CN105861358A - Fomesafen degrading bacterium and application thereof - Google Patents
Fomesafen degrading bacterium and application thereof Download PDFInfo
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- CN105861358A CN105861358A CN201610214645.4A CN201610214645A CN105861358A CN 105861358 A CN105861358 A CN 105861358A CN 201610214645 A CN201610214645 A CN 201610214645A CN 105861358 A CN105861358 A CN 105861358A
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- Prior art keywords
- fomesafen
- degradation
- degradation bacteria
- bacterium
- bacterial strain
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- BGZZWXTVIYUUEY-UHFFFAOYSA-N fomesafen Chemical compound C1=C([N+]([O-])=O)C(C(=O)NS(=O)(=O)C)=CC(OC=2C(=CC(=CC=2)C(F)(F)F)Cl)=C1 BGZZWXTVIYUUEY-UHFFFAOYSA-N 0.000 title claims abstract description 111
- 241000894006 Bacteria Species 0.000 title claims abstract description 56
- 230000000593 degrading effect Effects 0.000 title abstract 4
- 230000015556 catabolic process Effects 0.000 claims abstract description 71
- 238000006731 degradation reaction Methods 0.000 claims abstract description 71
- 239000002689 soil Substances 0.000 claims abstract description 26
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 claims abstract description 4
- 230000007935 neutral effect Effects 0.000 claims abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 239000002068 microbial inoculum Substances 0.000 claims description 7
- 240000008042 Zea mays Species 0.000 claims description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 3
- 241000193755 Bacillus cereus Species 0.000 claims description 2
- 239000002054 inoculum Substances 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims 1
- 235000009973 maize Nutrition 0.000 claims 1
- 230000001580 bacterial effect Effects 0.000 abstract description 23
- 238000005067 remediation Methods 0.000 abstract description 4
- 238000004321 preservation Methods 0.000 abstract description 2
- 239000002609 medium Substances 0.000 description 15
- 241000196324 Embryophyta Species 0.000 description 13
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 12
- 230000000813 microbial effect Effects 0.000 description 11
- 238000000034 method Methods 0.000 description 10
- 244000025254 Cannabis sativa Species 0.000 description 9
- 239000012530 fluid Substances 0.000 description 8
- 210000000582 semen Anatomy 0.000 description 8
- 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 7
- 239000008103 glucose Substances 0.000 description 7
- 239000001963 growth medium Substances 0.000 description 7
- 238000001514 detection method Methods 0.000 description 6
- USIUVYZYUHIAEV-UHFFFAOYSA-N diphenyl ether Chemical compound C=1C=CC=CC=1OC1=CC=CC=C1 USIUVYZYUHIAEV-UHFFFAOYSA-N 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 230000002363 herbicidal effect Effects 0.000 description 5
- 239000004009 herbicide Substances 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 239000012071 phase Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- YCKRFDGAMUMZLT-UHFFFAOYSA-N Fluorine atom Chemical compound [F] YCKRFDGAMUMZLT-UHFFFAOYSA-N 0.000 description 3
- 244000068988 Glycine max Species 0.000 description 3
- 235000010469 Glycine max Nutrition 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 3
- 229910052782 aluminium Inorganic materials 0.000 description 3
- 150000001412 amines Chemical class 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 229960002413 ferric citrate Drugs 0.000 description 3
- 229910052731 fluorine Inorganic materials 0.000 description 3
- 239000011737 fluorine Substances 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- NPFOYSMITVOQOS-UHFFFAOYSA-K iron(III) citrate Chemical compound [Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NPFOYSMITVOQOS-UHFFFAOYSA-K 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000005070 sampling Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- 201000004384 Alopecia Diseases 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 102000016938 Catalase Human genes 0.000 description 2
- 108010053835 Catalase Proteins 0.000 description 2
- 102000004316 Oxidoreductases Human genes 0.000 description 2
- 108090000854 Oxidoreductases Proteins 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- XGCDHPDIERKJPT-UHFFFAOYSA-N [F].[S] Chemical compound [F].[S] XGCDHPDIERKJPT-UHFFFAOYSA-N 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 239000000090 biomarker Substances 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- -1 methylsulfonyl-2-Nitrobenzol formyl Amine Chemical class 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 230000000607 poisoning effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- FDDDEECHVMSUSB-UHFFFAOYSA-N sulfanilamide Chemical compound NC1=CC=C(S(N)(=O)=O)C=C1 FDDDEECHVMSUSB-UHFFFAOYSA-N 0.000 description 2
- 229940124530 sulfonamide Drugs 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- IPPAUTOBDWNELX-UHFFFAOYSA-N (2-ethoxy-2-oxoethyl) 5-[2-chloro-4-(trifluoromethyl)phenoxy]-2-nitrobenzoate Chemical group C1=C([N+]([O-])=O)C(C(=O)OCC(=O)OCC)=CC(OC=2C(=CC(=CC=2)C(F)(F)F)Cl)=C1 IPPAUTOBDWNELX-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 208000035126 Facies Diseases 0.000 description 1
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 description 1
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241000255964 Pieridae Species 0.000 description 1
- 241000589755 Pseudomonas mendocina Species 0.000 description 1
- DICDYWOAKAGJLU-UHFFFAOYSA-N S(=O)(C1=CC=C(C=C1)N)(=O)N.[F] Chemical compound S(=O)(C1=CC=C(C=C1)N)(=O)N.[F] DICDYWOAKAGJLU-UHFFFAOYSA-N 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 241000607762 Shigella flexneri Species 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000122971 Stenotrophomonas Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229930002875 chlorophyll Natural products 0.000 description 1
- 235000019804 chlorophyll Nutrition 0.000 description 1
- 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 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 125000001033 ether group Chemical group 0.000 description 1
- 244000037666 field crops Species 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- CONWAEURSVPLRM-UHFFFAOYSA-N lactofen Chemical compound C1=C([N+]([O-])=O)C(C(=O)OC(C)C(=O)OCC)=CC(OC=2C(=CC(=CC=2)C(F)(F)F)Cl)=C1 CONWAEURSVPLRM-UHFFFAOYSA-N 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000007269 microbial metabolism Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 1
- XITQUSLLOSKDTB-UHFFFAOYSA-N nitrofen Chemical compound C1=CC([N+](=O)[O-])=CC=C1OC1=CC=C(Cl)C=C1Cl XITQUSLLOSKDTB-UHFFFAOYSA-N 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000032696 parturition Effects 0.000 description 1
- 230000000505 pernicious effect Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
Classifications
-
- 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
-
- 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
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/04—Pesticides, e.g. insecticides, herbicides, fungicides or nematocides
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/28—Organic substances containing oxygen, sulfur, selenium or tellurium, i.e. chalcogen
Abstract
The invention discloses bacillus sp. FE-1 degrading fomesafen with the preservation number of CCTCC M 2016044. The fomesafen degrading bacterium has maximum fomesafen degradation rate under neutral pH condition, and the degradation rate is gradually improved while culture temperature gradually rises; meanwhile, the bacterial strain has good contaminated soil remediation potential. Therefore, the fomesafen degrading bacterium has positive meaning on degradation of fomesafen residue in polluted environment.
Description
Technical field
The present invention relates to a kind of antibacterial, particularly relate to a kind of fomesafen degradation bacteria and application thereof.
Background technology
Introduce technology as far back as domestic the beginning to of 20 century 70s, produce, use diphenyl ether herbicide.Mainly exist
Rice terrace uses nitrofen and Mo 9 granular；Middle and late stage mainly starts to use prowers, gram wealthy pleasure etc. at Soybean Field.Along with domestic this removes
The progress of grass agent research and development technology, and land for growing field crops uses the skilled grasp of technology, enters between several years of the beginning of this century, due to Semen sojae atricolor,
The further expansion of the oil crop cultivated areas such as Semen arachidis hypogaeae, weeds in field grass changes mutually, pernicious based on broad leaved weed
Weeds are on the rise, and both production and marketing thrive to cause diphenyl ether herbicide market.
Fomesafen [chemical name 5-(chloro-4 one 4-trifluoromethylphenopendants of 2-)-N one methylsulfonyl-2-Nitrobenzol formyl
Amine], it is a kind of diphenyl ether contact killing type herbicide, is mainly used in preventing and kill off the broad leaved weed of Soybean Field.Leaf is may result in after dispenser
Sheet is withered rapidly, Lipid peroxidation metabolism, destroys chlorophyll, is widely used because of the characteristic of its high-efficiency low-toxicity.Along with Semen sojae atricolor
The expansion of cultivated area, the application area of fomesafen expands year by year.Soybean Field perennial broadleaf weed the most in recent years
Resistance increases, and the increase of malignant weed and weather reason cause the consumption of fomesafen to sharply increase, and make in soil residual
Having stayed substantial amounts of fomesafen, not only polluted soil environment, sensitive crop multiple to rear stubble causes key, after causing simultaneously
The stubble sensitive crop underproduction is even had no harvest, and has had a strong impact on the adjustment of agricultural planting structure and the safety of agricultural production.Therefore, research
How to reduce fomesafen residual quantity in soil, reduce it and pollution that succession crop and environment are caused is had important
Theory and practice meaning.
Research shows, microbial degradation is to reduce fomesafen main path of residual quantity in soil, therefore, fluorine sulphur
The screening of amine grass ether degradation bacteria and one of soil remediation Hot Contents becoming Chinese scholar research in recent years.Currently, with respect to fluorine
The existing some reports of the degradation bacteria of sulfanilamide grass ether.Liang Bo etc. drop to fomesafen from by big Tanaka's separation screening of pollution by pesticides
Solve bacterium ZB-1, this bacterium the most also degradable lactofen and fluoroglycofen-ethyl.Wu Caiqiu etc. utilize enrichment culture technique from
The antibacterial that in the soil of chronic administration fomesafen, isolated 1 strain can grow with fomesafen for sole carbon source, life
Entitled F-12, this bacterium is under optimal condition, and after cultivating 2 days, the degradation efficiency to fomesafen reaches more than 80%, and having should
Use the ability of fomesafen contaminated soil biological restoration.Additionally, it is shigella flexneri FB5, pseudomonas mendocina FB8, micro-
The effect with degraded fomesafen it is accredited as addicted to acid Stenotrophomonas BX3 etc..The excavation of these microbial resources is to fluorine
Sulfanilamide grass ether contaminated soil reparation has great importance.Therefore, isolated and purified go out a high-efficiency degradation bacterium, it is intended to for solve
Fomesafen herbicide carryover problem in soil, the microbial resources of abundant degraded fomesafen, for this herbicide
Biological restoration research work provides more effective way.
Summary of the invention
The invention provides a kind of degradation efficiency fomesafen degradation bacteria high, fireballing and application thereof.
A kind of fomesafen degradation bacteria, Classification And Nomenclature is bacillus cereus (Bacillus sp.) FE-1, is preserved in and is positioned at
The China typical culture collection center (CCTCC) of Wuchang District, Wuhan City, Hubei Province No. 299 Wuhan Universitys of Bayi Road, preservation date
Being on January 18th, 2016, preserving number is CCTCC M 2016044.
This bacterial strain Main Morphology and biological property be: gram positive, thalline is shaft-like, atrichia, size
Being about (0.75 μm～0.8 μm) × (2.7 μm～3.7 μm), bacterium colony is milk yellow, circle, neat in edge, smooth moistening, is giving birth to
Being difficult to dispersion in reason saline, catalase is positive, oxidase positive.The GenBank number of logging in of this bacterial strain 16S rDNA is
KU145775。
The invention provides a kind of microbial inoculum containing above-mentioned fomesafen degradation bacteria, this microbial inoculum can be by by above-mentioned fluorine sulphur
Amine grass ether degradation bacteria makes water preparation after mixing with conventional additives or powder obtains.
The present invention also provides for described fomesafen degradation bacteria and remains the application in fomesafen in removing water body.
Specifically include: described fomesafen degradation bacteria is inoculated in pending water body, cultivate a period of time.
Fomesafen degradation bacteria of the present invention can be degraded the fomesafen in water body fast and efficiently, above-mentioned microbial inoculum with
Strain final concentration of 107Cfu/mL (trains containing 2.5g/L LB at the minimal medium containing 0.5,1 or 10mg/L fomesafen
Support base and 3.6g/L glucose) in cultivate 14h after, the fomesafen residual quantity of 0.5 or 1mg/L group has been less than Monitoring lower-cut
(0.01mg/L), the degradation rate of 10mg/L group is 82.9%, and degradation rate is respectively 0.036,0.071 and 0.592mg/ (h
L), corresponding degradation half life is respectively 5.36h, 5.66h and 5.57h.
Fomesafen degradation bacteria of the present invention is best to the degradation effect of fomesafen under conditions of pH is neutrality.
Fomesafen degradation bacteria of the present invention is when temperature is 15 DEG C, 25 DEG C or 35 DEG C, and the degraded to fomesafen is imitated
Fruit improve successively, wherein 35 DEG C time degradation rate reach 93.3%.
Invention further provides described fomesafen degradation bacteria in the reparation to fomesafen contaminated soil
Application.
Use fomesafen degradation bacteria of the present invention to process fomesafen contaminated soil, can substantially recover sensitive crop
Every Biological indicators (plant height and fresh weight) of Semen Maydis, and to Semen Maydis without poisoning effect.
Fomesafen degradation bacteria of the present invention can be degraded the residual fomesafen in water body and soil fast and efficiently,
This strain culturing technique is simple, and low production cost, use is convenient to, and has broad application prospects.
Accompanying drawing explanation
Fig. 1 is the Electronic Speculum figure of fomesafen degradation bacteria of the present invention；
Fig. 2 is that the fomesafen degradation bacteria of the present invention degraded to variable concentrations fomesafen under the conditions of pure culture is bent
Line chart；A:0.5mg/L；B:1mg/L；C:10mg/L
Fig. 3 is the fomesafen degradation bacteria of the present invention degradation curve figure to fomesafen under different pH condition；
Fig. 4 is the fomesafen degradation bacteria of the present invention degradation curve figure to fomesafen under condition of different temperatures；
Fig. 5 is after fomesafen degradation bacteria of the present invention is used corn growth to be affected result figure.A:1mg/L；B:
2mg/L；C:3mg/L
Detailed description of the invention
Culture medium:
LB culture medium is purchased from Sangon Biotech (Shanghai) Co., Ltd..
Minimal medium: MgS04·7H2O 0.1g、K2HPO4 1.0g、(NH4)2SO4 1.0g、KH2PO41.0g, steaming
Distilled water supplies 1000mL, stirs after mixing, and regulation pH value is to 7.0, and high pressure steam sterilization (121 DEG C, 20min) prepares afterwards.
Additionally, additionally add 2.5g LB culture medium and 3.6g glucose as carbon source and other nutrient substance.Glucose solution is using
Front degerming through 0.22 μm membrane filtration.
The detection of fomesafen in fluid medium:
Proceed to, in 250mL separatory funnel, use 50mL dichloromethane respectively by the 20mL liquid medium containing fomesafen
Extracting 3 times, lower floor's organic facies, in anhydrous sodium sulfate layer is incorporated in 250mL boiling flask, reduces pressure dense on the rotary evaporator
It is reduced to about 1mL, then dries up with nitrogen stream, by chromatograph methanol constant volume to 10mL for liquid-phase chromatographic analysis.
Chromatograph of liquid: use Agilent 1200 chromatograph of liquid；Chromatographic column: Agilent 4.6mm (id) ×
150mm, Eclipse XDB-C18；Flowing phase: acetonitrile: phosphoric acid (0.1%)=60: 40 (v/v)；Flow velocity: 1.0mL/min；Detection
Wavelength: 290nm；Sample size: 10 μ L.
Fomesafen residual quantity computing formula is as follows:
Wherein: X is the concentration (mg/L) of fomesafen in testing sample；AxFor the face, peak of fomesafen in sample
Long-pending；A0For fomesafen standard sample peak area；VxFor sample volume (mL)；V0For last constant volume (mL)；CsFor fluorine sulphur
The concentration (mg/L) of amine grass ether standard sample.
Embodiment 1
(1) strains separation purification
Pedotheque picks up from Zhejiang Province's Jiaxing Paddy Soil.Weigh for examination pedotheque 10.00g in 100mL serum bottle, add
Enter the Fe (OH) of 1mL synthetic3Suspension (being 15.4mg/mL containing Fe amount) and 50mL deionized water, stand half-light at 30 DEG C
Cultivate 1 week.Soil supension after cultivating is centrifuged 10min in 700rpm, takes supernatant as microbial inoculant liquid.By inoculation liquid
Diluting 100 times, take 100 μ L and be coated with on LB solid medium flat board, culture dish sealed membrane seals.Put in 30 DEG C of incubators and keep away
Light cultivates 2d.Select the flat board that bacterium colony is evenly distributed, whole picking list bacterium colonies, be respectively connected in ferric citrate fluid medium training
Support.Culture bottle uses the serum bottle of 10mL, every bottle of adding citric acid ferrum fluid medium 5mL, rushes nitrogen 5min, seals.If
In incubation, ferric citrate fluid medium is gradually become white by yellow green, shows that ferric citrate is reduced, this bacterial strain
There is the function of reduction Fe (III).Separation screening has the purpose bacterial strain of reduction accordingly.
(2) identification of strains
Isolated and purified bacterial strain is carried out morphological characteristic and molecular biology identification, the electromicroscopic photograph of this bacterial strain such as Fig. 1 institute
Show.This bacterial strain Main Morphology and biological property be: gram positive, and thalline is shaft-like, atrichia, and size is about
(0.75 μm～0.8 μm) × (2.7 μm～3.7 μm), bacterium colony is milk yellow, circle, neat in edge, smooth moistening, at physiology salt
Being difficult to dispersion in water, catalase is positive, oxidase positive.This bacterium the most suitable growth pH value is 7.0, and optimum growth temperature is 35 DEG C.
This bacterium can reduce the nitro of fomesafen, is changed into amino.The 16S rDNA of this bacterial strain is checked order, sequence such as SEQ
Shown in ID No:1, identify the unknown species that this bacterial strain is bacillus, named spore bar through 16S rDNA sequence analysis
Bacterium (Bacillus sp.) FE-1.
Embodiment 2
The impact that fomesafen is degraded by pesticide concentration:
In order to study the impact on himself microbial degradation of the fomesafen concentration, respectively to the blood of three 25mL sterilizings
Clear bottle adds 20mL pH 7.0 minimal medium (containing 2.5g/L LB culture medium and 3.6g/L glucose), the most each bottle
Interpolation fomesafen concentration is to 0.5,1 and 10mg/L respectively, is inoculated in by the appropriate bacterial strain FE-1 being in exponential phase
In fluid medium, strain quantity is made to reach 107Cfu/ml, is subsequently placed in shaking table (30 DEG C, 150rpm) dark shaken cultivation,
The correspondingly configuration 3 blank without this bacterium, matched group is cultivated the most under these conditions.
It is 0,2,4,6,8,10,12 and 14h timing sampling at incubation time, residual according to said method detection fomesafen
Allowance.Test group and matched group are respectively three repetitions.Bacterial strain of the present invention under the conditions of pure culture to variable concentrations fomesafen
Degradation curve as shown in Figure 2.
Observation Fig. 2 finds, after cultivating 14h, in process group, the fomesafen residual quantity of 0.5 and 1mg/L is less than detection
Limit (0.01mg/L).Degradation bacteria is 82.9% to the degradation rate of the fomesafen of 10mg/L.The fomesafen of variable concentrations
(0.5,1 and 10mg/L) degradation rate is respectively 0.036,0.071 and 0.592mg/ (h L), and corresponding degradation half life is respectively
For 5.36h, 5.66h and 5.57h.The degradation rate of fomesafen becomes preferable linear relationship (R respectively with its concentration2Respectively
It is 0.998,0.970 and 0.998), show that their degraded meets Quiroga-Sales second-order reaction kinetics equation.Not
Adding in the comparison of bacterium, the percent hydrolysis of fomesafen is respectively less than 4.8%.Show that this bacterial strain has the strongest degraded to fomesafen
Ability.
Embodiment 3
The pH value impact on fomesafen microbial degradation:
In order to study the different pH value impact on fomesafen microbial degradation, respectively to the serum of three 25mL sterilizings
20mL pH 6.0,7.0 and 8.0 buffer (containing 2.5g/L LB culture medium and 3.6g/L glucose) is added in Ping, the most each
Bottle adds 10mg/L fomesafen respectively.The appropriate bacterial strain FE-1 being in exponential phase is inoculated in fluid medium
In, make strain quantity reach 107Cfu/mL, is subsequently placed in shaking table (30 DEG C, 150rpm) dark shaken cultivation, correspondingly configures
3 blanks without this bacterium, matched group is cultivated the most under these conditions.
It is 0,2,4,6,8,10,12 and 14h timing sampling detection fomesafen residual quantity at incubation time.Test group with
Matched group is respectively three repetitions.Bacterial strain of the present invention under different pH condition to the degradation curve of fomesafen as shown in Figure 3.
Shown in observation Fig. 3, under conditions of pH 6.0,7.0 and 8.0, the microbial degradation of fomesafen meets
Quiroga-Sales second-order reaction kinetics feature.Under conditions of pH 6.0,7.0 and 8.0, the degradation rate of fomesafen
Being 65.7%, 84.0% and 74.5% respectively, the corresponding half-life is respectively 11.57h, 6.19h and 8.29h.It is being not added with bacterium
In comparison, fomesafen percent hydrolysis under conditions of pH 6.0,7.0 and 8.0 is respectively less than 2.8%.Fomesafen is at pH
Degradation half life under conditions of 6.0 is its 1.87,1.40 times under conditions of pH 7.0, pH 8.0 respectively.Fluorine sulfanilamide grass
Ether degradation effect under condition of different pH is pH 7.0 ＞ pH 8.0 ＞ pH 6.0.Result shows that degradation bacteria FE-1 is at neutral bar
Under part, the degradation effect to fomesafen is best.
Embodiment 4
The temperature impact on fomesafen microbial degradation:
In order to study the different temperatures impact on fomesafen microbial degradation, respectively to the serum of three 25mL sterilizings
The minimal medium (containing 2.5g/L LB culture medium and 3.6g/L glucose) of 20mL pH 7.0, the most each bottle is added in Ping
Add the fomesafen of 10mg/L respectively.The appropriate bacterial strain FE-1 being in exponential phase is inoculated in fluid medium
In, make strain quantity reach 107Cfu/mL, is then respectively placed in the shaking table (150rpm) of 15,25 and 35 DEG C dark vibration training
Supporting, be not added with the blank of microbial inoculum simultaneously, matched group is cultivated the most under these conditions.
Incubation time is 0,2,4,6,8,10,12 and 14h timing sampling detection fomesafen residual quantity.Test group is with right
It is respectively three repetitions according to group.Bacterial strain of the present invention under condition of different temperatures to the degradation curve of fomesafen as shown in Figure 4.
Shown in observation Fig. 4, under conditions of 25 and 35 DEG C, the microbial degradation of fomesafen all meets Quiroga-
Sales second-order reaction kinetics feature, and under the conditions of 15 DEG C, the degradation rate of fomesafen and matched group no significant difference (P
≤0.05).Showing that microbial metabolism is slow under the conditions of 15 DEG C, the ability of degraded fomesafen is more weak.At the bar of 25,35 DEG C
Under part, the degradation rate of the microorganism of fomesafen is 69.3% and 93.3% respectively, and the corresponding half-life is 10.65h respectively
And 4.79h.In the comparison being not added with bacterium, fomesafen percent hydrolysis under conditions of 15,25 and 35 DEG C is respectively less than 4.3%.
Bacterial strain of the present invention degradation effect to fomesafen under condition of different temperatures is 25 DEG C of ＞ of 35 DEG C of ＞ 15 DEG C.Result show with
The rising of cultivation temperature, the degradation rate of fomesafen becomes big.
Embodiment 5
The soil remediation test of bacterial strain FE-1
In order to verify the bacterial strain FE-1 repair ability to fomesafen contaminated soil, late stubble sensitive crop is chosen in this experiment
Semen Maydis carries out soil inoculation pot experiment.The fresh soil of Zhejiang University Hua Jiachi school district 0～20cm is picked up from for examination soil, indoor
Air-dry, after removing foreign material, cross 2.0mm sieve.Under room temperature, (20-25 DEG C) is placed standby.Aluminum box (diameter it is sub-packed in after soil sample is sterilized
6cm, high 5.5cm).The interpolation concentration of fomesafen is 1,2,3mg/kg.By the appropriate bacterial strain FE-being in exponential phase
1 is inoculated in fluid medium, makes strain quantity reach 107After cfu/g, fully mixing, add minimal medium (containing 2.5g/L
LB culture medium and 3.6g/L glucose) to waterflooding 1cm.Arrange without fomesafen and do not connect bacterium for blank group
(blank), arrange add fomesafen and connect bacterium for process group (treatment), interpolation fomesafen is set but not
Connect bacterium for CK-1, arrange without fomesafen but connect bacterium for CK-2.Often group is repeated 6 times.All it is placed in 35 DEG C of cultivations
Case stands dark processing 5 days (d).Move to 40 DEG C of baking ovens after 5 days make excessive moisture evaporate into soil moisture content about maximum to hold
The 70% of the water yield.3 corn seeds are planted equably in aluminum box.After aluminum box is positioned in artificial climate incubator, plant
Plant height and fresh weight is measured after planting 14 days.
Shown in observation Fig. 5, in CK-1 matched group, 1,2,3mg/kg variable concentrations fomesafen process after milpa
Short and small withered and yellow, plant height suppression ratio is respectively 55%, 80.1% and 86.3%, and substantially less than blank group level (P≤
0.05).After degradation bacteria FE-1 is used, plant height inhibitory action significantly reduces, and plant height reaches blank group level.Meanwhile, different fluorine sulphurs
After amine grass ether residue concentration processes, plant fresh weight is subject to bigger suppression, and performance downtrod to root length is similar.Matched group CK-1,
Plant fresh weight and blank group also exist notable and extremely significant difference.(P≤0.05 or P≤0.01).Degradation bacteria FE-1 is executed
With rear, after the process of variable concentrations fomesafen, the fresh weight of milpa is close to blank group level.Potted plant raw survey is tested
Showing, bacterial strain FE-1 processes fomesafen contaminated soil, can substantially recover every Biological indicators (strain of sensitive crop Semen Maydis
Height and fresh weight), and to Semen Maydis without poisoning effect, show preferable repairing effect.
Can obtain from above-mentioned test, fomesafen degradation bacteria of the present invention neutrality exist under conditions of to fomesafen
Degradation rate is maximum, and being stepped up along with cultivation temperature, and its degradation rate becomes larger.And this bacterial strain has well pollution
Soil remediation potential.Therefore, fomesafen degradation bacteria of the present invention has polluting the degraded of fomesafen residual in environment
Positive meaning.
Claims (10)
1. a fomesafen degradation bacteria, it is characterised in that named bacillus cereus (Bacillus sp.) FE-1, preserving number
For: CCTCC M 2016044.
2. the microbial inoculum comprising fomesafen degradation bacteria as claimed in claim 1.
3. microbial inoculum as claimed in claim 2, it is characterised in that described microbial inoculum is water preparation or powder.
4. fomesafen degradation bacteria as claimed in claim 1 remains the application in fomesafen in removing water body.
Apply the most as claimed in claim 4, it is characterised in that including: described fomesafen degradation bacteria is inoculated in and waits to locate
In the water body of reason, cultivate a period of time.
Apply the most as claimed in claim 5, it is characterised in that the pH value of water body is neutral.
Apply the most as claimed in claim 5, it is characterised in that the temperature of cultivation is 35 DEG C.
Apply the most as claimed in claim 5, it is characterised in that the inoculum density of fomesafen degradation bacteria is 107cfu/mL。
9. fomesafen degradation bacteria application in the reparation to fomesafen contaminated soil as claimed in claim 1.
Apply the most as claimed in claim 9, it is characterised in that soil maize planting to be repaired.
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| CN110358717A (en) * | 2019-09-02 | 2019-10-22 | 黑龙江大学 | A kind of microbial bacterial agent and its preparation method and application for fomesafen of degrading |
| CN110904005A (en) * | 2019-12-09 | 2020-03-24 | 江苏省农业科学院 | Fomesafen degrading bacterium and application thereof |
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| CN101525584A (en) * | 2009-01-05 | 2009-09-09 | 南京农业大学 | Degradation bacteria for removing fomesafen pesticide residue and microbial inoculum prepared by same |
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Non-Patent Citations (2)
| Title |
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| BO LIANG等: "Biodegradation of fomesafen by strain Lysinibacillus sp. ZB-1 isolated from soil", 《CHEMOSPHERE》 * |
| 赵欢欢等: "微生物降解二苯醚类除草剂的研究进展", 《植物保护》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110358717A (en) * | 2019-09-02 | 2019-10-22 | 黑龙江大学 | A kind of microbial bacterial agent and its preparation method and application for fomesafen of degrading |
| CN110358717B (en) * | 2019-09-02 | 2020-12-04 | 黑龙江大学 | Microbial agent for degrading fomesafen and preparation method and application thereof |
| CN110904005A (en) * | 2019-12-09 | 2020-03-24 | 江苏省农业科学院 | Fomesafen degrading bacterium and application thereof |
| CN110904005B (en) * | 2019-12-09 | 2022-07-29 | 江苏省农业科学院 | Fomesafen degrading bacterium and application thereof |
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