CN117327609A - Saline-alkali tolerant growth promoting bacterium and application thereof in improvement of heavy metal contaminated soil in saline-alkali environment - Google Patents
Saline-alkali tolerant growth promoting bacterium and application thereof in improvement of heavy metal contaminated soil in saline-alkali environment Download PDFInfo
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- CN117327609A CN117327609A CN202310879118.5A CN202310879118A CN117327609A CN 117327609 A CN117327609 A CN 117327609A CN 202310879118 A CN202310879118 A CN 202310879118A CN 117327609 A CN117327609 A CN 117327609A
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- soil
- saline
- solid preparation
- alkali
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- 239000002689 soil Substances 0.000 title claims abstract description 85
- 239000003513 alkali Substances 0.000 title claims abstract description 29
- 229910001385 heavy metal Inorganic materials 0.000 title claims abstract description 25
- 230000012010 growth Effects 0.000 title abstract description 11
- 241000894006 Bacteria Species 0.000 title abstract description 7
- 230000006872 improvement Effects 0.000 title abstract description 7
- 230000001737 promoting effect Effects 0.000 title abstract description 5
- 241000193747 Bacillus firmus Species 0.000 claims abstract description 14
- 229940005348 bacillus firmus Drugs 0.000 claims abstract description 14
- 150000003839 salts Chemical class 0.000 claims abstract description 14
- 230000000694 effects Effects 0.000 claims abstract description 9
- 244000005700 microbiome Species 0.000 claims abstract description 7
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- 238000009629 microbiological culture Methods 0.000 claims abstract description 4
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- 239000005017 polysaccharide Substances 0.000 claims abstract description 3
- 230000003248 secreting effect Effects 0.000 claims abstract 2
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- 239000002609 medium Substances 0.000 claims description 27
- 238000002360 preparation method Methods 0.000 claims description 21
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 18
- 239000002068 microbial inoculum Substances 0.000 claims description 17
- 239000001963 growth medium Substances 0.000 claims description 15
- 239000000843 powder Substances 0.000 claims description 15
- 238000012258 culturing Methods 0.000 claims description 14
- 238000000855 fermentation Methods 0.000 claims description 14
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- 239000001888 Peptone Substances 0.000 claims description 9
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- 230000000813 microbial effect Effects 0.000 claims description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 6
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- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 6
- 229910052799 carbon Inorganic materials 0.000 claims description 6
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 240000004658 Medicago sativa Species 0.000 claims description 5
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 claims description 5
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- 238000012216 screening Methods 0.000 claims description 5
- 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 4
- 229910052793 cadmium Inorganic materials 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 229910052745 lead Inorganic materials 0.000 claims description 4
- 230000005012 migration Effects 0.000 claims description 4
- 238000013508 migration Methods 0.000 claims description 4
- -1 salt ions Chemical class 0.000 claims description 4
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 3
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 3
- 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 3
- 240000008042 Zea mays Species 0.000 claims description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 3
- 229910052785 arsenic Inorganic materials 0.000 claims description 3
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- 239000004202 carbamide Substances 0.000 claims description 3
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- 239000005720 sucrose Substances 0.000 claims description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 2
- 229910019142 PO4 Inorganic materials 0.000 claims description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 claims description 2
- 229940041514 candida albicans extract Drugs 0.000 claims description 2
- 235000012343 cottonseed oil Nutrition 0.000 claims description 2
- 235000013312 flour Nutrition 0.000 claims description 2
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 claims description 2
- 239000010452 phosphate Substances 0.000 claims description 2
- 238000005067 remediation Methods 0.000 claims description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L sulfate group Chemical group S(=O)(=O)([O-])[O-] QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 claims description 2
- 239000012138 yeast extract Substances 0.000 claims description 2
- 239000010902 straw Substances 0.000 abstract description 6
- 239000002028 Biomass Substances 0.000 abstract 1
- 230000002411 adverse Effects 0.000 abstract 1
- 239000002699 waste material Substances 0.000 abstract 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 16
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- QJZYHAIUNVAGQP-UHFFFAOYSA-N 3-nitrobicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid Chemical compound C1C2C=CC1C(C(=O)O)C2(C(O)=O)[N+]([O-])=O QJZYHAIUNVAGQP-UHFFFAOYSA-N 0.000 description 5
- 240000008415 Lactuca sativa Species 0.000 description 5
- 235000003228 Lactuca sativa Nutrition 0.000 description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 5
- 239000004021 humic acid Substances 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- PAJPWUMXBYXFCZ-UHFFFAOYSA-N 1-aminocyclopropanecarboxylic acid Chemical compound OC(=O)C1(N)CC1 PAJPWUMXBYXFCZ-UHFFFAOYSA-N 0.000 description 4
- 235000011299 Brassica oleracea var botrytis Nutrition 0.000 description 4
- 240000003259 Brassica oleracea var. botrytis Species 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- YKYOUMDCQGMQQO-UHFFFAOYSA-L cadmium dichloride Chemical compound Cl[Cd]Cl YKYOUMDCQGMQQO-UHFFFAOYSA-L 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 3
- 238000005273 aeration Methods 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000003306 harvesting Methods 0.000 description 3
- 239000005416 organic matter Substances 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 238000011218 seed culture Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000003113 dilution method Methods 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- HWSZZLVAJGOAAY-UHFFFAOYSA-L lead(II) chloride Chemical compound Cl[Pb]Cl HWSZZLVAJGOAAY-UHFFFAOYSA-L 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000035800 maturation Effects 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- PTLRDCMBXHILCL-UHFFFAOYSA-M sodium arsenite Chemical compound [Na+].[O-][As]=O PTLRDCMBXHILCL-UHFFFAOYSA-M 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 108091000130 1-aminocyclopropane-1-carboxylate deaminase Proteins 0.000 description 1
- AEQDJSLRWYMAQI-UHFFFAOYSA-N 2,3,9,10-tetramethoxy-6,8,13,13a-tetrahydro-5H-isoquinolino[2,1-b]isoquinoline Chemical compound C1CN2CC(C(=C(OC)C=C3)OC)=C3CC2C2=C1C=C(OC)C(OC)=C2 AEQDJSLRWYMAQI-UHFFFAOYSA-N 0.000 description 1
- 102100032252 Antizyme inhibitor 2 Human genes 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 235000017647 Brassica oleracea var italica Nutrition 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 101000798222 Homo sapiens Antizyme inhibitor 2 Proteins 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- 108010048581 Lysine decarboxylase Proteins 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 102000052812 Ornithine decarboxylases Human genes 0.000 description 1
- 108700005126 Ornithine decarboxylases Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000010881 fly ash Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000003673 groundwater Substances 0.000 description 1
- 125000001475 halogen functional group Chemical group 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 239000010813 municipal solid waste Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 239000000176 sodium gluconate Substances 0.000 description 1
- 235000012207 sodium gluconate Nutrition 0.000 description 1
- 229940005574 sodium gluconate Drugs 0.000 description 1
- 239000004016 soil organic matter Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01B—SOIL WORKING IN AGRICULTURE OR FORESTRY; PARTS, DETAILS, OR ACCESSORIES OF AGRICULTURAL MACHINES OR IMPLEMENTS, IN GENERAL
- A01B79/00—Methods for working soil
-
- 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
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C2101/00—In situ
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2101/00—Agricultural use
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Soil Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Mechanical Engineering (AREA)
- Environmental Sciences (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Materials Engineering (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Environmental & Geological Engineering (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
A salt-tolerant growth-promoting bacterium and its application in improving heavy metal contaminated soil in salt-tolerant environment are disclosed, wherein bacillus firmus L71 is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) with a preservation date of 2023, 3 months and 23 days, and a preservation number of CGMCC No.26877. The strain can endure high-concentration salt stress, quickly degrade biomass waste such as straw and the like into organic matters, relieve adverse effects of salt stress on plants by secreting extracellular polysaccharide, promote the growth of plants in saline-alkali soil by utilizing the growth promoting characteristic of the strain, and realize double effects of soil improvement and plant growth.
Description
Technical Field
The invention relates to a technology in the field of environmental engineering, in particular to a saline-alkali resistant growth promoting bacterium and an application thereof in improving heavy metal contaminated soil in a saline-alkali environment.
Background
The saline-alkali soil area of the beach of China is about 9000 mu, and beach reclamation is an important mode for relieving land resource shortage and increasing the cultivated area. In the artificially reclaimed beach area, due to short land forming time, low land form and high groundwater level, soil body shows obvious characteristics of coastal or offshore salinized soil, namely, the characteristics of high salt content, insignificant basification, low organic matter content and the like, the effective utilization of the reclaimed beach land and soil resources is severely restricted. Meanwhile, in early reclamation, domestic sludge, urban solid waste and the like which are not decomposed are mostly adopted, and heavy metal such As Cd, as, pb and the like is put into soil, so that the risk of heavy metal pollution of the soil is caused.
Disclosure of Invention
Aiming at the defect that the existing strain cannot be applied to the improvement of the saline-alkali soil with heavy metal pollution risk, the invention provides a saline-alkali resistant growth promoting bacterium and application thereof in the improvement of the heavy metal pollution soil in a saline-alkali environment.
The invention is realized by the following technical scheme:
the invention relates to bacillus firmus (bacillus sp.) L71 which is preserved in the China general microbiological culture Collection center, with the preservation address: beijing, chaoyang area, north Chenxi way No.1, no. 3, post code: 100101. the preservation date is 2023, 3 and 23 days, and the preservation number is CGMCC No.26877.
The bacillus firmus L71 is obtained by screening and culturing soil in a severe salinized region of the beach, and 16S rDNA is shown as SEQ ID No. 1.
The invention relates to an application of bacillus firmus L71 to the soil of a tidal flat saline-alkali soil to realize soil restoration, which comprises the following steps: the bacillus firmus L71 is subjected to expansion culture to obtain a solid preparation, the solid preparation is applied to soil to be repaired, vegetable plants are planted in the soil, the microbial inoculum absorbs salt ions in the soil through secretion of extracellular polysaccharide, the salt damage effect of salt stress on the plants is relieved, the migration risk of heavy metals in the soil is reduced, and the plant growth is promoted, so that the double purposes of improving the soil and guaranteeing the safety of the vegetables are achieved.
The expansion culture is as follows: inoculating bacillus firmus L71 strain into a sterile fermentation culture medium containing a carbon source, a nitrogen source, inorganic salt and water, and culturing for 8-24 hours under the environment that the C/N ratio is more than or equal to 5, the initial pH is 6.0-7.5 and the culture temperature is 20-37 ℃.
The aseptic fermentation medium comprises the following components: 2-20 g/L of carbon source, 0.1-10 g/L of nitrogen source, 0.01-10 g/L of inorganic salt and the balance of water, and the pH value is 6.0-7.5; wherein the carbon source is glucose, sucrose, maltose, bran, glycerol, corn flour,Molasses or a combination thereof; the nitrogen source is bean cake powder, cottonseed cake powder, yeast extract, peptone, beef extract, urea, (NH) 4 ) 2 SO 4 、KNO 3 、NH 4 NO 3 Or a combination thereof; the inorganic salt is sulfate, phosphate, dihydrogen phosphate, hydrochloride or a combination thereof.
The solid preparation contains more than 2 hundred million spores/gram of bacillus firmus L71 strain and organic materials.
The organic materials include, but are not limited to, humic acid, fly ash, straw powder and the like.
The vegetable plant comprises: leaf vegetables, rhizome vegetables, black nightshade, alfalfa.
The administration includes:
1) Aiming at the soil of the tidal flat saline-alkali soil, 50-100 kg of the microbial solid preparation is applied to each mu of soil, leaf vegetables, root vegetables and alfalfa are directly planted after 1-3 days, and then the microbial solid preparation is applied at an amount of 25-40 kg/mu every 30 days;
2) Aiming at the soil which is polluted by the saline-alkali soil and at least one heavy metal of Cd, as and Pb in the beach area, 50-100 kg of microorganism solid preparation is applied to each mu of soil, the black nightshade is planted after 1 day, the black nightshade is harvested after 30 days, 25-40 kg of microorganism solid preparation is applied again, the leaf vegetables, the rhizome vegetables and the alfalfa are planted after 2-3 days, and then 15kg of microorganism solid preparation is applied every 30 days.
The mud flat saline-alkali soil refers to: the pH value of the soil is 7.5-8.8, and the total salt content is less than or equal to 10g/kg.
The heavy metal contaminated soil is as follows: cd. The total Cd content in the As and Pb contaminated soil is less than or equal to 3.0mg/kg, the total As content is less than or equal to 100mg/kg, and the total Pb content is less than or equal to 400mg/kg.
Technical effects
The Cytobacillus sp.L71 culture conditions obtained by screening are quite extensive, the operation is convenient and simple, the saline-alkali soil can be effectively restored, the organic matter content of the soil is improved, the salt content of the soil is reduced, the migration risk of heavy metals in the soil can be obviously reduced, the absorption of super-accumulated plants to the heavy metals is improved, the plant growth is promoted, and the restoration of the saline-alkali soil polluted by the heavy metals is effectively realized.
Drawings
FIG. 1 is a schematic representation of the colony morphology of Cytobacillus sp.L71 of the present invention;
FIG. 2 is a schematic diagram of a scanning electron microscope of the strain Cytobacillus sp.L71 of the invention.
Detailed Description
Example 1
Separation and purification of CytoBacillus sp.L71
Enrichment medium: yeast powder 5g/L, peptone 10g/L, naCl 100g/L, pH 7.0.
Collecting soil samples from the Chongming beach severe salinization area of Shanghai, adding 5g of soil at a sampling point into an enrichment culture medium, culturing for 2 days at 28 ℃ and 150r/min, inoculating into a new enrichment culture medium with 10% of inoculum size, continuously culturing according to the method, and repeatedly domesticating for 4 times to enrich salt-tolerant bacteria.
Placing the enrichment culture solution in 80 ℃ water bath for 10min, and sucking the enrichment culture solution after 1mL water bath into 9mL sterile physiological saline to obtain 10 -1 Diluting the diluted bacterial suspension to 10 in sequence by a 10-fold dilution method -8 . Absorbing 0.1mL of each dilution suspension to the enrichment solid culture medium, uniformly coating, inversely culturing for 3 days at 28 ℃, picking single bacterial colony, and carrying out streak separation on the solid culture medium to obtain the salt-tolerant bacteria.
The bacillus sp.L71 obtained by screening in this example was cultured on peptone agar medium in which the vegetative cells were 0.5 to 0.6X1.3 to 2.0. Mu.m, at 37℃for 2 to 3 days to form spores in the shape of oblong or cylindrical shape. The cells can grow in large quantities by culturing for 8 hours at 30 ℃ in the culture medium. The colony is white, has complete edge, is opaque and does not flash.
The culture temperature of the Cytobacillus sp.L71 is 20-37 ℃, and the optimal temperature is 32 ℃; growing in the pH range of 5.8-7.5; gram staining detection is positive; positive in contact enzyme experiment, V-P experiment and gelatin liquefaction detection, and arginine decarboxylase experiment and arginine doubleHydrolase test, phenylalanine deaminase test, ornithine decarboxylase test, lysine decarboxylase test, methyl red test, H 2 S production experiment, indole production experiment, starch hydrolysis experiment and oxidase experiment detection are negative.
The Cytobacillus sp.L71 is measured by a 16S rDNA sequence, wherein most of the 16S rDNA sequence 1484bp is shown as SEQ ID No.1, and the homology between the sequence and a gene sequence in GenBank is found to reach 98.98 percent by comparing the sequence with the gene sequence in Cytobacillusfirmus NBRC 15306, and the Cytobacillus sp.L71 is used in the embodiment.
The detection of the growth promotion characteristics of the Cytobacillus sp.L71 specifically comprises the following steps:
i. identification of Nitrogen fixation Capacity
Inoculating Cytobacillus sp.L71 into LB medium, shake culturing at 30deg.C under 200r/min for 24 hr, and sucking the above 1mL culture solution into 9mL sterile physiological saline to obtain 10 -1 Diluting the diluted bacterial suspension to 10 in sequence by a 10-fold dilution method -8 . And respectively sucking 0.1mL to Abbe's nitrogen-free culture medium of each dilution suspension, and carrying out inverted culture at 28 ℃ for 5 days, wherein bacterial colonies are generated, which shows that Cytobacillus sp.L71 has nitrogen fixation capability.
identification of iron-producing Carrier
CAS solid assay plate: weighing 2.0g/L sucrose and 3.0g/L acid casein, caCl 2 0.001 g/L,
MgSO 4 ·7H 2 O0.001 g/L, agar 20.0g/L, natural pH. 5% CAS dye was mixed in before pouring the plate.
CAS dye liquor: chrome azure 0.60g/L FeCl 3 0.016 g/L, hexadecyl trimethyl ammonium bromide HDTMA 1.46g/L, na 2 HPO 4 ·12H 2 O 2.427g/L,NaH 2 PO 4 ·2H 2 O 0.59g/L,KH 2 PO 4 0.075 g/L,NH 4 Cl 0.25g/L,NaCl 0.125g/L,pH 6.8。
Cytobacillus sp.L71 strain was picked and inoculated on CAS detection plates and incubated at 28℃for 48h, and yellow halos were found around the colonies indicating that Cytobacillus sp.L71 had the ability to produce iron carriers.
identification of 1-aminocyclopropane-1-carboxylic Acid (ACC) deaminase-producing ability
Inoculating Cytobacillus sp.L71 into 5mL of liquid nitrogen-free medium, culturing for 24h at 30 ℃ under shaking at 200r/min, inoculating 0.1mL of the culture solution into 5mL of DF medium, culturing for 24h under shaking, inoculating 0.1mL of the culture solution into 5mLADF medium, culturing for 24-48h under shaking, repeatedly transferring and culturing strains growing in ADF, and taking the ADF medium as a negative control, wherein the result shows that Cytobacillus sp.L71 can grow by taking ACC as a unique nitrogen source, namely the strain has the capability of producing ACC deaminase.
The heavy metal tolerance characteristic detection of the Cytobacillus sp.L71 specifically comprises the following steps:
LB medium: yeast powder 5g/L, peptone 10g/L and NaCl 10g/L.
1) Respectively preparing cadmium chloride, lead chloride and sodium arsenite solution, adding the cadmium chloride, lead chloride and sodium arsenite solution into LB culture medium, and respectively preparing Cd 2+ Final concentration of 0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0mg/L, pb 2+ Final concentrations of 0, 200, 400, 600, 800, 1000, 1200, 1400mg/L, and final concentrations of As (III) of 0, 0.5, 1.0, 5.0, 10.0, 20.0, 40.0, 60.0, 80.0mg/L.
2) Culturing Cytobacillus sp.L71 in LB medium at 30deg.C under shaking table of 200r/min for 12 hr, inoculating 2% (v/v) seed solution to seed solution containing Cd with different concentration 2+ 、Pb 2+ After 48h of incubation in As (III) medium, the OD of the broth was measured at 600nm in an ELISA reader. The results show that lower concentrations of Cd 2+ 、Pb 2+ As (III) promotes the growth of the strain Cytobacillus sp.L71, but the growth of the strain is significantly inhibited after reaching a certain concentration, and in general, the strain can tolerate Cd 2+ 、Pb 2+ As (III) concentrations were 3.5mg/L, 1200mg/L, 60mg/L, respectively.
The components of the medium involved in this example are specifically as follows:
the composition of the LB medium (per liter) was: yeast powder 5g/L, peptone 10g/L and NaCl 10g/L.
Ababetes (Abstract.) KuntzeNitrogen-free medium: mannitol 10g/L MgSO 4 ·7H 2 O 0.2g/L,CaSO 4 ·2H 2 O 0.1g/L,
KH 2 PO 4 0.2 g/L,CaCO 3 5g/L, 20g/L of agar and pH 7.2-7.4.
The DF medium (per liter) had the following composition: component one: h 3 BO 3 1 g/L,MnSO 4 ·4H 2 O 1.12g/L,ZnSO 4 ·7H 2 O 12.46g/L,CuSO 4 ·5H 2 O 7.82g/L,MnO 3 1 g/L. And a second component: feSO 4 ·7H 2 O10 g/L, constant volume to 10mL. 0.01% of component one, 0.01% of component two, KH 2 PO 4 4 g/L,Na 2 HPO 4 6 g/L,MgSO 4 ·7H 2 O0.2 g/L, glucose 2g/L, sodium gluconate 2g/L, citric acid 2g/L, (NH) 4 ) 2 SO 4 2 g/L,pH7.2。
The ADF enriched medium (per liter) had the following composition: DF medium was not added (NH) 4 ) 2 SO 4 Sterilizing, preparing 3.0 mmol/L1-aminocyclopropane-1-carboxylic Acid (ACC) by ultrapure water, filtering, sterilizing, and pH7.2.
Example 2
This example relates to the preparation of a solid preparation from Cytobacillus sp.L71 obtained in example 1, comprising in particular:
Cytobacillussp.L71 is cultivated for 12 hours under the conditions of a seed culture medium, 30 ℃ and a shaking table of 200r/min, seed liquid is inoculated into a 1000L fermentation tank preloaded with 600L of sterilized fermentation culture medium according to the seed quantity of 2% (v/v) for cultivation, and the cultivation conditions are as follows: the aeration rate is 1.5L/min at 32 ℃ at 100r/min, and the pH is not required to be regulated in the fermentation process. Fermenting for 10 hr until the number of spores reaches 100×10 8 cfu/mL. Adsorbing the culture solution with humic acid, wherein the culture solution comprises: the ratio of humic acid is 1:40 (L: kg), and the solid microbial inoculum containing the Cytobacillus sp.L71 is obtained after stirring and mixing. The solid preparation contains more than 2 hundred million spores per gram formed by Cytobacillus sp.L71 strain.
The components of the medium involved in this example are specifically as follows:
the seed medium (per liter) had the following composition: yeast powder 5g/L, peptone 10g/L and NaCl 10g/L.
The components content of the fermentation medium (per liter) were: molasses 20g/L, bean cake powder 10g/L, dipotassium hydrogen phosphate 1.0g/L, naCl 50g/L and pH 7.0.
Example 3
The embodiment relates to restoration of soil in tidal flat saline-alkali soil by using the Cytobacillus sp.L71 obtained in the embodiment 1, and specifically comprises the following steps:
step (1), cytobacillus sp.L71 is cultivated for 12 hours under the conditions of a seed culture medium, 30 ℃ and a shaking table of 200r/min, seed liquid is inoculated into a 500L fermentation tank preloaded with 300L of sterilized fermentation culture medium according to the seed amount of 2% (v/v) for cultivation, and the cultivation conditions are as follows: the aeration rate is 1.5L/min at 35 ℃ and 150r/min, and the pH is not required to be regulated in the fermentation process. Fermenting for 12 hr until the number of spores reaches 110×10 8 cfu/mL. Adsorbing the culture solution with straw powder, wherein the culture solution comprises: the ratio of the straw powder is 1:50 (L: kg), and the solid microbial inoculum containing Cytobacillus sp.L71 is obtained after stirring and mixing. The solid preparation contains more than 2 hundred million spores per gram formed by Cytobacillus sp.L71 strain.
And (2) uniformly throwing and broadcasting a solid microbial inoculum and straw powder containing Cytobacillussp.L71 into the soil (pH 8.3, total salt amount 6.8 g/kg) of the tidal flat saline-alkali soil, uniformly mixing the soil with shallow ploughing with the soil, normally irrigating to moisten the soil, and planting lettuce (Lactucasativa Linn. Var. Ramosa Hort.) after 1 day. Lettuce was harvested after 35d and soil and plant samples were collected for testing. And setting a blank control, namely, not applying straw powder or microbial inoculum for treatment. As can be seen from Table 1, the application of the microbial inoculum significantly reduced the total salt content of the soil (removal rate 33.8%), especially Na + 、Cl - The content of (a) is reduced by 42.6 percent and 55.6 percent respectively), and the content of organic matters in soil is improved (the improvement rate is 13.4 percent). Can also effectively promote the growth of vegetables, and the growth promotion rate of the average single plant fresh weight of lettuce reaches 32.4 percent (table 2).
Table 1 restoration effect of microbial inoculum on soil of tidal flat saline-alkali soil
TABLE 2 Effect of microbial agents on lettuce yield
The components of the medium involved in this example are specifically as follows:
the seed medium (per liter) had the following composition: glucose 5g/L, beef extract 3g/L, peptone 5g/L, naCl 10g/L, pH 7.0.
The components content of the spore-producing medium (per liter) are: bran 20g/L, ammonium sulfate 10g/L, monopotassium phosphate 0.5g/L, manganese sulfate 0.1g/L, naCl 20g/L and pH 7.0.
Example 4
The embodiment relates to restoration of heavy metal contaminated beach saline-alkali soil by using Cytobacillus sp.L71 obtained in the embodiment 1, and specifically comprises the following steps:
step (1), cytobacillus sp.L71 is cultivated for 12 hours under the conditions of a seed culture medium, 30 ℃ and a shaking table of 200r/min, seed liquid is inoculated into a 50L fermentation tank preloaded with 30L of sterilized fermentation culture medium according to the seed amount of 2% (v/v) for cultivation, and the cultivation conditions are as follows: the aeration rate is 2.0L/min at 32 ℃ and 180r/min, and the pH value is not required to be regulated in the fermentation process. Fermenting for 12 hr until the number of spores reaches 110×10 8 cfu/mL. Mixing the culture solution with humic acid, wherein the culture solution is as follows: the ratio of humic acid is 1:50 (L: kg), and the solid microbial inoculum containing Cytobacillus sp.L71 is obtained after stirring, mixing and airing. The solid preparation contains more than 2 hundred million spores per gram formed by Cytobacillus sp.L71 strain.
Uniformly throwing a solid microbial inoculum containing Cytobacillus sp.L71 on the surface of heavy metal polluted beach saline-alkali soil (pH 8.0, total salt content 5.3g/kg, cd 1.41mg/kg, pb 188mg/kg and As 54.77 mg/kg), applying 80 kg/mu, shallow plowing to uniformly mix the microbial inoculum with the soil, normally irrigating to moisten the soil, planting black nightshade after 1 day, and harvesting after row spacing 50cm and plant spacing 40cm for 30 days; then applying a microbial inoculum at a rate of 40 kg/mu, planting cauliflowers after 2 days, wherein the row spacing is 45cm, the plant spacing is 40cm, then applying the microbial inoculum at intervals of 30 days according to a rate of 15 kg/mu, and harvesting after maturation; and applying a microbial inoculum at a rate of 50 kg/mu, planting black nightshade after 1 day, and harvesting after 60 days, wherein the row spacing is 50cm and the plant spacing is 40 cm. And setting a control group, namely planting the black nightshade and the cauliflower in the same way, and applying no microbial inoculum. And (5) detecting the soil after maturation. The result shows that the application of the microbial inoculum obviously reduces the total salt content of the soil (the removal rate is 47.2%), improves the organic matter content of the soil (the improvement rate is 11.2%), and can also effectively reduce the Cd, pb and As contents of the soil. Can also effectively promote the growth of vegetables, and the average per mu yield of the cauliflower reaches 14.6 percent.
TABLE 3 remediation effect of heavy metal contaminated beach saline-alkali soil by microbial inoculum
TABLE 4 Effect of microbial agents on broccoli yield
The components of the medium involved in this example are specifically as follows:
the seed medium (per liter) had the following composition: yeast powder 5g/L, peptone 10g/L and NaCl 10g/L.
The components content of the spore-producing medium (per liter) are: 10g/L of bran, 10g/L of corn cake, 10g/L of urea, 0.5g/L of magnesium sulfate, 0.1g/L of calcium carbonate, 20g/L of NaCl and pH 7.0.
Compared with the prior art, the Cytobacillus sp.L71 obtained by screening can effectively repair the saline-alkali soil with heavy metal pollution risk, the soil salinity is reduced by improving the microbial activity of the soil, the soil organic matter content is improved, the migration risk of heavy metal in the soil can be obviously reduced, the absorption of super-accumulated plants to heavy metal is improved, and the repair of the saline-alkali soil with heavy metal pollution is effectively realized.
The foregoing embodiments may be partially modified in numerous ways by those skilled in the art without departing from the principles and spirit of the invention, the scope of which is defined in the claims and not by the foregoing embodiments, and all such implementations are within the scope of the invention.
Claims (6)
1. A bacillus firmus is characterized by comprising the following steps: cytobacillus sp.L71 is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) with a preservation date of 2023, 3 and 23 days and a preservation number of CGMCC No.26877;
the bacillus firmus L71 is obtained by screening and culturing soil in a severe salinized region of the beach, and 16SrDNA of the bacillus firmus L71 is shown as SEQ ID No. 1.
2. Use of bacillus firmus L71 according to claim 1, characterized in that it is applied to the soil of tidal flat saline-alkali soil to achieve soil remediation, in particular: the bacillus firmus L71 is subjected to expansion culture to obtain a solid preparation, the solid preparation is applied to soil to be repaired, vegetable plants are planted in the soil, and the microbial inoculum is used for absorbing salt ions in the soil through secreting extracellular polysaccharide, so that the salt damage effect of salt stress on the plants is relieved, the migration risk of heavy metals in the soil is reduced, the plant growth is promoted, and the double purposes of improving the soil and guaranteeing the safety of the vegetables are achieved;
the mud flat saline-alkali soil refers to: the pH value of the soil is 7.5-8.8, and the total salt content is less than or equal to 10g/kg;
the heavy metal contaminated soil is as follows: cd. The total Cd content in the As and Pb contaminated soil is less than or equal to 3.0mg/kg, the total As content is less than or equal to 100mg/kg, and the total Pb content is less than or equal to 400mg/kg.
3. The use according to claim 2, wherein said expansion culture means: inoculating bacillus firmus L71 strain into a sterile fermentation culture medium containing a carbon source, a nitrogen source, inorganic salt and water, and culturing for 8-24 hours under the environment that the C/N ratio is more than or equal to 5, the initial pH is 6.0-7.5 and the culture temperature is 20-37 ℃.
4. The use according to claim 2, wherein the sterile fermentation medium comprises the following components: 2-20 g/L of carbon source, 0.1-10 g/L of nitrogen source, 0.01-10 g/L of inorganic salt and the balance of water, and the pH value is 6.0-7.5;
the carbon source is glucose, sucrose, maltose, bran, glycerol, corn flour, molasses or a combination thereof;
the nitrogen source is bean cake powder, cottonseed cake powder, yeast extract, peptone, beef extract, urea, (NH) 4 ) 2 SO 4 、KNO 3 、NH 4 NO 3 Or a combination thereof;
the inorganic salt is sulfate, phosphate, dihydrogen phosphate, hydrochloride or a combination thereof.
5. The use according to claim 2, wherein said solid preparation comprises more than 2 hundred million spores/g of bacillus firmus strain L71 and organic material.
6. The use according to claim 2, wherein said applying comprises:
1) Aiming at the soil of the tidal flat saline-alkali soil, 50-100 kg of the microbial solid preparation is applied to each mu of soil, leaf vegetables, root vegetables and alfalfa are directly planted after 1-3 days, and then the microbial solid preparation is applied at an amount of 25-40 kg/mu every 30 days;
2) Aiming at the soil which is polluted by the saline-alkali soil and at least one heavy metal of Cd, as and Pb in the beach area, 50-100 kg of microorganism solid preparation is applied to each mu of soil, the black nightshade is planted after 1 day, the black nightshade is harvested after 30 days, 25-40 kg of microorganism solid preparation is applied again, the leaf vegetables, the rhizome vegetables and the alfalfa are planted after 2-3 days, and then 15kg of microorganism solid preparation is applied every 30 days.
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