CN117568332A - Microbial agent for treating soil and preparation method thereof - Google Patents
Microbial agent for treating soil and preparation method thereof Download PDFInfo
- Publication number
- CN117568332A CN117568332A CN202311522517.2A CN202311522517A CN117568332A CN 117568332 A CN117568332 A CN 117568332A CN 202311522517 A CN202311522517 A CN 202311522517A CN 117568332 A CN117568332 A CN 117568332A
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- Prior art keywords
- bacillus
- liquid
- seed
- culture medium
- microbial agent
- Prior art date
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 56
- 239000002689 soil Substances 0.000 title claims abstract description 56
- 239000003795 chemical substances by application Substances 0.000 title claims abstract description 51
- 238000002360 preparation method Methods 0.000 title claims abstract description 28
- 238000000855 fermentation Methods 0.000 claims abstract description 59
- 230000004151 fermentation Effects 0.000 claims abstract description 59
- 238000002156 mixing Methods 0.000 claims abstract description 58
- 241000187395 Streptomyces microflavus Species 0.000 claims abstract description 51
- 241000223261 Trichoderma viride Species 0.000 claims abstract description 51
- 241000193755 Bacillus cereus Species 0.000 claims abstract description 49
- 241000194107 Bacillus megaterium Species 0.000 claims abstract description 49
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 48
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 47
- 241000193417 Brevibacillus laterosporus Species 0.000 claims abstract description 47
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical class O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims abstract description 40
- 238000000034 method Methods 0.000 claims abstract description 31
- 244000005700 microbiome Species 0.000 claims abstract description 29
- 229920001661 Chitosan Polymers 0.000 claims abstract description 23
- 239000002131 composite material Substances 0.000 claims abstract description 19
- 239000011162 core material Substances 0.000 claims abstract description 19
- 229920000858 Cyclodextrin Polymers 0.000 claims abstract description 12
- 239000001116 FEMA 4028 Substances 0.000 claims abstract description 12
- 108010010803 Gelatin Proteins 0.000 claims abstract description 12
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 claims abstract description 12
- 235000011175 beta-cyclodextrine Nutrition 0.000 claims abstract description 12
- 229960004853 betadex Drugs 0.000 claims abstract description 12
- 239000008273 gelatin Substances 0.000 claims abstract description 12
- 229920000159 gelatin Polymers 0.000 claims abstract description 12
- 235000019322 gelatine Nutrition 0.000 claims abstract description 12
- 235000011852 gelatine desserts Nutrition 0.000 claims abstract description 12
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims abstract description 9
- 230000003213 activating effect Effects 0.000 claims abstract description 3
- 239000007788 liquid Substances 0.000 claims description 100
- 239000001963 growth medium Substances 0.000 claims description 81
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 46
- 238000011218 seed culture Methods 0.000 claims description 36
- 239000000243 solution Substances 0.000 claims description 30
- 238000009630 liquid culture Methods 0.000 claims description 29
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 28
- 239000011780 sodium chloride Substances 0.000 claims description 23
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 22
- 238000010438 heat treatment Methods 0.000 claims description 20
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 19
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 18
- 239000001888 Peptone Substances 0.000 claims description 17
- 108010080698 Peptones Proteins 0.000 claims description 17
- 235000015278 beef Nutrition 0.000 claims description 17
- 239000000284 extract Substances 0.000 claims description 17
- 235000019319 peptone Nutrition 0.000 claims description 17
- 239000007864 aqueous solution Substances 0.000 claims description 16
- 238000001816 cooling Methods 0.000 claims description 15
- 238000001035 drying Methods 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 15
- 238000005406 washing Methods 0.000 claims description 15
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 14
- 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 claims description 14
- 229920002472 Starch Polymers 0.000 claims description 14
- 240000008042 Zea mays Species 0.000 claims description 14
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 14
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 14
- 235000005822 corn Nutrition 0.000 claims description 14
- 239000008103 glucose Substances 0.000 claims description 14
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 14
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 14
- 239000008107 starch Substances 0.000 claims description 14
- 235000019698 starch Nutrition 0.000 claims description 14
- 239000008367 deionised water Substances 0.000 claims description 12
- 229910021641 deionized water Inorganic materials 0.000 claims description 12
- 229910052757 nitrogen Inorganic materials 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 11
- 229930006000 Sucrose Natural products 0.000 claims description 10
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 10
- 229910021536 Zeolite Inorganic materials 0.000 claims description 10
- 229940041514 candida albicans extract Drugs 0.000 claims description 10
- 238000001914 filtration Methods 0.000 claims description 10
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 10
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 10
- 238000005067 remediation Methods 0.000 claims description 10
- 239000005720 sucrose Substances 0.000 claims description 10
- 239000012138 yeast extract Substances 0.000 claims description 10
- 239000010457 zeolite Substances 0.000 claims description 10
- 244000061456 Solanum tuberosum Species 0.000 claims description 9
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 9
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 9
- 239000004202 carbamide Substances 0.000 claims description 9
- CNFDGXZLMLFIJV-UHFFFAOYSA-L manganese(II) chloride tetrahydrate Chemical compound O.O.O.O.[Cl-].[Cl-].[Mn+2] CNFDGXZLMLFIJV-UHFFFAOYSA-L 0.000 claims description 9
- 239000004323 potassium nitrate Substances 0.000 claims description 9
- 235000010333 potassium nitrate Nutrition 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 9
- 229960000583 acetic acid Drugs 0.000 claims description 7
- 239000012362 glacial acetic acid Substances 0.000 claims description 7
- RUTXIHLAWFEWGM-UHFFFAOYSA-H iron(3+) sulfate Chemical compound [Fe+3].[Fe+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O RUTXIHLAWFEWGM-UHFFFAOYSA-H 0.000 claims description 7
- 229910000360 iron(III) sulfate Inorganic materials 0.000 claims description 7
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 claims description 7
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 7
- 239000010902 straw Substances 0.000 claims description 6
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 claims description 5
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 5
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 5
- 235000019764 Soybean Meal Nutrition 0.000 claims description 5
- 230000032683 aging Effects 0.000 claims description 5
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 5
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 5
- 238000001354 calcination Methods 0.000 claims description 5
- 235000012054 meals Nutrition 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 5
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 5
- 230000001105 regulatory effect Effects 0.000 claims description 5
- 238000007873 sieving Methods 0.000 claims description 5
- 239000004455 soybean meal Substances 0.000 claims description 5
- 238000001291 vacuum drying Methods 0.000 claims description 5
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 4
- 239000001110 calcium chloride Substances 0.000 claims description 4
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 4
- 238000012258 culturing Methods 0.000 abstract description 15
- 230000000694 effects Effects 0.000 abstract description 11
- 238000001179 sorption measurement Methods 0.000 abstract description 8
- 230000015556 catabolic process Effects 0.000 abstract description 6
- 238000006731 degradation reaction Methods 0.000 abstract description 6
- 235000015097 nutrients Nutrition 0.000 abstract description 6
- 239000002994 raw material Substances 0.000 abstract description 5
- 239000000969 carrier Substances 0.000 abstract description 4
- 239000003431 cross linking reagent Substances 0.000 abstract description 3
- 244000052616 bacterial pathogen Species 0.000 abstract description 2
- 230000009466 transformation Effects 0.000 abstract description 2
- 230000000052 comparative effect Effects 0.000 description 13
- 229920001817 Agar Polymers 0.000 description 12
- 239000008272 agar Substances 0.000 description 12
- 230000001954 sterilising effect Effects 0.000 description 12
- 230000008569 process Effects 0.000 description 6
- 241000233866 Fungi Species 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 230000002285 radioactive effect Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 241001446247 uncultured actinomycete Species 0.000 description 4
- 230000009471 action Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 238000009629 microbiological culture Methods 0.000 description 3
- 230000008635 plant growth Effects 0.000 description 3
- 238000003900 soil pollution Methods 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 240000008415 Lactuca sativa Species 0.000 description 2
- 235000003228 Lactuca sativa Nutrition 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 235000019797 dipotassium phosphate Nutrition 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- 239000000017 hydrogel Substances 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000005416 organic matter Substances 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 229910052748 manganese Inorganic materials 0.000 description 1
- 239000011572 manganese Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000002957 persistent organic pollutant Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000000197 pyrolysis Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- 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
-
- 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
- B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
-
- 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
- C09K17/40—Soil-conditioning materials or soil-stabilising materials containing mixtures of inorganic and organic compounds
-
- 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
- C12N1/14—Fungi; Culture media therefor
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- 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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
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- 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
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/085—Bacillus cereus
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/11—Bacillus megaterium
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- 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
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- 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
- C12R2001/465—Streptomyces
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- 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
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/885—Trichoderma
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- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
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Abstract
The invention relates to the technical field of microbial agents, in particular to a microbial agent for treating soil and a preparation method thereof. The invention prepares mixed microorganism fermentation liquor by activating bacillus subtilis, bacillus megaterium, bacillus cereus, bacillus laterosporus, trichoderma viride and streptomyces microflavus, culturing seeds, fermenting and culturing and uniformly mixing; then the modified biochar and the modified zeolite are used as composite carriers, and the mixed microorganism fermentation broth is fixed by an adsorption method to prepare a core material; finally, chitosan, gelatin and beta-cyclodextrin are used as raw materials, glutaraldehyde is used as a cross-linking agent, and the core material is further coated and fixed to prepare the microbial agent. The microbial agent prepared by the invention can promote the transformation of soil nutrients, the degradation of organic matters, improve the soil structure and inhibit pathogenic bacteria of soil, thereby improving the soil quality and the crop yield and realizing the effect of soil treatment.
Description
Technical Field
The invention relates to the technical field of microbial agents, in particular to a microbial agent for treating soil and a preparation method thereof.
Background
With the increasing severity of environmental pollution, soil pollution treatment is a highly-needed problem. Compared with the traditional physical or chemical method, the biological repair method has the characteristics of ecology, wide application, simplicity, high efficiency, sustainability and the like.
However, most of the current research is directed to bacteria, and less is directed to fungi and actinomycetes. These microbial species play an important role in soil pollution remediation, as they are capable of decomposing more complex organic pollutants. In addition, the current research on the physiological and biochemical characteristics and degradation mechanisms of microbial strains is mostly the research result of single pollutants, and the actual situation is often a complex environment in which multiple pollutants coexist. Therefore, it is necessary to develop a microbial agent with a multi-strain mixture to obtain a more stable treatment effect in practical use. In addition, conventional microbial agent carriers do not facilitate mass transfer between microorganisms and contaminants, which limits the effectiveness of the microbial agent.
Therefore, we propose a microbial agent for treating soil and a preparation method thereof.
Disclosure of Invention
The invention aims to provide a microbial agent for treating soil and a preparation method thereof, which are used for solving the problems in the background technology.
In order to solve the technical problems, the invention provides the following technical scheme:
a preparation method of a microbial agent for treating soil comprises the following steps:
step S1: activating bacillus subtilis, bacillus megaterium, bacillus cereus, bacillus laterosporus, trichoderma viride and streptomyces microflavus, and then respectively inoculating the activated bacillus subtilis, bacillus megaterium, bacillus cereus, bacillus laterosporus, trichoderma viride and streptomyces microflavus into a seed culture medium for seed culture to obtain bacillus subtilis seed liquid, bacillus megaterium seed liquid, bacillus cereus seed liquid, trichoderma viride seed liquid and streptomyces microflavus seed liquid;
step S2: inoculating bacillus subtilis seed liquid, bacillus megaterium seed liquid, bacillus cereus seed liquid, bacillus laterosporus seed liquid, trichoderma viride seed liquid and streptomyces microflavus seed liquid into a liquid culture medium respectively for liquid fermentation to form bacillus subtilis fermentation liquid, bacillus megaterium fermentation liquid, bacillus cereus fermentation liquid, bacillus laterosporus fermentation liquid, trichoderma viride fermentation liquid and streptomyces microflavus fermentation liquid, and uniformly mixing to obtain mixed microorganism fermentation liquid;
step S3: uniformly mixing the modified biochar and the modified zeolite to prepare a composite carrier; uniformly mixing the mixed microorganism fermentation liquor and the composite carrier to prepare a core material;
step S4: uniformly mixing chitosan and glacial acetic acid solution to prepare chitosan solution; uniformly mixing chitosan solution, gelatin aqueous solution and beta-cyclodextrin aqueous solution, adding core material, uniformly mixing, stirring at 30-40 ℃ for 0.5-1h, adding glutaraldehyde, uniformly mixing, reacting at room temperature for 2-3h, washing, suction filtering, and drying to obtain the microbial agent.
In the technical scheme, the comprehensive effect of soil treatment can be improved by mixing a plurality of different microorganism strains, and the different strains have different metabolic characteristics and ecological functions, can act synergistically, and can enhance the nutrient conversion and organic matter degradation capacity of the soil; the modified biochar and the modified zeolite are used as composite carriers, and the mixed microorganism fermentation broth is fixed by an adsorption method to prepare a core material, so that a good adhesion surface can be provided, adhesion and growth of microorganism strains are facilitated, and the survival rate and the use efficiency of the microorganism strains are improved; and then chitosan, gelatin and beta-cyclodextrin are used as raw materials, glutaraldehyde is used as a cross-linking agent, so that hydrogel with a three-dimensional network porous structure is formed, a core material is further coated and fixed, and the stability of the microbial agent in the storage and application processes is improved, so that the soil treatment effect is enhanced.
Further, the specific steps of activation in the step S1 are as follows: inoculating bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus respectively, streaking to beef extract peptone culture medium, and culturing at 30-37 ℃ for 24-48h; inoculating Trichoderma viride, streaking on PDA culture medium, and culturing at 28-30deg.C for 3-5d; inoculating Streptomyces microflavus, streaking to actinomycete culture medium, and culturing at 28-30deg.C for 5-7d.
Further, the beef extract peptone culture medium is as follows: 3-5g/L of beef extract, 10-15g/L of peptone, 3-5g/L of sodium chloride, 15-20g/L of agar, pH=7.2-7.4, and sterilizing at 121 ℃ for 20-30min; the PDA culture medium is as follows: peeling potato 200-202g/L, glucose 18-20g/L, and agar 15-20g/L, sterilizing at 121deg.C for 20-30min; the radioactive fungus culture medium is as follows: 18-20g/L of soluble starch, 1.0-1.5g/L of potassium nitrate, 0.3-0.5g/L of sodium chloride, 0.3-0.5g/L of magnesium sulfate, 0.3-0.5g/L of dipotassium hydrogen phosphate and 18-20g/L of agar, wherein the pH value is 7.2-7.4, and sterilizing at 121 ℃ for 20-30min.
Further, the specific steps of seed culture in the step S1 are as follows: respectively inoculating and streaking the activated bacillus subtilis, bacillus megatherium, bacillus cereus and bacillus laterosporus to a seed culture medium, and performing shake culture at the temperature of 30-37 ℃ and the speed of 160-180r/min for 1-2d to form bacillus subtilis seed liquid, bacillus megatherium seed liquid, bacillus cereus seed liquid and bacillus laterosporus seed liquid; inoculating and streaking activated trichoderma viride to a seed culture medium, and carrying out shake culture at 28-30 ℃ and 160-180r/min for 2-3d to form trichoderma viride seed liquid; inoculating and streaking the activated streptomyces microflavus to a seed culture medium, and carrying out shake culture at the temperature of 28-30 ℃ and the speed of 160-180r/min for 5-7d to form streptomyces microflavus seed liquid.
Further, the seed culture medium of bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus in the step S1 is as follows: 3-5g/L of beef extract, 10-15g/L of peptone, 3-5g/L of sodium chloride and pH=7.2-7.4; the seed culture medium of the trichoderma viride is: peeling potato 200-202g/L and glucose 18-20g/L; the seed culture medium of the streptomyces microflavus is as follows: 18-20g/L of soluble starch, 1.0-1.5g/L of potassium nitrate, 0.3-0.5g/L of sodium chloride, 0.3-0.5g/L of magnesium sulfate, 0.3-0.5g/L of dipotassium hydrogen phosphate and pH=7.2-7.4.
Further, the specific steps of the fermentation culture in the step S2 are as follows: inoculating the bacillus subtilis seed liquid, the bacillus megaterium seed liquid, the bacillus cereus seed liquid and the bacillus laterosporus seed liquid into liquid culture mediums respectively, and performing shake culture for 3-5d at the temperature of 30-37 ℃ and the speed of 180-200r/min to form bacillus subtilis fermentation liquid, bacillus megaterium fermentation liquid, bacillus cereus fermentation liquid and bacillus laterosporus fermentation liquid; inoculating the trichoderma viride seed solution into a liquid culture medium, and performing shake culture at the temperature of 28-30 ℃ and the speed of 160-180r/min for 7-8d to form trichoderma viride fermentation liquor; inoculating Streptomyces microflavus into liquid culture medium, and shake culturing at 28-30deg.C and 160-180r/min for 7-10d to obtain Streptomyces microflavus fermentation broth.
Further, the liquid culture medium of bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus in the step S2 is as follows: 10-15g/L of glucose, 2-3g/L of yeast extract powder, 1-2g/L of ammonium sulfate, 2-3g/L of dipotassium hydrogen phosphate, 0.1-0.2g/L of magnesium sulfate heptahydrate and 0.1-0.2g/L, pH =7.2-7.4 of calcium chloride; the liquid culture medium of the trichoderma viride is: 10-15g/L of sucrose, 12-13g/L of corn meal, 1-2g/L of ammonium nitrate, 4-5g/L of soybean meal, 1-2g/L of magnesium sulfate and 3-4g/L of dipotassium hydrogen phosphate; the liquid culture medium of the streptomyces microflavus is as follows: 4-5g/L of soluble starch, 2-3g/L of sucrose, 2-3g/L of bean cake powder, 0.4-0.5g/L of monopotassium phosphate, 0.3-0.5g/L of yeast extract, 0.25g/L of magnesium sulfate heptahydrate, 0.1-0.2g/L of sodium chloride and pH=7.2-7.4.
Further, in the step S2, the mass ratio of each strain in the mixed microorganism fermentation broth is bacillus subtilis: bacillus megaterium: bacillus cereus: bacillus laterosporus: trichoderma viride: streptomyces microflavus= (4-5): (2-3): (2-3): (1-2): (1-2): 1.
further, the preparation process of the modified biochar comprises the following steps:
mixing manganese chloride tetrahydrate, urea and deionized water uniformly, adding corn straw, mixing uniformly, ultrasonically dissolving for 30-50min, stirring for 12-24h, vacuum drying at 70-80 ℃ for 22-24h to obtain a mixture, placing the mixture into a muffle furnace, heating to 700-800 ℃ at a heating rate of 8-10 ℃/min under the protection of nitrogen, pyrolyzing for 2-3h, cooling to room temperature, sieving with a 150-200 mesh sieve, washing and drying to obtain the modified biochar.
In the technical scheme, corn straw, manganese chloride tetrahydrate and urea are used as raw materials, mn and N co-doped biochar composite materials, namely modified biochar, are prepared by a pyrolysis method, and the modified biochar has a larger specific surface area and a pore structure, so that the adsorption capacity of the modified biochar on nutrients can be improved, and the soil water retention property is improved; the doping of the manganese chloride tetrahydrate and urea can increase the active site on the surface of the biochar and enhance the adsorption effect on nutrients, wherein the doping of the manganese chloride tetrahydrate can increase the manganese content in the biochar and provide trace elements required by plants.
Further, the mass ratio of the manganese chloride tetrahydrate to the urea to the deionized water is 1: (0.5-0.8): (10-12).
Further, the corn stalks are crushed, dried, ground and sieved by a 40-50 mesh sieve in sequence.
Further, the mass of the corn stalk is 1-2 times of that of urea.
Further, the preparation process of the modified zeolite comprises the following steps:
mixing manganese sulfate monohydrate, ferric sulfate and zeolite uniformly, adding deionized water, mixing uniformly, introducing nitrogen, dropwise adding sodium hydroxide solution, regulating the pH to 12-13, continuing stirring for 30-60min, ageing at 80-90 ℃ for 24-48h, cooling to room temperature, washing, filtering, drying, putting into a muffle furnace, heating to 550-650 ℃ at a heating rate of 8-10 ℃/min, calcining for 2-4h, preserving heat for 30-40min, and cooling to room temperature to obtain the modified zeolite.
In the technical scheme, manganese sulfate monohydrate, ferric sulfate and zeolite are used as raw materials to prepare the modified zeolite, so that the surface active sites and adsorption sites of the modified zeolite can be increased, and the adsorption capacity of heavy metals in soil is improved.
Further, the mass ratio of the manganese sulfate monohydrate to the ferric sulfate to the zeolite is 1: (2.0-2.5): (2.0-2.5).
Further, the mass of the deionized water is 10-15 times of that of zeolite.
Further, the concentration of the sodium hydroxide solution is 3-5mol/L.
Further, in the step S3, the mass ratio of the modified biochar to the modified zeolite is 1: (0.5-1.0).
Further, in the step S3, the mass ratio of the mixed microbial fermentation broth to the composite carrier is 1: (3-4).
Further, in the step S4, the mass ratio of the chitosan to the glacial acetic acid solution is 1: (50-55), the mass fraction of the glacial acetic acid solution is 2-3%.
Further, in the step S4, the mass ratio of the chitosan solution, the gelatin aqueous solution and the beta-cyclodextrin aqueous solution is 1: (1-1.2): (1-1.5), the mass fraction of the gelatin aqueous solution is 25-30%, and the mass fraction of the beta-cyclodextrin aqueous solution is 10-12%.
Further, the mass of glutaraldehyde in the step S4 is 0.3-0.5% of the total mass of the chitosan solution, the gelatin aqueous solution and the beta-cyclodextrin aqueous solution.
Further, the effective viable count in the microbial agent in the step S4 is 1×10 9 -1×10 10 CFU/g。
Compared with the prior art, the invention has the following beneficial effects:
1. according to the microbial agent for treating soil and the preparation method thereof, disclosed by the invention, the comprehensive effect of soil treatment can be improved by mixing a plurality of different microbial strains, and the different strains have different metabolic characteristics and ecological functions, can act synergistically, and can enhance the nutrient conversion and organic matter degradation capability of soil; the modified biochar and the modified zeolite are used as composite carriers, and the mixed microorganism fermentation broth is fixed by an adsorption method to prepare a core material, so that a good adhesion surface can be provided, adhesion and growth of microorganism strains are facilitated, and the survival rate and the use efficiency of the microorganism strains are improved; and then chitosan, gelatin and beta-cyclodextrin are used as raw materials, glutaraldehyde is used as a cross-linking agent, so that hydrogel with a three-dimensional network porous structure is formed, and the core material is further coated and fixed, so that the stability of the microbial agent in the storage and application processes is improved. The microbial agent prepared by the invention can promote the transformation of soil nutrients, the degradation of organic matters, improve the soil structure, inhibit soil pathogenic bacteria and promote the growth of plants through various ways, thereby improving the soil quality and the crop yield and realizing the effect of soil treatment.
2. According to the microbial agent for treating soil and the preparation method thereof, disclosed by the invention, the organic carrier modified biochar and the inorganic carrier modified zeolite are compounded, so that the respective characteristics of the organic carrier modified biochar and the inorganic carrier modified zeolite can be fully exerted, and a synergistic effect is formed, so that the soil restoration effect and the crop yield are improved. The modified biochar has a good pore structure and a high specific surface area, can adsorb organic matters and provide microbial habitat, and promotes the microbial activity of soil and the degradation of organic matters; the modified zeolite has excellent ion exchange capacity and adsorption performance, can adsorb heavy metals and harmful substances in soil, and can purify the soil environment. Through the compound use, the organic carrier modified biochar and the inorganic carrier modified zeolite can be mutually supplemented and jointly act on the soil restoration process. The application of the composite carrier is beneficial to improving the soil quality, reducing the soil pollution risk, promoting the plant growth and increasing the crop yield, and has important significance for sustainable agricultural development.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In this example, bacillus subtilis Bacillus subtilis: the purchase number CGMCC1.821 is derived from China general microbiological culture collection center; bacillus megaterium Bacillus megaterium: the purchase number is CGMCC1.16094, which is derived from China general microbiological culture Collection center; bacillus cereus: purchase number ACCC 10405 is derived from China center for type culture Collection of agricultural microorganisms; bacillus laterosporus Brevibacillus laterosporus: the product number GOYJ10638 is from Shanghai Gu research industries, inc.; trichoderma viride: the product number GIM 3.602 is from Shanghai household pharmaceutical technology Co., ltd; streptomyces microflavus Streptomyces microflavus: the purchase number is CGMCC4.6556, which is derived from China general microbiological culture Collection center; corn stalk: derived from Ningxia Huidi agriculture and animal husbandry limited; zeolite: the granularity is 325 meshes, and the particle size is from Hebei Heng Guangdong mineral products Co., ltd; chitosan: the water-soluble chitosan is from Shaanxi Runfeng biotechnology Co., ltd; gelatin: food grade, 160-200 freezing force, is derived from Guangzhou Tonghai biotechnology limited company; beta-cyclodextrin: food grade is from Hubei Shiteng chemical technology Co.
Example 1: the preparation method of the microbial agent for treating the soil comprises the following steps:
step S1: respectively inoculating bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus to a beef extract peptone culture medium, and culturing at 30 ℃ for 24 hours; inoculating Trichoderma viride, streaking to PDA culture medium, and culturing at 28deg.C for 3d; inoculating streptomyces microflavus, streaking to an actinomycete culture medium, and culturing for 5d at 28 ℃; respectively inoculating and streaking the activated bacillus subtilis, bacillus megatherium, bacillus cereus and bacillus laterosporus to a seed culture medium, and carrying out shake culture for 1d at the temperature of 30 ℃ and 160r/min to form bacillus subtilis seed liquid, bacillus megatherium seed liquid, bacillus cereus seed liquid and bacillus laterosporus seed liquid; inoculating and streaking the trichoderma viride to a seed culture medium, and carrying out shake culture for 2d at the temperature of 28 ℃ and the speed of 160r/min to form trichoderma viride seed liquid; inoculating and streaking streptomyces microflavus to a seed culture medium, and carrying out shake culture at 28 ℃ and 160r/min for 5d to form streptomyces microflavus seed liquid;
the beef extract peptone culture medium is as follows: beef extract 3g/L, peptone 10g/L, sodium chloride 3g/L, agar 15g/L, pH=7.2, sterilizing at 121deg.C for 20min; the PDA culture medium is as follows: peeling potato 200g/L, glucose 18g/L, and agar 15g/L, sterilizing at 121deg.C for 20min; the radioactive fungus culture medium is as follows: 18g/L of soluble starch, 1.0g/L of potassium nitrate, 0.3g/L of sodium chloride, 0.3g/L of magnesium sulfate, 0.3g/L of dipotassium hydrogen phosphate and 18g/L of agar, wherein the pH value is=7.2, and sterilizing for 20min at 121 ℃;
the seed culture medium of bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus is as follows: beef extract 3g/L, peptone 10g/L, sodium chloride 3g/L, pH=7.2; the seed culture medium of the trichoderma viride is: peeled potato 200g/L and glucose 18g/L; the seed culture medium of the streptomyces microflavus is as follows: soluble starch 18g/L, potassium nitrate 1.0g/L, sodium chloride 0.3g/L, magnesium sulfate 0.3g/L, dipotassium hydrogen phosphate 0.3g/L, pH=7.2;
step S2: inoculating the bacillus subtilis seed liquid, the bacillus megaterium seed liquid, the bacillus cereus seed liquid and the bacillus laterosporus seed liquid into liquid culture mediums respectively, and carrying out shake culture for 3d at the temperature of 30 ℃ and the speed of 180r/min to form bacillus subtilis fermentation liquid, bacillus megaterium fermentation liquid, bacillus cereus fermentation liquid and bacillus laterosporus fermentation liquid; inoculating the trichoderma viride seed solution into a liquid culture medium, and performing shaking culture at 28 ℃ and 160r/min for 7d to form trichoderma viride fermentation liquor; inoculating streptomyces microflavus into a liquid culture medium, and carrying out shaking culture for 7d at 28 ℃ and 160r/min to form a streptomyces microflavus fermentation broth, and uniformly mixing (the mass ratio of each strain is bacillus subtilis to bacillus megaterium to bacillus cereus to bacillus laterosporus to trichoderma viride to streptomyces microflavus=4:2:2:1:1:1) to prepare a mixed microorganism fermentation broth;
the liquid culture medium of bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus is as follows: glucose 10g/L, yeast extract 2g/L, ammonium sulfate 1g/L, dipotassium phosphate 2g/L, magnesium sulfate heptahydrate 0.1g/L, calcium chloride 0.1g/L, pH =7.2; the liquid culture medium of the trichoderma viride is: 10g/L of sucrose, 12g/L of corn meal, 1g/L of ammonium nitrate, 4g/L of soybean meal, 1g/L of magnesium sulfate and 3g/L of dipotassium hydrogen phosphate; the liquid culture medium of the streptomyces microflavus is as follows: 4g/L of soluble starch, 2g/L of sucrose, 2g/L of bean cake powder, 0.4g/L of monopotassium phosphate, 0.3g/L of yeast extract, 0.2g/L of magnesium sulfate heptahydrate, 0.1g/L of sodium chloride and pH=7.2;
step S3: uniformly mixing 20g of modified biochar and 10g of modified zeolite to prepare a composite carrier; uniformly mixing 10g of mixed microorganism fermentation liquor and 30g of composite carrier to prepare a core material;
step S4: uniformly mixing 2g of chitosan and 100g of 2wt% glacial acetic acid solution to prepare a chitosan solution; uniformly mixing 100g of chitosan solution, 100g of 25wt% gelatin aqueous solution and 100g of 10wt% beta-cyclodextrin aqueous solution, adding a core material, uniformly mixing, stirring at 30 ℃ for 0.5h, adding 0.9g of glutaraldehyde, uniformly mixing, reacting at room temperature for 2h, washing, suction filtering, and drying to obtain a microbial agent;
the preparation process of the modified biochar comprises the following steps:
uniformly mixing 40g of manganese chloride tetrahydrate, 20g of urea and 400g of deionized water, adding 20g of corn straw, uniformly mixing, ultrasonically dissolving for 30min, stirring for 12h, vacuum drying at 70 ℃ for 22h to obtain a mixture, placing the mixture into a muffle furnace, heating to 700 ℃ at a heating rate of 8 ℃/min under the protection of nitrogen, pyrolyzing for 2h, cooling to room temperature, sieving with a 150-mesh sieve, washing and drying to obtain modified biochar;
the preparation process of the modified zeolite comprises the following steps:
mixing 5g of manganese sulfate monohydrate, 10g of ferric sulfate and 10g of zeolite uniformly, adding 100g of deionized water, mixing uniformly, introducing nitrogen, dropwise adding sodium hydroxide solution, regulating the pH to 12, continuously stirring for 30min, aging at 80 ℃ for 24h, cooling to room temperature, washing, filtering, drying, putting into a muffle furnace, heating to 550 ℃ at a heating rate of 8 ℃/min, calcining for 2h, preserving heat for 30min, and cooling to room temperature to obtain the modified zeolite.
Example 2: the preparation method of the microbial agent for treating the soil comprises the following steps:
step S1: respectively inoculating bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus to a beef extract peptone culture medium, and culturing at 36 ℃ for 36 hours; inoculating and streaking Trichoderma viride to PDA culture medium, and culturing at 29 deg.C for 4d; inoculating streptomyces microflavus, streaking to an actinomycete culture medium, and culturing for 6d at 29 ℃; respectively inoculating and streaking the activated bacillus subtilis, bacillus megatherium, bacillus cereus and bacillus laterosporus to a seed culture medium, and carrying out shake culture at 36 ℃ and 170r/min for 1.5d to form bacillus subtilis seed liquid, bacillus megatherium seed liquid, bacillus cereus seed liquid and bacillus laterosporus seed liquid; inoculating and streaking the trichoderma viride to a seed culture medium, and carrying out shaking culture for 2.5d at 29 ℃ and 170r/min to form trichoderma viride seed liquid; inoculating and streaking streptomyces microflavus to a seed culture medium, and carrying out shaking culture for 6d at 29 ℃ and 170r/min to form streptomyces microflavus seed liquid;
the beef extract peptone culture medium is as follows: beef extract 4g/L, peptone 13g/L, sodium chloride 4g/L, agar 18g/L, pH=7.3, sterilizing at 121deg.C for 25min; the PDA culture medium is as follows: peeling potato 201g/L, glucose 19g/L, and agar 18g/L, sterilizing at 121deg.C for 25min; the radioactive fungus culture medium is as follows: 19g/L of soluble starch, 1.4g/L of potassium nitrate, 0.4g/L of sodium chloride, 0.4g/L of magnesium sulfate, 0.4g/L of dipotassium hydrogen phosphate and 19g/L of agar, wherein pH=7.3, and sterilizing at 121 ℃ for 25min;
the seed culture medium of bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus is as follows: beef extract 4g/L, peptone 13g/L, sodium chloride 4g/L, pH=7.3; the seed culture medium of the trichoderma viride is: peeled potato 201g/L and glucose 19g/L; the seed culture medium of the streptomyces microflavus is as follows: soluble starch 19g/L, potassium nitrate 1.4g/L, sodium chloride 0.4g/L, magnesium sulfate 0.4g/L, dipotassium hydrogen phosphate 0.4g/L, pH=7.3;
step S2: inoculating the bacillus subtilis seed liquid, the bacillus megaterium seed liquid, the bacillus cereus seed liquid and the bacillus laterosporus seed liquid into liquid culture mediums respectively, and carrying out shake culture for 4d at 36 ℃ and 190r/min to form bacillus subtilis fermentation liquid, bacillus megaterium fermentation liquid, bacillus cereus fermentation liquid and bacillus laterosporus fermentation liquid; inoculating the trichoderma viride seed solution into a liquid culture medium, and performing shaking culture at 29 ℃ and 170r/min for 7.5 days to form trichoderma viride fermentation liquor; inoculating streptomyces microflavus into a liquid culture medium, and carrying out shaking culture for 9d at 29 ℃ and 170r/min to form a streptomyces microflavus fermentation broth, and uniformly mixing (the mass ratio of each strain is bacillus subtilis to bacillus megaterium to bacillus cereus to bacillus laterosporus to trichoderma viride to streptomyces microflavus=4.5:2.5:2.5:1.5:1.5:1) to prepare a mixed microorganism fermentation broth;
the liquid culture medium of bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus is as follows: 13g/L glucose, 2.5g/L yeast extract powder, 1.5g/L ammonium sulfate, 2.5g/L dipotassium phosphate, 0.15g/L magnesium sulfate heptahydrate, 0.15g/L, pH =7.3; the liquid culture medium of the trichoderma viride is: 13g/L of sucrose, 12.5g/L of corn meal, 1.5g/L of ammonium nitrate, 4.5g/L of soybean meal, 1.2g/L of magnesium sulfate and 3.5g/L of dipotassium hydrogen phosphate; the liquid culture medium of the streptomyces microflavus is as follows: 4.5g/L of soluble starch, 2.5g/L of sucrose, 2.5g/L of bean cake powder, 0.45g/L of monopotassium phosphate, 0.4g/L of yeast extract, 0.25g/L of magnesium sulfate heptahydrate, 0.15g/L of sodium chloride and pH=7.3;
step S3: uniformly mixing 20g of modified biochar and 15g of modified zeolite to prepare a composite carrier; uniformly mixing 10g of mixed microorganism fermentation liquor and 35g of composite carrier to prepare a core material;
step S4: uniformly mixing 2g of chitosan and 108g of 2.5wt% glacial acetic acid solution to prepare a chitosan solution; uniformly mixing 100g of chitosan solution, 110g of 28wt% gelatin aqueous solution and 120g of 11wt% beta-cyclodextrin aqueous solution, adding a core material, uniformly mixing, stirring at 35 ℃ for 0.8h, adding 1.32g of glutaraldehyde, uniformly mixing, reacting at room temperature for 2.5h, washing, suction filtering, and drying to obtain a microbial agent;
the preparation process of the modified biochar comprises the following steps:
uniformly mixing 50g of manganese chloride tetrahydrate, 30g of urea and 550g of deionized water, adding 20g of corn straw, uniformly mixing, ultrasonically dissolving for 40min, stirring for 18h, vacuum drying at 75 ℃ for 23h to obtain a mixture, placing the mixture into a muffle furnace, heating to 750 ℃ at a heating rate of 9 ℃/min under the protection of nitrogen, pyrolyzing for 2.5h, cooling to room temperature, sieving with a 180-mesh sieve, washing and drying to obtain modified biochar;
the preparation process of the modified zeolite comprises the following steps:
mixing 6.5g of manganese sulfate monohydrate, 15g of ferric sulfate and 15g of zeolite uniformly, adding 180g of deionized water, mixing uniformly, introducing nitrogen, dropwise adding sodium hydroxide solution, regulating the pH to 12, continuously stirring for 50min, aging at 85 ℃ for 36h, cooling to room temperature, washing, filtering, drying, putting into a muffle furnace, heating to 600 ℃ at a heating rate of 9 ℃/min, calcining for 3h, preserving heat for 35min, and cooling to room temperature to obtain the modified zeolite.
Example 3: the preparation method of the microbial agent for treating the soil comprises the following steps:
step S1: inoculating bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus respectively, streaking to a beef extract peptone culture medium, and culturing at 37 ℃ for 48 hours; inoculating Trichoderma viride, streaking to PDA culture medium, and culturing at 30deg.C for 5d; inoculating streptomyces microflavus, streaking to an actinomycete culture medium, and culturing for 7d at 30 ℃; respectively inoculating and streaking the activated bacillus subtilis, bacillus megatherium, bacillus cereus and bacillus laterosporus to a seed culture medium, and carrying out shake culture for 2d at 37 ℃ and 180r/min to form bacillus subtilis seed liquid, bacillus megatherium seed liquid, bacillus cereus seed liquid and bacillus laterosporus seed liquid; inoculating and streaking the trichoderma viride to a seed culture medium, and carrying out shake culture for 3d at the temperature of 30 ℃ and the speed of 180r/min to form trichoderma viride seed liquid; inoculating and streaking streptomyces microflavus to a seed culture medium, and carrying out shaking culture at 30 ℃ and 180r/min for 7d to form streptomyces microflavus seed liquid;
the beef extract peptone culture medium is as follows: beef extract 5g/L, peptone 15g/L, sodium chloride 5g/L, agar 20g/L, pH=7.4, sterilizing at 121deg.C for 30min; the PDA culture medium is as follows: peeling potato 202g/L, glucose 20g/L, and agar 20g/L, sterilizing at 121deg.C for 30min; the radioactive fungus culture medium is as follows: 20g/L of soluble starch, 1.5g/L of potassium nitrate, 0.5g/L of sodium chloride, 0.5g/L of magnesium sulfate, 0.5g/L of dipotassium hydrogen phosphate and 20g/L of agar, wherein the pH value is=7.4, and sterilizing for 30min at 121 ℃;
the seed culture medium of bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus is as follows: beef extract 5g/L, peptone 15g/L, sodium chloride 5g/L, pH=7.4; the seed culture medium of the trichoderma viride is: peeling potato 202g/L and glucose 20g/L; the seed culture medium of the streptomyces microflavus is as follows: soluble starch 20g/L, potassium nitrate 1.5g/L, sodium chloride 0.5g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, pH=7.4;
step S2: inoculating the bacillus subtilis seed liquid, the bacillus megaterium seed liquid, the bacillus cereus seed liquid and the bacillus laterosporus seed liquid into liquid culture mediums respectively, and carrying out shake culture for 5d at 37 ℃ and 200r/min to form bacillus subtilis fermentation liquid, bacillus megaterium fermentation liquid, bacillus cereus fermentation liquid and bacillus laterosporus fermentation liquid; inoculating the trichoderma viride seed solution into a liquid culture medium, and performing shaking culture at 30 ℃ and 180r/min for 8d to form trichoderma viride fermentation liquor; inoculating streptomyces microflavus into a liquid culture medium, and carrying out shaking culture for 10d at the temperature of 30 ℃ and 180r/min to form a streptomyces microflavus fermentation broth, and uniformly mixing (the mass ratio of each strain is bacillus subtilis to bacillus megaterium to bacillus cereus to bacillus laterosporus to trichoderma viride to streptomyces microflavus=5:3:3:2:2:1) to prepare a mixed microorganism fermentation broth;
the liquid culture medium of bacillus subtilis, bacillus megaterium, bacillus cereus and bacillus laterosporus is as follows: 15g/L glucose, 3g/L yeast extract powder, 2g/L ammonium sulfate, 3g/L dipotassium hydrogen phosphate, 0.2g/L magnesium sulfate heptahydrate, 0.2g/L, pH =7.4 calcium chloride; the liquid culture medium of the trichoderma viride is: 15g/L of sucrose, 13g/L of corn meal, 2g/L of ammonium nitrate, 5g/L of soybean meal, 2g/L of magnesium sulfate and 4g/L of dipotassium hydrogen phosphate; the liquid culture medium of the streptomyces microflavus is as follows: 5g/L of soluble starch, 3g/L of sucrose, 3g/L of bean cake powder, 0.5g/L of monopotassium phosphate, 0.5g/L of yeast extract, 0.3g/L of magnesium sulfate heptahydrate, 0.2g/L of sodium chloride and pH=7.4;
step S3: uniformly mixing 20g of modified biochar and 20g of modified zeolite to prepare a composite carrier; uniformly mixing 10g of mixed microorganism fermentation liquor and 40g of composite carrier to prepare a core material;
step S4: uniformly mixing 2g of chitosan and 110g of 3wt% glacial acetic acid solution to prepare a chitosan solution; uniformly mixing 100g of chitosan solution, 120g of 30wt% gelatin aqueous solution and 150g of 12wt% beta-cyclodextrin aqueous solution, adding a core material, uniformly mixing, stirring at 40 ℃ for 1h, adding 1.85g of glutaraldehyde, uniformly mixing, reacting at room temperature for 3h, washing, suction filtering and drying to obtain a microbial agent;
the preparation process of the modified biochar comprises the following steps:
uniformly mixing 50g of manganese chloride tetrahydrate, 40g of urea and 600g of deionized water, adding 20g of corn straw, uniformly mixing, ultrasonically dissolving for 50min, stirring for 24h, vacuum drying at 80 ℃ for 24h to obtain a mixture, placing the mixture into a muffle furnace, heating to 800 ℃ at a heating rate of 10 ℃/min under the protection of nitrogen, pyrolyzing for 3h, cooling to room temperature, sieving with a 200-mesh sieve, washing and drying to obtain modified biochar;
the preparation process of the modified zeolite comprises the following steps:
mixing 8g of manganese sulfate monohydrate, 20g of ferric sulfate and 20g of zeolite uniformly, adding 300g of deionized water, mixing uniformly, introducing nitrogen, dropwise adding sodium hydroxide solution, regulating the pH to 13, continuously stirring for 60min, aging at 90 ℃ for 48h, cooling to room temperature, washing, filtering, drying, putting into a muffle furnace, heating to 650 ℃ at a heating rate of 10 ℃/min, calcining for 4h, preserving heat for 40min, and cooling to room temperature to obtain the modified zeolite.
Comparative example 1: the preparation method of the microbial agent for treating the soil comprises the following steps:
in comparison with example 2, comparative example 1 does not include the process for preparing the modified biochar or the modified zeolite, and the modified biochar or the modified zeolite in step S3 is replaced with the normal biochar or the normal zeolite, and the other steps are the same as in example 2.
Comparative example 2: the preparation method of the microbial agent for treating the soil comprises the following steps:
compared with example 2, comparative example 2 uses only modified zeolite as carrier, and the mixed microorganism fermentation broth and the modified zeolite are uniformly mixed to prepare a core material; the other steps were the same as in example 2.
Comparative example 3: the preparation method of the microbial agent for treating the soil comprises the following steps:
comparative example 3 does not include Trichoderma viride and Streptomyces microflavus as in example 2, and the other steps are the same as in example 2.
Comparative example 4: the preparation method of the microbial agent for treating the soil comprises the following steps:
in comparison with example 2, comparative example 4 does not include step S4, and the other steps are the same as in example 2 except that the core material is used as the microbial agent.
Experiment
Experiment 1: the microbial agents prepared in example 2 and comparative examples 1 to 3 were placed in 0.9% physiological saline, and after the microbial agents were completely dissolved, the microbial agents were filtered to obtain a filtrate. Subsequently, the cells in the filtrate were counted using a hemocytometer, and the detection results were recorded:
from the data in the above table, the following conclusions can be clearly drawn:
1. compared with the example 2, the effective viable bacteria number of the product obtained in the comparative examples 1-2 is reduced, and the invention can realize the synergistic effect by taking the modified biochar and the modified zeolite as the composite carrier, and adsorb and fix the mixed microorganism fermentation broth, thereby providing a good adhesion surface, being beneficial to the adhesion and growth of microorganism strains and improving the survival rate and the use efficiency of the microorganism strains.
2. Compared with the example 2, the effective viable bacteria number of the products obtained in the comparative examples 3-4 is reduced, which shows that the addition of trichoderma viride and streptomyces microflavus and the coating fixation of the core material in the invention can obviously improve the viable bacteria number and activity in the composite carrier.
Experiment 2: physical and chemical property test of soil
In the experiment, 5 treatment groups and 1 control group are arranged, and 1 mu of experimental field is selected for planting lettuce. The test fields were divided into 6 groups, wherein 5 treatment groups were respectively applied with the microbial agent prepared in example 2 and the microbial agents prepared in comparative examples 1 to 4, the fertilization amount was 25 kg/mu, and the final group was a blank control group without any microbial agent added. Before lettuce is planted, common farmyard manure is applied to 5 groups of test fields, and the fertilizing amount is 350 kg/mu. After planting for 30 days, regular quantitative watering management is carried out in the growth process, and no compound fertilizer or pesticide is applied. After 30 days, soil samples were collected and subjected to physical and chemical property test.
Test results
From the data in the above table, the following conclusions can be clearly drawn:
compared with the example 2, the products obtained in the comparative examples 1-4 have reduced soil treatment effects, which shows that the microbial agent prepared by the preparation method has better soil treatment effects, and can increase the contents of organic matters, nitrogen, phosphorus and potassium in the soil, thereby improving the soil fertility, promoting the plant growth and improving the soil texture.
It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process method article or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process method article or apparatus.
Finally, it should be noted that: the foregoing description is only a preferred embodiment of the present invention, and the present invention is not limited thereto, but it is to be understood that modifications and equivalents of some of the technical features described in the foregoing embodiments may be made by those skilled in the art, although the present invention has been described in detail with reference to the foregoing embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A preparation method of a microbial agent for treating soil is characterized by comprising the following steps: the method comprises the following steps:
step S1: activating bacillus subtilis, bacillus megaterium, bacillus cereus, bacillus laterosporus, trichoderma viride and streptomyces microflavus, and then respectively inoculating the activated bacillus subtilis, bacillus megaterium, bacillus cereus, bacillus laterosporus, trichoderma viride and streptomyces microflavus into a seed culture medium for seed culture to obtain bacillus subtilis seed liquid, bacillus megaterium seed liquid, bacillus cereus seed liquid, trichoderma viride seed liquid and streptomyces microflavus seed liquid;
step S2: inoculating bacillus subtilis seed liquid, bacillus megaterium seed liquid, bacillus cereus seed liquid, bacillus laterosporus seed liquid, trichoderma viride seed liquid and streptomyces microflavus seed liquid into a liquid culture medium respectively for liquid fermentation to form bacillus subtilis fermentation liquid, bacillus megaterium fermentation liquid, bacillus cereus fermentation liquid, bacillus laterosporus fermentation liquid, trichoderma viride fermentation liquid and streptomyces microflavus fermentation liquid, and uniformly mixing to obtain mixed microorganism fermentation liquid;
step S3: uniformly mixing the modified biochar and the modified zeolite to prepare a composite carrier; uniformly mixing the mixed microorganism fermentation liquor and the composite carrier to prepare a core material;
step S4: uniformly mixing chitosan and glacial acetic acid solution to prepare chitosan solution; uniformly mixing chitosan solution, gelatin aqueous solution and beta-cyclodextrin aqueous solution, adding core material, uniformly mixing, stirring at 30-40 ℃ for 0.5-1h, adding glutaraldehyde, uniformly mixing, reacting at room temperature for 2-3h, washing, suction filtering, and drying to obtain the microbial agent.
2. The method for preparing the microbial agent for soil remediation according to claim 1, wherein the method comprises the following steps: the specific steps of seed culture in the step S1 are as follows: respectively inoculating and streaking the activated bacillus subtilis, bacillus megatherium, bacillus cereus and bacillus laterosporus to a seed culture medium, and performing shake culture at the temperature of 30-37 ℃ and the speed of 160-180r/min for 1-2d to form bacillus subtilis seed liquid, bacillus megatherium seed liquid, bacillus cereus seed liquid and bacillus laterosporus seed liquid; inoculating and streaking activated trichoderma viride to a seed culture medium, and carrying out shake culture at 28-30 ℃ and 160-180r/min for 2-3d to form trichoderma viride seed liquid; inoculating and streaking the activated streptomyces microflavus to a seed culture medium, and carrying out shake culture at the temperature of 28-30 ℃ and the speed of 160-180r/min for 5-7d to form streptomyces microflavus seed liquid.
3. The method for preparing the microbial agent for soil remediation according to claim 1, wherein the method comprises the following steps: the seed culture medium of the bacillus subtilis, the bacillus megaterium, the bacillus cereus and the bacillus laterosporus in the step S1 is as follows: 3-5g/L of beef extract, 10-15g/L of peptone, 3-5g/L of sodium chloride and pH=7.2-7.4; the seed culture medium of the trichoderma viride is: peeling potato 200-202g/L and glucose 18-20g/L; the seed culture medium of the streptomyces microflavus is as follows: 18-20g/L of soluble starch, 1.0-1.5g/L of potassium nitrate, 0.3-0.5g/L of sodium chloride, 0.3-0.5g/L of magnesium sulfate, 0.3-0.5g/L of dipotassium hydrogen phosphate and pH=7.2-7.4.
4. The method for preparing the microbial agent for soil remediation according to claim 1, wherein the method comprises the following steps: the liquid culture medium of the bacillus subtilis, the bacillus megaterium, the bacillus cereus and the bacillus laterosporus in the step S2 is as follows: 10-15g/L of glucose, 2-3g/L of yeast extract powder, 1-2g/L of ammonium sulfate, 2-3g/L of dipotassium hydrogen phosphate, 0.1-0.2g/L of magnesium sulfate heptahydrate and 0.1-0.2g/L, pH =7.2-7.4 of calcium chloride; the liquid culture medium of the trichoderma viride is: 10-15g/L of sucrose, 12-13g/L of corn meal, 1-2g/L of ammonium nitrate, 4-5g/L of soybean meal, 1-2g/L of magnesium sulfate and 3-4g/L of dipotassium hydrogen phosphate; the liquid culture medium of the streptomyces microflavus is as follows: 4-5g/L of soluble starch, 2-3g/L of sucrose, 2-3g/L of bean cake powder, 0.4-0.5g/L of monopotassium phosphate, 0.3-0.5g/L of yeast extract, 0.2-0.3g/L of magnesium sulfate heptahydrate, 0.1-0.2g/L of sodium chloride and pH=7.2-7.4.
5. The method for preparing the microbial agent for soil remediation according to claim 1, wherein the method comprises the following steps: the mass ratio of each strain in the mixed microorganism fermentation liquor in the step S2 is as follows: bacillus megaterium: bacillus cereus: bacillus laterosporus: trichoderma viride: streptomyces microflavus= (4-5): (2-3): (2-3): (1-2): (1-2): 1.
6. the method for preparing the microbial agent for soil remediation according to claim 1, wherein the method comprises the following steps: the preparation process of the modified biochar in the step S3 comprises the following steps:
mixing manganese chloride tetrahydrate, urea and deionized water uniformly, adding corn straw, mixing uniformly, ultrasonically dissolving for 30-50min, stirring for 12-24h, vacuum drying at 70-80 ℃ for 22-24h to obtain a mixture, placing the mixture into a muffle furnace, heating to 700-800 ℃ at a heating rate of 8-10 ℃/min under the protection of nitrogen, pyrolyzing for 2-3h, cooling to room temperature, sieving with a 150-200 mesh sieve, washing and drying to obtain the modified biochar.
7. The method for preparing the microbial agent for soil remediation according to claim 1, wherein the method comprises the following steps: the preparation process of the modified zeolite in the step S3 comprises the following steps:
mixing manganese sulfate monohydrate, ferric sulfate and zeolite uniformly, adding deionized water, mixing uniformly, introducing nitrogen, dropwise adding sodium hydroxide solution, regulating the pH to 12-13, continuing stirring for 30-60min, ageing at 80-90 ℃ for 24-48h, cooling to room temperature, washing, filtering, drying, putting into a muffle furnace, heating to 550-650 ℃ at a heating rate of 8-10 ℃/min, calcining for 2-4h, preserving heat for 30-40min, and cooling to room temperature to obtain the modified zeolite.
8. The method for preparing the microbial agent for soil remediation according to claim 1, wherein the method comprises the following steps: in the step S3, the mass ratio of the mixed microorganism fermentation liquor to the composite carrier is 1: (3-4).
9. The method for preparing a microbial agent for soil remediation according to claim 1, wherein the effective viable count of the microbial agent in step S4 is 1×10 9 -1×10 10 CFU/g。
10. The method according to any one of claims 1 to 9, wherein a microbial agent for soil remediation is produced.
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| CN117862195A (en) * | 2024-03-12 | 2024-04-12 | 山西青联农业科技有限公司 | Method for carrying out iron tailing soil formation by utilizing ectopic ore-decomposing biological fermentation bed |
| CN118489698A (en) * | 2024-07-16 | 2024-08-16 | 山东劲脉植物细胞信息技术有限公司 | Plant disease-resistant agent containing antibacterial peptide and preparation method thereof |
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| CN117862195A (en) * | 2024-03-12 | 2024-04-12 | 山西青联农业科技有限公司 | Method for carrying out iron tailing soil formation by utilizing ectopic ore-decomposing biological fermentation bed |
| CN117862195B (en) * | 2024-03-12 | 2024-05-14 | 山西青联农业科技有限公司 | Method for carrying out iron tailing soil formation by utilizing ectopic ore-decomposing biological fermentation bed |
| CN118489698A (en) * | 2024-07-16 | 2024-08-16 | 山东劲脉植物细胞信息技术有限公司 | Plant disease-resistant agent containing antibacterial peptide and preparation method thereof |
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