CN108048360A - A kind of bacillus subtilis with degrading organic phosphor and diseases prevention double action - Google Patents
A kind of bacillus subtilis with degrading organic phosphor and diseases prevention double action Download PDFInfo
- Publication number
- CN108048360A CN108048360A CN201711455361.5A CN201711455361A CN108048360A CN 108048360 A CN108048360 A CN 108048360A CN 201711455361 A CN201711455361 A CN 201711455361A CN 108048360 A CN108048360 A CN 108048360A
- Authority
- CN
- China
- Prior art keywords
- wkphy
- bacterial strain
- eggplant
- bacillus subtilis
- present
- 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
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 244000063299 Bacillus subtilis Species 0.000 title claims abstract description 36
- 235000014469 Bacillus subtilis Nutrition 0.000 title claims abstract description 36
- 201000010099 disease Diseases 0.000 title claims abstract description 34
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 34
- 230000000593 degrading effect Effects 0.000 title claims abstract description 27
- 230000002265 prevention Effects 0.000 title claims abstract description 18
- 230000009471 action Effects 0.000 title abstract description 8
- 230000001580 bacterial effect Effects 0.000 claims abstract description 95
- 244000061458 Solanum melongena Species 0.000 claims abstract description 74
- 235000002597 Solanum melongena Nutrition 0.000 claims abstract description 68
- 241000894006 Bacteria Species 0.000 claims abstract description 49
- 241000082085 Verticillium <Phyllachorales> Species 0.000 claims abstract description 34
- 241000196324 Embryophyta Species 0.000 claims abstract description 26
- 230000000813 microbial effect Effects 0.000 claims abstract description 14
- 240000008067 Cucumis sativus Species 0.000 claims abstract description 11
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 claims abstract description 11
- 241000223221 Fusarium oxysporum Species 0.000 claims abstract description 11
- 229920000742 Cotton Polymers 0.000 claims abstract description 9
- 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 abstract description 6
- 241000223218 Fusarium Species 0.000 claims abstract description 6
- 241000813090 Rhizoctonia solani Species 0.000 claims description 10
- 241001123668 Verticillium dahliae Species 0.000 claims description 8
- 241000879806 Fusarium oxysporum f. sp. vasinfectum Species 0.000 claims description 5
- 238000010353 genetic engineering Methods 0.000 claims description 2
- 239000002689 soil Substances 0.000 abstract description 14
- 239000003814 drug Substances 0.000 abstract description 10
- 230000002401 inhibitory effect Effects 0.000 abstract description 10
- 239000003795 chemical substances by application Substances 0.000 abstract description 9
- 230000003449 preventive effect Effects 0.000 abstract description 7
- 244000061456 Solanum tuberosum Species 0.000 abstract description 6
- 235000002595 Solanum tuberosum Nutrition 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 230000000845 anti-microbial effect Effects 0.000 abstract description 4
- 239000003337 fertilizer Substances 0.000 abstract description 4
- 230000008635 plant growth Effects 0.000 abstract description 4
- 238000001228 spectrum Methods 0.000 abstract description 4
- 229940079593 drug Drugs 0.000 abstract description 2
- 230000002045 lasting effect Effects 0.000 abstract description 2
- 239000002686 phosphate fertilizer Substances 0.000 abstract 1
- 239000011574 phosphorus Substances 0.000 description 33
- 229910052698 phosphorus Inorganic materials 0.000 description 33
- 238000002360 preparation method Methods 0.000 description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 21
- 238000012360 testing method Methods 0.000 description 20
- 241000193830 Bacillus <bacterium> Species 0.000 description 18
- 244000005700 microbiome Species 0.000 description 18
- 239000007788 liquid Substances 0.000 description 17
- 239000002609 medium Substances 0.000 description 16
- 230000012010 growth Effects 0.000 description 15
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 14
- 239000012530 fluid Substances 0.000 description 13
- 239000000843 powder Substances 0.000 description 13
- 239000002068 microbial inoculum Substances 0.000 description 11
- 244000068988 Glycine max Species 0.000 description 10
- 235000010469 Glycine max Nutrition 0.000 description 10
- 240000008042 Zea mays Species 0.000 description 10
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 10
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 10
- 235000005822 corn Nutrition 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 238000000855 fermentation Methods 0.000 description 10
- 230000004151 fermentation Effects 0.000 description 10
- 235000013312 flour Nutrition 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 244000052769 pathogen Species 0.000 description 10
- 230000001717 pathogenic effect Effects 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 101150013736 gyrB gene Proteins 0.000 description 8
- WPEXVRDUEAJUGY-UHFFFAOYSA-B hexacalcium;(2,3,4,5,6-pentaphosphonatooxycyclohexyl) phosphate Chemical compound [Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])(=O)OC1C(OP([O-])([O-])=O)C(OP([O-])([O-])=O)C(OP([O-])([O-])=O)C(OP([O-])([O-])=O)C1OP([O-])([O-])=O WPEXVRDUEAJUGY-UHFFFAOYSA-B 0.000 description 8
- 241000726221 Gemma Species 0.000 description 7
- 238000012545 processing Methods 0.000 description 7
- 239000011780 sodium chloride Substances 0.000 description 7
- 108020004465 16S ribosomal RNA Proteins 0.000 description 6
- 108020004414 DNA Proteins 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 239000000470 constituent Substances 0.000 description 6
- 238000010790 dilution Methods 0.000 description 6
- 239000012895 dilution Substances 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 239000011159 matrix material Substances 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 6
- 238000007747 plating Methods 0.000 description 6
- 230000001954 sterilising effect Effects 0.000 description 6
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 5
- 229910019142 PO4 Inorganic materials 0.000 description 5
- 230000004913 activation Effects 0.000 description 5
- 235000016709 nutrition Nutrition 0.000 description 5
- 230000035764 nutrition Effects 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 238000010998 test method Methods 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 4
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 4
- 238000012408 PCR amplification Methods 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 4
- 230000015556 catabolic process Effects 0.000 description 4
- 238000006731 degradation reaction Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000000877 morphologic effect Effects 0.000 description 4
- 239000010452 phosphate Substances 0.000 description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 239000004563 wettable powder Substances 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- WYWFMUBFNXLFJK-UHFFFAOYSA-N [Mo].[Sb] Chemical compound [Mo].[Sb] WYWFMUBFNXLFJK-UHFFFAOYSA-N 0.000 description 3
- 244000144987 brood Species 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000009977 dual effect Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 3
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000000474 nursing effect Effects 0.000 description 3
- 239000000825 pharmaceutical preparation Substances 0.000 description 3
- 230000003405 preventing effect Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000002791 soaking Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 239000012137 tryptone Substances 0.000 description 3
- 241000193417 Brevibacillus laterosporus Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 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 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 208000035199 Tetraploidy Diseases 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 230000008485 antagonism Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 229910052564 epsomite Inorganic materials 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 229910001629 magnesium chloride Inorganic materials 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000006916 nutrient agar Substances 0.000 description 2
- 101150012629 parE gene Proteins 0.000 description 2
- 230000007918 pathogenicity Effects 0.000 description 2
- 238000004080 punching Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 239000004576 sand Substances 0.000 description 2
- 230000003381 solubilizing effect Effects 0.000 description 2
- 238000009331 sowing Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 230000009897 systematic effect Effects 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- UFBJCMHMOXMLKC-UHFFFAOYSA-N 2,4-dinitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1[N+]([O-])=O UFBJCMHMOXMLKC-UHFFFAOYSA-N 0.000 description 1
- WFJIVOKAWHGMBH-UHFFFAOYSA-N 4-hexylbenzene-1,3-diol Chemical compound CCCCCCC1=CC=C(O)C=C1O WFJIVOKAWHGMBH-UHFFFAOYSA-N 0.000 description 1
- 241000588624 Acinetobacter calcoaceticus Species 0.000 description 1
- 241000588625 Acinetobacter sp. Species 0.000 description 1
- 244000153158 Ammi visnaga Species 0.000 description 1
- 235000010585 Ammi visnaga Nutrition 0.000 description 1
- 241000193755 Bacillus cereus Species 0.000 description 1
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 206010011732 Cyst Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001145289 Paenibacillus elgii Species 0.000 description 1
- 241000983368 Pantoea sp. Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241000219000 Populus Species 0.000 description 1
- 241000589774 Pseudomonas sp. Species 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 229940037003 alum Drugs 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000443 biocontrol Effects 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 208000031513 cyst Diseases 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 230000002554 disease preventive effect Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 239000010977 jade Substances 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000002366 mineral element Substances 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 244000039328 opportunistic pathogen Species 0.000 description 1
- 150000002903 organophosphorus compounds Chemical class 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- -1 phytate Chemical class 0.000 description 1
- 235000002949 phytic acid Nutrition 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000002881 soil fertilizer Substances 0.000 description 1
- 244000000000 soil microbiome Species 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000001038 titanium pigment Substances 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 239000003643 water by type Substances 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
-
- 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
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Soil Sciences (AREA)
- Materials Engineering (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of bacillus subtilis (Bacillus subtilis) bacterial strain WKPHY 54, deposit number is CGMCC No.14951.Also disclose the microbial bacterial agent containing the bacterial strain.Bacterial strain WKPHY 54 of the present invention has degrading organic phosphor and diseases prevention double action, can both improve the utilization rate of organophosphor in phosphate fertilizer or soil, promotes plant growth, reduces fertilizers input, reduces production cost；Also there is higher preventive effect to plant diseases such as eggplant verticillium wilts simultaneously, average preventive effect is more than 70.0%；Secondly, 54 antimicrobial spectrums of bacterial strain WKPHY of the present invention are wide, and the bacterium that withers to eggplant Huang, cucumber Fusarium oxysporum, cotton wilt fusarium and potato black mole bacterium etc. all have good inhibiting effect；In addition, 54 bacterial strains of WKPHY of the present invention are strong to the plant diseases specialization such as eggplant verticillium wilt；The resistance to the action of a drug is not likely to produce, lasting medicine is good.
Description
Technical field
The invention belongs to field of agricultural microorganism, and in particular to a kind of withered with degrading organic phosphor and diseases prevention double action
Careless bacillus, further relates to the microbial bacterial agent containing the bacillus subtilis and they make in degrading organic phosphor and prevention
Application in terms of object disease.
Background technology
Phosphorus is one of mineral element necessary to maintaining plant normal growth, be plant nucleic acid in vivo and a variety of enzymes, coenzyme,
The important component of ATP etc..The required phosphorus of growth and development of plants is essentially from the supply of fertilizer and soil.Phosphorus in being manured into soil
Fertilizer easily forms the phosphate of slightly solubility and is difficult to be absorbed by crops utilization, therefore the phosphorus nutrition key for improving plant is that conversion is sharp
With the insoluble phosphorus in soil.And the conversion of phosphorus be unable to do without the participation of phosphate solubilizing bacteria in soil, phosphate solubilizing bacteria is divided into solution inorganic phosphorus bacteria
Conciliate two kinds of organic phosphobacteria.Organic phosphorus degrading bacterium main decomposition organic phosphorus compound, including phytate, phosphatide, organophosphor
Hydrochlorate etc., the mechanism of action are inorganic form mainly by means of generated enzyme degrading organic phosphor in bacterium vital movement, from
And it is absorbed and used by plants.
At present, the bacterium of degraded in soil organophosphor mainly have bacillus (Bacillus sp., CN106978347A),
General bacterium (Pantoea sp., CN104498403A, CN104962500A), acinetobacter calcoaceticus (Acinetobacter sp.,
CN106544303A, CN104974962A) and pseudomonad (Pseudomonas sp., CN105112319A) etc..Wherein gemma
Bacillus has bacillus subtilis (Bacillus subtilis, CN106244504A), bacillus amyloliquefaciens
(CN106434455A, CN105420156A) etc..In terms of the effect principal degradation organophosphor of these bacterial strains, for prevention phytopathy
Evil aspect has not been reported.
Eggplant verticillium wilt is a kind of soil-borne disease as caused by verticillium dahliae (Verticillium dahliae), mainly
Cause harm blade, can fall ill in eggplant each growth period, fallen ill with fruiting period most heavy.With the development of facility cultivation, eggplant connects
It is more and more common to make cultivation, eggplant verticillium wilt is caused to aggravate year by year, the yield and quality of eggplant is greatly reduced.
It is relied primarily in production and largely prevents eggplant verticillium wilt using chemical pesticide, not only enhance the anti-medicine of pathogen
Property, destroy balance between edaphon, and cause the pesticide residue on vegetables and environmental pollution.Therefore, biological control conduct
One important measures of control of plant disease, have caused the great attention of people.
At present prevention eggplant verticillium wilt bacillus mainly have bacillus laterosporus (Bacillus laterosporus,
CN107151641A), bacillus amyloliquefaciens (CN105238723A), bacillus subtilis (Bacillus subtilis,
CN102925394A；CN102154186A；Shandong studies of the Hongloumeng etc., hubei agricultural science, 2013,52 (21)：5199-5202；Cheng Lei etc.,
Changjiang University's journal, natural science edition, 06 phase in 2012；Sun Yi etc., Jiangsu's agriculture journal, 2008,24 (4)：425-430；Woods
Tinkling of pieces of jade etc., Chinese biological preventing and treating journal, 2010 (s1)：40-46), series bacillus (Paenibacillus elgii,
CN102086444B), waxy Bacillus (Bacillus cereus, Yang Wei etc., microbiology circulate a notice of 05 phase in 2011) etc..
Above-mentioned known bacillus function is relatively single or only has phosphate solubilization or only has controlling disease effect；
Secondly, with the change of environmental condition or the evolution of pathogen, the function of bacillus may also evolve therewith, change
Become, therefore, constantly separation, screening have the function of efficient phosphorus-dissolution, and the bacillus that antimicrobial spectrum is wide from soil, for adjusting
The imbalance between supply and demand of soil phosphorus is improved to plant disease preventive effect, is reduced production cost, is preserved the ecological environment, and promote agricultural can
Sustainable development is of great significance.
The content of the invention
Present invention aims at a kind of bacillus subtilis strain is provided, which has degrading organic phosphor and controlling disease
Dual function.
The present invention second is designed to provide the purposes of above-mentioned bacillus subtilis.
The present invention the 3rd is designed to provide a kind of microbial bacterial agent containing above-mentioned bacillus subtilis.
The present invention the 4th is designed to provide the preparation method of mentioned microorganism microbial inoculum.
The present invention the 5th is designed to provide the purposes of mentioned microorganism microbial inoculum.
The invention is realized by the following technical scheme：
A kind of bacillus subtilis (Bacillus subtilis) bacterial strain WKPHY-54, deposit number CGMCC
No.14951。
Applications of the above-mentioned bacterial strains WKPHY-54 on degrading organic phosphor.
Applications of the above-mentioned bacterial strains WKPHY-54 in soil of degrading on organophosphor
Applications of the above-mentioned bacterial strains WKPHY-54 on plant growth is promoted.
Applications of the above-mentioned bacterial strains WKPHY-54 in controlling plant diseases.
Plant disease described in above application refers to that eggplant verticillium wilt (Verticillium dahliae), cucumber are withered
Sick (F.oxysporum f.sp.cucumebrium), cotton wilt (F.oxysporum f.sp.vasinfectum) or horse
Bell potato tar spot (R.solani) etc..
Applications of the above-mentioned bacterial strains WKPHY-54 on degrading organic phosphor and prevention eggplant verticillium wilt.
The present invention also provides a kind of microbial bacterial agents containing above-mentioned bacterial strains WKPHY-54.
Mentioned microorganism microbial inoculum can be liquid preparation or solid pharmaceutical preparation.
Viable count containing bacterial strain WKPHY-54 in mentioned microorganism microbial inoculum is 2.0 × 106~2.0 × 108Cfu/mL or
2.0×106~2.0 × 108cfu/g。
The preparation method of the liquid preparation of mentioned microorganism microbial inoculum, includes the following steps：
(1) actication of culture：The WKPHY-54 bacterial strains of Cord blood are activated on LB plating mediums, picking single bacterium colony exists
When 25~35 DEG C of cultures 10~16 are small on LB slant mediums, the bacterial strain of activation is obtained；
(2) prepared by seed liquor：The bacterial strain of activation in a ring step (1) is scraped with sterile oese, is then seeded into
In 100mL LB fluid nutrient mediums, when culture 10~16 is small under conditions of 25~35 DEG C, shaking speed is 150~220rpm,
Obtain seed liquor；
(3) fermented and cultured：The seed liquor of step (2) is linked into corn flour soya bean according to the ratio that volume ratio is 1~3%
In powder culture medium (pH value 7.2), the fermented and cultured under conditions of temperature is 25~35 DEG C, shaking speed is 150~220rpm
36~40h obtains zymotic fluid；Thalline and Number of spores in zymotic fluid are detected every 30min, treats that grown spore accounts for bud in zymotic fluid
Spore and thalline sum 90% when stop fermented and cultured；Gained is the liquid preparation of WKPHY-54 bacterial strains.
The constituent and its weight of LB plating mediums or LB slant mediums described in above-mentioned preparation method step (1)
Than for：8~12g of tryptone, 4~6g of yeast extract, 4~6g of sodium chloride, 12~18g of agar powder, water 1000mL.
The constituent of LB fluid nutrient mediums and its weight ratio are described in above-mentioned preparation method step (2)：Tryptone 8
~12g, 4~6g of yeast extract, 4~6g of sodium chloride, water 1000mL.
LB plating mediums, LB slant mediums and the LB fluid nutrient mediums is conventionally prepared.
Corn flour soybean powder medium described in above-mentioned preparation method step (3), constituent and its weight percent
Than for：Corn flour 1.0~3.0%, analysis for soybean powder 1.0~3.0%, NaCl 0.1~1.0%, MnSO4·H2O 0.5~1.0%,
Remaining is water.
The preparation method of the corn flour soybean powder medium, according to weight percent by corn flour, analysis for soybean powder, NaCl
And MnSO4·H2O is mixed, and is added water, and is adjusted pH and is stirred evenly.
Mentioned microorganism microbial inoculum is in the application on degrading organic phosphor.
Mentioned microorganism microbial inoculum is in the application in soil of degrading on organophosphor.
Application of the mentioned microorganism microbial inoculum on plant growth is promoted.
Mentioned microorganism microbial inoculum is in the application in controlling plant diseases.
Plant disease described in above application refers to that eggplant verticillium wilt (Verticillium dahliae), cucumber are withered
Sick (F.oxysporum f.sp.cucumebrium), cotton wilt (F.oxysporum f.sp.vasinfectum) or horse
Bell potato tar spot (R.solani) etc..
Application of the mentioned microorganism microbial inoculum on degrading organic phosphor and prevention eggplant verticillium wilt.
The application method of mentioned microorganism microbial inoculum：Above-mentioned gained microbial bacterial agent is diluted with water to viable bacteria body number is
107Cfu/mL sows preceding seed soaking half an hour in eggplant or adsorbs above-mentioned gained microbial bacterial agent with calcium carbonate, is fabricated to Xie Dian
Afnyloliquefaciens WKPHY-54 pulvis is spreaded manuer in holes before eggplant sowing.
The present invention also provides more than one to state the genetic engineering bacterium that bacterial strain WKPHY-54 is recipient bacterium.The gene work
Journey bacterium improves the ability of degrading organic phosphor or improves the preventive effect of the plant diseases such as prevention eggplant verticillium wilt；Or degradation
The dual function of the plant diseases such as organophosphor and prevention eggplant verticillium wilt is all improved.
The present invention has the advantage that and advantageous effect：(1) bacterial strain WKPHY-54 of the present invention is with degrading organic phosphor and prevents
Control the microorganism of eggplant verticillium wilt double action；The ability of the existing stronger degrading organic phosphors of bacterial strain WKPHY-54 of the present invention is right
In raising utilization rate of fertilizer, plant growth, reduction fertilizers input, reduction production cost is promoted to play an important role, while the present invention
Bacterial strain WKPHY-54 is good to eggplant verticillium wilt control effect, and average preventive effect provides more than 70.0% for prevention eggplant verticillium wilt
One efficient microorganism；(2) bacterial strain WKPHY-54 antimicrobial spectrums of the present invention are wide, and wither to eggplant Huang bacterium, cucumber Fusarium oxysporum, cotton
Fusarium oxysporum and potato black mole bacterium etc. all have good inhibiting effect.(3) WKPHY-54 bacterial strains of the present invention are to eggplant verticillium wilt
Specialization is strong；The resistance to the action of a drug is not likely to produce, lasting medicine is good；(4) microbial bacterial agent of the present invention is to people, animal safety, and there is no rings
Border pollution problem；(5) microorganism formulation preparation method of the present invention it is simple, it is at low cost, using simple.
Biological deposits
Bacillus subtilis (Bacillus subtilis) bacterial strain WKPHY-54 of the present invention be the present inventor voluntarily
Screening obtains, the bacterial strain oneself on November 22nd, 2017 be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms
Center, preservation address are：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, deposit number
For：CGMCC No.14951.
Description of the drawings
Fig. 1 are the WKPHY-54 bacterial strain systematic growth tree graphs obtained according to 16S rDNA sequences.
Fig. 2 are the WKPHY-54 bacterial strain systematic growth tree graphs obtained according to gyrB gene orders.
Specific embodiment
The present invention is further explained and illustrated below by embodiment, but any restrictions are not formed to the present invention.
Experimental method in following embodiments is conventional method unless otherwise instructed；Percentage composition in following embodiments, such as nothing
It illustrates, is weight percentage.
The screening separation process of 1 bacterial strain WKPHY-54 of the present invention of embodiment and taxonomic identification
(1) acquisition, separation and the screening process of WKPHY-54 bacterial strains
In September, 2015 is that Baoding micro-control bio tech ltd tries to win the champion Hebei province of county alum mountain phosphorus ore area five in Hebei province
Point 5 parts of soil sample of acquisition, every part of 200g.1.0g is weighed respectively to air-dry in triangular flask of the soil sample addition with sterilizing bead, is added
99mL sterile waters stand 20min, 30 DEG C on shaking table, 180r/min fully vibrate 30min, then carried out by 10 times of dilution methods
Gradient dilution takes 10 respectively-3, 10-4, 10-5100 μ L of dilution, be coated on solution organophosphor culture medium on, each 3 weights of concentration
It is multiple；5-10min is stood after coating in clean bench, treats that bacterium solution is adsorbed into culture medium, in 35 DEG C of constant temperature incubation 5-7d.And with saturating
Bright circle method, molybdenum antimony resistance colorimetric method filter out the bacterial strain with phosphorus decomposing, while with tablet face-off method, pot experiment method to selected
Bacterial strain screens the ability for preventing eggplant verticillium wilt, finally filters out with phosphorus decomposing and prevention eggplant verticillium wilt dual function
Bacterial strain, name as WKPHY-54.
(2) taxonomic identification of WKPHY-54 bacterial strains：
(1) identification by morphological characters
Thalline is cultivated on LB culture mediums to be rod-shaped, cultivates and brood cell is generated after 10h, raw, ellipse in brood cell, cyst is swollen
Greatly, acid-fast stain is negative, and no parasporal crystal can move, flagellum Zhousheng.On nutrient agar panel, the light breast of Initial stage of culture bacterium colony
White, membranaceous, circular, late stage of culture bacterium colony is faint yellow, and edge is irregular；It rules and cultivates on nutrient agar slopes, linearly
Shape；Static gas wave refrigerator in liquid medium, surface form white mycoderm.These morphological features with《Common bacteria system identification hand
Volume》Bacillus morphological feature described in (east show pearl etc. writes Science Press .2001) is basically identical, tentatively sentences
Disconnected bacterial strain WKPHY-54 belongs to bacillus.
(2) classified using 16S rDNA Sequence Identifications
Using the genomic DNA of WKPHY-54 as template, PCR amplification, institute are carried out by primer of universal primer F27 and R1492
The primer sequence stated is：F27：5’-AGAGTTTGATCATGGCTCAG-3’；R1492：5’-GGCTACCTTGTTACGACTT-
3’；The reaction system (50 μ L) of PCR is:10×PCR Buffer(Mg2+) 5 μ L, dNTP Mixture (2.5mM) 5 μ L, Taq
(5U/ μ L) 1 μ L, F27 (10 μm of ol/L) 1 μ L, R1492 (10 μm of ol/L) 1 μ L, WKPHY-54 genomic DNA 50ng, ddH2O
Complement to 50 μ L.The reaction condition of PCR is 95 DEG C of 5min；95 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 1.5min, 30 Xun Huans；72℃
10min.Gained pcr amplification product is subjected to gel electrophoresis, the sequencing of Shanghai Sheng Gong bioengineering Co., Ltd is delivered, obtains
The 16S rDNA sequences of WKPHY-54 are (see SEQ ID No:1).By the 16S rDNA sequences of gained WKPHY-54 in Genbank
Tetraploid rice is carried out, it turns out that the 16S rDNA homologys of bacterial strain WKPHY-54 and bacillus are up to 99%；Profit simultaneously
With MEGA software building phylogenetic trees, together with as a result WKPHY-54 is aggregated to bacillus (see Fig. 1), illustrate WKPHY-
54 belong to bacillus (Bacillus).
(3) the identification classification of gyrB gene orders is utilized
Using WKPHY-54 genomic DNAs template, using bacillus gyrB genes degenerate primer gyrB-F and gyrB-R as
Primer carries out PCR amplification；The wherein described primer sequence is：gyrB-F：5’-TTGRCGGHRGYGGHTATAAAGT-3’；
gyrB-R：5’-TCCDCCSTCAGARTCWCCCTC-3’.The pcr amplification reaction system (50 μ L) of gyrB is:10×PCR
Buffer(Mg2+) 5 μ L, dNTP Mixture (2.5mM) 5 μ L, Taq (5U/ μ L) 1 μ L, gyrB-F (10 μm of ol/L) 1 μ L, gyrB-
R (10 μm of ol/L) 1 μ L, WKPHY-54 genomic DNAs 50ng, ddH2O complements to 50 μ L.The reaction condition of PCR is 95 DEG C
5min；95 DEG C of 30s, 55 DEG C of 45s, 72 DEG C of 1min, 30 Xun Huans；72℃10min.Amplified production is delivered into Shanghai life work biology work
Journey Co., Ltd is sequenced, and obtains the gyrB gene orders of WKPHY-54 bacterial strains (see SEQ ID No:2).By gained gyrB gene orders
Tetraploid rice is carried out in Genbank, it turns out that WKPHY-54 and the gyrB gene homologies of bacillus subtilis
Highest reaches 99%；Simultaneously using MEGA software building phylogenetic trees, as a result found (see Fig. 2) WKPHY-54 bacterial strains with it is withered
Careless bacillus is aggregated to together, illustrates WKPHY-54 for bacillus subtilis (Bacillus subtilis), and is one
New strains.
In summary morphological feature, 16S rDNA and gyrB gene homology comparative analyses as a result, understanding
WKPHY-54 belongs to bacillus subtilis (Bacillus subtilis), and different with existing Bacillus strain, is one
A new bacillus subtilis strain.
The preparation of the microbial bacterial agent of the invention containing WKPHY-54 bacterial strains of embodiment 2
It carries out in accordance with the following steps：
(1) actication of culture：- 80 DEG C of bacillus subtilis strain WKPHY-54 (deposit number CGMCC will be stored in
No.14951 (30 DEG C)) are activated on LB plating mediums, picking single bacterium colony is trained on LB slant mediums at 30 DEG C
Support 12 it is small when, obtain the bacterial strain of activation；The constituent and its weight ratio of wherein LB plating mediums or LB slant mediums be：Pancreas
Peptone 10g, yeast extract 5g, sodium chloride 5g, agar powder 15g, water 1000mL；
(2) preparation of seed liquor：LB fluid nutrient medium (its constituent and its weight ratio are packed into 250mL triangular flasks
For：Tryptone 10g, yeast extract 5g, sodium chloride 5g, water 1000mL) 100mL, high pressure moist heat sterilization treats that temperature drops to room
Wen Hou accesses the bacterial strain activated in an oese step (1) in every bottle, under conditions of 30 DEG C, shaking speed 180rpm into
When row shaken cultivation 12 is small, seed liquor is obtained；
(3) preparation of corn flour soybean powder medium：According to weight percent by corn flour 2.5%, analysis for soybean powder 2.5%,
NaCl 0.6%, MnSO4·H2O 0.6% is added to the water, and is uniformly mixed to get corn flour soybean powder medium；It is sub-packed in
In 500mL triangular flasks, every bottle of 200mL；Sterilizing 30 minutes is carried out to corn flour soybean powder medium at 121 DEG C, then cools to 30
It is DEG C spare；
(4) fermented and cultured：Into every bottle of corn flour soybean powder medium 200mL obtained by step (3) obtained by inoculation step (2)
Seed liquor 2mL；Carried out under the conditions of 30 DEG C, shaking speed 180rpm fermented and cultured 36 it is small when, later every 30 minutes from triangle
Bottle in sampling carry out microscopy, the gemma in the visual field and total thalline number are counted, and calculate gemma rate (gemma rate (%)=into
Ripe gemma number/(grown spore number+thalline number) × 100)；Gemma rate stops fermented and cultured when reaching 90%；Common fermentation culture 48
Hour obtains the liquid preparation of bacillus subtilis WKPHY-54.
It is measured using the method for plate culture count, as a result the viable bacteria of the bacillus amyloliquefaciens WKPHO-12 liquid preparations of gained
Content is 2.6 × 108cfu/mL。
3 bacterial strain WKPHY-54 degrading organic phosphor abilities qualitative determination of the present invention of embodiment is tested
It carries out as follows：
The WKPHY-54 bacterial strains point that 2 step of embodiment (1) has activated is seeded in solution organophosphor tablet with sterilizing toothpick
(its constituent and its weight ratio are culture medium：Glucose 10.0g, (NH4)2SO40.2g, MgSO4·7H2O 0.5g, KCl
0.1g, MgCl2·6H2O 5.0g, phytic acid calcium 2.0g, agar 20.0g, distilled water 1000mL, pH：On 7.0-8.0), 30 DEG C are placed in
When constant incubator culture 72 is small, the diameter of transparent loop diameter and bacterium colony is measured.
It turns out that WKPHY-54 bacterial strains of the present invention generate diameter on the organophosphor plating medium containing phytic acid calcium
13.4 millimeters of transparent circle, illustrate bacterial strain WKPHY-54 of the present invention can degrading organic phosphor phytic acid calcium very well, there is degradation soil
The potentiality of middle organophosphor.
The quantitative determination experiment of 4 WKPHY-54 strains for degrading organophosphor ability of the present invention of embodiment
It carries out as follows：
(1) prepared by fermentation medium：According to weight ratio by glucose 10.0g, (NH4)2SO4 0.2g、MgSO4·7H2O
0.5g、KCl 0.1g、MgCl2·6H2O 5.0g, phytic acid calcium 2.0g and agar 20.0g are added in distilled water 1000mL, mixing
Uniformly, fermentation medium (pH is obtained：7.0-8.0)；Fermentation medium is encased in conical flask according to 100mL/300mL bottles of amount
In, autoclave sterilization, for use.
(2) prepared by zymotic fluid：By the bacterial strain WKPHY-54 seed liquors of gained in 2 step of embodiment (2) and blank control training
Nutrient solution (the LB fluid nutrient mediums for not being inoculated with WKPHY-54 bacterial strains) is inoculated in step according to the inoculum concentration that weight percent is 2% respectively
Suddenly in the fermentation medium that prepared by (1), every group of 3 repetitions in 30 DEG C, 180r/min culture 6d, obtain zymotic fluid.
(3) OD is drawn720Nm- phosphorus standard curves：The accurate KH for drawing 5mg/L respectively2PO4Standard solution 0.0mL, 1.0mL,
2.0mL, 3.0mL, 4.0mL, 5.0mL add 1~2 drop 2,4-DNP to make indicator, with 10% in 50mL colorimetric cylinders
NaOH and 5% dilution heat of sulfuric acid adjust pH value, and the total volume that adding water makes each colorimetric cylinder reaches 30mL, shakes up；It is eventually adding molybdenum antimony
Anti- reagent 5.0mL mixings colour developing, constant volume.After 30min, colorimetric is carried out under 720nm wavelength, using phosphorus concentration value as abscissa, phase
The OD values answered are ordinate, plot standard curve.Obtain phosphorus standard curve regression equation y=0.3457x-0.00176 (R2=
0.99927, y=OD value, x=phosphorus concentrations).
(4) fermentation liquor treatment：The zymotic fluid of culture gained in step (2) is transferred in sterile Centrifuge Cup, is used
KQ5200DE types numerical control excusing from death ripple washer carries out ultrasonic cell-break (broken condition：200-240V, 2A, 50/60Hz, when
Between 20min), be allowed to release intracellular available phosphorus.10min is centrifuged with the rotating speed of 8000r/min, then takes 2.5mL supernatants
Liquid adds 2 drop 2,4-DNPs to make indicator in 50mL colorimetric cylinders, and pH is adjusted with 10%NaOH and 5% dilution heat of sulfuric acid
Value has been in just yellowish to solution, adds the anti-color developing agent 5mL of molybdenum antimony, constant volume, after reacting 30min.It is divided with T6 new centuries UV, visible light
OD value of the photometric determination supernatant at 720nm.The available phosphorus content in supernatant is drawn according to standard curve.
(5) result calculates：Sample solution colorimetric is absorbed value, then according to working curve (y=0.3457x-
0.00176) phosphorus content (mg/L) of corresponding colorimetric solution is calculated, then available phosphorus content in zymotic fluid is calculated as follows：Effectively
Phosphorus (mg/L) × extension rate of phosphorus (mg/L)=color solution.
1 WKPHY-54 strains for degrading organophosphor ability of the present invention of table quantitative determines result of the test
| Strain number | OD720(nm) | Titanium pigment content (mg/L) |
| WKPHY-54 | 0.693 | 40.19 |
| Blank control | 0.170 | 9.86 |
As a result (1 is shown in Table) compared with blank control, be inoculated with soluble in the fermentation culture of WKPHY-54 bacterial strains of the present invention
Phosphorus content increases to 40.19mg/L, there are significant difference between blank control, illustrates that WKPHY-54 bacterial strains of the present invention have very
The ability of good degrading organic phosphor phytic acid calcium.
5 bacterial strain WKPHY-54 of the present invention of embodiment tests the growth promoting function of eggplant plant
(1) test material:
(1) matrix：Sand+substrate
Wherein：Sand is rinsed with water 3 times in advance, air-dries spare, pH：6.0 left and right；
Substrate：Phytic acid calcium, 1g/kg matrix.
(2) Eggplant Varieties：Agricultural university 601
(2) test process：
(1) handle：Phytic acid calcium+WKPHY-54 zymotic fluids+scarce phosphorus nutrition liquid
(2) compare：Phytic acid calcium+original fermentation medium+scarce phosphorus nutrition liquid
(3) test method：This experiment is in early January, 2016 in Baoding micro-control bio tech ltd laboratory
It carries out.Eggplant seedling is cultivated in nutritive cube, is transplanted when rough leaf is unfolded (high into the flowerpot of the husky amount 3.5kg/ basins of dress：
20cm, basin mouth diameter：21cm, basin bottom diameter：14cm), per 2 plants of basin, it is placed in greenhouse and cultivates, start to test after slow seedling.Experiment
Two processing are set, handle to pour WKPHY-54 bacterial strain fermentation liquor dilution (concentration prepared by 4 step of 250mL embodiments (2)
For 1 × 107CFU/mL) the former fermented and cultured prepared in crop root, blank control for 4 step of embodiment (1) of pouring equivalent
Base dilution.Often processing is repeated 3 times, often repeatedly 3 basin.Period normal management, keeps the skin wet in due course, each 400mL/ basins.7d is poured
Once lack phosphorus nutrition liquid (ingredient for lacking phosphorus nutrition liquid is shown in patent application 201110107663X), each 250mL/ basins.It is surveyed after 50d
Determine the fingers such as phosphorus content in the plant height of eggplant, overground part fresh weight and available phosphorus in underground part fresh weight, matrix and eggplant plant body
Mark.
The influence result of the test that 2 WKPHY-54 bacterial strains of the present invention of table grow eggplant
As a result (2 are shown in Table) compared with blank control, by the eggplant strain of bacillus subtilis WKPHY-54 fermentation liquor treatments
Height increases 1.66%, with compareing that there was no significant difference；Overground part, underground part fresh weight growth rate are respectively 27.98% and
There are significant differences between 42.23%, with blank control.The above results illustrate that WKPHY-54 bacterial strains of the present invention can be notable
Promote the growth of eggplant.
3 WKPHY-54 bacterial strains of the present invention of table are to the influence result of the test of matrix and eggplant plant available phosphorus
From table 3 it is observed that after WKPHY-54 bacterial strains of the present invention processing, the growth rate of available phosphorus is in matrix
12.26%, and available phosphorus growth rate is 105.36% in eggplant plant.Illustrate that bacterial strain WKPHY-54 of the present invention can effectively degrade
Organophosphor in matrix, and the absorption of the available phosphorus after the degradation of eggplant plant pair is promoted, so as to promote the growth of eggplant plant.
6 bacterial strain WKPHY-54 of the present invention of embodiment tests the inhibitory action of Verticillium Wilt Pathogen of Eggplant
(1) for examination cause of disease bacteria strain：Eggplant Huang withers bacterium EVD-1：It is isolated from Baoding Qingyuan County Zhang Deng towns Zhang Dengdian villages eggplant
Sub- disease fruit isolates and purifies through Baoding micro-control bio tech ltd, verticillium dahliae is accredited as through Agricultural University Of Hebei
(Verticillium dahaliae), Pathogenic Tests show as High pathogenicity.
(2) test method：
Eggplant Huang is withered bacterium EVD-1 activation culture 3-7 days on PDA plate first, then uses card punch
Bacterium piece is made in the punching of colony edge region；By the switching of bacterium piece in another PDA plate center, then will be living in 2 step of embodiment (1)
The WKPHY-54 bacterial strains point of change is connected on away from 2.0 centimeters of indicator bacteria bacterium piece, if blank control (do not put and connect WKPHY-54 bacterial strains).
25 DEG C of constant temperature incubations 3-10 days observe WKPHY-54 bacterial strains and the growing state for examination disease fungus day by day, treat blank control disease
During opportunistic pathogen length to culture dish edge, measure the control increment (colony radius) of pathogen and through bacterial strain WKPHY-54 of the present invention at
The increment (the inhibition growth radius after inoculation WKPHY-54) of reason, antagonism is represented with bacteriostasis rate.Bacteriostasis rate calculation formula
For：
Bacteriostasis rate (%)=(control increment-processing increment)/control increment × 100).
As a result wither to the eggplant Huang bacteriostasis rate of bacterium of the bacterial strain WKPHY-54 of the present invention that (is shown in Table 4) is 63.12%, illustrates WKPHY-
54 bacterial strains wither to eggplant Huang bacterium with apparent inhibitory action.
4 WKPHY-54 bacterial strains of the present invention of table wither to eggplant Huang the bacteriostasis result of the test of bacterium
| Pathogen | Normal growth (mm) | Inhibit growth (mm) | Bacteriostasis rate (%) |
| Eggplant Huang withers bacterium (V.dahliae) | 32.0 | 11.8 | 63.12 |
The liquid preparation of 7 bacterial strain WKPHY-54 of the present invention of embodiment is to the efficiency test of eggplant verticillium wilt
(1) test material：
(1) for examination Eggplant Varieties：Agricultural university 601；The Eggplant Varieties being bred as Agricultural University Of Hebei.
(2) for examination cause of disease bacteria strain：Eggplant Huang withers bacterium EVD-1.
(2) test process：
(1) WKPHY-54 liquid preparations：The WKPHY-54 liquid preparations for being prepared embodiment 2 with water dilute 100 times.
(2) medicament compares：1000000000 living spores/gram bacillus subtilis wettable powder (the green rich biochemical technology of Baoding section
Co., Ltd)；It is diluted with water 100 times.
(3) blank control：Clear water
(3) test method：
100 times of water diluent seed soaking half an hour of WKPHY-54 liquid preparations that before sowing prepared by Application Example 2；With " 10
100 times of water diluent seed soaking half an hour of hundred million living spores/gram bacillus subtilis wettable powder " compare as medicament；With clear water
Half an hour soak seed as blank control.After planting normal culture.When eggplant seedling grows the 4th~5 true leaf, using Qie Genfa
Inoculation eggplant Huang withers the spore suspension (10 of bacterium7A spore/mL).Routine Management continues culture and fully falls ill to blank control
When investigate disease index, calculate control effect.
As a result (table 5) WKPHY-54 bacterial strains of the present invention compare prevention with medicament to the preventive effect of eggplant verticillium wilt up to 72.60%
The effect (70.29%) of eggplant verticillium wilt is suitable.Illustrate that WKPHY-54 and its liquid preparation of the present invention have eggplant verticillium wilt
Good control effect.
5 WKPHY-54 of the present invention of table is to eggplant verticillium wilt efficiency test result
The solid pharmaceutical preparation of 8 bacterial strain WKPHY-54 of the present invention of embodiment is to the efficiency test of eggplant verticillium wilt
(1) test material：
(1) Eggplant Varieties：Agricultural university 601.
(2) for examination cause of disease bacteria strain：Eggplant Huang withers bacterium EVD-1.
(2) test process：
(1) WKPHY-54 pulvis：According to 1：1 ratio adds carbonic acid into WKPHY-54 liquid preparations prepared by embodiment 2
Calcium stirs evenly, and obtains WKPHY-54 pulvis.
(2) medicament compares：1000000000 living spores/gram bacillus subtilis wettable powder (the green rich biochemical technology of Baoding section
Co., Ltd).
(3) blank control：It is untreated
(3) test method：
By nursery soil high pressure steam sterilization 2 it is small when put to room temperature, be paved with seedlings nursing plate, be colonized 1 plant of eggplant seedling in seedlings nursing plate per cave,
It is spare when culture is to 4~5 true leaves.Eggplant Huang is withered into bacterium EVD-1 inoculated by hypha block in PDB culture solutions, at 25 DEG C,
180rpm/min is cultivated 5 days, is filtered off mycelia and is obtained conidium, conidium is mixed into sterile seedling medium, makes wherein eggplant
Sub- Huang withers the conidial concentration of bacterium EVD-1 for 106A/gram soil.The mixed seedling medium that carries disease germs is packed into flowerpot (diameter
15cm), the position for being colonized eggplant seedling wherein applies 2.0 grams of WKPHY-54 pulvis, and eggplant transplantation of seedlings (is held up into flowerpot during transplanting
Seedlings nursing plate makes the eggplant seedling root tip break in favor of pathogen infection) and band soil bacteria is covered, it is colonized one plant of eggplant seedling in every basin.To spread manuer in holes
1000000000 gemma/2.0 grams of gram bacillus subtilis wettable powder be medicament control, using do not process the eggplant seedling directly transplanted as
Blank control.Often processing is repeated 3 times, often repeatedly 5 basin.It is normally cultivated after transplanting, until investigating the state of an illness when blank control is fully fallen ill
Index calculates control effect.
As a result (table 6) bacterial strain WKPHY-54 of the present invention to the preventive effect of eggplant verticillium wilt up to 71.79%, with compareing microorganism
The effect (74.44%) of bactericidal agent for preventing and treating eggplant verticillium wilt is suitable.Illustrate bacterial strain WKPHY-54 and its solid pharmaceutical preparation pair of the present invention
Eggplant verticillium wilt has good control effect.
6 bacterial strain WKPHY-54 of the present invention of table is to eggplant verticillium wilt efficiency test result
| Processing | Disease index | Preventive effect (%) |
| WKPHY-54 pulvis | 22.69b | 71.79 |
| 1000000000 work brood cell/gram bacillus subtilis wettable powders | 20.56b | 74.44 |
| Blank control | 80.43a | -- |
9 bacterial strain WKPHY-54 of the present invention of embodiment tests the inhibitory action of three kinds of pathogens
(1) for examination cause of disease bacteria strain
(1)：Cucumber fusarium axysporum FOC-1：Dong Luobao townshiies of Baoding Dingxing County Dong Ce villages cucumber diseased plant is isolated from, through Hebei
Agriculture university is accredited as Fusarium oxysporum cucumber transformant (Fusarium oxysporum f.sp.cucumebrium)
(2)：Cotton-wilt fusarium FOV-7：Xingtai City Wei County cotton diseased plant is isolated from, point is accredited as through Agricultural University Of Hebei
Fusarium oxysporum wilting specialized form (Fusarium oxysporum f.sp.vasinfectum)
(3)：Black scurf of potato RS-3：It is isolated from the western poplar ditch village potato disease in Zhangjiakou City Shangyi County first stone river township
Strain, Rhizoctonia solani Kuhn (Rhizoctonia solani) is accredited as through Agricultural University Of Hebei
Three above strain pathogenic strength measure shows as High pathogenicity.
(2) test method：
This experiment is carried out in early June, 2017 in Baoding micro-control bio tech ltd laboratory.It first will be for
Pathogen activation culture 4 days on PDA plate are tried, then use card punchIn colony edge region, bacterium is made in punching
Disk, then by bacterium disk transfer another PDA plate center, then by 2 step of embodiment (1) activate after bacillus amyloliquefaciens
WKPHY-54 points are connected on away from 2.0 centimeters of indicator bacteria bacterium disk, if blank control (do not put and connect WKPHY-54 bacterial strains).In 25 DEG C of constant temperature
Culture when blank control will cover with entire culture dish, measures control increment (colony radius) and the place of tomato gray mould bacterium
Increment (the inhibition growth radius after inoculation WKPHY-54) is managed, antagonism is represented with bacteriostasis rate.The calculation formula of bacteriostasis rate
For：
Bacteriostasis rate (%)=(control increment-processing increment)/control increment × 100.
7 bacterial strain WKPHY-54 of the present invention of table is to the inhibitory action result of the test of three kinds of pathogens
As a result the bacterial strain WKPHY-54 of the present invention that (is shown in Table 7) is 74.55% to the inhibiting rate of cucumber Fusarium oxysporum, is withered to cotton
The inhibiting rate of bacterium is 74.63%, and the inhibiting rate to black scurf of potato bacterium is 77.59%, illustrates bacillus subtilis WKPHY-
54 pairs of these three pathogens have apparent inhibitory action, and antimicrobial spectrum is wide, have prevention cucumber fusarium axysporum, cotton wilt and horse
The Biocontrol Potential of the plant diseases such as bell potato tar spot.
Sequence table
<110>Baoding micro-control bio tech ltd
<120>A kind of bacillus subtilis with degrading organic phosphor and diseases prevention double action
<130> 2017S1138INH
<141> 2017-12-27
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1437
<212> DNA
<213> Bacillus subtilis
<400> 1
atacatgcaa gtcgagcgga cagatgggag cttgctccct gatgttagcg gcggacgggt 60
gagtaacacg tgggtaacct gcctgtacga ctgggataac tccgggaaac cggggctaat 120
accggatggt tgtttgaacc gcatggttca aacataaaag gtggcttcgg ctaccactta 180
cagatggacc cgcggcgcat tagctagttg gtgaggtaac ggctcaccaa ggcaacgatg 240
cgtagccgac ctgagagggt gatcggccac actgggactg agacacggcc cagactcctt 300
cgggaggcag cagtagggaa tcttccgcaa tggacgaaag tctgacggag caacgccgcg 360
tgagtgatga aggttttcgg atcgtaaagc tctgttgtta gggaagaaca agtaccgttc 420
gaatagggcg gtaccttgac ggtacctaac cagaaagcca cggctaacta cgtgccagca 480
gccgcggtaa tacgtaggtg gcaagcgttg tccggaatta ttggccgtaa agggctcgca 540
ggcggtttct taagtctgat gtgaaagccc ccggctcaac cggggagggt cattggaaac 600
tggggaactt gagtgcagaa gaggagagtg gaattccacg tgtagcggtg aaatgcgtag 660
agatgtggag gaacaccagt ggcgaaggcg actctctggt ctgtaactga cgctgaggag 720
cgaaagcgtg gggtgcgaac aggattagat accctggtag tccacgccgt aaacgatgag 780
tgctaagtgt tagggggttt ccgcccctta gtgctgcagc taacgcatta agcactccgc 840
ctggggagta cggtcgcaag actgaaactc aaaggaattg acgggggccc gcacaagcgg 900
tggagcatgt ggtttaattc gaagcaacgc gaagaacctt accaggtctt gacatcctct 960
gacaatccta gagataggac gtccccttcg ggggcagagt gacaggtggt gcatggatgt 1020
cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgatctt 1080
agttgccagc attcagttgg gcactctaag gtgactgccg gtgacaaacc ggaggaaggt 1140
ggggatgacg tcaaatcatc atgcccctta tgacctgggc tacacacgtg ctacaatgga 1200
cagaacaaag ggcagcgaaa ccgcgaggtt aagccaatcc cacaaatctg ttctcagttc 1260
ggatcgcagt ctgcaactcg actgcgtgaa gctggaatcc ctagtaatcg cggatcagca 1320
tgccgcggtg aatacgttcc cgggccttgt acacaccgcc cgtcacacca cgagagtttg 1380
taacacccga agtcggtgag gtaacctttt aggagccagc cgccgaaggt gggacag 1437
<210> 2
<211> 808
<212> DNA
<213> Bacillus subtilis
<400> 2
cgtgacggta gaattcaccg ccaaacctat aaacgcggag ttccggttac agaccttgaa 60
atcattggcg aaacggatca tacaggaacg acgacacaat ttgtcccgga ccctgaaatt 120
ttctcagaaa caaccgagta tgattacgat ctgcttgcca accgcgtgcg tgaattagcc 180
tttttaacaa agggcgtaaa catcacgatt gaagataaac gtgaaggaca agagcgcaca 240
aatgaatacc attacgaagg cggaattaaa agttatgtag agtatttaaa ccgctctaaa 300
gaggttgtcc atgaagagcc gatttacatt gaaggcgaaa aggacggcat tacggttgaa 360
gtggctttgc aatacaatga cagctacaca agcaacattt actcgtttac aaacaacatt 420
aacacgtacg aaggcggtac ccatgaagct cgcttcaaaa cgggcctgac tcgtgttatc 480
aacgattacg ccagaaaaaa agggcttatt aaagaaaatg atccaaacct aagcggagat 540
gacgtaaggg aagggctgac agcgattatt tcaatcaaac accctgatcc gcattttgag 600
ggccaaacaa aaacaaagct gggcaactca gaagcacgga cgatcaccga tacgttattt 660
tctacggcga tggaaacatt tatgctggaa aatccagatg cagccaaaaa aattgtcgat 720
aacggtttaa tggcggcaag agcaagaatg gctgcgaaaa aagcgcgtga actaacacgc 780
cgtaagagtg ctttggaaat ttcaaacc 808
Claims (10)
1. a kind of bacillus subtilis (Bacillus subtilis) bacterial strain WKPHY-54, deposit number CGMCC
No.14951。
2. applications of the bacterial strain WKPHY-54 described in claim 1 on degrading organic phosphor.
3. applications of the bacterial strain WKPHY-54 described in claim 1 in controlling plant diseases；The wherein described plant disease is
Refer to eggplant verticillium wilt (Verticillium dahliae), cucumber fusarium axysporum (F.oxysporum f.sp.cucumebrium),
Cotton wilt (F.oxysporum f.sp.vasinfectum) or black scurf of potato (R.solani).
4. applications of the bacterial strain WKPHY-54 described in claim 1 on degrading organic phosphor and prevention eggplant verticillium wilt.
5. a kind of microbial bacterial agent containing bacterial strain WKPHY-54 described in claim 1.
6. microbial bacterial agent according to claim 5, it is characterised in that contain bacterial strain in the microbial bacterial agent
The viable count of WKPHY-54 is 2.0 × 106~2.0 × 108Cfu/mL or 2.0 × 106~2.0 × 108cfu/g。
7. the microbial bacterial agent described in claim 5 is in the application on degrading organic phosphor.
8. the microbial bacterial agent described in claim 5 is in the application in controlling plant diseases；The wherein described plant disease is
Refer to eggplant verticillium wilt (Verticillium dahliae), cucumber fusarium axysporum (F.oxysporum f.sp.cucumebrium),
Cotton wilt (F.oxysporum f.sp.vasinfectum) or black scurf of potato (R.solani).
9. application of the microbial bacterial agent on degrading organic phosphor and prevention eggplant verticillium wilt described in claim 5.
It is 10. a kind of using bacterial strain WKPHY-54 described in claim 1 as the genetic engineering bacterium of recipient bacterium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711455361.5A CN108048360B (en) | 2017-12-28 | 2017-12-28 | Bacillus subtilis with dual functions of degrading organic phosphorus and preventing diseases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711455361.5A CN108048360B (en) | 2017-12-28 | 2017-12-28 | Bacillus subtilis with dual functions of degrading organic phosphorus and preventing diseases |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108048360A true CN108048360A (en) | 2018-05-18 |
| CN108048360B CN108048360B (en) | 2021-01-01 |
Family
ID=62128772
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711455361.5A Active CN108048360B (en) | 2017-12-28 | 2017-12-28 | Bacillus subtilis with dual functions of degrading organic phosphorus and preventing diseases |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108048360B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110079484A (en) * | 2019-05-28 | 2019-08-02 | 青岛力力惠生物科技股份有限公司 | A kind of bacillus subtilis and its application in agricultural production |
| CN110079483A (en) * | 2019-05-28 | 2019-08-02 | 黄河三角洲京博化工研究院有限公司 | A kind of bacillus subtilis and its application |
| CN114231444A (en) * | 2021-11-29 | 2022-03-25 | 河北省农林科学院植物保护研究所 | Salt-tolerant bacillus and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105238723A (en) * | 2015-11-03 | 2016-01-13 | 保定微控生物科技有限公司 | Bacillus amyloliquefaciens strain for control of crop greensickness and microbial agent thereof |
| CN106244504A (en) * | 2016-10-08 | 2016-12-21 | 河北省农林科学院植物保护研究所 | There is degrading organic phosphor and the bacillus subtilis of bacteriostasis and microbial inoculum thereof |
| CN106399178A (en) * | 2016-10-08 | 2017-02-15 | 河北省农林科学院植物保护研究所 | Bacillus amyloliquefaciens with inorganic phosphorus degrading and bacteria inhibiting functions and application of bacillus amyloliquefaciens |
-
2017
- 2017-12-28 CN CN201711455361.5A patent/CN108048360B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105238723A (en) * | 2015-11-03 | 2016-01-13 | 保定微控生物科技有限公司 | Bacillus amyloliquefaciens strain for control of crop greensickness and microbial agent thereof |
| CN106244504A (en) * | 2016-10-08 | 2016-12-21 | 河北省农林科学院植物保护研究所 | There is degrading organic phosphor and the bacillus subtilis of bacteriostasis and microbial inoculum thereof |
| CN106399178A (en) * | 2016-10-08 | 2017-02-15 | 河北省农林科学院植物保护研究所 | Bacillus amyloliquefaciens with inorganic phosphorus degrading and bacteria inhibiting functions and application of bacillus amyloliquefaciens |
Non-Patent Citations (1)
| Title |
|---|
| SHAOMIN YAN等: "Bottleneck in secretion of α-amylase in Bacillus subtilis", 《MICROB CELL FACT》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110079484A (en) * | 2019-05-28 | 2019-08-02 | 青岛力力惠生物科技股份有限公司 | A kind of bacillus subtilis and its application in agricultural production |
| CN110079483A (en) * | 2019-05-28 | 2019-08-02 | 黄河三角洲京博化工研究院有限公司 | A kind of bacillus subtilis and its application |
| CN110079483B (en) * | 2019-05-28 | 2021-07-16 | 黄河三角洲京博化工研究院有限公司 | Bacillus subtilis and application thereof |
| CN110079484B (en) * | 2019-05-28 | 2022-07-08 | 青岛力力惠生物科技股份有限公司 | Bacillus subtilis and application thereof in agricultural production |
| CN114231444A (en) * | 2021-11-29 | 2022-03-25 | 河北省农林科学院植物保护研究所 | Salt-tolerant bacillus and application thereof |
| CN114231444B (en) * | 2021-11-29 | 2023-06-23 | 河北省农林科学院植物保护研究所 | Salt-tolerant bacillus and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108048360B (en) | 2021-01-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104928212B (en) | Bacillus megaterium X3 and preparation method thereof, application | |
| CN103146586B (en) | Trichoderma harzianum strain for controlling plant fungus diseases and application thereof | |
| CN106399178B (en) | Bacillus amyloliquefaciens and its application with degradation Phos and bacteriostasis | |
| CN106399177B (en) | Bacillus amyloliquefaciens and its microbial inoculum with degradation Phos and bacteriostasis | |
| CN102586142B (en) | Bacillus subtilis for preventing and curing cucumber downy mildew and microbial inoculants thereof | |
| CN106939290B (en) | Bacillus subtilis HMB26553 and application thereof | |
| CN106244504B (en) | Bacillus subtilis and its microbial inoculum with degrading organic phosphor and bacteriostasis | |
| CN108192838B (en) | Bacillus amyloliquefaciens with dual functions of inorganic phosphorus degradation and disease prevention | |
| CN104164394A (en) | Antagonistic phytopathogen strain and application thereof | |
| CN111073825B (en) | Bacterium with plant soil-borne disease resistance effect and application thereof | |
| CN108841744A (en) | A kind of bacillus subtilis with diseases prevention and degradation Phos double action | |
| CN105543132A (en) | Bacillus methylotrophicus YB-F7 and application thereof in preventing plant diseases | |
| CN105176894B (en) | A kind of bacillus amyloliquefaciens and its microbial bacterial agent of prevention graw mold of tomato | |
| CN105199996B (en) | A kind of bacillus amyloliquefaciens and its application for preventing graw mold of tomato | |
| CN105238723B (en) | A kind of bacillus amyloliquefaciens and its microbial bacterial agent of prevention crop verticillium wilt | |
| CN103109870B (en) | Application of fermentation supernatant liquid of pseudomonas | |
| CN108048360A (en) | A kind of bacillus subtilis with degrading organic phosphor and diseases prevention double action | |
| CN105176893B (en) | A kind of bacillus amyloliquefaciens and its application for preventing crop damping-off | |
| CN103146609B (en) | Pseudomonas fluorescens and method for preventing phytophthora capsici thereby | |
| CN108795794A (en) | A kind of preparation method and application of bacillus pumilus and its screening technique, microbial inoculum | |
| CN110317747A (en) | A kind of bacillus amyloliquefaciens JT68 and its application in prevention and treatment tea anthracnose | |
| CN107043719B (en) | bacillus amyloliquefaciens HMB28353 and application thereof | |
| CN107043720B (en) | Bacillus amyloliquefaciens HMB28388 and application thereof | |
| CN105018393A (en) | Bacillus megaterium strain and application thereof | |
| CN114874953B (en) | Peanut rhizosphere biocontrol bacterium-Bacillus beleisi SW-1 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 | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20200415 Address after: No. 302, unit 2, building 2, No. 40, Xianfeng street, Xinshi District, Baoding City, Hebei Province 071000 Applicant after: Qi Yongxin Address before: 071000 No. 118 Fuxing West Road, Hebei, Baoding Applicant before: BAODING WEIKONG BIOTECHNOLOGY Co.,Ltd. |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20231215 Address after: Banqiao Community Industrial Park, Kunming 650000 Patentee after: Yunnan Sijiaotian Agricultural Technology Co.,Ltd. Address before: 071000 No. 302, unit 2, building 2, No. 40, Xianfeng street, Xinshi District, Baoding City, Hebei Province Patentee before: Qi Yongxin |
|
| TR01 | Transfer of patent right |