CN114058518A - Culture method of beauveria bassiana - Google Patents
Culture method of beauveria bassiana Download PDFInfo
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- CN114058518A CN114058518A CN202111421594.XA CN202111421594A CN114058518A CN 114058518 A CN114058518 A CN 114058518A CN 202111421594 A CN202111421594 A CN 202111421594A CN 114058518 A CN114058518 A CN 114058518A
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- 241000751139 Beauveria bassiana Species 0.000 title claims abstract description 69
- 238000012136 culture method Methods 0.000 title description 4
- 239000001963 growth medium Substances 0.000 claims abstract description 62
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 56
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims abstract description 28
- 235000019341 magnesium sulphate Nutrition 0.000 claims abstract description 28
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims abstract description 28
- 229910000368 zinc sulfate Inorganic materials 0.000 claims abstract description 28
- 229960001763 zinc sulfate Drugs 0.000 claims abstract description 28
- 239000000843 powder Substances 0.000 claims abstract description 26
- 229920001817 Agar Polymers 0.000 claims abstract description 18
- 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 18
- 239000001888 Peptone Substances 0.000 claims abstract description 18
- 108010080698 Peptones Proteins 0.000 claims abstract description 18
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 18
- 239000008272 agar Substances 0.000 claims abstract description 18
- 239000008103 glucose Substances 0.000 claims abstract description 18
- 235000019319 peptone Nutrition 0.000 claims abstract description 18
- 238000012258 culturing Methods 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
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims abstract description 16
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 229910001868 water Inorganic materials 0.000 claims abstract description 16
- 235000010419 agar Nutrition 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 14
- 238000009630 liquid culture Methods 0.000 claims abstract description 7
- IPCXNCATNBAPKW-UHFFFAOYSA-N zinc;hydrate Chemical compound O.[Zn] IPCXNCATNBAPKW-UHFFFAOYSA-N 0.000 claims abstract description 4
- 235000013312 flour Nutrition 0.000 claims description 24
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 12
- 244000068988 Glycine max Species 0.000 claims description 12
- 235000010469 Glycine max Nutrition 0.000 claims description 12
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 12
- 229930006000 Sucrose Natural products 0.000 claims description 12
- 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 12
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 12
- 240000008042 Zea mays Species 0.000 claims description 12
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 12
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 12
- 239000001110 calcium chloride Substances 0.000 claims description 12
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 12
- 239000004202 carbamide Substances 0.000 claims description 12
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 12
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 12
- 235000005822 corn Nutrition 0.000 claims description 12
- 239000008101 lactose Substances 0.000 claims description 12
- 239000005720 sucrose Substances 0.000 claims description 12
- 235000015099 wheat brans Nutrition 0.000 claims description 12
- 244000061456 Solanum tuberosum Species 0.000 claims description 8
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 8
- 229920001592 potato starch Polymers 0.000 claims description 4
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 abstract description 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 abstract description 2
- 229910052749 magnesium Inorganic materials 0.000 abstract description 2
- 239000011777 magnesium Substances 0.000 abstract description 2
- 235000016709 nutrition Nutrition 0.000 abstract description 2
- 230000035764 nutrition Effects 0.000 abstract description 2
- 239000011573 trace mineral Substances 0.000 abstract description 2
- 235000013619 trace mineral Nutrition 0.000 abstract description 2
- 239000011701 zinc Substances 0.000 abstract description 2
- 229910052725 zinc Inorganic materials 0.000 abstract description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003053 toxin Substances 0.000 description 2
- 231100000765 toxin Toxicity 0.000 description 2
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 1
- 241000238876 Acari Species 0.000 description 1
- 241000235349 Ascomycota Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- GYSCAQFHASJXRS-FFCOJMSVSA-N beauvericin Chemical compound C([C@H]1C(=O)O[C@@H](C(N(C)[C@@H](CC=2C=CC=CC=2)C(=O)O[C@@H](C(=O)N(C)[C@@H](CC=2C=CC=CC=2)C(=O)O[C@@H](C(=O)N1C)C(C)C)C(C)C)=O)C(C)C)C1=CC=CC=C1 GYSCAQFHASJXRS-FFCOJMSVSA-N 0.000 description 1
- GYSCAQFHASJXRS-UHFFFAOYSA-N beauvericin Natural products CN1C(=O)C(C(C)C)OC(=O)C(CC=2C=CC=CC=2)N(C)C(=O)C(C(C)C)OC(=O)C(CC=2C=CC=CC=2)N(C)C(=O)C(C(C)C)OC(=O)C1CC1=CC=CC=C1 GYSCAQFHASJXRS-UHFFFAOYSA-N 0.000 description 1
- 108010079684 beauvericin Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- QXDMQSPYEZFLGF-UHFFFAOYSA-L calcium oxalate Chemical compound [Ca+2].[O-]C(=O)C([O-])=O QXDMQSPYEZFLGF-UHFFFAOYSA-L 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 210000003705 ribosome Anatomy 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000004763 spore germination Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N3/00—Spore forming or isolating processes
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- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
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Abstract
The invention discloses a method for culturing beauveria bassiana, which comprises inoculating beauveria bassiana into a first-stage culture medium, wherein the first-stage culture medium comprises glucose, yeast powder, peptone, agar, monopotassium phosphate, magnesium sulfate, zinc sulfate and water, carrying out shake culture on the beauveria bassiana in a liquid culture medium for 30-40h at 20-22 ℃ to obtain beauveria bassiana mycelium, inoculating the beauveria bassiana mycelium in a second-stage culture medium, carrying out culture for 120-130h at 24-25 ℃ to obtain beauveria bassiana conidia, the beauveria bassiana grows in the first-stage culture medium and the second-stage culture medium in sequence, the first-stage culture medium contains glucose, yeast powder, peptone, agar, trace elements such as magnesium and zinc, thereby increasing the spore rate, and then providing nutrition through the second-stage culture medium, so that the beauveria bassiana conidia grow and continuously germinate, and the content of the finally obtained beauveria bassiana conidia is improved.
Description
Technical Field
The invention relates to the technical field of beauveria bassiana culture, in particular to a culture method of beauveria bassiana.
Background
Beauveria bassiana is an entomogenous fungus of ascomycetes, the main species comprise Beauveria bassiana, Beauveria bassiana and the like, conidia are usually generated through asexual propagation, and hypha has transverse septa and branches. The distribution range of the beauveria bassiana is wide, the beauveria bassiana is found in mountains with the altitude of several meters to more than 2000 meters, the beauveria bassiana can invade into the bodies of more than 200 insects and mites of 15 families of 6 orders to breed in a large quantity, simultaneously, beauvericin (non-ribosomal polypeptide toxin), oosporine (benzoquinone toxin) and calcium oxalate crystals are generated, and the substances can cause insect poisoning and disturb metabolism to cause death.
When the existing beauveria bassiana is cultured, most of the beauveria bassiana are placed in a culture medium for simple single culture, and the conidium content of the finally obtained beauveria bassiana is low due to poor culture effect.
Disclosure of Invention
The invention aims to provide a method for culturing beauveria bassiana, which aims to solve the problems that most of beauveria bassiana in the prior art are placed in a culture medium for simple single culture, and the conidium content of the finally obtained beauveria bassiana is low due to poor culture effect when the prior beauveria bassiana is cultured.
In order to achieve the purpose, the invention provides the following technical scheme: a method for culturing beauveria bassiana comprises the following steps:
s1: inoculating beauveria bassiana in a primary culture medium, wherein the primary culture medium comprises glucose, yeast powder, peptone, agar, monopotassium phosphate, magnesium sulfate, zinc sulfate and water;
wherein the components are calculated according to the weight percentage as follows:
40-50 parts of glucose, 10-20 parts of yeast powder, 10-20 parts of peptone, 20-40 parts of agar, 1-2 parts of monopotassium phosphate, 1-2 parts of magnesium sulfate, 0.1-0.3 part of zinc sulfate and 1200 parts of water;
s2: performing shake culture on beauveria bassiana in a liquid culture medium at 20-22 ℃ for 30-40h to obtain beauveria bassiana mycelium;
s3: inoculating beauveria bassiana mycelium into a secondary culture medium, wherein the secondary culture medium comprises, by weight, 20-30 parts of potato powder, 20-30 parts of corn flour, 15-20 parts of soybean flour, 50-70 parts of wheat bran, 6-8 parts of urea, 8-10 parts of sucrose, 8-12 parts of lactose, 0.03-0.05 part of zinc sulfate, 0.03-0.05 part of magnesium sulfate, 0.03-0.05 part of copper sulfate and 0.1-0.2 part of calcium chloride;
s4: culturing at 24-25 deg.c for 120-130 hr to obtain conidia of beauveria bassiana.
Preferably, the primary culture medium comprises, by weight, 40 parts of glucose, 10 parts of yeast powder, 10 parts of peptone, 20 parts of agar, 1 part of monopotassium phosphate, 1 part of magnesium sulfate, 0.1 part of zinc sulfate and 1000 parts of water.
Preferably, the primary culture medium comprises, by weight, 45 parts of glucose, 15 parts of yeast powder, 15 parts of peptone, 30 parts of agar, 1.5 parts of monopotassium phosphate, 1.5 parts of magnesium sulfate, 0.2 part of zinc sulfate and 1100 parts of water.
Preferably, the primary culture medium comprises, by weight, 50 parts of glucose, 20 parts of yeast powder, 20 parts of peptone, 40 parts of agar, 2 parts of monopotassium phosphate, 2 parts of magnesium sulfate, 0.3 part of zinc sulfate and 1200 parts of water.
Preferably, the second-stage culture medium comprises, by weight, 20 parts of potato flour, 20 parts of corn flour, 15 parts of soybean flour, 50 parts of wheat bran, 6 parts of urea, 8 parts of sucrose, 8 parts of lactose, 0.03 part of zinc sulfate, 0.03 part of magnesium sulfate, 0.03 part of copper sulfate and 0.1 part of calcium chloride.
Preferably, the secondary culture medium comprises, by weight, 25 parts of potato flour, 25 parts of corn flour, 17.5 parts of soybean flour, 60 parts of wheat bran, 7 parts of urea, 9 parts of sucrose, 10 parts of lactose, 0.04 part of zinc sulfate, 0.04 part of magnesium sulfate, 0.04 part of copper sulfate and 0.15 part of calcium chloride.
Preferably, the second-stage culture medium comprises, by weight, 30 parts of potato flour, 30 parts of corn flour, 20 parts of soybean flour, 70 parts of wheat bran, 8 parts of urea, 10 parts of sucrose, 12 parts of lactose, 0.05 part of zinc sulfate, 0.05 part of magnesium sulfate, 0.05 part of copper sulfate and 0.2 part of calcium chloride.
Compared with the prior art, the invention has the beneficial effects that:
1) the beauveria bassiana grows in a first-stage culture medium and a second-stage culture medium in sequence, the first-stage culture medium contains glucose, yeast powder, peptone, agar, trace elements magnesium and zinc, so that the spore germination rate can be improved, then the second-stage culture medium provides nutrition to enable the spores to grow and continuously germinate, and the conidium content of the beauveria bassiana finally obtained is improved;
2) the culture method provided by the invention is simple to operate, has a short period, and can effectively improve the content of conidia.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it should be noted that the terms "upper", "lower", "inner", "outer", "top/bottom", and the like, indicate orientations or positional relationships for convenience in describing the present invention and simplifying the description, but do not indicate or imply that the referred device or element must have a particular orientation, be constructed and operated in a particular orientation, and thus should not be construed as limiting the present invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "disposed," "sleeved/connected," "connected," and the like are to be construed broadly, e.g., "connected," which may be fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
The invention provides a technical scheme that: a method for culturing beauveria bassiana comprises the following steps:
s1: inoculating beauveria bassiana in a primary culture medium, wherein the primary culture medium comprises glucose, yeast powder, peptone, agar, monopotassium phosphate, magnesium sulfate, zinc sulfate and water;
wherein the components are calculated according to the weight percentage as follows:
40-50 parts of glucose, 10-20 parts of yeast powder, 10-20 parts of peptone, 20-40 parts of agar, 1-2 parts of monopotassium phosphate, 1-2 parts of magnesium sulfate, 0.1-0.3 part of zinc sulfate and 1200 parts of water;
s2: performing shake culture on beauveria bassiana in a liquid culture medium at 20-22 ℃ for 30-40h to obtain beauveria bassiana mycelium;
s3: inoculating beauveria bassiana mycelium into a secondary culture medium, wherein the secondary culture medium comprises, by weight, 20-30 parts of potato powder, 20-30 parts of corn flour, 15-20 parts of soybean flour, 50-70 parts of wheat bran, 6-8 parts of urea, 8-10 parts of sucrose, 8-12 parts of lactose, 0.03-0.05 part of zinc sulfate, 0.03-0.05 part of magnesium sulfate, 0.03-0.05 part of copper sulfate and 0.1-0.2 part of calcium chloride;
s4: culturing at 24-25 deg.c for 120-130 hr to obtain conidia of beauveria bassiana.
Example 1:
the method comprises the steps of inoculating beauveria bassiana in a primary culture medium, wherein the primary culture medium comprises 40g of glucose, 10g of yeast powder, 10g of peptone, 20g of agar, 1g of monopotassium phosphate, 1g of magnesium sulfate, 0.1g of zinc sulfate and 1000g of water, carrying out shake culture on the beauveria bassiana in a liquid culture medium for 30-40h at the temperature of 20-22 ℃ to obtain beauveria bassiana mycelia, inoculating the beauveria bassiana mycelia in a secondary culture medium, wherein the secondary culture medium comprises 20g of potato powder, 20g of corn flour, 15g of soybean flour, 50g of wheat bran, 6g of urea, 8g of sucrose, 8g of lactose, 0.03g of zinc sulfate, 0.03g of magnesium sulfate, 0.03g of copper sulfate and 0.1g of calcium chloride, and culturing the beauveria bassiana conidia at the temperature of 24-25 ℃ for 130h to obtain the beauveria bassiana conidia.
Example 2:
the preparation method comprises the steps of inoculating beauveria bassiana into a primary culture medium, wherein the primary culture medium comprises 45g of glucose, 15g of yeast powder, 15g of peptone, 30g of agar, 1.5g of monopotassium phosphate, 1.5g of magnesium sulfate, 0.2g of zinc sulfate and 1100g of water, carrying out shake culture on the beauveria bassiana in a liquid culture medium at the temperature of 20-22 ℃ for 30-40h to obtain beauveria bassiana mycelia, inoculating the beauveria bassiana mycelia into a secondary culture medium, wherein the secondary culture medium comprises 25g of potato powder, 25g of corn flour, 17.5g of soybean flour, 60g of wheat bran, 7g of urea, 9g of sucrose, 10g of lactose, 0.04g of zinc sulfate, 0.04g of magnesium sulfate, 0.04g of copper sulfate and 0.15g of calcium chloride, and carrying out culture for 120-plus 130h at the temperature of 24-25 ℃ to obtain the beauveria bassiana conidia.
Example 3:
the method comprises the steps of inoculating beauveria bassiana into a primary culture medium, wherein the primary culture medium comprises 50g of glucose, 20g of yeast powder, 20g of peptone, 40g of agar, 2g of monopotassium phosphate, 2g of magnesium sulfate, 0.3g of zinc sulfate and 1200g of water, carrying out shake culture on the beauveria bassiana in a liquid culture medium for 30-40h at the temperature of 20-22 ℃ to obtain beauveria bassiana mycelia, inoculating the beauveria bassiana mycelia into a secondary culture medium, wherein the secondary culture medium comprises 30g of potato powder, 30g of corn flour, 20g of soybean flour, 70g of wheat bran, 8g of urea, 10g of sucrose, 12g of lactose, 0.05g of zinc sulfate, 0.05g of magnesium sulfate, 0.05g of copper sulfate and 0.2g of calcium chloride, and culturing for 130h at the temperature of 24-25 ℃ to obtain the conidia of the beauveria bassiana conidia.
The conidium contents finally obtained in the above examples were measured as follows, wherein the contents of the inoculated beauveria bassiana were all 107spore/mL;
| numbering | Example 1 | Example 2 | Example 3 |
| Content (wt.) | 128*108Spores/g | 136*108Spores/g | 142*108Spores/g |
While there have been shown and described the fundamental principles and essential features of the invention and advantages thereof, it will be apparent to those skilled in the art that the invention is not limited to the details of the foregoing exemplary embodiments, but is capable of other specific forms without departing from the spirit or essential characteristics thereof; the present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (7)
1. A method for culturing beauveria bassiana is characterized in that: the method comprises the following steps:
s1: inoculating beauveria bassiana in a primary culture medium, wherein the primary culture medium comprises glucose, yeast powder, peptone, agar, monopotassium phosphate, magnesium sulfate, zinc sulfate and water;
wherein the components are calculated according to the weight percentage as follows:
40-50 parts of glucose, 10-20 parts of yeast powder, 10-20 parts of peptone, 20-40 parts of agar, 1-2 parts of monopotassium phosphate, 1-2 parts of magnesium sulfate, 0.1-0.3 part of zinc sulfate and 1200 parts of water;
s2: performing shake culture on beauveria bassiana in a liquid culture medium at 20-22 ℃ for 30-40h to obtain beauveria bassiana mycelium;
s3: inoculating beauveria bassiana mycelium into a secondary culture medium, wherein the secondary culture medium comprises, by weight, 20-30 parts of potato powder, 20-30 parts of corn flour, 15-20 parts of soybean flour, 50-70 parts of wheat bran, 6-8 parts of urea, 8-10 parts of sucrose, 8-12 parts of lactose, 0.03-0.05 part of zinc sulfate, 0.03-0.05 part of magnesium sulfate, 0.03-0.05 part of copper sulfate and 0.1-0.2 part of calcium chloride;
s4: culturing at 24-25 deg.c for 120-130 hr to obtain conidia of beauveria bassiana.
2. The method for culturing beauveria bassiana according to claim 1, wherein the culture medium comprises: the first-stage culture medium comprises, by weight, 40 parts of glucose, 10 parts of yeast powder, 10 parts of peptone, 20 parts of agar, 1 part of monopotassium phosphate, 1 part of magnesium sulfate, 0.1 part of zinc sulfate and 1000 parts of water.
3. The method for culturing beauveria bassiana according to claim 1, wherein the culture medium comprises: the first-stage culture medium comprises, by weight, 45 parts of glucose, 15 parts of yeast powder, 15 parts of peptone, 30 parts of agar, 1.5 parts of monopotassium phosphate, 1.5 parts of magnesium sulfate, 0.2 part of zinc sulfate and 1100 parts of water.
4. The method for culturing beauveria bassiana according to claim 1, wherein the culture medium comprises: the first-stage culture medium comprises, by weight, 50 parts of glucose, 20 parts of yeast powder, 20 parts of peptone, 40 parts of agar, 2 parts of monopotassium phosphate, 2 parts of magnesium sulfate, 0.3 part of zinc sulfate and 1200 parts of water.
5. The method for culturing beauveria bassiana according to claim 1, wherein the culture medium comprises: the second-stage culture medium comprises, by weight, 20 parts of potato powder, 20 parts of corn flour, 15 parts of soybean flour, 50 parts of wheat bran, 6 parts of urea, 8 parts of sucrose, 8 parts of lactose, 0.03 part of zinc sulfate, 0.03 part of magnesium sulfate, 0.03 part of copper sulfate and 0.1 part of calcium chloride.
6. The method for culturing beauveria bassiana according to claim 1, wherein the culture medium comprises: the second-stage culture medium comprises, by weight, 25 parts of potato powder, 25 parts of corn flour, 17.5 parts of soybean flour, 60 parts of wheat bran, 7 parts of urea, 9 parts of sucrose, 10 parts of lactose, 0.04 part of zinc sulfate, 0.04 part of magnesium sulfate, 0.04 part of copper sulfate and 0.15 part of calcium chloride.
7. The method for culturing beauveria bassiana according to claim 1, wherein the culture medium comprises: the second-stage culture medium comprises, by weight, 30 parts of potato flour, 30 parts of corn flour, 20 parts of soybean flour, 70 parts of wheat bran, 8 parts of urea, 10 parts of sucrose, 12 parts of lactose, 0.05 part of zinc sulfate, 0.05 part of magnesium sulfate, 0.05 part of copper sulfate and 0.2 part of calcium chloride.
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| CN106497805A (en) * | 2016-12-30 | 2017-03-15 | 四川省农业科学院蚕业研究所 | A kind of cultural method of beauveria bassiana |
| CN106834123A (en) * | 2016-11-30 | 2017-06-13 | 四川省农业科学院蚕业研究所 | A kind of muscardine separates expanding propagation method |
| CN106906148A (en) * | 2017-04-25 | 2017-06-30 | 四川省农业科学院蚕业研究所 | A kind of culture medium of beauveria bassiana, the cultural method of beauveria bassiana |
| CN111676142A (en) * | 2020-04-14 | 2020-09-18 | 中国烟草总公司陕西省公司 | Preparation and application method of beauveria bassiana powder |
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2021
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106834123A (en) * | 2016-11-30 | 2017-06-13 | 四川省农业科学院蚕业研究所 | A kind of muscardine separates expanding propagation method |
| CN106497805A (en) * | 2016-12-30 | 2017-03-15 | 四川省农业科学院蚕业研究所 | A kind of cultural method of beauveria bassiana |
| CN106906148A (en) * | 2017-04-25 | 2017-06-30 | 四川省农业科学院蚕业研究所 | A kind of culture medium of beauveria bassiana, the cultural method of beauveria bassiana |
| CN111676142A (en) * | 2020-04-14 | 2020-09-18 | 中国烟草总公司陕西省公司 | Preparation and application method of beauveria bassiana powder |
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