CN114456986B - Broad-spectrum antibacterial bacillus aryabhattai and application thereof - Google Patents
Broad-spectrum antibacterial bacillus aryabhattai and application thereof Download PDFInfo
- Publication number
- CN114456986B CN114456986B CN202210230797.9A CN202210230797A CN114456986B CN 114456986 B CN114456986 B CN 114456986B CN 202210230797 A CN202210230797 A CN 202210230797A CN 114456986 B CN114456986 B CN 114456986B
- Authority
- CN
- China
- Prior art keywords
- germ
- bacillus aryabhattai
- brown spot
- bacillus
- fermentation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000976738 Bacillus aryabhattai Species 0.000 title claims abstract description 47
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 13
- 230000001717 pathogenic effect Effects 0.000 claims abstract description 27
- 241001347962 Lecanosticta pini Species 0.000 claims abstract description 18
- 244000052616 bacterial pathogen Species 0.000 claims abstract description 15
- 244000053095 fungal pathogen Species 0.000 claims abstract description 8
- 230000002265 prevention Effects 0.000 claims abstract description 6
- 238000000855 fermentation Methods 0.000 claims description 31
- 230000004151 fermentation Effects 0.000 claims description 31
- 230000005764 inhibitory process Effects 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 14
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 238000012258 culturing Methods 0.000 claims description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 5
- 108010080698 Peptones Proteins 0.000 claims description 5
- 235000019319 peptone Nutrition 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 244000241235 Citrullus lanatus Species 0.000 claims description 4
- 235000012828 Citrullus lanatus var citroides Nutrition 0.000 claims description 4
- 240000007049 Juglans regia Species 0.000 claims description 4
- 235000009496 Juglans regia Nutrition 0.000 claims description 4
- 244000061176 Nicotiana tabacum Species 0.000 claims description 4
- 235000002637 Nicotiana tabacum Nutrition 0.000 claims description 4
- 238000004321 preservation Methods 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 4
- 235000020234 walnut Nutrition 0.000 claims description 4
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 claims description 3
- 241000251468 Actinopterygii Species 0.000 claims description 3
- 241000207199 Citrus Species 0.000 claims description 3
- 241000223218 Fusarium Species 0.000 claims description 3
- 241000223195 Fusarium graminearum Species 0.000 claims description 3
- 206010027146 Melanoderma Diseases 0.000 claims description 3
- 241000219000 Populus Species 0.000 claims description 3
- 206010037660 Pyrexia Diseases 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 3
- 244000273928 Zingiber officinale Species 0.000 claims description 3
- 235000006886 Zingiber officinale Nutrition 0.000 claims description 3
- 235000020971 citrus fruits Nutrition 0.000 claims description 3
- 235000008397 ginger Nutrition 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 241000233866 Fungi Species 0.000 claims description 2
- 241000233647 Phytophthora nicotianae var. parasitica Species 0.000 claims description 2
- 235000007688 Lycopersicon esculentum Nutrition 0.000 claims 2
- 241001533598 Septoria Species 0.000 claims 2
- 240000003768 Solanum lycopersicum Species 0.000 claims 2
- 235000009754 Vitis X bourquina Nutrition 0.000 claims 2
- 235000012333 Vitis X labruscana Nutrition 0.000 claims 2
- 240000006365 Vitis vinifera Species 0.000 claims 2
- 235000014787 Vitis vinifera Nutrition 0.000 claims 2
- 244000144730 Amygdalus persica Species 0.000 claims 1
- 235000017060 Arachis glabrata Nutrition 0.000 claims 1
- 244000105624 Arachis hypogaea Species 0.000 claims 1
- 235000010777 Arachis hypogaea Nutrition 0.000 claims 1
- 235000018262 Arachis monticola Nutrition 0.000 claims 1
- 235000002566 Capsicum Nutrition 0.000 claims 1
- 235000016623 Fragaria vesca Nutrition 0.000 claims 1
- 240000009088 Fragaria x ananassa Species 0.000 claims 1
- 235000011363 Fragaria x ananassa Nutrition 0.000 claims 1
- 239000006002 Pepper Substances 0.000 claims 1
- 235000016761 Piper aduncum Nutrition 0.000 claims 1
- 235000017804 Piper guineense Nutrition 0.000 claims 1
- 244000203593 Piper nigrum Species 0.000 claims 1
- 235000008184 Piper nigrum Nutrition 0.000 claims 1
- 235000006040 Prunus persica var persica Nutrition 0.000 claims 1
- 241000918585 Pythium aphanidermatum Species 0.000 claims 1
- 235000021307 Triticum Nutrition 0.000 claims 1
- 244000098338 Triticum aestivum Species 0.000 claims 1
- 240000006677 Vicia faba Species 0.000 claims 1
- 235000010749 Vicia faba Nutrition 0.000 claims 1
- 235000002098 Vicia faba var. major Nutrition 0.000 claims 1
- 235000020232 peanut Nutrition 0.000 claims 1
- 238000001228 spectrum Methods 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 29
- 235000008331 Pinus X rigitaeda Nutrition 0.000 abstract description 27
- 235000011613 Pinus brutia Nutrition 0.000 abstract description 27
- 241000018646 Pinus brutia Species 0.000 abstract description 27
- 244000052769 pathogen Species 0.000 abstract description 27
- 238000011282 treatment Methods 0.000 abstract description 18
- 230000002401 inhibitory effect Effects 0.000 abstract description 10
- 230000003042 antagnostic effect Effects 0.000 abstract description 5
- 239000003085 diluting agent Substances 0.000 abstract description 4
- 238000009629 microbiological culture Methods 0.000 abstract description 4
- 239000007788 liquid Substances 0.000 description 19
- 201000010099 disease Diseases 0.000 description 18
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 18
- 230000001580 bacterial effect Effects 0.000 description 16
- 241000894006 Bacteria Species 0.000 description 15
- 238000002360 preparation method Methods 0.000 description 14
- 239000001963 growth medium Substances 0.000 description 13
- 239000002689 soil Substances 0.000 description 11
- 238000012360 testing method Methods 0.000 description 11
- 230000012010 growth Effects 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 8
- 239000002609 medium Substances 0.000 description 6
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 description 5
- 239000006013 carbendazim Substances 0.000 description 5
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 description 5
- 238000009630 liquid culture Methods 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- 235000005638 Austrian pine Nutrition 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 235000008565 Pinus banksiana Nutrition 0.000 description 3
- 244000019397 Pinus jeffreyi Species 0.000 description 3
- 235000013264 Pinus jeffreyi Nutrition 0.000 description 3
- 235000008578 Pinus strobus Nutrition 0.000 description 3
- 235000008585 Pinus thunbergii Nutrition 0.000 description 3
- 235000014030 Podocarpus spicatus Nutrition 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000002207 metabolite Substances 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 235000017985 rocky mountain lodgepole pine Nutrition 0.000 description 3
- 238000005070 sampling Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000008223 sterile water Substances 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 241000223221 Fusarium oxysporum Species 0.000 description 2
- 241000220286 Sedum Species 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- NWFNSTOSIVLCJA-UHFFFAOYSA-L copper;diacetate;hydrate Chemical compound O.[Cu+2].CC([O-])=O.CC([O-])=O NWFNSTOSIVLCJA-UHFFFAOYSA-L 0.000 description 2
- 230000000593 degrading effect Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 239000012669 liquid formulation Substances 0.000 description 2
- 230000002906 microbiologic effect Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 239000004563 wettable powder Substances 0.000 description 2
- 241000228197 Aspergillus flavus Species 0.000 description 1
- 229930192334 Auxin Natural products 0.000 description 1
- 239000005946 Cypermethrin Substances 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 241000235527 Rhizopus Species 0.000 description 1
- 206010039509 Scab Diseases 0.000 description 1
- 239000000589 Siderophore Substances 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 239000002363 auxin Substances 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- KAATUXNTWXVJKI-UHFFFAOYSA-N cypermethrin Chemical compound CC1(C)C(C=C(Cl)Cl)C1C(=O)OC(C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 KAATUXNTWXVJKI-UHFFFAOYSA-N 0.000 description 1
- 229960005424 cypermethrin Drugs 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- -1 polyethylene Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- 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
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Virology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a broad-spectrum antibacterial bacillus aryabhattai and application thereof. The strain is bacillus aryabhattai AU0102, has been preserved in China general microbiological culture Collection center (CGMCC) for 15 days in 2018, has broad-spectrum antibacterial property and antagonistic effect on 18 pathogenic fungi and 4 pathogenic bacteria including brown spot pathogen of pine needle, especially has an inhibitory effect on brown spot pathogen of pine needle up to 93.90%, and the prevention and treatment effect of 300 times and 500 times of diluent on brown spot of pine needle up to 95.42% and 92.86%, and can effectively control the occurrence of brown spot of pine needle.
Description
Technical Field
The invention relates to a broad-spectrum antibacterial bacillus aryabhattai and application thereof, belonging to the technical field of microorganisms.
Background
Bacillus aryabhattai (Bacillus aryabhattai) belongs to the bacterial kingdom, the phylum Thick-walled bacteria, the phylum Paenium, the class of Bacillus, the order of Bacillus, and the family of Bacillus, and is a novel class of reporter bacteria of the genus Bacillus. The bacillus albopictus has wide application, is reported to be used for preventing and controlling diseases such as tobacco black shank (CN 201310559441.0), rice blast, fusarium wilt, potato scab (CN 201910375594.7) and the like, can also be used for inhibiting rhizopus albopictus (201910836765.1) and aspergillus flavus (CN 201510296912.2), generating auxin, producing siderophores, producing ammonia, producing protease, salt tolerance, antagonizing and phosphate solubilizing (CN 201610036163.4 and CN 202111159072.7), degrading cypermethrin (CN20161136508. X), effectively degrading polyethylene plastics (CN 202111159072.7) and the like, but has no report of preventing and controlling pine needle brown spot by the bacillus albopictus.
The pine needle brown spot is a common disease on pine trees caused by the sedum pini (Lecunosticta acicola), is mostly found in new needles, can not only cause the great reduction of the pine forest area, but also cause great ecological losses. At present, chemical agent prevention and treatment, cultivation measures and other prevention and treatment methods are mainly adopted for preventing and treating the brown spot of pine needles. Although the chemical control has better control effect, the chemical control has the problems of environmental pollution, easy drug resistance generation and the like. But the prevention and control effect of the adoption of cultivation measures is poor. Biological control is increasingly favored because of the advantages of high control effect, strong specificity, good drug effect durability, environmental friendliness, difficult generation of drug resistance and the like. But few reports on biological control of the brown spot of pine needles are presented at present.
Disclosure of Invention
Aiming at the problems, the invention provides a broad-spectrum antibacterial bacillus albosis (Bacillus aryabhattai) AU0102 and application thereof, wherein the bacillus albosis AU0102 has an inhibition effect on plant diseases of different families, has antibacterial broad-spectrum property, particularly has the best inhibition effect on brown spot pathogens of pine needles, can develop microbial pesticides aiming at brown spot pathogens of pine needles, and realizes green and efficient control on brown spot pathogens of pine needles.
The bacillus albopictus (Bacillus aryabhattai) AU0102 which has a strong inhibition effect on the brown spot of pine needles is screened from pine tree farm soil in the verdigris district of WiHai city of Shandong, and the strain is preserved in the China general microbiological culture collection center (CGMCC) with the preservation number of 15218 in the 01 month 15 days of 2018. Experiments show that: the strain AU0102 and the metabolite thereof can inhibit the growth of brown spot pathogen of pine needle with high efficiency, and have wide application prospect in preventing and treating brown spot of pine needle.
The invention also provides application of the bacillus aryabhattai AU0102 in inhibiting pathogenic fungi such as brown spot pathogen, fusarium graminearum, poplar rot pathogen, apple ring rot pathogen and pathogenic bacteria such as citrus canker pathogen, ginger fever pathogen, walnut black spot pathogen and watermelon soft rot pathogen.
The invention also provides a fermentation method of the bacillus aryabhattai (Bacillus aryabhattai) AU0102, which is characterized in that after the bacillus aryabhattai AU0102 prepares seed liquid, the seed liquid is inoculated into a fermentation culture medium for fermentation culture for 20-30 h at the temperature of 26-30 ℃ to obtain fermentation liquor.
The formula (mass ratio) of the seed liquid culture medium is as follows: 1% of peptone, 0.3% of beef powder, 0.5% of sodium chloride and the balance of water, and regulating the pH value to be 7.3+/-0.1.
The formula (mass ratio) of the liquid fermentation medium is as follows: 2.0 to 2.5 percent of soluble starch, 3.0 to 3.5 percent of fish peptone, 0.1 to 0.2 percent of magnesium sulfate, 0.5 to 0.8 percent of sodium chloride and the balance of water, and adjusting the pH value to be 7.1 plus or minus 0.1.
The invention also provides application of the bacillus aryabhattai AU0102 fermentation liquor in preventing and treating pine needle brown spot.
The beneficial effects of the invention are as follows:
1. broad spectrum of antibacterial activity
The bacillus aryasis AU0102 has antagonistic broad spectrum and has different degrees of inhibition effects on the growth of 18 pathogenic fungi (brown spot pathogen, fusarium graminearum, poplar rot pathogen, apple ring rot pathogen and the like) and 4 pathogenic bacteria (citrus canker, ginger fever pathogen, walnut black spot pathogen and watermelon soft rot pathogen); particularly has remarkable inhibition effect on brown spot pathogen and fusarium oxysporum, which are 93.90 percent and 91.19 percent respectively.
2. Has good effect of preventing and treating the brown spot of pine needles
Experiments show that: the strain AU0102 and the metabolite thereof can inhibit the growth of brown spot bacteria with high efficiency. The liquid preparation (fermentation liquor) prepared by taking the strain as a raw material is directly sprayed after being diluted 300 times and 500 times, has a control effect as high as 95.42 percent and 92.86 percent, and has wide application prospect in the control of the brown spot of pine needles.
3. The strain is easy to ferment and produce, has a simple use mode, and is suitable for large-area popularization and use.
Drawings
FIG. 1 shows the inhibitory effect of Bacillus aryabhattai AU0102 on brown spot pine needle bacteria;
FIG. 2 shows the inhibitory effect of fermentation supernatant of Bacillus aryabhattai AU0102 on brown spot pine needle bacteria;
FIG. 3 shows the morphology of single colonies of Bacillus aryabhattai AU 0102;
FIG. 4 shows the form of the strain Bacillus aryabhattai AU0102 (simple staining);
FIG. 5 shows gram stain results of Bacillus aryabhattai AU 0102;
FIG. 6 shows the results of staining of Bacillus aryabhattai AU0102 spores.
Detailed Description
The effects thereof are described below with reference to examples.
Example 1: isolation and purification of Bacillus aryabhattai (Bacillus aryabhattai) Strain AU0102
The bacillus albopictus (Bacillus aryabhattai) AU0102 is separated from soil by a dilution plate method and a plate streaking method, and the specific separation method is as follows:
(1) Soil sample collection
The sampling place is the black pine forest land of the verdigris district of the Wisea city in Shandong province; the sampling method is five-point sampling method, namely respectively collecting proper amounts of soil around the land and within the depth range of 10-15cm in the center, uniformly mixing the soil with equal amounts, putting the soil into a fresh-keeping bag, marking the collecting place and time and immediately separating the soil from the collecting person or putting the soil into a refrigerator at 4 ℃ for preservation.
(2) Preparation of soil suspension
Accurately weighing 1g of soil sample in a triangular flask containing 99mL of sterile water, placing the triangular flask containing glass beads in a shaking table at 28 ℃ and uniformly shaking at 120rpm for 20-30 min, and then placing the triangular flask in a water bath at 55-60 ℃ for incubation for 20-30 min to obtain soil suspension.
(3) Dilution coating
Sequentially diluting the bacterial sample into 10 by 10-fold dilution method -1 ～10 -6 A diluted bacterial sample; respectively from 10 -4 、10 -5 And 10 -6 100. Mu.L of the diluted bacterial sample was pipetted onto a plate in NA solid medium, each ladderThree parallel coats were applied.
(4) Culture purification
Culturing at 28deg.C for 36 hr, picking out microbial colonies with different forms on NA culture medium, streaking on NA culture medium plate, and observing colony growth condition at regular time; and then adopting a plate streaking method to select single bacteria for purification, and respectively numbering and storing.
By the method, 12 strains are initially separated according to different colony morphologies, and the strains are respectively numbered AU 0101-AU 0112.
Example 2: screening of efficient antagonistic bacterial strain for brown spot of pine needle
(1) Primary screen
The antagonistic strain of the pathogenic sedum pinolense (Lecunosticta acicola) which is highly effective against the brown spot disease of pine needle is screened by using a flat plate opposite culture method.
Culturing 12 strains to be detected separated and purified in the embodiment 1 on an NA culture medium in a streaking way, and culturing at the constant temperature of 28 ℃ for 24 hours to obtain bacterial liquid of the strains to be selected; selecting a strain of brown spot of pine needle, performing activation culture on a PDA culture medium, and performing constant-temperature culture at 25 ℃ for 60 hours; a puncher is used for punching a bacterial cake with the diameter of 5mm at the edge of the activated brown spot bacterial, and the bacterial cake is inoculated in the center of a flat plate; inoculating the bacterial liquid of the strain to be selected with an inoculating loop at the periphery 1.5cm away from the center; the control group only inoculates 5mm brown spot bacteria on the center of the PDA plate; each treatment was repeated 3 times; after the treatment is finished, culturing at the constant temperature of 28 ℃, observing the inhibition effect of the selected strain on pathogenic bacteria day by day, measuring the size of the disease spots of the experimental group after the pathogenic bacteria of the control group grow on the flat plate, and calculating the bacteriostasis rate.
Antibacterial ratio = [ (control colony diameter-cake diameter) - (treated colony diameter-cake diameter) ]/(control colony diameter-cake diameter) ×100%.
The results showed that, of the 12 strains isolated in example 1, 5 strains had inhibitory effects on the growth of brown spot pine needle bacteria, AU0102, AU0104, AU0107, AU0109 and AU0112 (Table 1); the statistical antibacterial rate shows that the bacterial strain AU0102 has a high inhibition effect on brown spot bacteria of 90.54 percent, and the antibacterial rate is obviously higher than that of other bacterial strains (table 1 and figure 1).
TABLE 1 inhibition effect of different strains on brown spot pine needle pathogen
(2) Double screen
And fermenting and cultivating 5 strains of bacteria obtained through preliminary screening to obtain AU0102, AU0104, AU0107, AU0109 and AU0112 with an inhibitory effect on the growth of brown spot bacteria. The specific method comprises the following steps: transferring a loop by using a fungus inoculating loop, placing the loop into a triangular flask (50 mL) filled with 10mL NA liquid culture medium, and carrying out constant-temperature shaking culture at 28 ℃ and 120r/min for 24 hours to obtain seed liquid; inoculating the seed solution into a triangular flask (250 mL) filled with 100mL NA liquid culture medium according to 1% inoculum size, and performing constant-temperature shaking culture at 28 ℃ and 120r/min for 24 hours to obtain fermentation liquor; filtering the fermentation broth with 0.22 μm bacterial filter to obtain fermentation supernatant; uniformly mixing the fermentation supernatant with PDA culture medium at 45-50deg.C at a ratio of 1:19, pouring into a culture dish, cooling, and placing pathogenic bacteria cake with diameter of 5mm in the center of the plate; sterile water is added into the control group according to the same proportion to replace fermentation supernatant; each treatment was repeated 3 times; after the treatment is finished, culturing at the constant temperature of 28 ℃, observing the inhibition effect of 5 strains fermentation supernatant on pathogenic bacteria day by day, measuring the size of the disease spots of the experimental group after the pathogenic bacteria of the control group grow on a flat plate, and calculating the bacteriostasis rate.
The results show that all of the AU0102, AU0104, AU0107, AU0109 and AU0112 fermentation supernatants have an inhibitory effect on the growth of brown spot pathogen, and the inhibition rate of the AU0102 fermentation supernatant is as high as 88.30% (see Table 2 and FIG. 2).
TABLE 2 inhibition effect of fermentation supernatants of different strains on brown spot pine needle bacteria
The results show that the strain AU0102 and the metabolite thereof can inhibit the growth of brown spot pathogen of pine needle with high efficiency, and have wide application prospect in preventing and treating brown spot disease of pine needle.
Example 3: identification of Strain AU0102
(1) Microbiological characteristics: the bacterial colony form of the strain on an NA culture medium or an LB culture medium is regular round, and the surface of the strain is moist, smooth, sticky and semitransparent; the later surface has folds and bulges, is milky white and has a diameter of 3-4 mm (refer to figure 3). Culturing on NA medium at 28deg.C for 24 hr, and performing simple staining with crystal violet, microscopic examination, and short bar shape of thallus (see FIG. 4); gram staining was positive (see figure 5). Spore staining showed that spores were oval.
(2) Molecular biological Properties
The 16S rDNA gene sequence of this strain was determined as follows (SEQ-1):
CTAGCTCCTTACGTGTATACTCTCCGACTTCGGGTGTTACAAACTCTCGTGGTGTGA CGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTAC TAGCGATTCCAGCTTCATGTAGGCGAGTTGCAGCCTACAATCCGAACTGAGAATGGTTTT ATGGGATTGGCTTGACCTCGCGGTCTTGCAGCCCTTTGTACCATCCATTGTAGCACGTGT GTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGT CACCGGCAGTCACCTTAGAGTGCCCAACTAAATGCTGGCAACTAAGATCAAGGGTTGCG CTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCAC CTGTCACTCTGTCCCCCGAAGGGGAACGCTCTATCTCTAGAGTTGTCAGAGGATGTCAA GACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGG GCCCCCGTCAATTCCTTTGAGTTTCAGTCTTGCGACCGTACTCCCCAGGCGGAGTGCTTA ATGCGTTAGCTGCAGCACTAAAGGGCGGAAACCCTCTAACACTTAGCACTCATCGTTTA CGGCGTGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCACGCTTTCGCGCCTCAGCGT CAGTTACAGACCAAAAAGCCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTC ACCGCTACACGTGGAATTCCGCTTTTCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGA CCCTCCACGGTTGAGCCGTGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGCGCGCT TTACGCCCAATAATTCCGGATAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACG TAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGTACGAGCAGTTACTCTCGTACTT GTTCTTCCCTAACAACAGAGTTTTACGACCCGAAAGCCTTCATCACTCACGCGGCGTTG CTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTG GGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTATGCATCGTTGCC TTGGTGAGCCGTTACCTCACCAACTAGCTAATGCACCGCGGGCCCATCTGTAAGTGATAG CCGAAACCATCTTTCAATCATCTCCCATGAAGGAGAAGATCCTATCCGGTATTAGCTTCG GTTTCCCGAAGTTATCCCAGTCTTACAGGCAGGTTGCCCACGTGTTACTCACCCGTCCGC CGCTAACGTCATAGAAGCAAGCTTCTAATCAGTTCGCTCGACTGCATTATAGTCTGC。
the strain AU0102 is identified as bacillus aryabhattai (Bacillus aryabhattai) by microbiological characteristics and molecular biological characteristics, and is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) with a preservation number of CGMCC NO.15218 in the year of 15 of 2018 and 01 month.
Example 4: bacteriostasis broad-spectrum research of bacillus aryabhattai AU0102
The bacteriostasis capability of the bacillus aryabhattai AU0102 on the tested pathogenic bacteria (18 pathogenic fungi and 4 pathogenic bacteria) is measured by adopting a plate counter method, and the specific method is as follows:
control group: the activated test pathogen was inoculated in the center of the PDA or NA plate, and 2. Mu.L of sterile water was inoculated at 1.5cm from the center.
Experimental group: inoculating the activated pathogen to the center of PDA or NA plate, and inoculating 2 μl of activated Bacillus aryabhattai AU0102 bacterial liquid (culturing Bacillus aryabhattai AU0102 on NA medium by streaking at 28deg.C for 24 hr) around 1.5cm from the center.
Each treatment was repeated three times. Culturing each treatment in a 28 ℃ incubator, photographing after the control group pathogenic bacteria grow on the flat plate, detecting the size of the disease spots of the experimental group, and calculating the bacteriostasis rate.
As a result of analysis, it was found that Bacillus aryabhattai AU0102 had inhibitory effects on the growth of 18 pathogenic fungi and 4 pathogenic bacteria to different degrees (refer to tables 3, 4); particularly has remarkable inhibition effect on brown spot pathogen and fusarium oxysporum, which are 93.90 percent and 91.19 percent respectively. The result shows that the bacillus aryabhattai AU0102 has inhibition effect on plant diseases of different families and has bacteriostasis broad spectrum; especially has the best effect of inhibiting the brown spot pathogen of pine needles, can develop microbial pesticides for the brown spot of pine needles, and realizes green and high-efficiency control of the brown spot of pine needles.
TABLE 3 inhibition of pathogenic fungi by AU0102
TABLE 4 inhibition of pathogenic bacteria by AU0102
Note that: "+" indicates that the inhibition effect is remarkable.
Example 5: preparation process of bacillus aryabhattai AU0102 liquid preparation
The formula of the seed liquid culture medium comprises: 1% of peptone, 0.3% of beef powder, 0.5% of sodium chloride and the balance of water, and regulating the pH value to be 7.3+/-0.1.
The formula of the fermentation medium comprises: 2.2% of soluble starch, 3.2% of fish peptone, 0.15% of magnesium sulfate, 0.6% of sodium chloride and the balance of water, and regulating the pH value to be 7.1+/-0.1.
The preparation process of the bacillus aryabhattai AU0102 liquid preparation comprises the following steps:
(1) Seed liquid obtaining: and (3) picking a small amount of bacteria from the inclined surface of the test tube by the bacillus arvensis AU0102, transferring the bacteria into a seed solution culture medium, and culturing the bacteria for 24 hours at the constant temperature of 28 ℃ and 120r/min by shaking, thereby obtaining seed solution.
(2) And (3) carrying out mass fermentation culture: sterilizing the fermentation medium material in a fermentation tank (the temperature is 121-124 ℃ for 30 min); under the aseptic condition, the cultured seed liquid (according to the inoculation volume ratio of 5-10%) is poured into a fermentation tank. Fermenting and culturing at 28 deg.c for 24-28 hr. The bacterial count can reach about 500 hundred million/ml, and the transformation rate can reach about 85%. The fermentation liquor is AU0102 liquid preparation.
Example 6: bacillus aryabhattai AU0102 liquid preparation for preventing and treating brown spot of pine needle
The example provides a related experiment of bacillus aryabhattai AU0102 liquid formulations against pine needle brown spot.
(1) Test agent
AU0102 liquid formulation (prepared in example 5); 75% carbendazim wettable powder (commercially available).
(2) Test crop and control object
The test crop is black pine; the control object is pine needle brown spot.
(3) Test ground conditions
The experimental land is arranged in the black pine forest farm of the round green area of the tobacco stand of Shandong province, and the brown spot of pine needles in the past year is serious in the land; the test for preventing and treating pine needle brown spot in woodland is carried out in 4 months and 15 days of 2019. And the cultivation conditions and management measures of all the test cells are consistent.
(4) Test design and arrangement
The test designs 500 times, 700 times and 1000 times dilution treatment of the liquid preparation of the bacillus aryabhattai AU0102, and 5 treatments of 75% carbendazim 700 times and no-drug-application clear water as comparison are carried out, wherein each cell is 30m 3 Repeating for 4 times, 20 cells, each randomly arranged. The medicine is applied (pine needles are sprayed) on the day of 4 months 15, the day of 4 months 30, the day of 5 months 15 and the day of 5 months 30 respectively, the medicine application rate (diluted) is 50 kg/cell, and the disease condition is investigated at the bottom of 6 months.
(5) Test investigation and calculation method
And counting the disease state after the last medicine application interval of 30d, and calculating the prevention and treatment effect.
See table 5 for the stage number of the onset of brown spot of pine needles.
TABLE 5 grading Standard for the incidence of brown spot on pine needles
Disease index DI (%) = [ Σ (disease grade) x number of plants in the grade) ]/[ (highest value of disease grade x total number of investigation) ] ×100 control effect (%) = (control grade index-treatment disease index)/control disease index x 100
The resulting data were statistically analyzed using DPS data processing software and Excel software.
(6) Results
TABLE 6 incidence index of pine needle brown spot after each treatment and control effect
After 30d treatment, the disease condition was investigated, and the disease index and disease prevention effect were counted, and the results are shown in table 6. The clear water control has a morbidity index of 52.67%, and the morbidity index of each treatment is lower than that of the clear water control. The morbidity indexes of the AU0102 liquid preparation after 300 times, 500 times and 700 times of diluent treatment are respectively 2.42%, 3.76% and 5.67%, and the morbidity index of the 75% carbendazim 700 times of diluent is 8.19%; the control effects of the liquid preparation of each multiple of AU0102 are 95.42%, 92.86% and 89.24%, wherein the control effect of the 300-fold diluted preparation is best, and the liquid preparation has obvious difference with other treatments on the level of P < 0.05; the control effect of the 75% carbendazim 700-time diluent is 84.45%. In conclusion, the liquid preparation containing the bacillus aryabhattai AU0102 can effectively control the harm of the brown spot of pine needles, and the control effect is better than that of 75% carbendazim wettable powder.
SEQUENCE LISTING
<110> Shandong national Wei forestry control Co., ltd
<120> broad-spectrum antibacterial bacillus aryabhattai and application thereof
<130> 0
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1427
<212> DNA
<213> 16S rDNA Gene sequence of Bacillus aryabhattai (Bacillus aryabhattai) AU0102
<400> 1
ctagctcctt acgtgtatac tctccgactt cgggtgttac aaactctcgt ggtgtgacgg 60
gcggtgtgta caaggcccgg gaacgtattc accgcggcat gctgatccgc gattactagc 120
gattccagct tcatgtaggc gagttgcagc ctacaatccg aactgagaat ggttttatgg 180
gattggcttg acctcgcggt cttgcagccc tttgtaccat ccattgtagc acgtgtgtag 240
cccaggtcat aaggggcatg atgatttgac gtcatcccca ccttcctccg gtttgtcacc 300
ggcagtcacc ttagagtgcc caactaaatg ctggcaacta agatcaaggg ttgcgctcgt 360
tgcgggactt aacccaacat ctcacgacac gagctgacga caaccatgca ccacctgtca 420
ctctgtcccc cgaaggggaa cgctctatct ctagagttgt cagaggatgt caagacctgg 480
taaggttctt cgcgttgctt cgaattaaac cacatgctcc accgcttgtg cgggcccccg 540
tcaattcctt tgagtttcag tcttgcgacc gtactcccca ggcggagtgc ttaatgcgtt 600
agctgcagca ctaaagggcg gaaaccctct aacacttagc actcatcgtt tacggcgtgg 660
actaccaggg tatctaatcc tgtttgctcc ccacgctttc gcgcctcagc gtcagttaca 720
gaccaaaaag ccgccttcgc cactggtgtt cctccacatc tctacgcatt tcaccgctac 780
acgtggaatt ccgcttttct cttctgcact caagttcccc agtttccaat gaccctccac 840
ggttgagccg tgggctttca catcagactt aagaaaccgc ctgcgcgcgc tttacgccca 900
ataattccgg ataacgcttg ccacctacgt attaccgcgg ctgctggcac gtagttagcc 960
gtggctttct ggttaggtac cgtcaaggta cgagcagtta ctctcgtact tgttcttccc 1020
taacaacaga gttttacgac ccgaaagcct tcatcactca cgcggcgttg ctccgtcaga 1080
ctttcgtcca ttgcggaaga ttccctactg ctgcctcccg taggagtctg ggccgtgtct 1140
cagtcccagt gtggccgatc accctctcag gtcggctatg catcgttgcc ttggtgagcc 1200
gttacctcac caactagcta atgcaccgcg ggcccatctg taagtgatag ccgaaaccat 1260
ctttcaatca tctcccatga aggagaagat cctatccggt attagcttcg gtttcccgaa 1320
gttatcccag tcttacaggc aggttgccca cgtgttactc acccgtccgc cgctaacgtc 1380
atagaagcaa gcttctaatc agttcgctcg actgcattat agtctgc 1427
Claims (7)
1. A plantBroad-spectrum antibacterial bacillus alrightBacillus aryabhattai) AU0102, wherein the preservation number of the strain is CGMCC NO.15218.
2. Use of bacillus aryabhattai AU0102 according to claim 1 for the inhibition of pathogenic fungi; the pathogenic fungi are pathogenic septoria pinnatifida of brown spot of pine needleLecunosticta acicolaAny one or more of fusarium graminearum, poplar rot germ, apple ring rot germ, tomato gray mold germ, tobacco red star germ, tomato early blight germ, peanut damping off germ, strawberry gray mold germ, tobacco black shank germ, wheat red mold germ, pepper anthracnose germ, broad bean fusarium wilt germ, watermelon fusarium wilt germ, grape anthracnose germ, peach brown germ, grape white rot germ and walnut brown spot germ.
3. Use of bacillus aryabhattai AU0102 according to claim 1 for inhibition of pathogenic bacteria; the pathogenic bacteria are one or more of citrus canker, ginger fever, walnut black spot and watermelon soft rot.
4. The fermentation method of bacillus aryabhattai AU0102 according to claim 1, wherein after preparing a seed solution from bacillus aryabhattai AU0102, inoculating the seed solution into a fermentation medium, and fermenting and culturing the seed solution at 26-30 ℃ for 20-30 hours to obtain a fermentation broth.
5. The fermentation method of bacillus aryabhattai AU0102 according to claim 4, wherein the fermentation medium is, by weight: 2.0-2.5% of soluble starch, 3.0-3.5% of fish peptone, 0.1-0.2% of magnesium sulfate, 0.5-0.8% of sodium chloride and the balance of water, and regulating pH 7.1+/-0.1.
6. A fermentation broth of Bacillus aryabhattai AU0102 prepared by the fermentation process of claim 4 or 5.
7. The process of claim 6, wherein the fermentation broth of the Bacillus aryabhattai AU0102 is used for treating the pathogenic septoria pinnatifida of brown spot of pine needleBacteria (fungus)Lecunosticta acicolaApplication in prevention and control.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210230797.9A CN114456986B (en) | 2022-03-09 | 2022-03-09 | Broad-spectrum antibacterial bacillus aryabhattai and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210230797.9A CN114456986B (en) | 2022-03-09 | 2022-03-09 | Broad-spectrum antibacterial bacillus aryabhattai and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114456986A CN114456986A (en) | 2022-05-10 |
| CN114456986B true CN114456986B (en) | 2023-06-02 |
Family
ID=81416709
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210230797.9A Active CN114456986B (en) | 2022-03-09 | 2022-03-09 | Broad-spectrum antibacterial bacillus aryabhattai and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114456986B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NL2031759B1 (en) * | 2022-05-02 | 2023-11-13 | Guangxi Academy Agricultural Sciences | Bacillus and its Application and Multi-strain Fermentation Compound Fermentation Plant Enzyme |
| CN114891679B (en) * | 2022-05-19 | 2023-06-20 | 山东省林业科学研究院 | Pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases |
| CN116218724B (en) * | 2023-02-02 | 2024-10-11 | 河北省科学院生物研究所 | Bacillus aryabhattai, microbial agent and application thereof |
| CN116574642B (en) * | 2023-04-19 | 2024-05-28 | 吉林农业大学 | Lecaniella albopictus YZ-151 fermentation preparation and preparation method and application thereof |
| CN117025444B (en) * | 2023-06-29 | 2024-04-02 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Polystyrene plastic degrading bacterium and method for degrading plastic by using same |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101942505A (en) * | 2010-04-28 | 2011-01-12 | 南京农业大学 | Pathogenic aeromonas hydrophila assay kit |
| CN101993836A (en) * | 2010-06-13 | 2011-03-30 | 河南省农业科学院 | Bacillus subtilis strain YB-81, fungicide and preparation method and application thereof |
| CN105993273A (en) * | 2016-05-31 | 2016-10-12 | 山东胜伟园林科技有限公司 | Method for carrying out original soil cultivation on mild saline-alkali soil through needle-leaved tree fallen leaf organic matter containing nutrient soil |
| CN108841744A (en) * | 2018-06-15 | 2018-11-20 | 保定微控生物科技有限公司 | A kind of bacillus subtilis with diseases prevention and degradation Phos double action |
-
2022
- 2022-03-09 CN CN202210230797.9A patent/CN114456986B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101942505A (en) * | 2010-04-28 | 2011-01-12 | 南京农业大学 | Pathogenic aeromonas hydrophila assay kit |
| CN101993836A (en) * | 2010-06-13 | 2011-03-30 | 河南省农业科学院 | Bacillus subtilis strain YB-81, fungicide and preparation method and application thereof |
| CN105993273A (en) * | 2016-05-31 | 2016-10-12 | 山东胜伟园林科技有限公司 | Method for carrying out original soil cultivation on mild saline-alkali soil through needle-leaved tree fallen leaf organic matter containing nutrient soil |
| CN108841744A (en) * | 2018-06-15 | 2018-11-20 | 保定微控生物科技有限公司 | A kind of bacillus subtilis with diseases prevention and degradation Phos double action |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114456986A (en) | 2022-05-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114456986B (en) | Broad-spectrum antibacterial bacillus aryabhattai and application thereof | |
| CN102433282B (en) | Bacillus subtilis NB12, as well as culture method and application thereof | |
| CN110066756B (en) | Paenibacillus kribbensis and preparation and application thereof | |
| CN108102929B (en) | Isaria javanica for resisting pymetrozine and application thereof | |
| CN113583888B (en) | Bacillus beleisi and application thereof in prevention and treatment of lycium bararum disease | |
| CN103756931A (en) | Paenibacillus kribbensis and its application | |
| CN112358984A (en) | Biocontrol strain YW-1 and preparation and application of biocontrol microbial inoculum thereof | |
| CN116445354A (en) | Pseudomonas friedrisburgh and application thereof in disease prevention | |
| CN105567600A (en) | Pathogen verticillium antagonistic bacterium and application thereof | |
| CN114806928A (en) | Pepper endophytic Bacillus beleisi PEB23 and application thereof | |
| CN113862160A (en) | Trichoderma pseudokoningii Tk905 strain with biocontrol and induced disease resistance effects and application thereof | |
| CN117106639A (en) | Streptomyces nojirimensis strain and application of fermentation liquor thereof in preventing and controlling pepper anthracnose | |
| CN102021131A (en) | Bacillus licheniformis strain and application thereof | |
| CN115851479B (en) | Bacteria with antagonism to botrytis cinerea and application thereof | |
| CN102719364B (en) | Trichoderma harzianum strain and application in prevention and control of phytophthora capsici Leonian thereof | |
| CN113832071B (en) | Brevibacillus halotolerans strain and application thereof in preparation of biocontrol microbial inoculum | |
| CN108998395A (en) | A kind of bacillus amyloliquefaciens and its application | |
| CN111394270A (en) | Nocardia gamboge and application thereof | |
| CN111349589B (en) | Bacillus amyloliquefaciens for preventing and treating stem rot of anoectochilus roxburghii and application thereof | |
| CN111454848B (en) | Actinomucor elegans GD48, fermentation product, microbial inoculum and application thereof | |
| CN114891679B (en) | Pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases | |
| CN117887589B (en) | Trichoderma FJ059 capable of parasitizing sclerotium of southern blight and application thereof | |
| CN114774288B (en) | Trichoderma harzianum and application thereof, and microbial agent for resisting anthracnose and promoting growth and development of strawberries | |
| CN116179640B (en) | Method for efficiently screening antagonistic bacteria of banana vascular wilt | |
| CN115010548B (en) | Biological compound fertilizer specially applied to antagonism and prevention of pear fire germ |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |