CN114410498B - Microbial preparation, preparation method and application thereof - Google Patents
Microbial preparation, preparation method and application thereof Download PDFInfo
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- CN114410498B CN114410498B CN202111124073.8A CN202111124073A CN114410498B CN 114410498 B CN114410498 B CN 114410498B CN 202111124073 A CN202111124073 A CN 202111124073A CN 114410498 B CN114410498 B CN 114410498B
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- 238000002360 preparation method Methods 0.000 title claims abstract description 33
- 230000000813 microbial effect Effects 0.000 title claims abstract description 21
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims abstract description 45
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 43
- 241000607479 Yersinia pestis Species 0.000 claims abstract description 27
- 230000001580 bacterial effect Effects 0.000 claims description 24
- 238000000855 fermentation Methods 0.000 claims description 23
- 230000004151 fermentation Effects 0.000 claims description 23
- 238000012258 culturing Methods 0.000 claims description 20
- 238000004321 preservation Methods 0.000 claims description 20
- 239000003795 chemical substances by application Substances 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 14
- 239000002068 microbial inoculum Substances 0.000 claims description 13
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- 241000488583 Panonychus ulmi Species 0.000 claims description 9
- 239000001963 growth medium Substances 0.000 claims description 8
- 238000004806 packaging method and process Methods 0.000 claims description 8
- 229920000881 Modified starch Polymers 0.000 claims description 7
- 239000004368 Modified starch Substances 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 7
- 235000019426 modified starch Nutrition 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 5
- 108010080698 Peptones Proteins 0.000 claims description 5
- 244000269722 Thea sinensis Species 0.000 claims description 5
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- 235000019319 peptone Nutrition 0.000 claims description 5
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims description 5
- 229930182490 saponin Natural products 0.000 claims description 5
- 150000007949 saponins Chemical class 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- 238000011218 seed culture Methods 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 239000005995 Aluminium silicate Substances 0.000 claims description 3
- 240000007594 Oryza sativa Species 0.000 claims description 3
- 235000007164 Oryza sativa Nutrition 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 3
- 230000004913 activation Effects 0.000 claims description 3
- 235000012211 aluminium silicate Nutrition 0.000 claims description 3
- 235000013312 flour Nutrition 0.000 claims description 3
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 claims description 3
- 229940057995 liquid paraffin Drugs 0.000 claims description 3
- 239000013028 medium composition Substances 0.000 claims description 3
- 235000009566 rice Nutrition 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims description 2
- 239000005909 Kieselgur Substances 0.000 claims description 2
- 239000000440 bentonite Substances 0.000 claims description 2
- 229910000278 bentonite Inorganic materials 0.000 claims description 2
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 claims description 2
- 239000011575 calcium Substances 0.000 claims description 2
- 229910052791 calcium Inorganic materials 0.000 claims description 2
- JZGWEIPJUAIDHM-UHFFFAOYSA-N chembl2007771 Chemical compound C1=CC=C2C(N=NC3=C4C(=CC(=CC4=CC=C3O)S(O)(=O)=O)S(O)(=O)=O)=CC=C(S(O)(=O)=O)C2=C1 JZGWEIPJUAIDHM-UHFFFAOYSA-N 0.000 claims description 2
- FPAFDBFIGPHWGO-UHFFFAOYSA-N dioxosilane;oxomagnesium;hydrate Chemical compound O.[Mg]=O.[Mg]=O.[Mg]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O FPAFDBFIGPHWGO-UHFFFAOYSA-N 0.000 claims description 2
- 238000005469 granulation Methods 0.000 claims description 2
- 230000003179 granulation Effects 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 241001092621 Sarucallis kahawaluokalani Species 0.000 claims 1
- 230000003213 activating effect Effects 0.000 claims 1
- 238000011081 inoculation Methods 0.000 claims 1
- 239000012188 paraffin wax Substances 0.000 claims 1
- 238000004659 sterilization and disinfection Methods 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 5
- 239000003094 microcapsule Substances 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 2
- 239000004480 active ingredient Substances 0.000 abstract 1
- 230000000694 effects Effects 0.000 description 25
- 241000238631 Hexapoda Species 0.000 description 18
- 230000000052 comparative effect Effects 0.000 description 16
- 238000002474 experimental method Methods 0.000 description 16
- 241001124076 Aphididae Species 0.000 description 13
- 230000001276 controlling effect Effects 0.000 description 13
- 230000009467 reduction Effects 0.000 description 11
- 230000000855 fungicidal effect Effects 0.000 description 10
- 239000000417 fungicide Substances 0.000 description 10
- 238000002156 mixing Methods 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 241000258937 Hemiptera Species 0.000 description 6
- 239000002609 medium Substances 0.000 description 5
- 239000003124 biologic agent Substances 0.000 description 4
- 239000002054 inoculum Substances 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 241001454295 Tetranychidae Species 0.000 description 3
- 238000012271 agricultural production Methods 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 239000000575 pesticide Substances 0.000 description 3
- 230000003405 preventing effect Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 241000238876 Acari Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 241000254137 Cicadidae Species 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 210000002615 epidermis Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 235000016623 Fragaria vesca Nutrition 0.000 description 1
- 240000009088 Fragaria x ananassa Species 0.000 description 1
- 235000011363 Fragaria x ananassa Nutrition 0.000 description 1
- 240000000161 Lagerstroemia indica Species 0.000 description 1
- 244000198896 Lagerstroemia speciosa Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 239000000447 pesticide residue Substances 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
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/12—Powders or granules
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
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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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Pest Control & Pesticides (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Plant Pathology (AREA)
- Environmental Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Dentistry (AREA)
- Inorganic Chemistry (AREA)
- Toxicology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a microbial preparation, which takes bacillus amyloliquefaciens and bacillus bailii as active ingredients, is prepared into a microcapsule type biological preparation with a quick-release and slow-release function through a microcapsule type process, and achieves the purpose of controlling piercing-sucking mouthpart pests by controlling the release time of the bacillus amyloliquefaciens and the bacillus bailii in the microbial preparation.
Description
Technical Field
The invention belongs to the field of biological pesticides, and particularly relates to a microbial preparation, a preparation method and application thereof.
Background
Piercing-sucking (sucking mouthparts) is a mouthpiece that insects that consume plant sap have to penetrate into the host and suck the host's body fluid. In the current agricultural pest control, piercing-sucking mouthpart pests are one of the plant pest groups in the world, and the piercing-sucking pests are very easy to generate drug resistance due to small individual, short generation period, fast growth and propagation, strong adaptability and high mutation rate, so that the agricultural production is extremely difficult to control, the agricultural production is greatly lost, the agricultural yield and the peasant income are seriously influenced, and the piercing-sucking pests become an important problem to be solved in the current agricultural production.
Chemical control is still the main countermeasure for controlling piercing-sucking mouthpart pests at present, but with the long-term and large-scale use of chemical agents, the drug resistance of the piercing-sucking pests is continuously enhanced, so that the control effect is quite unstable, and meanwhile, the serious pesticide residues and environmental safety problems are brought.
Disclosure of Invention
The invention screens and cultures the microorganism strain with specific high-efficient preventing and controlling activity to piercing-sucking mouthpart pests, and prepares the biological pesticide with quick-release and slow-release functions by adopting a microcapsule formulation process, so that the biological pesticide is released with time difference.
A microbial preparation comprises at least one of Bacillus amyloliquefaciens and Bacillus bailii, wherein one strain of Bacillus amyloliquefaciens (strain preservation number CCTCC M2021846) and one strain of Bacillus bailii (strain preservation number CCTCC M2021864) are adopted.
The active enzymes, protein polypeptides and other substances produced by the bacillus amyloliquefaciens and the bacillus belicus are utilized to dissolve the pest body wall, so that the pest body wall is thinned, the protection function is lost, and the pest body is easy to infect by beneficial bacteria, so that the pest body is atrophic and dead, and the effect of preventing and controlling piercing-sucking mouthpart pests is achieved. Among the sucking mouthparts pests include, but are not limited to: aphids, bugs, scale insects, mites, whiteflies, cicadas and the like, more preferably spider mites or aphids.
Furthermore, good control effect is obtained by controlling the release time of the bacillus amyloliquefaciens and the bacillus bailii; the bacillus amyloliquefaciens is a quick-release bacterial agent and utilizes biological enzymes generated by the bacillus amyloliquefaciens to firstly break the epidermis of piercing-sucking mouthpart pests; then the bacillus belicus is a slow-release microbial inoculum, and secondary action is carried out on the piercing-sucking mouth parts with damaged surfaces;
further, the control effect on piercing-sucking mouthpart pests is obtained by adjusting the proportion of bacillus amyloliquefaciens and bacillus bailii in the microbial inoculum.
Preferably, when aphids are prevented, the ratio of the bacillus amyloliquefaciens to the bacillus beijerinus is 3:2; when the spider mites are prevented, the ratio of the bacillus amyloliquefaciens to the bacillus bailii is 3:2, and the microbial preparation can obtain more efficient preventing and controlling effects.
Further, bacillus belicus is deposited in China center for type culture collection, and a strain deposit number is: cctccc M2021864, with a preservation time of 2021, 7 months and 12 days, named: bacillus belicus TP-06Bacillus velezensis TP-06, address: chinese armed chinese.
Further, the bacillus amyloliquefaciens is preserved in China center for type culture collection, and the strain preservation number is: cctccc M2021846, with a preservation time of 2021, 7 months and 8 days, named: bacillus amyloliquefaciens TP-08Bacillus amyloliquefaciens TP-08, address: chinese armed chinese.
The strain has the characteristics of mutual coordination when bacillus amyloliquefaciens and bacillus belicus play a role in mixing piercing-sucking mouthpart pests, and is a cultured strain with high-efficiency control activity on the piercing-sucking mouthpart pests.
The application of the microbial preparation comprises that the microbial preparation acts on piercing-sucking mouthpart pests simultaneously.
Further, the piercing-sucking mouthpart pests include, but are not limited to: aphids, bugs, scale insects, mites, whiteflies, cicadas, and the like.
Further, the piercing-sucking mouthpart pests are spider mites and aphids.
The preparation method of the bacillus amyloliquefaciens quick-release microbial inoculum comprises the steps of taking a preserved strain, performing activation culture on an LB culture medium, culturing for 24-48 hours at 25-40 ℃, inoculating the strain to an LB culture solution triangular flask, culturing for 24-48 hours at 25-40 ℃, inoculating the strain to a seed tank, sterilizing the seed tank by using the LB culture solution, inoculating a microbial inoculum, and culturing for 72 hours at 25-40 ℃;
inoculating the seed culture solution into a fermentation tank, and culturing at 25-40deg.C in TP medium for 72 hr.
Adding the bacillus amyloliquefaciens cultured by fermentation into the modified starch according to the mass of the fermentation liquid in a proportion of 1-5%, fully and uniformly stirring, and detecting that the bacterial content of the microbial inoculum reaches 1 multiplied by 10 8 The CFU/mL reaches the packaging requirement; or adding modified starch at a ratio of 1-5% by mass of fermentation broth, stirring thoroughly, granulating with carrier comprising one or more of Kaolin, diatomaceous earth, light calcium, pulvis Talci, and bentonite, and making into solid preparation with bacterial content of 1×10 8 And the CFU/g reaches the packaging requirement.
The preparation method of the bacillus belicus slow-release microbial inoculum comprises the steps of taking a preserved strain, performing activation culture on an LB culture medium, culturing for 24-48 hours at 25-40 ℃, inoculating the strain to an LB culture solution triangular shake flask, culturing for 24-48 hours at 25-40 ℃, inoculating the strain to a seed tank, sterilizing the seed tank by using the LB culture solution, inoculating a microbial inoculum, and culturing for 72 hours at 25-40 ℃;
inoculating the seed culture solution into a fermentation tank, and culturing at 25-40deg.C in TP medium for 72 hr.
Adding the bacillus belicus cultured by fermentation into liquid paraffin according to the mass of the fermentation liquid in a proportion of 5-10%, fully and uniformly stirring, and detecting that the bacterial content of the microbial inoculum reaches 1 multiplied by 10 8 Spray granulation is carried out when the CFU/mL reaches the packaging requirement.
Further, the TP medium composition is as follows in g/L: 5-10g of rice flour, 5-10g of peptone, 1-5g of yeast extract, 5-10g of sodium chloride, 2.5-5.0g of tea saponin and 1000 ml of water, and the pH value is regulated to 7.0+/-0.3.
The invention has the beneficial effects that: according to the invention, bacillus amyloliquefaciens and bacillus belicus are mixed and acted on piercing-sucking mouthpart pests for the first time, the bacillus amyloliquefaciens is used as a quick release bacterial agent and the bacillus belicus is used as a slow release bacterial agent, so that the release time difference of the two bacterial agents is formed, the epidermis of the pests is firstly destroyed by using active enzymes, protein polypeptides and the like secreted by the bacillus amyloliquefaciens, and then the bacillus belicus is released, so that the piercing-sucking mouthpart pests can be implemented once, the effect of the two bacterial agents on different time periods is obtained, and the pest control effect of the piercing-sucking mouthpart is remarkably improved.
Drawings
FIG. 1 shows the effect of clear water on aphid control in comparative experiments of the examples of the present invention;
FIG. 2 shows the aphid control effect of a 100-fold solution of Bacillus bailii in a comparative experiment according to an embodiment of the present invention;
FIG. 3 shows the aphid control effect of a 300-fold solution of Bacillus bailii in a comparative experiment according to an embodiment of the present invention;
FIG. 4 shows the aphid control effect of a 500-fold solution of Bacillus bailii in a comparative experiment according to an embodiment of the present invention;
FIG. 5 shows the effect of the solution of the invention in a ratio of 3:2 of the bacillus amyloliquefaciens fast-release fungicide to the bacillus bailii slow-release fungicide in the comparative experiment for controlling aphids;
FIG. 6 shows the aphid control effect of a solution of 300 times of the 3:2 mixture of the bacillus amyloliquefaciens fast-release bacterial agent and the bacillus bailii slow-release bacterial agent in the comparative experiment of the embodiment of the invention;
FIG. 7 shows the aphid control effect of a 600-fold solution of a bacillus amyloliquefaciens fast-release fungicide and a bacillus bailii slow-release fungicide in a comparative experiment of the embodiment of the invention;
FIG. 8 shows the effect of clear water on controlling red spiders in comparative experiments in accordance with the examples of the invention;
FIG. 9 shows the effect of the 100-fold solution of Bacillus amyloliquefaciens on controlling red spiders in the comparative experiment of the embodiment of the invention;
FIG. 10 shows the effect of the Bacillus amyloliquefaciens 300 times solution on controlling red spiders in the comparative experiment of the embodiment of the invention;
FIG. 11 shows the effect of the 600-fold solution of Bacillus amyloliquefaciens on controlling red spiders in the comparative experiment of the embodiment of the invention;
FIG. 12 shows the effect of controlling red spiders by a 150-fold solution of a bacillus amyloliquefaciens fast-release fungicide and a bacillus bailii slow-release fungicide in a 3:2 mixture in a comparative experiment of the embodiment of the invention;
FIG. 13 shows the effect of controlling red spiders by a 300-fold solution of a bacillus amyloliquefaciens fast-release fungicide and a bacillus bailii slow-release fungicide in a 3:2 mixture in a comparative experiment of the embodiment of the invention;
FIG. 14 shows the effect of controlling red spiders by a 600-fold solution of a bacillus amyloliquefaciens fast-release fungicide and a bacillus bailii slow-release fungicide in a 3:2 mixture in a comparative experiment of the embodiment of the invention.
Detailed Description
The invention is further illustrated in the following, in conjunction with the accompanying drawings and specific embodiments.
Bacillus bailii of the invention is deposited in China center for type culture collection, and the strain deposit number is: cctccc M2021864, with a preservation time of 2021, 7 months and 12 days, named: bacillus belicus TP-06Bacillus velezensis TP-06, address: chinese armed chinese.
The bacillus amyloliquefaciens is preserved in China center for type culture collection, and the strain preservation number is: cctccc M2021846, with a preservation time of 2021, 7 months and 8 days, named: bacillus amyloliquefaciens TP-08Bacillus amyloliquefaciens TP-08, address: chinese armed chinese.
Example 1
1. Fermentation broth preparation
Inoculating Bacillus bailii by aseptic technique with LB as culture medium
(Bacillus bailii, deposited in China center for type culture collection, with strain deposit number of CCTCC M2021864, deposit time of 2021, 7 months and 12 days, named: bacillus bailii TP-06Bacillus velezensis TP-06, address: wuhan, china.) after culturing for 48 hours at 30 ℃, inoculating the culture broth into a fermenter, and culturing for 72 hours at 30 ℃ in a TP medium with a composition of g/L: 10g of peptone, 5g of yeast extract, 10g of sodium chloride, 3.0g of tea saponin and 1000 ml of water, and the pH value is regulated to 7.0+/-0.3.
2. Preparation of liquid microbial inoculant products
Taking 10 liters of fermentation liquor, adding 250g and changingHeating the starch in a stirring and mixing tank to 40 ℃, stirring at constant speed for 3 hours, and detecting the bacterial content of the microbial inoculum to be 1 multiplied by 10 8 And (3) bottling the solution with CFU/mL above.
3. Preparation of solid microbial inoculant products
Taking 10L of fermentation liquor, adding 350g of modified starch, heating to 40 ℃ in a stirring and mixing tank, uniformly stirring for 3 hours, transferring to a granulator, mixing and granulating with 8kg of kaolin carrier, and preparing a solid preparation, wherein the bacterial content of the preparation reaches 1X 10 8 And (5) packaging the materials with CFU/g or more.
Example 2
1. Fermentation broth preparation
Inoculating Bacillus amyloliquefaciens (China center for type culture collection, strain collection number: CCTCC M2021846, collection date: 2021, 7 months and 8 days) by taking LB as a culture medium, culturing at 37 ℃ for 36h, inoculating the culture broth into a fermentation tank, culturing at 37 ℃ in TP medium for 72h, wherein the composition of the TP medium is expressed in g/L: 10g of peptone, 5g of yeast extract, 10g of sodium chloride, 4.0g of tea saponin and 1000 ml of water, and the pH value is regulated to 7.0+/-0.3.
2. Preparation of liquid microbial inoculant products
Taking 10L of fermentation liquor, adding 250g of modified starch, heating to 40 ℃ in a stirring and mixing tank, stirring at constant speed for 3 hours, and detecting that the bacterial content of the microbial inoculum is 1 multiplied by 10 8 And (3) bottling the solution with CFU/mL above.
3. Preparation of solid microbial inoculant products
Taking 10 liters of fermentation liquor, adding 300g of modified starch, heating to 40 ℃ in a stirring and mixing tank, uniformly stirring for 3 hours, transferring to a granulator, mixing and granulating with 5kg of carrier diatomite and 3kg of light calcium carbonate to prepare a solid preparation, wherein the bacterial content of the preparation reaches 1 multiplied by 10 8 And (5) packaging the materials with CFU/g or more.
Experimental example 1
As shown in FIGS. 1-7, comparative experiment 1 Lagerstroemia speciosa aphid
Experimental crops: lagerstroemia indica (L.) DC
Experimental temperature: normal temperature
Biological agent 1: the bacillus amyloliquefaciens quick-release bacterial agent is mixed with bacillus bailii slow-release bacterial agent in a ratio of 3:2;
the bacillus belicus is a strain of China center for type culture collection, and the strain is provided with a preservation number: CCTCC M2021864, date of preservation: a strain of 2021, 7, 12;
the bacillus amyloliquefaciens, china center for type culture collection, strain preservation number: CCTCC M2021846, date of preservation: a strain of 2021, 7, 8;
biological agent 2: bacillus beliae microbial inoculum, bacillus amyloliquefaciens, china center for type culture collection, strain deposit number: CCTCC M2021846, date of preservation: a strain of 2021, 7, 8;
the administration method comprises the following steps: spraying and using once. Calculating the experimental data pair of the insect population reduction rate (insect population reduction rate= (number of live insects before administration-number of live insects after administration)/number of live insects before administration x 100) and the control effect (control effect= (treatment insect population reduction rate-control insect population reduction rate)/(100-control insect population reduction rate), and the data are shown in table 1;
TABLE 1
Experimental example 2
Comparative experiment 2 strawberry red spider
Experimental crops: strawberry
Experimental temperature: normal temperature
Biological agent 1: the bacillus amyloliquefaciens quick-release bacterial agent is mixed with bacillus bailii slow-release bacterial agent in a ratio of 3:2;
the bacillus belicus is a strain of China center for type culture collection, and the strain is provided with a preservation number: CCTCC M2021864, date of preservation: a strain of 2021, 7, 12;
the bacillus amyloliquefaciens, china center for type culture collection, strain preservation number: CCTCC M2021846, date of preservation: a strain of 2021, 7, 8;
biological agent 2: a bacillus amyloliquefaciens bacterial agent, wherein the bacillus amyloliquefaciens is deposited with the China center for type culture collection, and the strain deposit number is as follows: CCTCC M2021846, date of preservation: a strain of 2021, 7, 8;
the administration method comprises the following steps: spraying and using once. The experimental data pair of the rate of reduction of the insect population (rate of reduction of the insect population = (number of live insects before administration-number of live insects after administration)/number of live insects before administration ×100) and the control effect (control effect = (rate of reduction of the insect population treated-rate of reduction of the insect population control)/(rate of reduction of the insect population control 100-rate of reduction of the insect population control) were calculated as shown in table 2:
TABLE 2
Those of ordinary skill in the art will recognize that the embodiments described herein are for the purpose of aiding the reader in understanding the principles of the present invention and should be understood that the scope of the invention is not limited to such specific statements and embodiments. Those of ordinary skill in the art can make various other specific modifications and combinations from the teachings of the present disclosure without departing from the spirit thereof, and such modifications and combinations remain within the scope of the present disclosure.
Claims (6)
1. A combination of microbial agents, characterized by: the bacillus amyloliquefaciens comprises bacillus amyloliquefaciens, and the strain preservation number is CCTCC M2021846 and bacillus belicus, and the strain preservation number is CCTCC M2021864; the bacillus amyloliquefaciens is a starch-embedded quick-release bacterial agent, and the bacillus bailii is a liquid paraffin-embedded slow-release bacterial agent; the ratio of the bacillus amyloliquefaciens to the bacillus belicus is 3:2.
2. The microbial preparation combination according to claim 1, wherein the starch embedding of the bacillus amyloliquefaciens involves suspending the bacillus amyloliquefaciens cultured by fermentation in a starch solution and granulating to prepare the quick-release microbial preparation.
3. The microbial preparation combination according to claim 1, wherein the liquid paraffin embedding of bacillus beliensis involves suspending bacillus beliensis cultured by fermentation in paraffin liquid and granulating to obtain a slow-release microbial preparation.
4. The microbial preparation combination according to claim 2, which is characterized by comprising the steps of taking a preserved strain for activating culture on an LB culture medium, culturing for 24-48 hours at 25-40 ℃, inoculating the strain to an LB culture solution triangular shake flask, culturing for 24-48 hours at 25-40 ℃ for inoculating a seed tank, inoculating a strain solution after sterilizing the seed tank, and culturing for 72 hours at 25-40 ℃;
inoculating the seed culture solution into a fermentation tank, and culturing for 72 hours at 25-40 ℃ in a TP culture medium; the TP medium composition is as follows in g/L: 5-10g of rice flour, 5-10g of peptone, 1-5g of yeast extract, 5-10g of sodium chloride, 2.5-5.0g of tea saponin and 1000 ml of water, and adjusting the pH to 7.0+0.3；
Adding the bacillus amyloliquefaciens cultured by fermentation into the modified starch according to the mass of the fermentation liquid in a proportion of 1-5%, fully and uniformly stirring, and detecting that the bacterial content of the microbial inoculum reaches 1 multiplied by 10 8 The CFU/mL reaches the packaging requirement; or adding modified starch at a ratio of 1-5% by mass of fermentation broth, stirring thoroughly, granulating with carrier comprising one or more of Kaolin, diatomaceous earth, light calcium, pulvis Talci, and bentonite, and making into solid preparation with bacterial content of 1×10 8 And the CFU/g reaches the packaging requirement.
5. The microbial preparation combination according to claim 3, which is characterized by comprising the steps of taking a preserved strain for activation culture on an LB culture medium, culturing for 24-48 hours at 25-40 ℃, inoculating the strain to an LB culture solution triangular shake flask, culturing for 24-48 hours at 25-40 ℃ for seed tank inoculation, inoculating a strain solution to the seed tank after sterilization of the LB culture solution, and culturing for 72 hours at 25-40 ℃;
inoculating the seed culture solution into a fermentation tank, and culturing for 72 hours at 25-40 ℃ in a TP culture medium; the TP medium composition is as follows in g/L: 5-10g of rice flour, 5-10g of peptone, 1-5g of yeast extract, 5-10g of sodium chloride, 2.5-5.0g of tea saponin and 1000 ml of water, and adjusting the pH to 7.0+0.3；
Adding Bacillus bailii cultured by fermentation into liquid paraffin solution at a ratio of 5-10% by mass of fermentation solution, stirring thoroughly, and detecting that the bacterial content of the microbial inoculum reaches 1×10 8 Spray granulation is carried out when the CFU/mL reaches the packaging requirement.
6. The combination of microbial agents of claim 1, wherein: acting on the control of sucking mouthparts pests including crape myrtle aphids and strawberry red spiders.
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| WO2023246208A1 (en) * | 2022-06-24 | 2023-12-28 | 华中农业大学 | Use of bacillus velezensis bv-6 in pest prevention |
| CN115161240B (en) * | 2022-07-25 | 2023-10-24 | 四川龙蟒福生科技有限责任公司 | Bacillus velezensis strain and application thereof |
| CN115161239B (en) * | 2022-07-25 | 2023-10-24 | 四川龙蟒福生科技有限责任公司 | Special fermentation medium for bacillus velezensis bacteria and application thereof |
| CN115125175B (en) * | 2022-07-25 | 2023-10-24 | 四川龙蟒福生科技有限责任公司 | Fermentation method of mite-killing strain and application thereof |
| CN115678813B (en) * | 2022-11-23 | 2023-06-30 | 湖北省生物农药工程研究中心 | Bacillus belicus with high poisoning activity on spider mites and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105331557A (en) * | 2015-11-18 | 2016-02-17 | 浙江省农业科学院 | Bacillus amyloliquefaciens HZ179 and application thereof in preparation of microbial insecticide |
| WO2018093199A2 (en) * | 2016-11-18 | 2018-05-24 | 주식회사 제일그린산업 | Novel microorganism bacillus oryzicola yc7011 producing bacillopeptin-series cyclic lipopeptide, and microbial formulation including same |
| CN110684683A (en) * | 2019-09-10 | 2020-01-14 | 北方民族大学 | A kind of Bacillus amyloliquefaciens, bacterial agent and application |
| CA3113257A1 (en) * | 2018-09-28 | 2020-04-02 | Fmc Corporation | Bacillus amyloliquefaciens fcc1256 compositions and methods of controlling plant pathogens |
| CN111411063A (en) * | 2020-05-15 | 2020-07-14 | 云南省微生物发酵工程研究中心有限公司 | A Bacillus compound inoculant with the function of preventing and treating tobacco soil-borne diseases and preparation method thereof |
| CN113271780A (en) * | 2018-09-28 | 2021-08-17 | Fmc公司 | Bacillus amyloliquefaciens FCC1256 composition and method for controlling plant pathogens |
-
2021
- 2021-09-24 CN CN202111124073.8A patent/CN114410498B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105331557A (en) * | 2015-11-18 | 2016-02-17 | 浙江省农业科学院 | Bacillus amyloliquefaciens HZ179 and application thereof in preparation of microbial insecticide |
| WO2018093199A2 (en) * | 2016-11-18 | 2018-05-24 | 주식회사 제일그린산업 | Novel microorganism bacillus oryzicola yc7011 producing bacillopeptin-series cyclic lipopeptide, and microbial formulation including same |
| CA3113257A1 (en) * | 2018-09-28 | 2020-04-02 | Fmc Corporation | Bacillus amyloliquefaciens fcc1256 compositions and methods of controlling plant pathogens |
| CN113271780A (en) * | 2018-09-28 | 2021-08-17 | Fmc公司 | Bacillus amyloliquefaciens FCC1256 composition and method for controlling plant pathogens |
| CN110684683A (en) * | 2019-09-10 | 2020-01-14 | 北方民族大学 | A kind of Bacillus amyloliquefaciens, bacterial agent and application |
| CN111411063A (en) * | 2020-05-15 | 2020-07-14 | 云南省微生物发酵工程研究中心有限公司 | A Bacillus compound inoculant with the function of preventing and treating tobacco soil-borne diseases and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 贝莱斯芽孢杆菌(Bacillus velezensis)研究进展;蔡高磊;张凡;欧阳友香;赵昌松;彭宣和;江爱明;;北方园艺(第12期);全文 * |
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