CN117000756A - Microorganism-mineral composite microbial agent and preparation method and application thereof - Google Patents
Microorganism-mineral composite microbial agent and preparation method and application thereof Download PDFInfo
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- CN117000756A CN117000756A CN202310820612.4A CN202310820612A CN117000756A CN 117000756 A CN117000756 A CN 117000756A CN 202310820612 A CN202310820612 A CN 202310820612A CN 117000756 A CN117000756 A CN 117000756A
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- 229910052500 inorganic mineral Inorganic materials 0.000 title claims abstract description 73
- 230000000813 microbial effect Effects 0.000 title claims abstract description 73
- 239000011707 mineral Substances 0.000 title claims abstract description 73
- 239000002131 composite material Substances 0.000 title claims abstract description 47
- 239000003795 chemical substances by application Substances 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title abstract description 8
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 claims abstract description 79
- 244000005700 microbiome Species 0.000 claims abstract description 46
- 230000003647 oxidation Effects 0.000 claims abstract description 39
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 39
- 229910002651 NO3 Inorganic materials 0.000 claims abstract description 35
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 claims abstract description 35
- 239000002734 clay mineral Substances 0.000 claims abstract description 28
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 27
- 239000002689 soil Substances 0.000 claims abstract description 27
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims abstract description 16
- 230000009467 reduction Effects 0.000 claims abstract description 15
- 229910001608 iron mineral Inorganic materials 0.000 claims abstract description 12
- 238000004519 manufacturing process Methods 0.000 claims abstract description 4
- 229910001448 ferrous ion Inorganic materials 0.000 claims description 12
- 239000002054 inoculum Substances 0.000 claims description 12
- 241000894006 Bacteria Species 0.000 claims description 9
- 239000001963 growth medium Substances 0.000 claims description 9
- 238000003501 co-culture Methods 0.000 claims description 5
- 230000008878 coupling Effects 0.000 claims description 4
- 238000010168 coupling process Methods 0.000 claims description 4
- 238000005859 coupling reaction Methods 0.000 claims description 4
- 230000001419 dependent effect Effects 0.000 claims description 4
- 241000363465 Acidovorax sp. BoFeN1 Species 0.000 claims description 3
- 241000588747 Klebsiella pneumoniae Species 0.000 claims description 2
- 239000003337 fertilizer Substances 0.000 claims description 2
- 239000002994 raw material Substances 0.000 claims description 2
- 230000009919 sequestration Effects 0.000 claims description 2
- GQPLMRYTRLFLPF-UHFFFAOYSA-N Nitrous Oxide Chemical compound [O-][N+]#N GQPLMRYTRLFLPF-UHFFFAOYSA-N 0.000 abstract description 24
- 238000011282 treatment Methods 0.000 abstract description 16
- 230000000694 effects Effects 0.000 abstract description 12
- 239000001272 nitrous oxide Substances 0.000 abstract description 12
- 230000001590 oxidative effect Effects 0.000 abstract description 12
- 230000009286 beneficial effect Effects 0.000 abstract description 10
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 abstract description 9
- 239000005431 greenhouse gas Substances 0.000 abstract description 8
- 238000009825 accumulation Methods 0.000 abstract description 5
- 230000007613 environmental effect Effects 0.000 abstract description 4
- 230000006872 improvement Effects 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- 239000011230 binding agent Substances 0.000 abstract description 3
- 239000000126 substance Substances 0.000 abstract description 3
- 230000007774 longterm Effects 0.000 abstract description 2
- 238000001308 synthesis method Methods 0.000 abstract description 2
- 235000010755 mineral Nutrition 0.000 description 36
- 239000000243 solution Substances 0.000 description 28
- 229930182555 Penicillin Natural products 0.000 description 26
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 26
- 229940049954 penicillin Drugs 0.000 description 26
- 238000006243 chemical reaction Methods 0.000 description 20
- 230000001580 bacterial effect Effects 0.000 description 15
- 229910052902 vermiculite Inorganic materials 0.000 description 13
- 239000010455 vermiculite Substances 0.000 description 13
- 235000019354 vermiculite Nutrition 0.000 description 13
- 239000002609 medium Substances 0.000 description 12
- 239000002068 microbial inoculum Substances 0.000 description 12
- 230000008859 change Effects 0.000 description 10
- 102000004169 proteins and genes Human genes 0.000 description 10
- 108090000623 proteins and genes Proteins 0.000 description 10
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 10
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 8
- 230000001954 sterilising effect Effects 0.000 description 8
- 239000007990 PIPES buffer Substances 0.000 description 7
- 229960002089 ferrous chloride Drugs 0.000 description 7
- NMCUIPGRVMDVDB-UHFFFAOYSA-L iron dichloride Chemical compound Cl[Fe]Cl NMCUIPGRVMDVDB-UHFFFAOYSA-L 0.000 description 7
- 239000005995 Aluminium silicate Substances 0.000 description 6
- 235000012211 aluminium silicate Nutrition 0.000 description 6
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 239000002244 precipitate Substances 0.000 description 6
- 239000000725 suspension Substances 0.000 description 6
- IHPYMWDTONKSCO-UHFFFAOYSA-N 2,2'-piperazine-1,4-diylbisethanesulfonic acid Chemical compound OS(=O)(=O)CCN1CCN(CCS(O)(=O)=O)CC1 IHPYMWDTONKSCO-UHFFFAOYSA-N 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 5
- 238000007789 sealing Methods 0.000 description 5
- 239000001632 sodium acetate Substances 0.000 description 5
- 235000017281 sodium acetate Nutrition 0.000 description 5
- 239000004317 sodium nitrate Substances 0.000 description 5
- 235000010344 sodium nitrate Nutrition 0.000 description 5
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 4
- 229910052782 aluminium Inorganic materials 0.000 description 4
- GUJOJGAPFQRJSV-UHFFFAOYSA-N dialuminum;dioxosilane;oxygen(2-);hydrate Chemical compound O.[O-2].[O-2].[O-2].[Al+3].[Al+3].O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O GUJOJGAPFQRJSV-UHFFFAOYSA-N 0.000 description 4
- 229910052742 iron Inorganic materials 0.000 description 4
- 229910052901 montmorillonite Inorganic materials 0.000 description 4
- 239000012452 mother liquor Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 229910021642 ultra pure water Inorganic materials 0.000 description 4
- 239000012498 ultrapure water Substances 0.000 description 4
- 238000005303 weighing Methods 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 239000004927 clay Substances 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- LNOPIUAQISRISI-UHFFFAOYSA-N n'-hydroxy-2-propan-2-ylsulfonylethanimidamide Chemical compound CC(C)S(=O)(=O)CC(N)=NO LNOPIUAQISRISI-UHFFFAOYSA-N 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 235000019270 ammonium chloride Nutrition 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- PPCXFTKZPBHXIW-UHFFFAOYSA-N ethyl ethanesulfonate Chemical compound CCOS(=O)(=O)CC PPCXFTKZPBHXIW-UHFFFAOYSA-N 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 229940050906 magnesium chloride hexahydrate Drugs 0.000 description 2
- DHRRIBDTHFBPNG-UHFFFAOYSA-L magnesium dichloride hexahydrate Chemical compound O.O.O.O.O.O.[Mg+2].[Cl-].[Cl-] DHRRIBDTHFBPNG-UHFFFAOYSA-L 0.000 description 2
- 238000007885 magnetic separation Methods 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- 241000589774 Pseudomonas sp. Species 0.000 description 1
- 241000907663 Siproeta stelenes Species 0.000 description 1
- 241000276425 Xiphophorus maculatus Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 230000004931 aggregating effect Effects 0.000 description 1
- HPTYUNKZVDYXLP-UHFFFAOYSA-N aluminum;trihydroxy(trihydroxysilyloxy)silane;hydrate Chemical compound O.[Al].[Al].O[Si](O)(O)O[Si](O)(O)O HPTYUNKZVDYXLP-UHFFFAOYSA-N 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229910001919 chlorite Inorganic materials 0.000 description 1
- 229910052619 chlorite group Inorganic materials 0.000 description 1
- QBWCMBCROVPCKQ-UHFFFAOYSA-N chlorous acid Chemical compound OCl=O QBWCMBCROVPCKQ-UHFFFAOYSA-N 0.000 description 1
- 238000000975 co-precipitation Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 229910001649 dickite Inorganic materials 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 229910052621 halloysite Inorganic materials 0.000 description 1
- 229910052900 illite Inorganic materials 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- VGIBGUSAECPPNB-UHFFFAOYSA-L nonaaluminum;magnesium;tripotassium;1,3-dioxido-2,4,5-trioxa-1,3-disilabicyclo[1.1.1]pentane;iron(2+);oxygen(2-);fluoride;hydroxide Chemical compound [OH-].[O-2].[O-2].[O-2].[O-2].[O-2].[F-].[Mg+2].[Al+3].[Al+3].[Al+3].[Al+3].[Al+3].[Al+3].[Al+3].[Al+3].[Al+3].[K+].[K+].[K+].[Fe+2].O1[Si]2([O-])O[Si]1([O-])O2.O1[Si]2([O-])O[Si]1([O-])O2.O1[Si]2([O-])O[Si]1([O-])O2.O1[Si]2([O-])O[Si]1([O-])O2.O1[Si]2([O-])O[Si]1([O-])O2.O1[Si]2([O-])O[Si]1([O-])O2.O1[Si]2([O-])O[Si]1([O-])O2 VGIBGUSAECPPNB-UHFFFAOYSA-L 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011946 reduction process Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/346—Iron bacteria
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
- C02F2101/163—Nitrates
-
- 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
-
- 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
- C12R2001/22—Klebsiella
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Environmental & Geological Engineering (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Medicinal Chemistry (AREA)
- Water Supply & Treatment (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Soil Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a microorganism-mineral composite microbial agent, a preparation method and application thereof, which are based on clay minerals and nitrate reduced ferrous oxidizing microorganisms, and iron minerals generated by ferrous oxidation are used as binders to tightly combine the clay minerals and the nitrate reduced ferrous oxidizing microorganisms together, so that the living space of the microorganisms is ensured, and the activity and the ferrous oxidizing capability of the microorganisms are improved continuously by the clay minerals. The microbial-mineral composite microbial agent has a stable structure, is beneficial to the long-term stable existence and function in soil, and realizes the sustainable improvement of the carbon fixation level of the soil. Meanwhile, the method can obviously promote nitrate reduction, reduce the accumulation of nitrite as a harmful substance and nitrous oxide as a greenhouse gas, and has positive significance on the treatment of the pollution of the source and the emission reduction of the greenhouse gas. The microorganism-mineral composite microbial agent has the advantages of simple synthesis method, low cost and environmental protection, and is beneficial to industrialized mass production and practical popularization and application.
Description
Technical Field
The invention relates to the technical field of environmental microorganisms, in particular to a microorganism-mineral composite microbial inoculum, a preparation method and application thereof.
Background
The land soil is a carbon reservoir with the largest earth surface, improves the carbon fixation level of the soil, is beneficial to realizing the aim of carbon-to-carbon neutralization of the peak of carbon in China, and simultaneously has important significance for the development of economic society in China while also taking into consideration the synergistic development of environmental protection and soil fertility improvement. The soil of the south paddy field is rich in iron elements, and high-concentration ferrous iron is often accumulated under the flooding condition, so that anaerobic iron oxidizing microorganisms are favorable for driving ferrous iron to oxidize into ore to fix carbon. Specifically, trivalent iron mineral formed in the ferrous oxidation process can fix or protect soil organic carbon in various modes such as adsorption, coprecipitation and the like, prevent migration and degradation of the organic carbon, and realize soil carbon fixation. Compared with a plurality of soil carbon fixation carburetion technologies based on material chemical synthesis and addition, the method for improving the soil carbon fixation level by oxidizing microorganism ferrous iron into ore has the advantages of high efficiency, low cost, sustainability and the like, and has wide application prospect.
Nitrate-reduced ferrous oxidizing bacteria are key microorganisms for driving anaerobic ferrous oxidation of paddy soil. The prior published patent relates to the use of such strains to drive ferrous oxidation to regulate soil carbon sequestration levels. However, these purified strains have limited activity and are easily inactive when applied directly to soil. Clay minerals are environment-friendly minerals with large specific surface area and rich surface groups, and can improve the environmental tolerance of microorganisms, but it is not clear whether the metabolic activity of nitrate-reduced ferrous oxidizing bacteria can be improved. In addition, since the surfaces of the clay mineral and the microbial cells are negatively charged, the microorganisms and the clay mineral are difficult to form a stable complex, which is not beneficial to the effective catalysis of the clay mineral and the preparation of the microorganism-mineral composite material. Therefore, although the use of nitrate-reducing ferrous oxidizing bacteria to drive anaerobic ferrous oxidation to realize soil carbon fixation has great application potential, technical bottlenecks still exist in the aspects of improving microbial activity and realizing effective combination of bacteria and ores.
Disclosure of Invention
The present invention aims to solve at least one of the above technical problems in the prior art. Therefore, the invention aims to provide a microorganism-mineral composite microbial inoculum, a preparation method and application thereof, which utilize clay minerals to improve the metabolic activity of nitrate-reduced ferrous oxidation bacteria, and combine negatively charged microorganism cells with the clay minerals through positively charged iron minerals formed in the ferrous oxidation process to form the microorganism mineral composite microbial inoculum, so that the microorganism activity improvement and the clay mineral long-acting catalysis can be achieved.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
the first aspect of the invention provides a microorganism-mineral composite microbial agent.
The second aspect of the invention provides a preparation method of the microorganism-mineral composite microbial agent.
The third aspect of the invention provides application of the microorganism-mineral composite microbial agent.
According to a first aspect of the present invention, a microorganism-mineral composite microbial agent is provided, wherein the microorganism is a microorganism having nitrate reduction coupling ferrous oxidation capability, the microorganism oxidizes ferrous ions to form an iron mineral, and the microorganism and the clay mineral are bonded through the iron mineral, and the raw materials for preparing the microorganism-mineral composite microbial agent comprise ferrous ions, microorganisms and clay minerals.
In some embodiments of the invention, the microorganism having nitrate reduction coupled ferrous oxidation capability comprises a nitrate dependent ferrous oxidizing bacteria.
In some preferred embodiments of the invention, the nitrate-dependent ferrooxidases are selected from at least one of Acidovorax sp.BoFeN1, pseudomonas sp.strain 2002, klebsiella pneumoniae strain L17.
In some preferred embodiments of the invention, the clay mineral comprises 1: type 1 mineral or 2: type 1 minerals.
In some preferred embodiments of the invention, the 1: the type 1 mineral comprises any one of kaolin, halloysite, nacreous layer clay and dickite; the method comprises the following steps: the type 1 mineral includes any one of montmorillonite, expanded vermiculite, illite and chlorite.
According to a second aspect of the present invention, a method for preparing a microorganism-mineral composite microbial agent is provided, comprising the steps of: and (3) carrying out anaerobic co-culture on the sterilized clay mineral and the microorganism with nitrate reduction coupling ferrous oxidation capability in an anaerobic culture medium containing ferrous ions, thus obtaining the microbial fertilizer.
In some embodiments of the invention, the anaerobic medium contains the following ingredients per liter: 0.42g of sodium acetate, 0.42g of sodium nitrate, 0.10g of calcium chloride, 0.14g of monopotassium phosphate, 0.42g of magnesium chloride hexahydrate, 0.30g of sodium chloride, 0.30g of ammonium chloride and 9.07g of 1-4-piperazine diethyl sulfonic acid (PIPES).
In some preferred embodiments of the invention, the anaerobic medium further comprises vitamins and trace elements.
In some preferred embodiments of the invention, the microorganism having nitrate reduction coupled ferrous oxidation capability has a biomass of 4.0X10 s in the culture system 8 ～1.5×10 9 And each mL.
In some preferred embodiments of the invention, the concentration of clay mineral in the culture system is 0.5g/L to 8.0g/L.
In some preferred embodiments of the invention, the concentration of the ferrous ion in the culture system is between 0.5mM and 10mM.
In some preferred embodiments of the invention, the anaerobic co-culture is a light-protected culture at a temperature of 25-35 ℃ for a time of 20-55 h.
In some more preferred embodiments of the invention, the method of making further comprises: and after the co-culture is finished, centrifuging the culture system liquid to obtain a precipitate, namely the microorganism-mineral composite microbial inoculum.
The invention provides an application of a microorganism-mineral composite microbial inoculum in soil carbon fixation.
The beneficial effects of the invention are as follows:
the microbial-mineral composite microbial agent is obtained by taking clay minerals and nitrate-reduced ferrous oxidizing bacteria as the basis and taking iron minerals generated by ferrous oxidation as a binder, can realize efficient ferrous oxidation in a simulated paddy field environment, and meets the actual requirement of improving the carbon fixation level of soil by regulating and controlling iron conversion.
The nitrate-reduced ferrous oxidizing bacteria and the clay minerals are tightly combined together through the iron minerals, so that the living space of microorganisms is ensured, and the clay minerals are beneficial to continuously improving the activity and ferrous oxidizing capability of the microorganisms. The microbial-mineral composite microbial agent has a stable structure, is beneficial to the long-term stable existence and function in soil, and realizes the sustainable improvement of the carbon fixation level of the soil. Meanwhile, the method can obviously promote nitrate reduction, reduce the accumulation of nitrite as a harmful substance and nitrous oxide as a greenhouse gas, and has positive significance on the treatment of the pollution of the source and the emission reduction of the greenhouse gas.
The microorganism-mineral composite microbial inoculum has the advantages of simple synthesis method, low cost and environment friendliness, is mainly made of natural clay minerals, and is favorable for industrialized mass production and practical popularization and application.
Drawings
FIG. 1 is a scanning electron microscope image of the microorganism-mineral complex microbial agent prepared in example 1;
FIG. 2 shows the change in ferrous iron concentration after the addition of the microbial-mineral composite microbial agent prepared in example 1 to system 1;
FIG. 3 is a graph showing the change in ferrous iron concentration after the addition of the microbial-mineral composite microbial agent prepared in example 1 to system 2;
FIG. 4 is a graph showing the change in nitrate concentration after the addition of the microorganism-mineral complex inoculant prepared in example 1 to system 1;
FIG. 5 is a graph showing the change in nitrate concentration after the addition of the microorganism-mineral complex inoculant prepared in example 1 to system 2;
FIG. 6 is a graph showing the change in nitrite concentration after adding the microbial-mineral composite microbial agent prepared in example 1 to system 1;
FIG. 7 is a graph showing the change in nitrous oxide concentration after addition of the microorganism-mineral complex inoculant prepared in example 1 to system 1;
FIG. 8 is a graph showing the change in nitrous oxide concentration after addition of the microorganism-mineral complex inoculant prepared in example 1 to System 2;
FIG. 9 shows the change in protein concentration of the microorganism-mineral complex microbial agent prepared in example 1 in system 1.
Detailed Description
The present invention will be described in further detail with reference to specific examples. The starting materials, reagents or apparatus used in the examples and comparative examples were either commercially available from conventional sources or may be obtained by prior art methods unless specifically indicated. Unless otherwise indicated, assays or testing methods are routine in the art.
The strain used below was Acidovorax sp.BoFeN1, a typical nitrate-reducing ferrous-oxidizing strain. Kaolin was purchased from Ara Ding Shiji (Shanghai) Inc. at the microorganism culture Collection (DSMZ; germany). Vermiculite is derived from Hebei Lingshu county and is provided by Hebei Yanxi mineral processing plant. Other embodiments have similar effects to embodiment 1 and will not be described again.
Since vermiculite contains certain impurities, it is first purified as follows:
(1) Cleaning: the vermiculite sample is stirred and washed for 5 times by ultrapure water, and impurities such as suspended matters are removed.
(2) Magnetic separation: the mixture is repeatedly stirred in vermiculite by using a magnet to remove magnetic substances such as magnetite.
(3) Acid washing: and (3) drying the vermiculite subjected to magnetic separation at 50-60 ℃ and mixing with dilute hydrochloric acid with the mass fraction of 1% (the mass ratio of the vermiculite to the hydrochloric acid is 1:15-1:20), stirring at 300rpm for 2 hours, and washing with ultrapure water.
(4) And (3) preserving: and (3) drying the pickled sample at the temperature of 95-105 ℃, grinding the sample into small pieces, and sealing and drying the small pieces for later use.
Expanded vermiculite: and 3.0 g-5.0 g of the purified vermiculite sample is taken, placed in a ceramic cup, placed in a muffle furnace, heated for 4 hours at the constant temperature of 300-500 ℃, and cooled to obtain the expanded vermiculite.
Microorganism proliferation culture:
the value-added culture nutrient of the microorganism is used for two stages of primary culture and transfer activation. The primary culture means that a sample of BoFeN1 strain (50% glycerol: bacterial liquid=1:1; v/v) stored at-80℃is taken out and thawed at room temperature. After thawing, the strain samples were transferred to penicillin bottles containing 500mL of sterile anaerobic medium at pH 7.0, and 0.1mL each of vitamins and trace elements was added.
The anaerobic medium was formulated as follows (1.0L): 0.42g of sodium acetate, 0.42g of sodium nitrate, 0.10g of calcium chloride, 0.14g of monopotassium phosphate, 0.42g of magnesium chloride hexahydrate, 0.30g of sodium chloride, 0.30g of ammonium chloride and 9.07g of 1-4-piperazine diethyl sulfonic acid (PIPES). Then the penicillin bottle is cultivated for 18 to 24 hours under the constant temperature and light-shielding condition at 30 ℃.
After the preliminary culture is finished, the microorganism with higher activity is obtained through transfer activation, and the specific steps are as follows: and transferring 10mL of the bacterial liquid after preliminary culture into a new culture medium, wherein the formula and the volume of the culture medium are the same as those of the culture medium. After transfer, the culture is continued for 12 hours under the constant temperature and light-shielding condition at 30 ℃, at this time, the microorganism grows to the logarithmic phase, and the microorganism activity is higher. Under aseptic conditions, transferring the bacterial liquid after the culture is finished into an aseptic centrifuge tube, centrifuging 8000 Xg to obtain bacterial sediment, re-suspending the bacterial sediment by using an aseptic PIPES buffer solution, and repeating the steps for 3 times. The cells obtained by the last centrifugation are used for the preparation ofResuspension with sterile PIPES buffer and OD adjustment 600 0.4 + -0.02. After the concentration is regulated, the bacterial liquid is aerated by high-purity sterile nitrogen to remove oxygen, and the bacterial liquid is placed in an anaerobic box for standby.
Example 1
Weighing a certain mass of kaolin, placing the kaolin into a 50mL penicillin bottle containing the anaerobic culture medium, enabling the final concentration of the kaolin to be 4.0g/L, sterilizing the penicillin bottle in a high-temperature high-pressure sterilizing pot, and placing the penicillin bottle in an anaerobic box. The bacterial solution and the anaerobic medium were mixed (bacterial solution: anaerobic medium=1:20; v/v), and the ferrous chloride mother liquor was added at the same time to give a ferrous ion concentration of about 5mM in the reaction system. Taking out the penicillin bottle after sealing by using a rubber plug and an aluminum cover, shaking uniformly, and placing the penicillin bottle in an incubator for constant temperature (30 ℃) light-shielding cultivation. After 6 hours of cultivation, clay minerals added into the cultivation system are gradually converted into light yellow or light yellow green, which indicates that ferrous oxidation occurs. After the culture is continued for 24 to 48 hours, the reaction system turns to yellow brown, and the culture is stopped at this time. And centrifuging the solution in the culture system to obtain a yellow brown precipitate, namely the microbial mineral composite microbial agent.
Fig. 1 is a scanning electron microscope photograph of a microbial mineral composite microbial agent prepared in the embodiment, and the result shows that a layer of irregularly shaped iron mineral is covered on the surface of microbial cells, and the microbial cells and platy kaolin are tightly combined together by taking iron mineral as a binder to form a stable microbial mineral polymer.
Example 2
Weighing montmorillonite of a certain mass, placing the montmorillonite into a 50mL penicillin bottle containing the anaerobic culture medium to ensure that the final concentration of the montmorillonite is 4.0g/L, sterilizing the penicillin bottle in a high-temperature high-pressure sterilizing pot, and placing the penicillin bottle into an anaerobic box. The bacterial solution and the anaerobic medium were mixed (bacterial solution: anaerobic medium=1:20; v/v), and the ferrous chloride mother liquor was added at the same time to give a ferrous ion concentration of about 5mM in the reaction system. Taking out the penicillin bottle after sealing by using a rubber plug and an aluminum cover, shaking uniformly, and placing the penicillin bottle in an incubator for constant temperature (30 ℃) light-shielding cultivation. After 6 hours of cultivation, clay minerals added into the cultivation system are gradually converted into light yellow or light yellow green, which indicates that ferrous oxidation occurs. After the culture is continued for 24 to 48 hours, the reaction system turns to yellow brown, and the culture is stopped at this time. And centrifuging the solution in the culture system to obtain a yellow brown precipitate, namely the microbial mineral composite microbial agent.
Example 3
Weighing a certain mass of nacreous layer pottery clay, placing in a 50mL penicillin bottle containing the anaerobic culture medium to make the final concentration of the nacreous layer pottery clay be 4.0g/L, sterilizing the penicillin bottle in a high-temperature high-pressure sterilizing pot, and placing in an anaerobic box. The bacterial solution and the anaerobic medium were mixed (bacterial solution: anaerobic medium=1:20; v/v), and the ferrous chloride mother liquor was added at the same time to give a ferrous ion concentration of about 5mM in the reaction system. Taking out the penicillin bottle after sealing by using a rubber plug and an aluminum cover, shaking uniformly, and placing the penicillin bottle in an incubator for constant temperature (30 ℃) light-shielding cultivation. After 6 hours of cultivation, clay minerals added into the cultivation system are gradually converted into light yellow or light yellow green, which indicates that ferrous oxidation occurs. After the culture is continued for 24 to 48 hours, the reaction system turns to yellow brown, and the culture is stopped at this time. And centrifuging the solution in the culture system to obtain a yellow brown precipitate, namely the microbial mineral composite microbial agent.
Example 4
Weighing a certain mass of (expanded) vermiculite, placing the (expanded) vermiculite into a 50mL penicillin bottle containing the anaerobic culture medium, enabling the final concentration of the (expanded) vermiculite to be 4.0g/L, sterilizing the penicillin bottle in a high-temperature high-pressure sterilizing pot, and placing the penicillin bottle into an anaerobic box. The bacterial solution and the anaerobic medium were mixed (bacterial solution: anaerobic medium=1:20; v/v), and the ferrous chloride mother liquor was added at the same time to give a ferrous ion concentration of about 5mM in the reaction system. Taking out the penicillin bottle after sealing by using a rubber plug and an aluminum cover, shaking uniformly, and placing the penicillin bottle in an incubator for constant temperature (30 ℃) light-shielding cultivation. After 6 hours of cultivation, clay minerals added into the cultivation system are gradually converted into light yellow or light yellow green, which indicates that ferrous oxidation occurs. After the culture is continued for 24 to 48 hours, the reaction system turns to yellow brown, and the culture is stopped at this time. And centrifuging the solution in the culture system to obtain a yellow brown precipitate, namely the microbial mineral composite microbial agent.
Test examples
Ferrous oxidation kinetics experiment
A penicillin bottle containing 50mL of sterile anaerobic PIPES solution is used as a microcosmic culture environment to simulate a soil solution system 1, which contains 5.0mM ferrous chloride, 5.0mM sodium nitrate and 2.0mM sodium acetate. A penicillin bottle containing 50mL of a sterile, anaerobic PIPES solution containing 5.0mM ferrous chloride, 5.0mM sodium nitrate and 5.0mM sodium acetate was used as the microcosm culture environment simulation soil solution system 2. The microbial mineral composite microbial agent prepared in the example 1 is added into an experimental group, the microbial mineral composite microbial agent with the same concentration (the concentration of the microbial mineral composite microbial agent is expressed by the concentration of protein) is added into a control group, the microbial mineral composite microbial agent is respectively cultivated at 30 ℃ in a dark place, then samples in a penicillin bottle are uniformly shaken to obtain suspension, ferrous oxidation, nitrate reduction and secondary product generation conditions in a system are measured, and 3 parallel experiments are set for each treatment group.
Extraction and determination of ferrous iron: 0.2mL of the above suspension was pipetted into a centrifuge tube containing 0.8mL of sulfamic acid solution (pH=1.8; 40 mM), respectively. The sulfamic acid solution can maintain acidity to prevent ferrous oxidation, can also quickly eliminate nitrite in a reaction system, and prevents ferrous and nitrite from reacting under acidic conditions so as to avoid affecting ferrous concentration measurement, and is more suitable for being used as a ferrous extractant in the experiment than hydrochloric acid. Shake the centrifuge tube on shaking table (180 rpm;60 min), after finishing, centrifugate, suck 0.1mL supernatant in 1.5mL centrifuge tube, add 0.4mL sulfamic acid solution as ferrous protection solution at the same time, add 0.5mL of malachite solution as color former, develop color for 5min after the room temperature is dark, solution become stable purple, add solution drop into enzyme-labeled plate and use enzyme-labeled instrument to measure sample absorbance under 562nm absorbance condition, finally combine ferrous standard curve to calculate final concentration of ferrous in the sample, result see figure 2 and figure 3.
As is clear from FIG. 2, in the system 1, the oxidation rate of ferrous iron was low in the treatment group to which only the seed strain (-M) was added as the reaction proceeded, and the ferrous iron consumption amount after 115 hours of cultivation was only 30% of the addition amount. In the treatment group to which the microbial mineral inoculant was added, the ferrous oxidation rate was initially slow, but the ferrous oxidation amount increased significantly with the extension of the culture time, and the ferrous consumption amount after the end of 115h of culture was about 60% of the addition amount, indicating that the use of the microbial mineral complex as the inoculant addition significantly improved the ferrous oxidation ability compared to the addition of the seed alone.
As can be seen from fig. 3, the composite microbial inoculum also has a promoting effect on ferrous oxidation and nitrate reduction in system 2 (high concentration organic carbon source addition) compared to system 1 (low concentration organic carbon source addition). As the reaction proceeds, the rate of ferrous oxidation is slower in the treatment group to which only the strain (-M) was added, and the ferrous consumption after 115 hours of culture was only 27% of the addition. In the group to which the microbial mineral inoculum was added (+M), the ferrous oxidation rate was initially the same as in the group to which no inoculum was added, but the ferrous oxidation amount increased significantly with the lapse of the culture time, and the ferrous consumption amount after the completion of the culture was about 41% of the addition amount. The microbial mineral composite microbial inoculum can adapt to carbon source environments with different concentrations, and has remarkable promotion effect on ferrous oxidation process under the conditions of low concentration and high concentration of carbon source.
Extraction and determination of nitrate, nitrite and nitrous oxide: accurately sucking 0.5mL of the suspension into a centrifuge tube containing 2.5mL of ultrapure water, and rapidly removing unreacted ferrous iron by using dry air or oxygen aeration. The sample was filtered using a 0.22 μm filter to remove iron mineral precipitate and microbial cells, and then placed in a-20℃refrigerator for use. After all samples were taken, the samples were thawed to a solution state at room temperature, and about 1mL of the samples were respectively sucked into sample bottles, and uniformly tested using an ion chromatograph. Nitrous oxide was measured by a gas chromatograph after extracting the vial headspace gas using a 1mL syringe, and the results are shown in fig. 4 to 8.
As is clear from FIG. 4, in the system 1, the nitrate was gradually decreased in the treatment group to which only the seed (-M) was added as the culture time increased, and the remaining amount of nitrate after 115 hours of culture was 3.8mM. In the treatment group (+M) to which the microbial mineral complex microbial agent was added, the residual amount of nitrate after the completion of the culture was 2.3mM, indicating that the complex microbial agent had a remarkable promoting effect on nitrate reduction.
As is clear from FIG. 5, in the system 2, the nitrate was gradually decreased in the treatment group to which only the seed strain (-M) was added as the culture time increased, and the residual amount of nitrate after 115 hours of culture was 1.0mM. In the treatment group (+M) added with the microbial mineral composite microbial agent, nitrate is completely consumed after the culture is finished, which shows that the microbial mineral composite microbial agent can adapt to carbon source environments with different concentrations, and has remarkable promotion effect on the nitrate reduction process under the conditions of low-concentration and high-concentration carbon sources.
As is clear from FIGS. 6 and 7, in the system 1, the nitrite concentration accumulated during the course of the culture in the treated group to which only the seed strain (-M) was added, and the maximum accumulated amount was 0.1mM, as the culture time was increased. Nitrous oxide (greenhouse gases) also accumulates, with a maximum accumulation of up to 0.68 μm. In the group of treatments with the addition of the microbial mineral inoculum, no nitrite accumulation was observed throughout the reaction, while nitrous oxide was always at very low concentration levels.
As is clear from FIG. 8, in the system 2, the maximum accumulation amount of nitrous oxide (greenhouse gas) was 0.5. Mu.M even in the treatment group to which only the seed strain (-M) was added, as the culture time increased. In the group treated with the addition of the mineral microbial inoculum, the cumulative amount of nitrous oxide during the whole reaction was only 0.2 μm at maximum. The microbial mineral composite inoculant has obvious effect on reducing the emission of greenhouse gas nitrous oxide.
Microbial number change assay
Since it is difficult to directly measure the change in concentration of microorganisms in a reaction system by a cell counting method by aggregating minerals and microorganisms together, the number of microorganisms is evaluated by measuring the concentration of proteins in the reaction system as follows: a penicillin bottle containing 50mL of sterile anaerobic PIPES solution is used as a microcosmic culture environment to simulate a soil solution system, which contains 5.0mM ferrous chloride, 5.0mM sodium nitrate and 2.0mM sodium acetate. The microbial mineral composite microbial agent prepared in example 1 was added to the experimental group, and the control group was added with only the microbial agent of the same concentration (the concentration of the strain is expressed as the protein concentration), and was cultivated at 30℃in a dark place. Then shaking the sample in the penicillin bottle uniformly to obtain a suspension, sucking 0.2mL of the suspension into a 1mL centrifuge tube, and centrifugally cleaning the suspension for three times by using sterile ultrapure water. After the last centrifugation was completed, 0.8ml of 0.2m NaOH solution was added to the pellet sample, the sample was transferred to a tube and heated and boiled in a autoclave for at least 10min to release the protein from the cells. After cooling to room temperature, the supernatant containing the protein was obtained by centrifugation, and the protein concentration was measured by the Bradford method, and the result is shown in fig. 9.
As is clear from FIG. 9, the number of microorganisms (in terms of protein concentration) gradually increased in the treated group to which only the seed strain (-M) was added with the increase in the culture time, and only 1.25mg/L was reached after the completion of the reaction. In the treatment group to which the mineral microbial inoculum was added (+M), the initial concentration of protein was substantially the same as that of the treatment group to which only the strain was added, indicating that the initial numbers of microorganisms were at the same level in both treatment groups. After the reaction, the protein concentration in the treated group to which the microbial mineral microbial inoculum was added was 1.89mg/L. The microbial mineral composite microbial agent is more beneficial to the proliferation and survival of microorganisms.
As can be seen from the above embodiments: the microbial mineral composite microbial agent obtained by the method can obviously improve the ferrous oxidation capacity in a reaction system, and can effectively regulate and control the soil iron circulation process so as to improve the carbon fixing capacity of soil. The microorganism is easier to proliferate in the form of a complex, which is beneficial to improving the ferrous oxidation capability of the whole microorganism. Meanwhile, the composite microbial inoculum can keep the promotion effect under various reaction conditions, and is beneficial to adapting to various soil environments. Minerals produced by ferrous oxidation can continue to fix microorganisms, and further form new microbial mineral composite materials, so that the ferrous oxidation capability and carbon fixation level of soil are continuously improved. Besides improving the ferrous oxidation effect, the microbial mineral complex obtained in the invention can cooperate with the rapid oxidation of ferrous and the efficient removal of nitrate, simultaneously avoids the generation of toxic substances nitrite and greenhouse gas nitrous oxide, realizes the synchronous treatment of carbon fixation and emission reduction and non-point source pollution, and has the capability of regulating and controlling a complex soil environment system.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (10)
1. The microbial-mineral composite microbial agent is characterized in that raw materials for preparing the microbial-mineral composite microbial agent comprise ferrous ions, microorganisms and clay minerals, wherein the microorganisms are microorganisms with nitrate reduction coupling ferrous oxidation capability, the microorganisms oxidize the ferrous ions to form iron minerals, and the microorganisms and the clay minerals are bonded through the iron minerals.
2. The microbial-mineral composite inoculant of claim 1, wherein the microorganism with nitrate reduction coupled ferrous oxidation capability comprises nitrate dependent ferrous oxidation bacteria.
3. The microorganism-mineral complex inoculant of claim 2, wherein the nitrate-dependent ferrooxidases are selected from at least one of Acidovorax sp.bofen1, pseudobulbenkiania sp.strain 2002, klebsiella pneumoniae strain L17.
4. The microbial-mineral composite microbial agent of claim 1, wherein the clay mineral comprises 1: type 1 mineral or 2: type 1 minerals.
5. The method for producing a microorganism-mineral complex microbial agent according to any one of claims 1 to 4, comprising the steps of: and (3) carrying out anaerobic co-culture on the sterilized clay mineral and the microorganism with nitrate reduction coupling ferrous oxidation capability in an anaerobic culture medium containing ferrous ions, thus obtaining the microbial fertilizer.
6. The method according to claim 5, wherein the microorganism having the ability to oxidize ferrous nitrate is 4.0X10% in the culture system 8 ～1.5×10 9 And each mL.
7. The method according to claim 6, wherein the clay mineral is present in the culture system at a concentration of 0.5g/L to 8.0g/L.
8. The method according to claim 7, wherein the concentration of the ferrous ion in the culture system is 0.5 mM-10 mM.
9. The method according to claim 5, wherein the anaerobic co-culture is a light-proof culture at 25 to 35 ℃ for 20 to 55 hours.
10. The use of the microorganism-mineral composite microbial agent according to any one of claims 1 to 4 in soil carbon sequestration.
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