CN117535169A - Bacillus licheniformis ZLP-81 and application thereof - Google Patents
Bacillus licheniformis ZLP-81 and application thereof Download PDFInfo
- Publication number
- CN117535169A CN117535169A CN202310964657.9A CN202310964657A CN117535169A CN 117535169 A CN117535169 A CN 117535169A CN 202310964657 A CN202310964657 A CN 202310964657A CN 117535169 A CN117535169 A CN 117535169A
- Authority
- CN
- China
- Prior art keywords
- bacteria
- zlp
- bacillus licheniformis
- cucumber
- tomato
- 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.)
- Granted
Links
- 241000194108 Bacillus licheniformis Species 0.000 title claims abstract description 40
- 239000002689 soil Substances 0.000 claims abstract description 11
- 238000004321 preservation Methods 0.000 claims abstract description 5
- 239000012681 biocontrol agent Substances 0.000 claims abstract description 4
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 4
- 230000000853 biopesticidal effect Effects 0.000 claims abstract description 3
- -1 biopesticides Substances 0.000 claims abstract description 3
- 241000894006 Bacteria Species 0.000 claims description 35
- 240000008067 Cucumis sativus Species 0.000 claims description 17
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 claims description 17
- 238000000855 fermentation Methods 0.000 claims description 17
- 230000004151 fermentation Effects 0.000 claims description 17
- 244000052616 bacterial pathogen Species 0.000 claims description 11
- 241001124076 Aphididae Species 0.000 claims description 10
- 235000007688 Lycopersicon esculentum Nutrition 0.000 claims description 10
- 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 8
- 241000223218 Fusarium Species 0.000 claims description 8
- 230000000844 anti-bacterial effect Effects 0.000 claims description 8
- 239000003899 bactericide agent Substances 0.000 claims description 7
- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 claims description 6
- 241001674044 Blattodea Species 0.000 claims description 5
- 206010027146 Melanoderma Diseases 0.000 claims description 5
- 241000233679 Peronosporaceae Species 0.000 claims description 5
- 241000500437 Plutella xylostella Species 0.000 claims description 5
- 241000985245 Spodoptera litura Species 0.000 claims description 5
- 244000000005 bacterial plant pathogen Species 0.000 claims description 5
- 241000395107 Cladosporium cucumerinum Species 0.000 claims description 4
- 241000813090 Rhizoctonia solani Species 0.000 claims description 4
- 241000607528 Aeromonas hydrophila Species 0.000 claims description 3
- 241000607574 Aeromonas veronii Species 0.000 claims description 3
- 241000223602 Alternaria alternata Species 0.000 claims description 3
- 229920000742 Cotton Polymers 0.000 claims description 3
- 241000607471 Edwardsiella tarda Species 0.000 claims description 3
- 244000299507 Gossypium hirsutum Species 0.000 claims description 3
- 235000014443 Pyrus communis Nutrition 0.000 claims description 3
- 241000191967 Staphylococcus aureus Species 0.000 claims description 3
- 240000008042 Zea mays Species 0.000 claims description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 3
- 235000005822 corn Nutrition 0.000 claims description 3
- 229940063673 spermidine Drugs 0.000 claims description 3
- 239000004094 surface-active agent Substances 0.000 claims description 3
- 239000005648 plant growth regulator Substances 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 241000227653 Lycopersicon Species 0.000 claims 3
- 244000052769 pathogen Species 0.000 claims 1
- 241000238631 Hexapoda Species 0.000 abstract description 10
- 201000010099 disease Diseases 0.000 abstract description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 9
- 230000002147 killing effect Effects 0.000 abstract description 8
- 230000008635 plant growth Effects 0.000 abstract description 4
- 230000001737 promoting effect Effects 0.000 abstract description 4
- 241000196324 Embryophyta Species 0.000 description 11
- 238000012216 screening Methods 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 8
- 230000000749 insecticidal effect Effects 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 240000003768 Solanum lycopersicum Species 0.000 description 7
- 235000010633 broth Nutrition 0.000 description 7
- 230000005764 inhibitory process Effects 0.000 description 7
- 241000193830 Bacillus <bacterium> Species 0.000 description 6
- 241000607479 Yersinia pestis Species 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 241000238814 Orthoptera Species 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 238000012258 culturing Methods 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000575 pesticide Substances 0.000 description 3
- 230000000243 photosynthetic effect Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 238000012271 agricultural production Methods 0.000 description 2
- 230000008485 antagonism Effects 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000006013 carbendazim Substances 0.000 description 2
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 description 2
- 229930002875 chlorophyll Natural products 0.000 description 2
- 235000019804 chlorophyll Nutrition 0.000 description 2
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 2
- 230000006806 disease prevention Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- PFAXACNYGZVKMX-UHFFFAOYSA-N fenethazine Chemical compound C1=CC=C2N(CCN(C)C)C3=CC=CC=C3SC2=C1 PFAXACNYGZVKMX-UHFFFAOYSA-N 0.000 description 2
- 229950007454 fenethazine Drugs 0.000 description 2
- 239000003337 fertilizer Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000223600 Alternaria Species 0.000 description 1
- 241000722823 Armadillidium Species 0.000 description 1
- 241000123650 Botrytis cinerea Species 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 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 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 206010039509 Scab Diseases 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 240000006677 Vicia faba Species 0.000 description 1
- 235000010749 Vicia faba Nutrition 0.000 description 1
- 235000002098 Vicia faba var. major Nutrition 0.000 description 1
- 238000003811 acetone extraction Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- XTLNYNMNUCLWEZ-UHFFFAOYSA-N ethanol;propan-2-one Chemical compound CCO.CC(C)=O XTLNYNMNUCLWEZ-UHFFFAOYSA-N 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000002655 kraft paper Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 238000000643 oven drying Methods 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 244000000003 plant pathogen Species 0.000 description 1
- 230000003652 pro-growth Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
- C12N1/205—Bacterial isolates
-
- 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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P3/00—Fungicides
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P7/00—Arthropodicides
- A01P7/04—Insecticides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2101/00—Agricultural use
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/10—Bacillus licheniformis
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Environmental Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- Mycology (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Public Health (AREA)
- General Engineering & Computer Science (AREA)
- Botany (AREA)
- Agronomy & Crop Science (AREA)
- Materials Engineering (AREA)
- Molecular Biology (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Epidemiology (AREA)
- Dentistry (AREA)
- Soil Sciences (AREA)
- Insects & Arthropods (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
Abstract
The invention relates to a bacillus licheniformis%Bacillus licheniformis) ZLP-81 with the preservation number of CGMCC No.25028 has the functions of preventing diseases, killing insects and promoting plant growth. The invention also discloses application of the strain in preparing biocontrol agents, biopesticides, soil restoration agents or biofertilizers.
Description
Technical Field
The invention relates to bacillus licheniformis ZLP-81 and application thereof.
Background
The plant diseases and insect pests bring serious loss to agricultural production, and the conventional chemical pesticide control not only kills a large number of natural enemies and beneficial microorganisms and destroys ecological balance, but also enables the drug resistance of the pests to be rapidly increased, and causes great harm to the environment and human health. Therefore, research and development of safe, efficient and environment-friendly products and chemical fertilizer and pesticide substitution technologies have become strategic demands for guaranteeing grain safety, food safety and environmental safety.
Bacillus licheniformis is an aerobic and spore-producing bacillus which has the characteristics of no toxicity and harm to human and livestock, environmental friendliness, strong stress resistance, high propagation speed and the like, and can secrete various active substances. Researches show that the microorganism has stronger inhibition effect on rhizoctonia solani, botrytis cinerea, fusarium wilt, early blight germ, leaf mold germ, black spot germ, anthracnose germ, downy mildew germ, cladosporium cucumerinum, alternaria leaf spot and the like, and can promote the growth of crops.
However, there is no report on the strains having the functions of preventing diseases, killing insects and promoting plant growth at the same time.
Disclosure of Invention
The invention aims to provide bacillus licheniformis ZLP-81 with functions of preventing diseases, killing insects and promoting plant growth and application thereof.
The invention adopts the following technical scheme:
bacillus licheniformis @Bacillus licheniformis) ZLP-81 with the preservation number of CGMCC No.25028, the preservation unit of China general microbiological culture Collection center with the address of: the collection date is 2022, 6, 9 days of the national institute of microbiology, national academy of sciences, 1, 3, north Chen, west Lu, the Korean region of Beijing, china.
The application of the bacillus licheniformis ZLP-81 in preventing and controlling aphids, plutella xylostella, prodenia litura, grubs, cricket, pillworm, cockroaches and other pests.
Application of the bacillus licheniformis ZLP-81 in preventing and treating plant pathogenic bacteria, aquatic pathogenic bacteria and staphylococcus aureus is provided.
Further, the plant pathogenic bacteria include: the plant growth regulator comprises the components of tomato leaf mold bacteria, tomato gray mold bacteria, pear black spot bacteria, fusarium, cotton fusarium wilt bacteria, cucumber gray mold bacteria, cucumber anthracnose bacteria, cucumber downy mildew bacteria, cucumber cladosporium cucumerinum, apple alternaria alternate bacteria, rhizoctonia solani, corn small spot bacteria and tomato early blight bacteria.
Further, the aquatic pathogenic bacteria include: aeromonas hydrophila, edwardsiella tarda and Aeromonas veronii.
Application of the bacillus licheniformis ZLP-81 in preparing biocontrol agents, biopesticides, soil restoration agents or biofertilizers.
Further, the biocontrol agent, the biological pesticide, the soil restoration agent or the biological fertilizer comprises bacillus licheniformis ZLP-81 thalli and/or fermentation liquor; or surfactant, iturin, fenethazine or spermidine separated from the fermentation broth.
A bactericide comprises bacillus licheniformis ZLP-81 thallus or fermentation broth.
A bactericide comprises surfactant, iturin, and fenethazine or spermidine separated from Bacillus licheniformis ZLP-81 fermentation broth.
The invention has the beneficial effects that:
the bacillus licheniformis ZLP-81 is an aerobic and spore-producing rod-shaped microorganism, has the characteristics of environmental friendliness, strong stress resistance, high propagation speed and the like, and can secrete various active substances to promote plant growth.
The bacillus licheniformis ZLP-81 has stronger insecticidal activity, can effectively kill pests such as plutella xylostella, prodenia litura, aphid, grub, cricket, pillworm and the like, has broad-spectrum antibacterial property, has obvious inhibition effect on various plant pathogenic bacteria such as gray mold, sickle disease, fusarium wilt, early blight, leaf mold, black spot, anthracnose, downy mildew, scab, ring spot, small spot and the like, and has good application prospect in the aspects of developing functional microbial preparations such as insecticidal or disease prevention and the like.
Drawings
FIG. 1 is a colony chart of Bacillus licheniformis ZLP-81.
FIG. 2 is a diagram of strain ZLP-81 of Bacillus licheniformis.
FIG. 3 shows the control and treatment groups (left) and (right) of Bacillus licheniformis ZLP-81 for killing cockroaches.
FIG. 4 shows the aphid killing treatment (left) and control (right) of Bacillus licheniformis ZLP-81.
Detailed Description
The technical scheme of the present invention will be described in detail with reference to the preferred embodiments. The following examples are only for the purpose of illustrating and explaining the present invention and are not to be construed as limiting the technical solution of the present invention.
EXAMPLE 1 isolation and screening of Bacillus licheniformis ZLP-81
(1) Sampling soil: collecting a soil sample from the Tibetan changchun of China, wherein each part is 50-100 g; placing into sterilized kraft paper bag, sealing the bag mouth, and recording sampling place, environment and date.
(2) Bacillus isolation and purification: soil samples were isolated on PB medium by plate dilution. The specific operation is as follows: accurately weighing 10g of soil sample, adding into 90mL 0.9% physiological saline conical flask filled with glass beads, shaking with 180r/min shaking table for 30min to uniformly distribute soil sample to obtain soil suspension with concentration of 10 -1 The method comprises the steps of carrying out a first treatment on the surface of the Sequentially diluting to a concentration of 10 -5 、10 -6 、10 -7 . Respectively carrying out constant-temperature water bath at 80 ℃ for 30min, respectively taking 100 mu L of diluent to coat a PB flat plate, and carrying out inverted culture on the flat plate in a constant-temperature incubator at 30 ℃ for 48h. Single colonies with different forms are selected, spore staining is used for identification, bacillus is purified by a streaking method and then is transferred to a PB culture medium test tube inclined plane, after being cultured at 30 ℃ for 48h, the bacillus is preserved in a refrigerator at 4 ℃ for standby.
(3) Screening of insecticidal disease-preventing strains
Screening of insecticidal strains: taking aphids as target organisms, carrying out liquid shake flask fermentation culture on the purified microorganisms, centrifuging the fermentation liquor at the speed of 12000r/min for 10min to remove thalli, and filtering the supernatant with a 0.22 mu m sterile filter membrane to prepare sterile fermentation filtrate. Putting the broad bean stem leaves with proper length into a conical flask. Spraying a proper amount of fermentation liquor to living aphids and stems and leaves, putting the living aphids and stems and leaves into a conical flask, covering the bottle mouth with thin gauze and fixing the bottle mouth with rubber bands. After 2d, counting the number of aphids, and screening out strains with good insecticidal effect.
Screening strains with disease prevention effect: the gray mold of cucumber is used as indicator bacteria, and a flat plate counter method is adopted for primary screening. The method comprises the steps of preparing a d=5mm bacterial cake of the gray mold pathogenic bacteria growing on a PDA culture medium by using a puncher in a sterile operation room, transferring the bacterial cake to the center of a PDA culture medium flat plate, respectively transferring sterile filtrate of each antagonistic bacteria obtained by primary screening to a position 2.5cm away from the pathogenic bacteria, setting 3 repetitions, simultaneously taking a flat plate only containing the pathogenic bacteria cake as a contrast, culturing at a constant temperature of 26 ℃, and measuring the colony diameter and calculating the average inhibition rate when CK grows to be full of the culture medium. Screening out strains with strong antagonism to pathogenic bacteria. The strain with strong insecticidal and disease-preventing effects is frozen and preserved at the temperature of minus 80 ℃.
(4) Identification of insecticidal disease-preventing strains
According to the experimental method in the common bacteria system identification manual, morphological and physiological biochemical identification is carried out on the strains with stronger insecticidal and disease-preventing effects obtained by screening. The strain is cultured on a PB culture medium plate at 32+/-1 ℃ for 14-16 hours, and a colony numbered ZLP-81 is observed to be irregularly round, white, matt and rough in surface, and the thallus is rod-shaped (shown in figures 1-2) and the spore is oval. The physiological and biochemical identification indexes are shown in table 1.
TABLE 1 physiological and biochemical characteristics of ZLP-81 strain
。
The strain with the number ZLP-81 is determined to be bacillus through morphological observation and physiological and biochemical experiments. And then carrying out molecular biology classification and identification, and adopting 16S rDNA sequence analysis to extract total genome DNA of the strain as a template to amplify the 16S rDNA sequence. The target fragment is amplified by adopting a universal primer 27F/1492R, and the amplified product is detected by 1% agarose gel electrophoresis and is subjected to gene sequencing by adopting a two-way sequencing method. Sequencing results BLAST sequence homology comparison is carried out through GenBank database, and then the strain with the number ZLP-81 is identified as bacillus licheniformisBacillus licheniformis) The strain is named as bacillus licheniformis ZLP-81.
The bacillus licheniformis ZLP-81 is sent to the China general microbiological culture Collection center for preservation, and the address is: the collection number is CGMCC No.25028, and the collection date is 2022, 6, 9 days.
Example 2 killing effect of Bacillus licheniformis ZLP-81 on aphids, plutella xylostella, prodenia litura, grubs, armadillidium, cockroaches and cricket
And (3) fermenting and culturing bacillus licheniformis ZLP-81. NB liquid medium was used, the formulation was: beef extract 5g/L, peptone 10g/L, naCl g/L, glucose 10g/L, pH7.0. Inoculating the strain into a culture medium according to the inoculum size of 5%, and culturing in a shake incubator at 180r/min and 32 ℃ for 48 hours to obtain fermentation broth. The fermentation broth level (viable count) was 1.18X10 7 cfu/mL。
And (3) a fermentation liquor (a treatment group) of bacillus licheniformis ZLP-81 and an NB liquid culture medium (a control group) are subjected to contact killing, aphids (50), plutella xylostella (30), grubs (30), prodenia litura (30), pillworm (30), cockroaches (50) and cricket (60) with the same insect body size are selected, after the larvae are raised for 24 hours in an environment with the temperature of 25-28 ℃, the fermentation liquor is uniformly sprayed on the surfaces of the insects by the treatment group, and the NB liquid culture medium is uniformly sprayed on the surfaces of the insects by the control group, so that the body surfaces of all insects can be contacted with liquid. And after spraying, the mixture is placed in a room for culture at normal temperature, and after 24 hours, the death number is counted and the corrected death rate is calculated, as shown in Table 2 and figures 3-4.
Wherein, correction mortality = (treatment group mortality-control group mortality)/(1-control group mortality) ×100%; mortality = number of dead insects/total number of insects x 100%.
TABLE 2 killing effect of Bacillus licheniformis ZLP-81 on pests
。
Example 3 determination of bacteriostasis spectra of Bacillus licheniformis ZLP-81
Common pathogenic bacteria such as: a plurality of plant pathogenic bacteria such as tomato leaf mold bacteria, tomato gray mold bacteria, pear black spot bacteria, fusarium, cotton fusarium wilt bacteria, cucumber gray mold bacteria, cucumber anthracnose bacteria, cucumber downy mildew bacteria, cucumber cladosporium cucumerinum bacteria, apple alternaria alternata, rhizoctonia solani, corn small spot bacteria, tomato early blight bacteria and the like; aquatic pathogenic bacteria such as aeromonas hydrophila, edwardsiella tarda, aeromonas veronii and staphylococcus aureus are used as targets, and a flat plate counter method is adopted to measure the bacteriostasis spectrum of the strain ZLP-81, so that the inhibition capability of the strain ZLP-81 on the growth of the pathogenic bacteria is determined.
As can be seen from the implementation results of Table 3, the strain ZLP-81 has strong antagonism to 18 pathogenic bacteria, wherein the inhibition to the tomato leaf mold bacteria is strongest, and the diameter of the inhibition zone can reach 25.20mm. The bactericide which takes the fermentation broth of the bacillus licheniformis ZLP-81 as an active ingredient has good application value in agricultural production.
TABLE 3 inhibition of Bacillus licheniformis ZLP-81 against different plant pathogens
。
Example 4 Protoffee of Bacillus licheniformis ZLP-81 bactericide
Cucumber is selected as a test crop, 20 seedlings/pot of the indoor potted cucumber are provided with 3 parallel plants, the treatment group adopts bacillus licheniformis ZLP-81 fermentation liquor (the method is the same as that of example 2, and the viable count is more than or equal to 10) 7 cfu/mL) and carbendazim solution (1 g of carbendazim is dissolved in 800mL of water) are uniformly mixed according to the volume ratio of 500:1 and sprayed to dry, and the number of viable bacteria is 10 7 cfu/kg soil application; the control group did not apply any pro-growth products. Then 10 cucumber seedlings of 28 days old are randomly selected, the plant height of the seedlings is measured by a ruler, and the stem thickness is measured by a vernier caliper. Deactivating enzyme at 105deg.C for 15min, oven drying at 75deg.C to constant weight, measuring dry mass of aerial part and underground part, measuring chlorophyll content by ethanol-acetone extraction method, and measuring photosynthetic parameters by portable photosynthetic measurement system. As can be seen from Table 4, the Bacillus licheniformis ZLP-81 bactericide has improved plant height, stem thickness, fresh quality of the aerial parts of the individual plants, dry quality of the aerial parts of the individual plants, leaf area, fresh quality of the root of the individual plants, dry quality of the root of the individual plants, total root volume of the individual plants, total chlorophyll amount and net photosynthetic rate by 3.68%, 10.99%, 18.687%, 15.41%, 0.49%, 77.33%, 8.51%, 119.57%, 63.54% and 89.03% compared with CK, and has reached significant levels of difference except plant height, leaf area and dry quality of the root of the individual plants. The invention shows that the bacillus licheniformis ZLP-81 bactericide of the invention can be used for young cucumbersThe seedlings have obvious growth promoting effect.
TABLE 4 Protoffee of Bacillus licheniformis ZLP-81 fermentation broths on cucumber seedlings
。
The above-mentioned examples and methods are the best embodiments of the present invention, and the present invention can be partially changed, modified, substituted and combined without departing from the technical principles of the present invention, and the present invention is also included in the scope of protection of the present invention.
Claims (7)
1. Bacillus licheniformis @Bacillus licheniformis) ZLP-81, characterized in that the preservation number is CGMCC No. 25028.
2. Use of bacillus licheniformis ZLP-81 according to claim 1 for controlling plutella xylostella, prodenia litura, aphid, grub, cricket, pillworm and cockroach.
3. Use of bacillus licheniformis ZLP-81 according to claim 1 for controlling phytopathogens, aquatic pathogens and staphylococcus aureus.
4. Use according to claim 3, wherein the plant pathogenic bacteria comprise: the plant growth regulator comprises the components of tomato leaf mold bacteria, tomato gray mold bacteria, pear black spot bacteria, fusarium, cotton fusarium wilt bacteria, cucumber gray mold bacteria, cucumber anthracnose bacteria, cucumber downy mildew bacteria, cucumber cladosporium cucumerinum, apple alternaria alternate bacteria, rhizoctonia solani, corn small spot bacteria and tomato early blight bacteria.
5. Use according to claim 3, wherein the aquatic pathogenic bacteria comprise: aeromonas hydrophila, edwardsiella tarda and Aeromonas veronii.
6. Use of bacillus licheniformis ZLP-81 according to claim 1 for the preparation of biocontrol agents, biopesticides, soil restoration agents or biofertilizers.
7. A bactericide is characterized by comprising bacillus licheniformis ZLP-81 thallus, fermentation liquor or surfactant, iturin, phetamin or spermidine separated from the fermentation liquor.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210954216.6A CN115369058A (en) | 2022-08-10 | 2022-08-10 | Bacillus licheniformis ZLP-81 and application thereof |
| CN2022109542166 | 2022-08-10 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117535169A true CN117535169A (en) | 2024-02-09 |
| CN117535169B CN117535169B (en) | 2024-06-11 |
Family
ID=84063026
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210954216.6A Pending CN115369058A (en) | 2022-08-10 | 2022-08-10 | Bacillus licheniformis ZLP-81 and application thereof |
| CN202310964657.9A Active CN117535169B (en) | 2022-08-10 | 2023-08-02 | Bacillus licheniformis ZLP-81 and application thereof |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210954216.6A Pending CN115369058A (en) | 2022-08-10 | 2022-08-10 | Bacillus licheniformis ZLP-81 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN115369058A (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115369058A (en) * | 2022-08-10 | 2022-11-22 | 河北省科学院生物研究所 | Bacillus licheniformis ZLP-81 and application thereof |
| CN117736877B (en) * | 2022-09-20 | 2024-06-11 | 慕恩（广州）生物科技有限公司 | Metarrhizium anisopliae strain and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103820349A (en) * | 2012-11-16 | 2014-05-28 | 中国科学院生态环境研究中心 | Bacillus licheniformis and preparation method and application of fungicide |
| CN109439563A (en) * | 2018-09-10 | 2019-03-08 | 青岛秾贝尔生物工程有限公司 | One bacillus licheniformis and its application in prevention and treatment grape root rot |
| CN113755381A (en) * | 2021-09-26 | 2021-12-07 | 青岛力力惠生物科技股份有限公司 | Bacillus licheniformis for preventing and treating plant diseases and application thereof |
| CN114717164A (en) * | 2022-05-19 | 2022-07-08 | 河北省科学院生物研究所 | Bacillus belgii ZLP-101 and application thereof |
| CN115369058A (en) * | 2022-08-10 | 2022-11-22 | 河北省科学院生物研究所 | Bacillus licheniformis ZLP-81 and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| UY36334A (en) * | 2014-12-29 | 2017-04-28 | Fmc Corp | COMPOSITIONS OF BACILLUS LICHENIFORMIS RT1184 AND USES METHODS TO BENEFIT THE GROWTH OF PLANTS |
-
2022
- 2022-08-10 CN CN202210954216.6A patent/CN115369058A/en active Pending
-
2023
- 2023-08-02 CN CN202310964657.9A patent/CN117535169B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103820349A (en) * | 2012-11-16 | 2014-05-28 | 中国科学院生态环境研究中心 | Bacillus licheniformis and preparation method and application of fungicide |
| CN109439563A (en) * | 2018-09-10 | 2019-03-08 | 青岛秾贝尔生物工程有限公司 | One bacillus licheniformis and its application in prevention and treatment grape root rot |
| CN113755381A (en) * | 2021-09-26 | 2021-12-07 | 青岛力力惠生物科技股份有限公司 | Bacillus licheniformis for preventing and treating plant diseases and application thereof |
| CN114717164A (en) * | 2022-05-19 | 2022-07-08 | 河北省科学院生物研究所 | Bacillus belgii ZLP-101 and application thereof |
| CN115369058A (en) * | 2022-08-10 | 2022-11-22 | 河北省科学院生物研究所 | Bacillus licheniformis ZLP-81 and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115369058A (en) | 2022-11-22 |
| CN117535169B (en) | 2024-06-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112680382B (en) | Bacillus belgii and application thereof | |
| CN117535169B (en) | Bacillus licheniformis ZLP-81 and application thereof | |
| CN102433282B (en) | Bacillus subtilis NB12, as well as culture method and application thereof | |
| CN114717164B (en) | Bacillus belgii ZLP-101 and application thereof | |
| CN117535170B (en) | Bacillus subtilis ZLP-121 and application thereof | |
| CN111040976B (en) | Bacillus amyloliquefaciens and application thereof | |
| CN108148794A (en) | A kind of the bacillus subtilis DYr3.3 and preparation method and application of broad-spectrum antibacterial activity | |
| KR20140071145A (en) | Novel Paenibacillus polymyxa AB-15 strain and use the same | |
| CN117004524B (en) | Bacillus amyloliquefaciens ZLP-01 and application thereof | |
| CN115261283B (en) | Bacillus cereus and application thereof in prevention and control of disease of dry farming potatoes | |
| CN108048357B (en) | Bacillus subtilis and culture method thereof | |
| CN107142226B (en) | Bacillus subtilis ATR2, preparation method and application | |
| CN107699526B (en) | Actinomycete strain for preventing and treating gray mold and application thereof | |
| CN113943670B (en) | Disease-preventing growth-promoting pseudomonas tolaciens and application thereof | |
| RU2440413C1 (en) | Strain of bacteria bacillus licheniformis (its versions), possessing bactericidal and fungicidal activity, and preparation based on said strain | |
| CN111808776B (en) | Saline-alkali-tolerant air bacillus and preparation method and application of viable bacteria preparation thereof | |
| CN105018395B (en) | One bacillus pumilus and its application in alternaria leaf spot of apple is prevented | |
| RU2701502C1 (en) | Crystal-forming bacterial strain brevibacillus laterosporus with a wide spectrum of antagonist activity and use thereof | |
| CN117511760B (en) | Bacillus polymyxa ZLP-61 and application thereof | |
| CN102925394A (en) | Endophytic bacillus subtilis with disease prevention and growth promotion effects | |
| CN115873770A (en) | Bacillus belgii and application thereof in prevention and treatment of tomato diseases | |
| CN117535168B (en) | Bacillus megaterium ZLP-21 and application thereof | |
| CN105907663B (en) | Bacillus pumilus and application thereof | |
| CN117511760A (en) | Bacillus polymyxa ZLP-61 and application thereof | |
| CN112940978B (en) | Bacillus methylotrophicus BAM-6 and application thereof |
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 |