CN110982738B - Composite microbial agent for treating soil and preparation method thereof - Google Patents
Composite microbial agent for treating soil and preparation method thereof Download PDFInfo
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 41
- 239000002689 soil Substances 0.000 title claims abstract description 28
- 239000002131 composite material Substances 0.000 title claims abstract description 19
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 239000003795 chemical substances by application Substances 0.000 title abstract description 22
- 239000007788 liquid Substances 0.000 claims abstract description 151
- 241000894006 Bacteria Species 0.000 claims abstract description 43
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 34
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 23
- 241000194107 Bacillus megaterium Species 0.000 claims abstract description 23
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 23
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 23
- 230000000243 photosynthetic effect Effects 0.000 claims abstract description 23
- 241000187395 Streptomyces microflavus Species 0.000 claims abstract description 22
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 14
- 241000605159 Nitrobacter Species 0.000 claims abstract description 12
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 29
- 239000008223 sterile water Substances 0.000 claims description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 28
- 235000013312 flour Nutrition 0.000 claims description 24
- 230000001580 bacterial effect Effects 0.000 claims description 21
- 239000001963 growth medium Substances 0.000 claims description 21
- 239000002054 inoculum Substances 0.000 claims description 18
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 16
- 240000008042 Zea mays Species 0.000 claims description 16
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 16
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 16
- 235000005822 corn Nutrition 0.000 claims description 16
- 239000011573 trace mineral Substances 0.000 claims description 16
- 235000013619 trace mineral Nutrition 0.000 claims description 16
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 15
- 229940041514 candida albicans extract Drugs 0.000 claims description 12
- 238000009630 liquid culture Methods 0.000 claims description 12
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 12
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 12
- 239000000843 powder Substances 0.000 claims description 12
- 239000012138 yeast extract Substances 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 11
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- 229910052796 boron Inorganic materials 0.000 claims description 10
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 10
- 229910052725 zinc Inorganic materials 0.000 claims description 10
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 9
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 9
- 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 8
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 8
- 244000068988 Glycine max Species 0.000 claims description 8
- 235000010469 Glycine max Nutrition 0.000 claims description 8
- 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 8
- 239000001888 Peptone Substances 0.000 claims description 8
- 108010080698 Peptones Proteins 0.000 claims description 8
- 229940099596 manganese sulfate Drugs 0.000 claims description 8
- 239000011702 manganese sulphate Substances 0.000 claims description 8
- 235000007079 manganese sulphate Nutrition 0.000 claims description 8
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 8
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 8
- 235000019319 peptone Nutrition 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- 230000003321 amplification Effects 0.000 claims description 7
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- 238000003199 nucleic acid amplification method Methods 0.000 claims description 7
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- 240000003768 Solanum lycopersicum Species 0.000 claims description 5
- 229920002472 Starch Polymers 0.000 claims description 5
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 5
- 229910052802 copper Inorganic materials 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 5
- 238000000265 homogenisation Methods 0.000 claims description 5
- 238000011081 inoculation Methods 0.000 claims description 5
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 5
- 229910052748 manganese Inorganic materials 0.000 claims description 5
- 239000011572 manganese Substances 0.000 claims description 5
- 239000008107 starch Substances 0.000 claims description 5
- 235000019698 starch Nutrition 0.000 claims description 5
- 238000003756 stirring 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
- 229910002651 NO3 Inorganic materials 0.000 claims description 4
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 claims description 4
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 4
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 4
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims description 4
- 235000019764 Soybean Meal Nutrition 0.000 claims description 4
- 235000019270 ammonium chloride Nutrition 0.000 claims description 4
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 4
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 4
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 4
- 239000011790 ferrous sulphate Substances 0.000 claims description 4
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 4
- 239000001632 sodium acetate Substances 0.000 claims description 4
- 235000017281 sodium acetate Nutrition 0.000 claims description 4
- JXKPEJDQGNYQSM-UHFFFAOYSA-M sodium propionate Chemical compound [Na+].CCC([O-])=O JXKPEJDQGNYQSM-UHFFFAOYSA-M 0.000 claims description 4
- 239000004324 sodium propionate Substances 0.000 claims description 4
- 229960003212 sodium propionate Drugs 0.000 claims description 4
- 235000010334 sodium propionate Nutrition 0.000 claims description 4
- 239000004455 soybean meal Substances 0.000 claims description 4
- 230000006641 stabilisation Effects 0.000 claims description 4
- 238000011105 stabilization Methods 0.000 claims description 4
- 235000021012 strawberries Nutrition 0.000 claims description 4
- 239000012137 tryptone Substances 0.000 claims description 4
- 238000000034 method Methods 0.000 claims 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims 1
- 239000007836 KH2PO4 Substances 0.000 claims 1
- 239000001110 calcium chloride Substances 0.000 claims 1
- 229910001628 calcium chloride Inorganic materials 0.000 claims 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims 1
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Inorganic materials [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims 1
- 230000002035 prolonged effect Effects 0.000 abstract description 4
- 239000003337 fertilizer Substances 0.000 description 13
- 239000000126 substance Substances 0.000 description 7
- 230000000694 effects Effects 0.000 description 5
- 239000000575 pesticide Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 3
- 241001148470 aerobic bacillus Species 0.000 description 3
- 238000013329 compounding Methods 0.000 description 3
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- 238000003860 storage Methods 0.000 description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- 230000003042 antagnostic effect Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
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- 241000282414 Homo sapiens Species 0.000 description 1
- 239000006002 Pepper Substances 0.000 description 1
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- 235000016761 Piper aduncum Nutrition 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- 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
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- 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
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Abstract
The invention relates to a composite microbial liquid microbial inoculum for treating soil and a preparation method thereof, belonging to the technical field of agricultural microbes, ecological planting and microbial agents. The composite microbial liquid microbial inoculum comprises the following microbial inoculum in parts by mass: liquid bacillus megaterium, liquid bacillus subtilis, liquid streptomyces microflavus, liquid bacillus licheniformis, liquid waxy bacillus, liquid nitrobacter and liquid photosynthetic bacteria. The composite microbial agent provided by the invention has a large number of effective viable bacteria and comprehensive functions; the various bacteria are coordinated and symbiotic with each other, and the shelf life of the bacteria is prolonged.
Description
Technical Field
The invention relates to a composite microbial liquid microbial inoculum for treating soil and a preparation method thereof, belonging to the technical field of agricultural microbes, ecological planting and microbial agents.
Background
Due to the long-term excessive application of the fertilizer, the soil in China has been largely hardened, acidified and unbalanced in nutrients, and beneficial microorganisms in the soil are sharply reduced. Part of elements such as nitrogen, phosphorus and potassium in the soil are lost, and part of elements are consolidated in the soil and cannot be absorbed and utilized by crops. Meanwhile, the increase of pesticide residues seriously threatens the health and safety of human beings.
In recent years, the national ministry of agriculture has proposed a zero-growth action scheme of fertilizer and pesticide and a soil pollution control scheme. To achieve zero growth of chemical fertilizers and pesticides, and to ensure both production and income increase of agricultural products and effective supply on the supply side, products capable of effectively replacing chemical fertilizers and pesticides are required. Therefore, microbial fertilizers capable of replacing chemical fertilizers and reducing pesticides are urgently needed for developing ecological agriculture at present, microbial preparations are used for improving soil structures, improving crop quality and increasing yield and income, and the microbial fertilizers become a necessary trend of modern agriculture and are necessary choices for developing green sustainable efficient ecological agriculture.
In recent years, various microbial fertilizers are abundant like bamboo shoots in spring after rain, but products with obvious fertilizer efficiency reaching or exceeding the fertilizer are rare, and the main reasons are as follows: firstly, the microbial strain is single, and various problems of crop growth cannot be solved; secondly, the storage life of the microbial inoculum is short, the fertilizer effect is not obvious, and the problems of putrefaction, death of effective bacteria and short storage life of the liquid microbial inoculum are difficult to solve in both single microbial inoculum and composite microbial inoculum.
Disclosure of Invention
The invention aims to provide a compound microbial liquid inoculant for treating soil and a preparation method thereof, and the microbial liquid inoculant and the preparation method thereof solve the problems of single microbial inoculant strain, short storage period and unobvious fertilizer effect.
In order to achieve the purpose, the invention provides a composite microbial liquid inoculant for treating soil, which comprises the following inocula in parts by mass: liquid bacillus megaterium, liquid bacillus subtilis, liquid streptomyces microflavus, liquid bacillus licheniformis, liquid waxy bacillus, liquid nitrobacter and liquid photosynthetic bacteria.
Further, the mixing proportion of the liquid bacillus megaterium, the liquid bacillus subtilis, the liquid streptomyces microflavus, the liquid bacillus licheniformis, the liquid waxy bacillus, the liquid nitrobacter and the liquid photosynthetic bacteria is as follows: 1-3: 1-5: 1: 1-3.
Furthermore, the content of the good-culture bacteria in each liquid microbial inoculum is more than or equal to 2 hundred million/ml.
Further, the microbial liquid inoculant also comprises trace elements.
Furthermore, the trace elements are selected from any one or more of B, Zn, Mn, Cu, Co and Fe.
The invention also provides a preparation method of the composite microbial liquid inoculant for treating soil, which comprises the following steps:
respectively culturing 7 bacteria of bacillus megatherium, bacillus subtilis, streptomyces microflavus, bacillus licheniformis, bacillus cereus, nitrobacteria and photosynthetic bacteria to prepare 7 liquid microbial agents;
mixing 7 liquid microbial inoculum, adding trace elements selected from any one or more of B, Zn, Mn, Cu, Co and Fe; stirring uniformly, standing, and obtaining the microbial liquid inoculum after homogenization is stable.
Further, the mixing proportion of the liquid bacillus megaterium, the liquid bacillus subtilis, the liquid streptomyces microflavus, the liquid bacillus licheniformis, the liquid waxy bacillus, the liquid nitrobacter and the liquid photosynthetic bacteria is as follows: 1-3: 1-5: 1: 1-3.
Further, the first-stage amplification culture of the strain is carried out at 30-37 ℃ and 100-140 r/min and under the condition of pH6.0-7.5.
Further, performing secondary amplification culture of strains at 30-37 ℃ at 100-140 r/min to obtain 7 secondary aerobic bacterial liquids.
Furthermore, the inoculation amount of each primary aerobic bacterial liquid is 0.7-1.3% of the mass of the liquid aerobic bacterial culture medium.
Advantageous effects
The photosynthetic flora belongs to independent nutrient microorganisms, can convert absorbed substances into bioactive substances such as saccharides, amino acids, vitamins and the like, and the synthesized substances can provide available substance nutrition for bacillus and promote the growth of the bacillus. Meanwhile, the independent culture of each flora ensures the normal propagation and development of the flora, so that the flora can reach the optimal growth node and then is compounded. After the compounding is finished, the flora is influenced by certain condition difference, the growth and the propagation of the single flora are slowed down, the non-antagonistic symbiotic environmental conditions are formed, and the preservation time under the normal temperature condition is greatly prolonged. Meanwhile, the compounding of the aerobic bacteria and the anaerobic bacteria can ensure that at least one kind of bacteria can play the normal proliferation and secretion functions under certain extreme conditions, thereby ensuring the normal effect.
The composite microbial agent provided by the invention has a large number of effective viable bacteria and comprehensive functions; the various bacteria are coordinated and symbiotic with each other, and the shelf life of the bacteria is prolonged.
The invention has more floras, the content of a single floras is not lower than 2 hundred million/ml, each floras can play different functions and coordinate with each other
The growth conditions of each flora are different, and the floras have excellent symbiosis under the condition that the floras do not mutually antagonize each other (certain antagonism exists among certain floras, and the problem can be solved by adjusting the proportion among strains). However, because the growth conditions are not optimal, the growth of each flora is inhibited to a certain extent, so that the development speed is slowed down, and the shelf life of the flora is prolonged from half a year to one year.
Drawings
FIGS. 1 and 2 are comparative diagrams of strawberry use, in which the microbial inoculum according to the present invention is used in the left and not used in the right.
Detailed Description
The following detailed description of embodiments of the invention refers to the accompanying drawings.
A composite microbial liquid inoculant for treating soil comprises the following inocula in parts by mass: 1-3 parts of a liquid bacillus megaterium agent; 1-5 parts of liquid bacillus subtilis preparation by mass; 1 part by mass of a liquid streptomyces microflavus microbial inoculum; 1-3 parts of a liquid bacillus licheniformis agent; 1-3 parts by mass of a liquid waxy bacillus agent; 1-3 parts by mass of a liquid nitrobacter agent; 1-3 parts of liquid photosynthetic bacteria agent.
The mixing proportion of the liquid bacillus megaterium, the liquid bacillus subtilis, the liquid streptomyces microflavus, the liquid bacillus licheniformis, the liquid waxy bacillus, the liquid nitrobacter and the liquid photosynthetic bacteria is as follows: 1-3: 1-5: 1: 1-3.
Wherein, the content of the good-culture bacteria in each liquid microbial inoculum is more than or equal to 2 hundred million/ml.
The microbial liquid microbial inoculum also comprises trace elements, wherein the trace elements are selected from any one or more of B, Zn, Mn, Cu, Co and Fe.
A preparation method of a composite microbial liquid microbial inoculum for treating soil comprises the following steps:
1. respectively culturing 7 kinds of bacillus megaterium, bacillus subtilis, streptomyces microflavus, bacillus licheniformis, bacillus cereus, nitrobacteria and photosynthetic bacteria to prepare 7 liquid microbial agents,
(1) respectively preparing liquid culture media of bacillus megatherium, bacillus subtilis, streptomyces microflavus, bacillus licheniformis, bacillus cereus, nitrobacteria and photosynthetic bacteria, respectively inoculating corresponding strains into 7 liquid aerobic bacteria culture media, and performing primary amplification culture on the strains under the conditions of 30-37 ℃, 100-140 r/min and pH 6.0-7.5 to obtain 7 primary aerobic bacteria liquids;
(2) then respectively taking 7 kinds of primary bacterial liquid, inoculating corresponding strains into 7 kinds of liquid culture media, and carrying out secondary expansion culture on the strains at the temperature of 30-37 ℃ and the speed of 100-140 r/min to obtain 7 kinds of secondary aerobic bacterial liquid, wherein the inoculation amount of each type of primary aerobic bacterial liquid is 0.7-1.3% of the mass of the liquid aerobic bacterial culture media;
2. mixing 7 liquid microbial agents according to the proportion of 1-3: 1-5: 1-3, and adding trace elements selected from any one or more of B, Zn, Mn, Cu, Co and Fe. Stirring uniformly, standing, and obtaining the microbial liquid inoculum after homogenization and stabilization.
The components of the culture medium:
b, bacillus megaterium: 20g of corn flour, 10g of soybean flour, K2HPO41.5g, MgSO41.5g, CaCO31.5g and 1000 mL of sterile water. The pH was 7.5.
B, bacillus subtilis: 40g of soybean meal, 20g of corn flour, 15g of glucose, 3g of dipotassium phosphate, 1.5g of monopotassium phosphate, 0.5g of magnesium sulfate, 0.35g of ammonium sulfate, 0.2g of yeast extract powder, 0.2g of manganese sulfate, 0.1g of ferrous sulfate, 0.1g of calcium carbonate and 1L of sterile water.
Streptomyces microflavus: KNO 31 g, 20g of soluble starch, 40.5g of K2HPO40, 40.5g of MgSO40, 0.5g of NaCl, 40.01g of FeSO40 and 1L of sterile water.
B, bacillus licheniformis: 5g of corn flour, 10g of peptone, 3g of bean cake powder, 5g of yeast extract, 0.1g of manganese sulfate, 1g of monopotassium phosphate, 0.2g of magnesium sulfate and 1L of sterile water.
Bacillus cereus: 2.5g of maltose, 5g of corn flour, 5g of soybean flour, 5g of tryptone, 20.5g of CaClO, 40.5g of MnSOO, 41g of K2HPO and 1L of sterile water.
Nitrate bacteria: 20g of maltose, 20g of yeast powder, KH2PO41.5g, MgSO41g and 1L of sterile water.
Photosynthetic bacteria: 1g of ammonium chloride, 1g of sodium bicarbonate, 1g of sodium acetate, 1g of sodium chloride, 0.2g of dipotassium hydrogen phosphate, 0.2g of sodium propionate, 0.2g of magnesium sulfate, 0.2g of peptone, 0.1g of yeast extract and 1L of sterile water.
Example 1
The liquid microbial inoculum consists of the following microbial inoculum in parts by mass:
3 parts of a liquid bacillus megaterium agent; 5 parts of liquid bacillus subtilis preparation; 1 part by mass of a liquid streptomyces microflavus microbial inoculum; 3 parts of liquid bacillus licheniformis agent; 3 parts of liquid waxy bacillus agent; 3 parts of liquid nitrobacter; 3 parts of liquid photosynthetic bacteria agent. The mixing proportion of the liquid bacillus megaterium, the liquid bacillus subtilis, the liquid streptomyces microflavus, the liquid bacillus licheniformis, the liquid waxy bacillus, the liquid nitrobacter and the liquid photosynthetic bacteria is as follows: 3:5:1:3:3:3:3.
Wherein, the content of the good-culture bacteria in each liquid microbial inoculum is more than or equal to 2 hundred million/ml.
The microbial liquid inoculum also comprises trace elements, and the trace elements are selected from 4 or any 3 of B, Zn, Co and Fe.
The preparation method comprises the following steps:
1. respectively culturing 7 kinds of bacteria to obtain 7 liquid microbial inoculum,
(1) respectively preparing liquid culture media of bacillus megatherium, bacillus subtilis, streptomyces microflavus, bacillus licheniformis, bacillus cereus, nitrobacteria and photosynthetic bacteria, respectively inoculating corresponding strains into 7 liquid culture media, and performing primary amplification culture on the strains under the conditions of 33-37 ℃, 120r/min and pH 6.5-7.5 to obtain 7 primary bacterial liquids;
(2) then respectively taking 7 kinds of primary bacterial liquid, inoculating corresponding strains into 7 kinds of liquid culture media, and carrying out secondary expansion culture on the strains under the conditions of 33-37 ℃, 120r/min and pH 6.5-7.5 to obtain 7 kinds of secondary bacterial liquid, wherein the inoculation amount of each kind of primary bacterial liquid is 1.0 percent of the mass of the liquid aerobic bacterial culture medium;
2. mixing the 7 liquid microbial inoculum according to the formula proportion, and adding trace elements selected from 4 or any 3 of B, Zn, Co and Fe. Stirring uniformly, standing, and obtaining the microbial liquid inoculum after homogenization and stabilization.
The components of the culture medium:
b, bacillus megaterium: 20g of corn flour, 10g of soybean flour, K2HPO41.5g, MgSO41.5g, CaCO31.5g and 1000 mL of sterile water. The pH was 7.5.
B, bacillus subtilis: 40g of soybean meal, 20g of corn flour, 15g of glucose, 3g of dipotassium phosphate, 1.5g of monopotassium phosphate, 0.5g of magnesium sulfate, 0.35g of ammonium sulfate, 0.2g of yeast extract powder, 0.2g of manganese sulfate, 0.1g of ferrous sulfate, 0.1g of calcium carbonate and 1L of sterile water.
Streptomyces microflavus: KNO 31 g, 20g of soluble starch, 40.5g of K2HPO40, 40.5g of MgSO40, 0.5g of NaCl, 40.01g of FeSO40 and 1L of sterile water.
B, bacillus licheniformis: 5g of corn flour, 10g of peptone, 3g of bean cake powder, 5g of yeast extract, 0.1g of manganese sulfate, 1g of monopotassium phosphate, 0.2g of magnesium sulfate and 1L of sterile water.
Bacillus cereus: 2.5g of maltose, 5g of corn flour, 5g of soybean flour, 5g of tryptone, 20.5g of CaClO, 40.5g of MnSOO, 41g of K2HPO and 1L of sterile water.
Nitrate bacteria: 20g of maltose, 20g of yeast powder, KH2PO41.5g, MgSO41g and 1L of sterile water.
Photosynthetic bacteria: 1g of ammonium chloride, 1g of sodium bicarbonate, 1g of sodium acetate, 1g of sodium chloride, 0.2g of dipotassium hydrogen phosphate, 0.2g of sodium propionate, 0.2g of magnesium sulfate, 0.2g of peptone, 0.1g of yeast extract and 1L of sterile water.
Example 2
The liquid microbial inoculum consists of the following microbial inoculum in parts by mass:
2 parts of liquid bacillus megaterium agent; 3 parts of liquid bacillus subtilis preparation; 1 part by mass of a liquid streptomyces microflavus microbial inoculum; 2 parts of liquid bacillus licheniformis agent; 2 parts of liquid waxy bacillus agent; 2 parts of liquid nitrobacter; 1 part by mass of liquid photosynthetic bacteria agent. The mixing proportion of the liquid bacillus megaterium, the liquid bacillus subtilis, the liquid streptomyces microflavus, the liquid bacillus licheniformis, the liquid waxy bacillus, the liquid nitrobacter and the liquid photosynthetic bacteria is as follows: 2:3:1:3:3:3:1.
Wherein, the content of the good-culture bacteria in each liquid microbial inoculum is more than or equal to 2 hundred million/ml.
The microbial liquid inoculum also comprises trace elements, and the trace elements are selected from 4 or any 3 of B, Zn, Co and Fe.
The preparation method comprises the following steps:
1. respectively culturing 7 kinds of bacteria to obtain 7 liquid microbial inoculum,
(1) respectively preparing liquid culture media of bacillus megatherium, bacillus subtilis, streptomyces microflavus, bacillus licheniformis, bacillus cereus, nitrobacteria and photosynthetic bacteria, respectively inoculating corresponding strains into 7 liquid culture media, and performing primary amplification culture on the strains under the conditions of 33-37 ℃, 120r/min and pH 6.5-7.5 to obtain 7 primary bacterial liquids;
(2) then respectively taking 7 kinds of primary bacterial liquid, inoculating corresponding strains into 7 kinds of liquid culture media, and carrying out secondary expansion culture on the strains under the conditions of 33-37 ℃, 120r/min and pH 6.5-7.5 to obtain 7 kinds of secondary bacterial liquid, wherein the inoculation amount of each kind of primary bacterial liquid is 1.0 percent of the mass of the liquid aerobic bacterial culture medium;
2. mixing the 7 liquid microbial inoculum according to the formula proportion, and adding trace elements selected from 4 or any 3 of B, Zn, Co and Fe. Stirring uniformly, standing, and obtaining the microbial liquid inoculum after homogenization and stabilization.
The components of the culture medium:
b, bacillus megaterium: 20g of corn flour, 10g of soybean flour, K2HPO41.5g, MgSO41.5g, CaCO31.5g and 1000 mL of sterile water. The pH was 7.5.
B, bacillus subtilis: 40g of soybean meal, 20g of corn flour, 15g of glucose, 3g of dipotassium phosphate, 1.5g of monopotassium phosphate, 0.5g of magnesium sulfate, 0.35g of ammonium sulfate, 0.2g of yeast extract powder, 0.2g of manganese sulfate, 0.1g of ferrous sulfate, 0.1g of calcium carbonate and 1L of sterile water.
Streptomyces microflavus: KNO 31 g, 20g of soluble starch, 40.5g of K2HPO40, 40.5g of MgSO40, 0.5g of NaCl, 40.01g of FeSO40 and 1L of sterile water.
B, bacillus licheniformis: 5g of corn flour, 10g of peptone, 3g of bean cake powder, 5g of yeast extract, 0.1g of manganese sulfate, 1g of monopotassium phosphate, 0.2g of magnesium sulfate and 1L of sterile water.
Bacillus cereus: 2.5g of maltose, 5g of corn flour, 5g of soybean flour, 5g of tryptone, 20.5g of CaClO, 40.5g of MnSOO, 41g of K2HPO and 1L of sterile water.
Nitrate bacteria: 20g of maltose, 20g of yeast powder, KH2PO41.5g, MgSO41g and 1L of sterile water.
Photosynthetic bacteria: 1g of ammonium chloride, 1g of sodium bicarbonate, 1g of sodium acetate, 1g of sodium chloride, 0.2g of dipotassium hydrogen phosphate, 0.2g of sodium propionate, 0.2g of magnesium sulfate, 0.2g of peptone, 0.1g of yeast extract and 1L of sterile water.
Example 3
Performing field test in 2019 spring, selecting 9 fields with the same terrain of 10m by 10m and similar soil conditions, dividing the fields into 3 groups, repeating the steps for 3 times, planting tomato seedlings, applying fertilizers to the two groups normally, adding no microbial inoculum to the first group, and applying the composite microbial liquid microbial inoculum to the second group; after harvest, the soil and tomato indices were measured separately and the data are as follows:
TABLE 1 soil indices
TABLE 2 tomato indices
As can be seen from the data in the table above, after the composite microbial liquid inoculant disclosed by the invention is applied, the content of organic matters in soil is increased, the content of nitrogen, phosphorus and potassium is increased, the pH value is closer to neutral, and the soil properties are greatly improved. The tomato plant height and stem thickness are increased, the SPAD, soluble sugar and soluble solid are increased, the quality is obviously improved, and obvious economic benefit can be generated.
Example 4
The strawberries planted in the greenhouse are divided into two groups, one group is applied with the composite microbial liquid inoculant, the other group is not applied, and the comparison of the figure shown in figure 1 and figure 2 shows that the soil property is greatly improved after the strawberries are used, and the strawberries grow well.
Example 5
In the invention, different floras are compounded to have multiple functions, for example, bacillus subtilis secretes antipathogenic bacteria such as polymyxin and the like, and soil-borne diseases caused by harmful floras in soil are inhibited; the bacillus megaterium can decompose organic phosphorus (pesticide ingredients) in soil; the bacillus licheniformis can inhibit the propagation of pathogenic bacteria in the soil and the invasion to plant roots, promote the decomposition of organic matters in the soil, release nutrients and the like, and plays a role in multiple functions while avoiding antagonistic action among different flora through reasonable proportioning and compounding. The results of comparing the formulated species with the control and the single species are shown in Table 1.
TABLE 1 comparison of the application effects of pepper planting
Claims (4)
1. A preparation method of a composite microbial liquid microbial inoculum for treating soil is characterized in that the composite microbial liquid microbial inoculum comprises the following steps: liquid bacillus megaterium, liquid bacillus subtilis, liquid streptomyces microflavus, liquid bacillus licheniformis, liquid waxy bacillus, liquid nitrobacter and liquid photosynthetic bacteria; the mixing proportion of the liquid bacillus megaterium, the liquid bacillus subtilis, the liquid streptomyces microflavus, the liquid bacillus licheniformis, the liquid waxy bacillus, the liquid nitrobacter and the liquid photosynthetic bacteria is as follows: 3:5:1:3:3:3: 3; the microbial liquid inoculum also comprises trace elements; the trace elements are selected from any one or more of B, Zn, Mn, Cu, Co and Fe; the content of the well-cultivated bacteria in each liquid microbial inoculum is more than or equal to 2 hundred million/ml;
the method comprises the following steps:
respectively preparing liquid culture media of bacillus megatherium, bacillus subtilis, streptomyces microflavus, bacillus licheniformis, bacillus cereus, nitrobacteria and photosynthetic bacteria, respectively inoculating corresponding strains into 7 liquid culture media, and performing primary amplification culture on the strains under the conditions of 33-37 ℃, 120r/min and pH 6.5-7.5 to obtain 7 primary bacterial liquids;
then respectively taking 7 kinds of primary bacterial liquid, inoculating corresponding strains into 7 liquid culture media, and carrying out secondary amplification culture on the strains under the conditions of 33-37 ℃, 120r/min and pH 6.5-7.5 to obtain 7 kinds of secondary bacterial liquids, wherein the inoculation amount of each type of primary bacterial liquid is 1.0 percent of the mass of the liquid culture media;
mixing the 7 liquid microbial inoculum according to the formula proportion, adding trace elements, wherein the trace elements are selected from 4 or any 3 of B, Zn, Co and Fe; stirring uniformly, standing, and obtaining the microbial liquid inoculum after homogenization and stabilization;
the components of the culture medium are as follows:
b, bacillus megaterium: corn flour 20g, soybean flour 10g, K2HPO4 1.5 g，MgSO4 1.5 g，CaCO31.5g, 1000 mL of sterile water; pH 7.5;
b, bacillus subtilis: 40g of soybean meal, 20g of corn flour, 15g of glucose, 3g of dipotassium phosphate, 1.5g of monopotassium phosphate, 0.5g of magnesium sulfate, 0.35g of ammonium sulfate, 0.2g of yeast extract powder, 0.2g of manganese sulfate, 0.1g of ferrous sulfate, 0.1g of calcium carbonate and 1L of sterile water;
streptomyces microflavus: KNO31g of soluble starch, 20g of soluble starch, K2HPO4 0.5g，MgSO4 0.5g，NaCl 0.5g，FeSO40.01g, 1L of sterile water;
b, bacillus licheniformis: 5g of corn flour, 10g of peptone, 3g of bean cake powder, 5g of yeast extract, 0.1g of manganese sulfate, 1g of monopotassium phosphate, 0.2g of magnesium sulfate and 1L of sterile water;
bacillus cereus: 2.5g of maltose, 5g of corn flour, 5g of soybean flour, 5g of tryptone and CaCl2 0.5g，MnSO40.5g，K2HPO41g, 1L of sterile water;
nitrate bacteria: maltose 20g, yeast powder 20g, KH2PO4 1.5g，MgSO41g, 1L of sterile water;
photosynthetic bacteria: 1g of ammonium chloride, 1g of sodium bicarbonate, 1g of sodium acetate, 1g of sodium chloride, 0.2g of dipotassium hydrogen phosphate, 0.2g of sodium propionate, 0.2g of magnesium sulfate, 0.2g of peptone, 0.1g of yeast extract and 1L of sterile water.
2. A composite microbial liquid inoculum for treating soil, which is prepared by the method of claim 1.
3. A method for culturing young tomato plants, characterized by applying the liquid microbial inoculum for soil improvement according to claim 2.
4. A method for cultivating strawberries in a greenhouse, which comprises applying the composite microbial liquid preparation for soil treatment according to claim 2.
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