract description 11
- 235000019341 magnesium sulphate Nutrition 0.000 claims abstract description 11
- 239000003102 growth factor Substances 0.000 claims abstract description 10
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 229960004172 pyridoxine hydrochloride Drugs 0.000 claims abstract description 10
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 claims abstract description 10
- 239000011764 pyridoxine hydrochloride Substances 0.000 claims abstract description 10
- RWVGQQGBQSJDQV-UHFFFAOYSA-M sodium;3-[[4-[(e)-[4-(4-ethoxyanilino)phenyl]-[4-[ethyl-[(3-sulfonatophenyl)methyl]azaniumylidene]-2-methylcyclohexa-2,5-dien-1-ylidene]methyl]-n-ethyl-3-methylanilino]methyl]benzenesulfonate Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C(=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C)C=2C(=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C)C=C1 RWVGQQGBQSJDQV-UHFFFAOYSA-M 0.000 claims abstract description 10
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims abstract description 7
- 150000003839 salts Chemical class 0.000 claims abstract description 7
- 229960001484 edetic acid Drugs 0.000 claims abstract description 5
- 239000002609 medium Substances 0.000 claims abstract description 5
- 159000000014 iron salts Chemical class 0.000 claims abstract description 3
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 claims description 7
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 claims description 7
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 5
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 5
- 239000004615 ingredient Substances 0.000 claims description 4
- 239000011684 sodium molybdate Substances 0.000 claims description 2
- 235000015393 sodium molybdate Nutrition 0.000 claims description 2
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims description 2
- 241000233866 Fungi Species 0.000 abstract description 11
- 235000010419 agar Nutrition 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 abstract description 2
- 241000223252 Rhodotorula Species 0.000 abstract 1
- 230000009286 beneficial effect Effects 0.000 abstract 1
- 229960002449 glycine Drugs 0.000 abstract 1
- 239000012153 distilled water Substances 0.000 description 20
- 230000001954 sterilising effect Effects 0.000 description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 20
- 238000005303 weighing Methods 0.000 description 20
- 238000007865 diluting Methods 0.000 description 19
- 239000011521 glass Substances 0.000 description 15
- 238000002372 labelling Methods 0.000 description 10
- 239000012452 mother liquor Substances 0.000 description 10
- 239000010413 mother solution Substances 0.000 description 10
- 239000000243 solution Substances 0.000 description 7
- 238000001816 cooling Methods 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 238000004806 packaging method and process Methods 0.000 description 4
- 238000007789 sealing Methods 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 241000222453 Boletaceae Species 0.000 description 1
- 241000917479 Brevicornu Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000206672 Gelidium Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 229940035544 magnesium sulfate 100 mg Drugs 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229940086387 pyridoxine hydrochloride 30 mg Drugs 0.000 description 1
- 229940053679 pyridoxine hydrochloride 50 mg Drugs 0.000 description 1
- 238000006748 scratching Methods 0.000 description 1
- 230000002393 scratching effect Effects 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
- A01G18/40—Cultivation 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
- A01G18/20—Culture media, e.g. compost
Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to a boletus serrulata isolation medium which comprises a large number of components, trace components, iron salts and growth factors. The components of the malt extract powder comprise malt extract powder with specific concentration, soytone, ammonium nitrate, potassium nitrate, calcium chloride, magnesium sulfate, potassium dihydrogen phosphate and agar; the trace components comprise potassium iodide, boric acid, manganese sulfate monohydrate, zinc sulfate heptahydrate, sodium molybdate dihydrate, copper sulfate pentahydrate and cobalt chloride hexahydrate with specific concentrations; the ferric salt comprises anhydrous ferrous sulfate and ethylene diamine tetraacetic acid with specific concentration; the growth factor contains inositol, nicotinic acid, pyridoxine hydrochloride, thiamine hydrochloride, and glycine at specific concentrations. The culture medium is beneficial to the growth of the rhodosporidium torvum strain, the strain activity is high, the growth speed is high, the produced aerial hypha is good, and a basic guarantee is provided for the molecular biology research of symbiotic fungi and the cultivation of ectomycorrhizal fungi.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a boletus brevicornus strain isolation medium.
Background
The boletus dunaliensis belongs to species of boletus dunaliensis of boletaceae, can form ectomycorrhiza with plants, is symbiotic fungi, is an ideal material for researching symbiotic relationship between plants and fungi, is favorable for carrying out subsequent molecular biology experimental operation by separating and culturing boletus dunaliensis strains, and provides possible basic guarantee for the cultivation of ectomycorrhiza fungi. The difficulty of the isolation and culture of the boletus dunaliensis is that hyphae are easy to age and sufficient aerial hyphae cannot be obtained.
It has been difficult to separate and culture symbiotic fungi, and different symbiotic fungi have different requirements on culture medium. The general symbiotic fungus culture medium is an MMN culture medium, the saprophytic fungus culture medium is a PDA culture medium, but experiments show that the two culture media are used for separating and culturing boletus sedentarius strains, the growth vigor is poor, hypha is easy to age, aerial hypha is less, and the culture media are not suitable for the growth of the symbiotic boletus strains, so that the development of subsequent research work is hindered. At present, no culture medium formula specially aiming at symbiotic boletus lanuginosus exists.
Disclosure of Invention
In order to solve the defects of the prior art, the invention provides the separating culture medium for the boletus dunaliensis strain, so that the boletus dunaliensis strain can keep the activity for a long time and generate good aerial hyphae.
In order to achieve the purpose, the invention adopts the technical scheme that:
an isolation medium for boletus serrulata comprises a large number of components, trace components, iron salts and growth factors, wherein the large number of components comprise 10000-20000mg/L malt extract powder, 1000-2000mg/L soytone, 800-850mg/L ammonium nitrate, 900-1000mg/L potassium nitrate, 155-175mg/L calcium chloride, 80-100mg/L magnesium sulfate, 75-95mg/L monopotassium phosphate and 6500-8500mg/L agar; the micro-ingredients comprise potassium iodide with the concentration of 0.400-0.430mg/L, boric acid with the concentration of 2.8-3.3mg/L, manganese sulfate monohydrate with the concentration of 8.30-8.60mg/L, zinc sulfate heptahydrate with the concentration of 4.1-4.5mg/L, sodium molybdate dihydrate with the concentration of 0.100-0.150mg/L, copper sulfate pentahydrate with the concentration of 0.0115-0.0135mg/L and cobalt chloride hexahydrate with the concentration of 0.0115-0.0135 mg/L; the ferric salt comprises anhydrous ferrous sulfate with the concentration of 7.0-8.0mg/L and ethylene diamine tetraacetic acid with the concentration of 18.4-18.8 mg/L; the growth factor contains inositol 40-60mg/L, nicotinic acid 0.15-0.35mg/L, pyridoxine hydrochloride 0.15-0.35mg/L, thiamine hydrochloride 0.03-0.07mg/L, and glycine 0.6-1.5 mg/L.
Preferably, the large amount of components comprise 12000-18000mg/L malt extract powder, 1200-1800mg/L soytone, 820-840mg/L ammonium nitrate, 920-980mg/L potassium nitrate, 160-170mg/L calcium chloride, 85-95mg/L magnesium sulfate, 80-90mg/L monopotassium phosphate and 7000-8000mg/L agar; the micro-ingredients comprise potassium iodide with the concentration of 0.410-0.420mg/L, boric acid with the concentration of 3.0-3.2mg/L, manganese sulfate monohydrate with the concentration of 8.40-8.50mg/L, zinc sulfate heptahydrate with the concentration of 4.2-4.4mg/L, sodium molybdate dihydrate with the concentration of 0.120-0.130mg/L, copper sulfate pentahydrate with the concentration of 0.0120-0.0130mg/L and cobalt chloride hexahydrate with the concentration of 0.0120-0.0130 mg/L; the ferric salt comprises anhydrous ferrous sulfate with the concentration of 7.4-7.8mg/L and ethylene diamine tetraacetic acid with the concentration of 18.6-18.7 mg/L; the growth factor contains inositol 45-55mg/L, nicotinic acid 0.20-0.30mg/L, pyridoxine hydrochloride 0.20-0.30mg/L, thiamine hydrochloride 0.04-0.06mg/L, and glycine 0.8-1.3 mg/L.
Preferably, the bulk component comprises malt extract at a concentration of 15000mg/L, soya peptone at 1500mg/L, ammonium nitrate at 825mg/L, potassium nitrate at 950mg/L, calcium chloride at 165mg/L, magnesium sulfate at 90mg/L, monopotassium phosphate at 85mg/L, agar-agar at 7500 mg/L; the micro-components comprise potassium iodide with the concentration of 0.415mg/L, boric acid with the concentration of 3.1mg/L, manganese sulfate monohydrate with the concentration of 8.45mg/L, zinc sulfate heptahydrate with the concentration of 4.3mg/L, sodium molybdate hydrate with the concentration of 0.125mg/L, copper sulfate pentahydrate with the concentration of 0.0125mg/L and cobalt chloride hexahydrate with the concentration of 0.0125 mg/L; the ferric salt comprises anhydrous ferrous sulfate with the concentration of 7.6mg/L and disodium ethylene diamine tetraacetate with the concentration of 18.63 mg/L; the growth factor comprises inositol with concentration of 50mg/L, nicotinic acid with concentration of 0.25mg/L, pyridoxine hydrochloride with concentration of 0.25mg/L, thiamine hydrochloride with concentration of 0.05mg/L and glycine with concentration of 1.0 mg/L.
The culture medium is used for carrying out separation culture on boletus fringensis strains, the strains have high activity and high growth speed, the diameter of the colony of the strains is large, and the produced aerial hyphae are good, thereby providing basic guarantee for molecular biology research of symbiotic fungi and cultivation of ectomycorrhizal fungi.
Detailed Description
Example 1
Weighing 80mg of potassium iodide, 0.56g of boric acid, 1.66g of manganese sulfate monohydrate, 0.82g of zinc sulfate heptahydrate, 20mg of sodium molybdate dihydrate, 2.3mg of copper sulfate pentahydrate and 2.3mg of cobalt chloride hexahydrate, dissolving with a small amount of distilled water, diluting to constant volume of 1L, storing in a brown glass bottle, and marking as mother solution
；
Weighing 1.4g of anhydrous ferrous sulfate and 3.68g of disodium ethylene diamine tetraacetate, dissolving with a small amount of distilled water, diluting to constant volume of 1L, storing in brown glass bottle, and labeling as mother solution
；
Weighing inositol 8g, nicotinic acid 30mg, pyridoxine hydrochloride 30mg, thiamine hydrochloride 6mg, and glycine 0.12g, dissolving with small amount of distilled water, diluting to constant volume of 1L, storing in brown glass bottle, and labeling as mother liquor
；
Weighing 10g of malt extract powder, 1g of soytone, 0.800g of ammonium nitrate, 0.900g of potassium nitrate, 0.155g of calcium chloride, 80mg of magnesium sulfate, 75mg of potassium dihydrogen phosphate and 6.5g of agar, placing in a 1L triangular flask, and collecting mother liquor
、
、
Taking 5ml of the solution, placing in the triangular flask, dissolving with a small amount of distilled water, diluting to constant volume of 1L, packaging into two 1L triangular flasks, sealing, sterilizing in a sterilizing pot (103.4 KPa, 121 deg.C) for 20 min, taking out, and coolingWhen the temperature is reduced to 40-50 ℃, pouring the culture medium into a culture dish, solidifying and sterilizing (the pressure is 0.105MPa, the temperature is 121 ℃, and the sterilization is 15-30 mIn), and then performing strain isolation culture operation. Example 2
Weighing 86mg of potassium iodide, 0.66g of boric acid, 1.72g of manganese sulfate monohydrate, 0.90g of zinc sulfate heptahydrate, 30mg of sodium molybdate dihydrate, 2.7mg of copper sulfate pentahydrate and 2.7mg of cobalt chloride hexahydrate, dissolving with a small amount of distilled water, diluting to constant volume of 1L, storing in a brown glass bottle, and marking as mother solution
；
Weighing 1.6g of anhydrous ferrous sulfate and 3.76g of disodium ethylene diamine tetraacetate, dissolving with a small amount of distilled water, diluting to constant volume of 1L, storing in brown glass bottle, and labeling as mother solution
；
Weighing inositol 12g, nicotinic acid 70mg, pyridoxine hydrochloride 70mg, thiamine hydrochloride 14mg, and glycine 0.3g, dissolving with small amount of distilled water, diluting to constant volume of 1L, storing in brown glass bottle, and labeling as mother liquor
；
Weighing malt extract powder 20g, soytone 2g, ammonium nitrate 0.850g, potassium nitrate 1.000g, calcium chloride 0.175g, magnesium sulfate 100mg, potassium dihydrogen phosphate 95mg, and agar 8.5g, placing in a 1L triangular flask, and collecting mother liquor
、
、
Sucking 5ml of the solution, placing in the above triangular flask, dissolving with a small amount of distilled water, diluting to constant volume of 1L, packaging into two 1L triangular flasks, sealing, and placing in a sterilizing pot (103.4 KPa, 121 deg.C)) Sterilizing for 20 min, taking out, cooling to 40-50 deg.C, pouring the culture medium into a culture dish, solidifying, and sterilizing (pressure of 0.105MPa, 121 deg.C, sterilization of 15-30 mIn) to separate and culture strain. Example 3
Weighing 82mg of potassium iodide, 0.60g of boric acid, 1.68g of manganese sulfate monohydrate, 0.84g of zinc sulfate heptahydrate, 24mg of sodium molybdate dihydrate, 2.4mg of copper sulfate pentahydrate and 2.4mg of cobalt chloride hexahydrate, dissolving with a small amount of distilled water, diluting to constant volume of 1L, storing in a brown glass bottle, and marking as mother solution
；
Weighing 1.48g of anhydrous ferrous sulfate and 3.72g of disodium ethylene diamine tetraacetate, dissolving with a small amount of distilled water, diluting to constant volume of 1L, storing in brown glass bottle, and labeling as mother solution
；
Weighing inositol 9g, nicotinic acid 40mg, pyridoxine hydrochloride 40mg, thiamine hydrochloride 8mg, and glycine 0.16g, dissolving with small amount of distilled water, diluting to constant volume of 1L, storing in brown glass bottle, and labeling as mother liquor
；
Weighing malt extract powder 12g, soytone 1.2g, ammonium nitrate 0.820g, potassium nitrate 0.920g, calcium chloride 0.160g, magnesium sulfate 85mg, potassium dihydrogen phosphate 80mg, and agar 7.0g, placing in a 1L triangular flask, and collecting mother liquor
、
、
Taking 5ml of the solution, placing in the triangular flask, dissolving with a small amount of distilled water, diluting to constant volume of 1L, packaging into two triangular flasks of 1L, sealing, and placingSterilizing in a sterilizing pot (103.4 KPa, 121 deg.C) for 20 min, taking out, cooling to 40-50 deg.C, pouring the culture medium into a culture dish, solidifying, and sterilizing (pressure of 0.105MPa, 121 deg.C, sterilizing 15-30 mIN) to obtain the final product. Example 4
Weighing 84mg of potassium iodide, 0.64g of boric acid, 1.70g of manganese sulfate monohydrate, 0.88g of zinc sulfate heptahydrate, 26mg of sodium molybdate dihydrate, 2.6mg of copper sulfate pentahydrate and 2.6mg of cobalt chloride hexahydrate, dissolving with a small amount of distilled water, diluting to constant volume of 1L, storing in a brown glass bottle, and marking as mother solution
；
Weighing 1.56g of anhydrous ferrous sulfate and 3.74g of disodium ethylene diamine tetraacetate, dissolving with a small amount of distilled water, diluting to constant volume of 1L, storing in brown glass bottle, and labeling as mother solution
；
Weighing inositol 11g, nicotinic acid 60mg, pyridoxine hydrochloride 60mg, thiamine hydrochloride 12mg, and glycine 0.26g, dissolving with small amount of distilled water, diluting to constant volume of 1L, storing in brown glass bottle, and labeling as mother liquor
；
Weighing 18g of malt extract powder, 1.8g of soytone, 0.840g of ammonium nitrate, 0.980g of potassium nitrate, 0.170g of calcium chloride, 95mg of magnesium sulfate, 90mg of monopotassium phosphate and 8.0g of agar, placing the malt extract powder, the peptone and the agar in a 1L triangular flask, and respectively extracting from mother liquor
、
、
Taking 5ml of the solution, placing in the triangular flask, dissolving with a small amount of distilled water, diluting to constant volume of 1L, and packaging into two containersSealing in a 1L triangular flask, sterilizing in a sterilizing pot (103.4 KPa, 121 deg.C) for 20 min, taking out, cooling to 40-50 deg.C, pouring culture medium into a culture dish, solidifying, and sterilizing (pressure of 0.105MPa, 121 deg.C, sterilizing 15-30 mIN) to obtain the final product. Example 5
Weighing 83mg of potassium iodide, 0.62g of boric acid, 1.69g of manganese sulfate monohydrate, 0.86g of zinc sulfate heptahydrate, 25mg of sodium molybdate dihydrate, 2.5mg of copper sulfate pentahydrate and 2.5mg of cobalt chloride hexahydrate, dissolving with a small amount of distilled water, diluting to constant volume of 1L, storing in a brown glass bottle, and marking as mother solution
；
Weighing anhydrous ferrous sulfate 1.52g and disodium ethylene diamine tetraacetate 3.726g, dissolving with small amount of distilled water, diluting to constant volume of 1L, storing in brown glass bottle, and labeling as mother solution
；
Weighing inositol 10g, nicotinic acid 50mg, pyridoxine hydrochloride 50mg, thiamine hydrochloride 10mg, and glycine 0.2g, dissolving with small amount of distilled water, diluting to constant volume of 1L, storing in brown glass bottle, and labeling as mother liquor
；
Weighing malt extract powder 15g, soytone 1.5g, ammonium nitrate 0.825g, potassium nitrate 0.950g, calcium chloride 0.165g, magnesium sulfate 90mg, potassium dihydrogen phosphate 85mg and agar 7.5g, placing in a 1L triangular flask, and collecting mother liquor
、
、
Taking 5ml of the solution, placing the solution in the triangular flask, dissolving the solution with a small amount of distilled water, and fixing the volumeAnd (3) adding the culture medium into 1L, subpackaging into two 1L triangular flasks, sealing, sterilizing in a sterilizing pot (103.4 KPa, 121 ℃) for 20 minutes, taking out, cooling to 40-50 ℃, pouring the culture medium into a culture dish, solidifying, and sterilizing (the pressure is 0.105MPa, and the temperature is 121 ℃, sterilizing 15-30 mIn) for later use.
Adopting PDA, MMN and the culture medium of the invention to carry out separation culture of boletus lanuginosus strain according to the following steps: collecting fresh sporophore of boletus brevicaulis, removing surface impurities, and dividing the sporophore into two parts by hand in an ultra-clean laboratory bench; sterilizing a scalpel and a pair of tweezers on an alcohol lamp, cooling, scratching tissue blocks with the size of about 2-3mm at the center of pileus flesh of a fruiting body by using the scalpel, putting the tissue blocks into culture dishes containing different culture media by using the tweezers, and putting 3 tissue blocks in each culture dish; thirdly, placing the culture dish in a constant temperature incubator at 22 ℃ for culturing for 4-5 weeks, then drawing culture medium blocks with the size of 2-3mm and growing hyphae, placing the culture medium blocks into new culture dishes, placing 3 culture medium blocks into each culture dish, placing the culture medium blocks into the constant temperature incubator at 22 ℃ for culturing, observing the growth condition of the hyphae and measuring the diameter of the hyphae. The growth of the mycelia of the three culture media cultured on boletus pendula is shown in Table 1. According to the comparison result, when the culture medium is used for culturing the boletus pendorf, the diameter of the mycelial group and the growth speed of hyphae are superior to those of other two culture media.
TABLE 1 different species isolation medium the growth of boletus flagellata hyphae (6 weeks)
Claims (3)
1. A culture medium for separating bolete serrulata comprises a large amount of components, trace components, iron salts and growth factors, and is characterized in that the large amount of components comprise 10000-20000mg/L malt extract powder, 1000-2000mg/L soytone, 800-850mg/L ammonium nitrate, 900-1000mg/L potassium nitrate, 155-175mg/L calcium chloride, 80-100mg/L magnesium sulfate, 75-95mg/L potassium dihydrogen phosphate and 6500-8500mg/L agar; the micro-ingredients comprise potassium iodide with the concentration of 0.400-0.430mg/L, boric acid with the concentration of 2.8-3.3mg/L, manganese sulfate monohydrate with the concentration of 8.30-8.60mg/L, zinc sulfate heptahydrate with the concentration of 4.1-4.5mg/L, sodium molybdate dihydrate with the concentration of 0.100-0.150mg/L, copper sulfate pentahydrate with the concentration of 0.0115-0.0135mg/L and cobalt chloride hexahydrate with the concentration of 0.0115-0.0135 mg/L; the ferric salt comprises anhydrous ferrous sulfate with the concentration of 7.0-8.0mg/L and ethylene diamine tetraacetic acid with the concentration of 18.4-18.8 mg/L; the growth factor comprises inositol with concentration of 40-60mg/L, nicotinic acid with concentration of 0.15-0.35mg/L, pyridoxine hydrochloride with concentration of 0.15-0.35mg/L, thiamine hydrochloride with concentration of 0.03-0.07mg/L, and glycine with concentration of 0.6-1.5 mg/L.
2. The culture medium of claim 1, wherein the bulk components comprise a concentration of 12000-18000mg/L malt extract powder, 1200-1800mg/L soytone, 820-840mg/L ammonium nitrate, 920-980mg/L potassium nitrate, 160-170mg/L calcium chloride, 85-95mg/L magnesium sulfate, 80-90mg/L monopotassium phosphate, 7000-8000mg/L agar; the micro-ingredients comprise potassium iodide with the concentration of 0.410-0.420mg/L, boric acid with the concentration of 3.0-3.2mg/L, manganese sulfate monohydrate with the concentration of 8.40-8.50mg/L, zinc sulfate heptahydrate with the concentration of 4.2-4.4mg/L, sodium molybdate dihydrate with the concentration of 0.120-0.130mg/L, copper sulfate pentahydrate with the concentration of 0.0120-0.0130mg/L and cobalt chloride hexahydrate with the concentration of 0.0120-0.0130 mg/L; the ferric salt comprises anhydrous ferrous sulfate with the concentration of 7.4-7.8mg/L and ethylene diamine tetraacetic acid with the concentration of 18.6-18.7 mg/L; the growth factor contains inositol 45-55mg/L, nicotinic acid 0.20-0.30mg/L, pyridoxine hydrochloride 0.20-0.30mg/L, thiamine hydrochloride 0.04-0.06mg/L, and glycine 0.8-1.3 mg/L.
3. The isolated medium of boletus brevicompactum as claimed in claim 1, wherein said plurality of components comprises malt extract powder at a concentration of 15000mg/L, soytone at 1500mg/L, ammonium nitrate at 825mg/L, potassium nitrate at 950mg/L, calcium chloride at 165mg/L, magnesium sulfate at 90mg/L, potassium dihydrogen phosphate at 85mg/L, agar at 7500 mg/L; the micro-components comprise potassium iodide with the concentration of 0.415mg/L, boric acid with the concentration of 3.1mg/L, manganese sulfate monohydrate with the concentration of 8.45mg/L, zinc sulfate heptahydrate with the concentration of 4.3mg/L, sodium molybdate hydrate with the concentration of 0.125mg/L, copper sulfate pentahydrate with the concentration of 0.0125mg/L and cobalt chloride hexahydrate with the concentration of 0.0125 mg/L; the ferric salt comprises anhydrous ferrous sulfate with the concentration of 7.6mg/L and disodium ethylene diamine tetraacetate with the concentration of 18.63 mg/L; the growth factor comprises inositol with concentration of 50mg/L, nicotinic acid with concentration of 0.25mg/L, pyridoxine hydrochloride with concentration of 0.25mg/L, thiamine hydrochloride with concentration of 0.05mg/L and glycine with concentration of 1.0 mg/L.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1338509A (en) * | 2000-08-15 | 2002-03-06 | 陈德良 | Process for preparing seeds of edible and medical ectomycorrhize fungus and its antificial cuture method |
| CN103181298A (en) * | 2013-04-09 | 2013-07-03 | 吉林农业大学 | Method for synthesizing suillus granulatus mycorrhiza |
| CN108260470A (en) * | 2018-01-14 | 2018-07-10 | 华中农业大学 | A kind of method for improving matsutake mycorrhizal seedling raising and application |
| CN110367102A (en) * | 2019-05-31 | 2019-10-25 | 中南林业科技大学 | A kind of method of chinquapin regeneration plant and Applying Ectomycorrhizal Fungi symbiosis |
| CN111647514A (en) * | 2020-06-18 | 2020-09-11 | 上海市农业科学院 | Preparation method and cooling program of novel edible fungus strain protective agent |
| CN112544342A (en) * | 2020-12-10 | 2021-03-26 | 广东省林业科学研究院 | Efficient special culture medium for amanita sinensis and preparation method and application thereof |
-
2021
- 2021-03-27 CN CN202110328895.1A patent/CN112772298A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1338509A (en) * | 2000-08-15 | 2002-03-06 | 陈德良 | Process for preparing seeds of edible and medical ectomycorrhize fungus and its antificial cuture method |
| CN103181298A (en) * | 2013-04-09 | 2013-07-03 | 吉林农业大学 | Method for synthesizing suillus granulatus mycorrhiza |
| CN108260470A (en) * | 2018-01-14 | 2018-07-10 | 华中农业大学 | A kind of method for improving matsutake mycorrhizal seedling raising and application |
| CN110367102A (en) * | 2019-05-31 | 2019-10-25 | 中南林业科技大学 | A kind of method of chinquapin regeneration plant and Applying Ectomycorrhizal Fungi symbiosis |
| CN111647514A (en) * | 2020-06-18 | 2020-09-11 | 上海市农业科学院 | Preparation method and cooling program of novel edible fungus strain protective agent |
| CN112544342A (en) * | 2020-12-10 | 2021-03-26 | 广东省林业科学研究院 | Efficient special culture medium for amanita sinensis and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 余丽 等: "保山野生美网柄牛肝菌的菌丝分离研究", 《食用菌》, vol. 39, no. 04, pages 20 - 23 * |
| 栾竹青;梁丽琨;龚雪琴;刘传林;由翠荣;初洋;: "组培条件下乳牛肝菌培养基的优化研究", 生物学杂志, no. 02, pages 106 - 109 * |
| 苏鸿雁 等: "《苍山常见大型真菌图谱》", 云南人民出版社, pages: 94 - 96 * |
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