CN117378629A - Rhizosphere colonization inducer, compound bacterial fertilizer and preparation method thereof - Google Patents
Rhizosphere colonization inducer, compound bacterial fertilizer and preparation method thereof Download PDFInfo
- Publication number
- CN117378629A CN117378629A CN202311272828.8A CN202311272828A CN117378629A CN 117378629 A CN117378629 A CN 117378629A CN 202311272828 A CN202311272828 A CN 202311272828A CN 117378629 A CN117378629 A CN 117378629A
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- China
- Prior art keywords
- bacillus
- colonization
- bacterial fertilizer
- promoting
- inducer
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- 239000003337 fertilizer Substances 0.000 title claims abstract description 79
- 230000001580 bacterial effect Effects 0.000 title claims abstract description 72
- 150000001875 compounds Chemical class 0.000 title claims abstract description 42
- 239000000411 inducer Substances 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title abstract description 12
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 128
- 239000000463 material Substances 0.000 claims abstract description 67
- 230000001737 promoting effect Effects 0.000 claims abstract description 35
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims abstract description 15
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 claims abstract description 14
- 241000194107 Bacillus megaterium Species 0.000 claims abstract description 11
- 239000002131 composite material Substances 0.000 claims abstract description 11
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 claims abstract description 10
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- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 claims abstract description 8
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 claims abstract description 8
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- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 claims abstract description 7
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 claims abstract description 5
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 claims abstract description 5
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 claims abstract description 5
- 229960003692 gamma aminobutyric acid Drugs 0.000 claims abstract description 5
- 239000001630 malic acid Substances 0.000 claims abstract description 5
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- 238000002156 mixing Methods 0.000 claims description 38
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 31
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- 238000000034 method Methods 0.000 claims description 26
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- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 18
- 235000019779 Rapeseed Meal Nutrition 0.000 claims description 12
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 12
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 12
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 12
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- 239000004455 soybean meal Substances 0.000 claims description 9
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- 235000019319 peptone Nutrition 0.000 claims description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 235000010216 calcium carbonate Nutrition 0.000 claims description 6
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- 235000007079 manganese sulphate Nutrition 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 235000013379 molasses Nutrition 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
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- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 8
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- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 239000005018 casein Substances 0.000 description 6
- 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 6
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- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000004471 Glycine Substances 0.000 description 4
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 4
- 244000046052 Phaseolus vulgaris Species 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
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- 229910000029 sodium carbonate Inorganic materials 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241001002356 Valeriana edulis Species 0.000 description 2
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- 239000006228 supernatant Substances 0.000 description 2
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 2
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 208000034309 Bacterial disease carrier Diseases 0.000 description 1
- AFCARXCZXQIEQB-UHFFFAOYSA-N N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CCNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 AFCARXCZXQIEQB-UHFFFAOYSA-N 0.000 description 1
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- 230000000443 biocontrol Effects 0.000 description 1
- 230000000853 biopesticidal effect Effects 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
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- OARRHUQTFTUEOS-UHFFFAOYSA-N safranin Chemical compound [Cl-].C=12C=C(N)C(C)=CC2=NC2=CC(C)=C(N)C=C2[N+]=1C1=CC=CC=C1 OARRHUQTFTUEOS-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/02—Saturated carboxylic acids or thio analogues thereof; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/36—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P3/00—Fungicides
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05C—NITROGENOUS FERTILISERS
- C05C3/00—Fertilisers containing other salts of ammonia or ammonia itself, e.g. gas liquor
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/10—Bacillus licheniformis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/11—Bacillus megaterium
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Pest Control & Pesticides (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Plant Pathology (AREA)
- Health & Medical Sciences (AREA)
- Environmental Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Agronomy & Crop Science (AREA)
- Dentistry (AREA)
- Biotechnology (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Virology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Mycology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fertilizers (AREA)
Abstract
The invention discloses a rhizosphere colonization inducer, a compound bacterial fertilizer and a preparation method thereof, and belongs to the technical field of agriculture. The composite bacterial fertilizer for promoting the colonization of bacillus comprises bacillus, rhizosphere colonization inducer, main materials and auxiliary materials; the bacillus comprises one or more of bacillus amyloliquefaciens, bacillus megaterium, bacillus licheniformis, bacillus bailii and bacillus mucilaginosus; the rhizosphere colonization inducer comprises one or more of stearic acid, palmitic acid, malic acid and gamma-aminobutyric acid. The bacillus in the compound bacterial fertilizer can grow at high density, and the rhizosphere colonization density of the bacillus in soil is also improved.
Description
Technical Field
The invention relates to the technical field of agriculture, in particular to a rhizosphere colonization inducer, a compound bacterial fertilizer and a preparation method thereof.
Background
At present, the farmland in China generally faces the problems of reduced soil quality, frequent occurrence of soil-borne diseases, pollution to agricultural lands and the like due to long-term excessive application of chemical fertilizers and pesticides, the yield and quality safety of crops are seriously affected, and the healthy and sustainable development of agriculture is hindered. The bacillus fertilizer which uses bacillus and the products thereof to generate the special fertilizer effect has the functions of activating root zone nutrients, promoting plant growth, enhancing plant stress resistance, inhibiting soil-borne diseases and the like, and has great application prospect in promoting fertilizer and pesticide application reduction and efficiency improvement. In the prior art, bacillus is the main plant rhizosphere beneficial bacteria applied in bacillus fertilizers and biopesticides, has disease-resistant and growth-promoting effects on plants, but is difficult to efficiently colonize the plant rhizosphere due to the influence of biological and non-biological factors in soil, so that the disease-resistant and growth-promoting effects of the bacillus fertilizers are unstable, and the field application and industrialized popularization of bacillus products are severely restricted. Therefore, it is necessary to invent a compound bacterial fertilizer for rhizosphere colonization induction and bacillus colonization promotion and a preparation method thereof.
Disclosure of Invention
Aiming at the defects of the prior art, on one hand, the invention provides a rhizosphere colonization inducer which can promote the colonization of bacillus in soil around plant roots and remarkably improve the colonization effect of bacillus. On the other hand, the invention provides a compound bacterial fertilizer for promoting the colonization of bacillus, bacillus in the compound bacterial fertilizer can grow at high density, and the colonization density of the rhizosphere of the bacillus in soil is also improved. The method is realized by the following specific techniques:
in a first aspect, the invention provides a rhizosphere colonization inducer comprising one or more of stearic acid, palmitic acid, malic acid, gamma-aminobutyric acid.
The rhizosphere colonization inducer has remarkable chemotactic effect on bacillus and can promote the colonization of bacillus in soil around plant roots.
The invention provides a compound bacterial fertilizer for promoting bacillus to colonize, which comprises the rhizosphere colonization inducer.
Preferably, the compound bacterial fertilizer further comprises bacillus, a main material and auxiliary materials; the bacillus includes one or more of Bacillus amyloliquefaciens, bacillus megaterium, bacillus licheniformis, bacillus bailii and Bacillus mucilaginosus.
The bacillus provided by the invention is matched with the main material and the auxiliary material for use, antagonism can be prevented, the main material and the auxiliary material provide carbon sources, nitrogen sources, inorganic salts, growth factors and the like for the bacillus to survive in the composting process, the bacillus cells can be supported for a long time to grow and reproduce after being applied into the soil, and an attachment place is provided for the bacillus, so that the high-density growth of the bacillus in the organic fertilizer is promoted, the use of the rhizosphere colonization inducer is combined, the colonization of the bacillus in soil around plant roots can be promoted, and the rhizosphere colonization inducer has a remarkable chemotaxis effect on the bacillus.
Preferably, the total viable count in the composite bacterial fertilizer for promoting the colonization of bacillus is more than or equal to 1.0X10 10 CFU/g。
Preferably, the main material comprises one or more of rapeseed meal, soybean meal, cottonseed meal, bran, edible fungus residues, chaff, crop straw, wood dust and molasses.
Preferably, the auxiliary materials comprise one or more of peptone, yeast powder, ammonium sulfate, manganese sulfate, ferric chloride and sodium chloride.
Preferably, the mass ratio of the main material to the rhizosphere colonization inducer is (1000-10000): 1.
Preferably, the mass ratio of the main material to the auxiliary material is (100-10000): 1.
The third aspect of the invention provides a preparation method of the compound bacterial fertilizer for promoting bacillus colonization, which comprises the following steps:
inoculating, activating and culturing bacillus strains, and then inoculating the bacillus strains into a seed culture medium for culturing to obtain bacillus seed liquid;
uniformly mixing the main material, the auxiliary material and water, piling to obtain a mixed material pile, adding the bacillus seed liquid into the mixed material pile, and uniformly mixing to obtain a pile body;
composting the pile to obtain initial bacterial manure;
adding the rhizosphere colonization inducer into the initial bacterial fertilizer, and uniformly mixing to obtain the composite bacterial fertilizer for promoting the colonization of bacillus.
Preferably, the seed medium comprises corn starch, soybean meal, calcium carbonate and ammonium sulfate. Further preferably, the mass ratio of the corn starch, the bean pulp, the calcium carbonate and the ammonium sulfate is (6.5-50): 10-70): 1-6): 1.
Preferably, the bacillus seed liquid is added to the mixture pile in the amount of (1-3) L/m 3 。
Preferably, the mass ratio of the main material to water is 1 (0.5-2).
Preferably, the internal temperature of the stack is between 35 and 50 ℃; the composting time is 10-20 d.
Preferably, the bacillus strain is activated on a solid LB culture medium by adopting a plate streaking method, a single colony is picked up after a single colony is grown, inoculated into a liquid LB culture medium, and shake-cultured for 8-12 h at 35-40 ℃.
Preferably, the bacillus species cultured in the liquid LB medium is inoculated into the seed medium in an inoculum size of 1 to 3% by volume of the seed medium (hereinafter, abbreviated as inoculum size of 1 to 3%) and cultured with shaking at 35 to 40 ℃ for 8 to 48 hours.
Preferably, the volume of the pile is more than or equal to 5m 3 The height of the pile is 0.5-1.0 m.
On the basis of analyzing the influence of factors such as soil properties, environmental conditions, interaction with host plant root systems and the like on the rhizosphere colonization behavior of the bacillus, the invention is compatible with the efficient decomposed bacillus to perform efficient decomposed fermentation on agricultural waste materials, optimizes the bacterial fertilizer production technology, aims at improving the rhizosphere colonization efficiency of the bacillus in field crops and the field disease resistance and growth promotion effect thereof, and has great significance for promoting the pesticide fertilizer to reduce application and increase efficiency and promoting the sustainable development of agriculture.
Compared with the prior art, the invention has the following advantages:
the rhizosphere colonization inducer has remarkable chemotactic effect on bacillus, can promote the bacillus to colonize in soil around plant roots in a trend of more than 3 times, and remarkably improves the colonization effect of the bacillus;
the compound bacterial fertilizer for promoting the colonization of bacillus optimizes the compatibility of bacillus species, main materials and auxiliary materials, and has the effects of preventing antagonism of each other so as to realize synergistic disease resistance and growth promotion of plants; meanwhile, the rhizosphere colonization inducer is combined, so that the prepared compound bacterial fertilizer can effectively inhibit bacterial and fungal soil-borne diseases from spreading, has the functions of promoting crop growth and improving crop disease resistance, effectively reduces the loss caused by the soil-borne diseases, solves the problems of low colonization density and poor stability of bacillus biocontrol bacillus in soil, and has great significance in promoting pesticide and fertilizer application reduction and synergy and promoting agricultural sustainable development; the invention promotes sporeIn the composite bacterial fertilizer for bacterial colonization, the total viable count is more than or equal to 1.0X10 10 CFU/g, when using a plurality of bacillus, the viable count of each bacillus is not less than 5.0X10 8 CFU/g, content of easily oxidized organic matters is more than or equal to 35%, and content of water-soluble amino acids is more than or equal to 40mg/g.
Detailed Description
In order to further illustrate the present invention, the following detailed description of the technical solutions provided by the present invention is provided in connection with the examples, but they should not be construed as limiting the scope of the present invention.
Unless otherwise specifically indicated, the various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or may be prepared by existing methods.
The preparation process of the seed culture medium comprises the steps of firstly dissolving corn starch by using boiled water, adding a proper amount of amylase for gelatinization when the corn starch is cooled to about 70 ℃ until the corn starch solution is water-like and is not sticky, then sequentially adding soybean meal, ammonium sulfate and calcium carbonate, subpackaging into triangular flasks, and sterilizing at 121 ℃ for 30 min.
After seed liquid of each strain is obtained, 10 mu L of seed liquid of each strain is respectively coated on a glass slide, safranine dye is used for dyeing for 2min, distilled water is used for washing out redundant dye, and after the water is dried, the glass slide is placed under a common optical microscope for microscopic examination, and whether the bacteria are polluted or not is analyzed through morphological characteristics. After no pollution is confirmed, the compound bacterial fertilizer for promoting bacillus colonization can be continuously prepared in the subsequent steps.
The invention discloses a preparation method of a compound bacterial fertilizer for promoting bacillus colonization, which comprises the following steps:
inoculating, activating and culturing bacillus strains, and then inoculating the bacillus strains into a seed culture medium for culturing to obtain bacillus seed liquid;
uniformly mixing the main material, the auxiliary material and water, piling to obtain a mixed material pile, adding the bacillus seed liquid into the mixed material pile, and uniformly mixing to obtain a pile body;
composting the pile to obtain initial bacterial manure;
adding the rhizosphere colonization inducer into the initial bacterial fertilizer, and uniformly mixing to obtain the composite bacterial fertilizer for promoting the colonization of bacillus.
Optionally, the bacillus species includes one or more of bacillus amyloliquefaciens, bacillus megaterium, bacillus licheniformis, bacillus bailii, bacillus mucilaginosus.
Optionally, the rhizosphere colonization inducer comprises one or more of stearic acid, palmitic acid, malic acid, gamma-aminobutyric acid.
Optionally, the main material comprises one or more of rapeseed meal, soybean meal, cottonseed meal, bran, edible fungus residues, chaff, crop straw, wood dust and molasses.
Optionally, the auxiliary materials comprise one or more of peptone, yeast powder, ammonium sulfate, manganese sulfate, ferric chloride and sodium chloride.
Optionally, the mass ratio of the main material to the rhizosphere colonization inducer is (1000-10000): 1.
Optionally, the mass ratio of the main material to the auxiliary material is (100-10000): 1.
Optionally, the mass ratio of the main material to the water is 1 (0.5-2).
Optionally, bacillus strains are activated on a solid LB culture medium by adopting a flat plate streaking method, single bacterial colonies are picked up after single bacterial colonies grow out and inoculated into a liquid LB culture medium, and shake culture is carried out for 8-12 h at 35-40 ℃.
Alternatively, the bacillus strain cultured in the liquid LB culture medium is inoculated into the seed culture medium according to the inoculum size of 1-3%, and is cultured for 8-48 h at 35-40 ℃.
Optionally, the seed medium includes corn starch, soybean meal, calcium carbonate, and ammonium sulfate.
Optionally, the mass ratio of corn starch, soybean meal, calcium carbonate and ammonium sulfate in the seed culture medium is (6.5-50): 10-70): 1-6): 1.
Optionally, the bacillus seed liquid is added into the mixed material pile in the amount of (1-3) L/m 3 。
Optionally, the volume of the pile body is more than or equal to 5m 3 The height of the pile is 0.5-1.0 m.
Optionally, the internal temperature of the pile body is 35-50 ℃; the composting time is 10-20 d.
The following description of the present invention will be made clearly and fully, and it is apparent that the embodiments described are only some, but not all, of the embodiments of the present invention. All other embodiments, which can be made by one of ordinary skill in the art without undue burden on the person of ordinary skill in the art based on embodiments of the present invention, are within the scope of the present invention.
According to the invention, different bacillus, main materials, auxiliary materials and rhizosphere colonization inducer are adopted for being matched for use, and a great number of experiments are carried out, so that satisfactory effects are obtained. In the following specific examples and comparative examples, bacillus was exemplified by Bacillus amyloliquefaciens and Bacillus megaterium, the main material was exemplified by rapeseed meal, and the auxiliary material was exemplified by peptone and ferric chloride.
Example 1
The embodiment provides a preparation method of a compound bacterial fertilizer for promoting bacillus colonization, which comprises the following steps:
s1, activating bacillus amyloliquefaciens on a solid LB (LB) culture medium by adopting a plate streaking method, picking a single colony after a single colony grows out, inoculating the single colony into a liquid LB culture medium, carrying out shake culture for 10 hours at a shaking table 37 ℃, then inoculating the single colony into a seed culture medium according to an inoculum size of 2%, and carrying out shake culture for 40 hours at the shaking table 37 ℃ to obtain bacillus amyloliquefaciens seed liquid; wherein the seed culture medium consists of corn starch, bean pulp, calcium carbonate and ammonium sulfate according to the mass ratio of 30:50:5:3; preparing bacillus megatherium seed liquid by the same method; uniformly mixing bacillus amyloliquefaciens seed liquid and bacillus megatherium seed liquid to obtain mixed bacillus seed liquid;
s2, uniformly mixing peptone and ferric chloride to obtain a mixture, uniformly mixing rapeseed meal and the mixture according to the mass ratio of 1000:1, then adding water according to the mass ratio of the rapeseed meal to the water of 1:1, uniformly mixing, piling to obtain a mixed material pile, and mixing bacillus seed liquid obtained in the step S1According to 2L/m 3 Adding the mixture into the mixture pile, uniformly mixing to obtain a pile body, wherein the volume of the pile body is 8m 3 The height of the pile body is 0.8m;
composting the pile for 18d, wherein the temperature of the pile is 38 ℃, the pile temperature is reduced after composting is finished, the materials are loose, and no odor exists, so that an initial bacterial fertilizer is obtained;
s3, adding stearic acid into the initial bacterial manure obtained in the step S2, and uniformly mixing to obtain a composite bacterial manure for promoting bacillus colonization; wherein the mass ratio of stearic acid to rapeseed meal is 1:5000.
Example 2
The embodiment provides a preparation method of a compound bacterial fertilizer for promoting bacillus colonization, which comprises the following steps:
s1, activating bacillus amyloliquefaciens on a solid LB (LB) culture medium by adopting a plate streaking method, picking a single colony after a single colony grows out, inoculating the single colony into a liquid LB culture medium, carrying out shake culture for 12 hours at a shaking table 35 ℃, then inoculating the single colony into a seed culture medium according to an inoculum size of 3%, and carrying out shake culture for 48 hours at the shaking table 35 ℃ to obtain bacillus amyloliquefaciens seed liquid; wherein the seed culture medium consists of corn starch, soybean meal, calcium carbonate and ammonium sulfate according to the mass ratio of 20:70:6:1; preparing bacillus megatherium seed liquid by the same method; uniformly mixing bacillus amyloliquefaciens seed liquid and bacillus megatherium seed liquid to obtain mixed bacillus seed liquid;
s2, uniformly mixing peptone and ferric chloride to obtain a mixture I, uniformly mixing rapeseed meal and the mixture I according to the mass ratio of 10000:1, then adding water according to the mass ratio of 1:2 of the rapeseed meal and the water, uniformly mixing, piling to obtain a mixed material pile, and mixing bacillus seed liquid obtained in the step S1 according to the mass ratio of 3L/m 3 Adding the mixture into the mixture pile, and uniformly mixing to obtain a pile body with the volume of 5m 3 The height of the pile body is 1.0m;
composting the pile for 20d, wherein the temperature of the pile is 50 ℃, the pile temperature is reduced after composting is finished, the materials are loose, and no odor exists, so that the initial bacterial manure is obtained;
s3, mixing stearic acid, malic acid and palmitic acid to obtain a mixture II, adding the mixture II into the initial bacterial fertilizer obtained in the step S2, and uniformly mixing to obtain the composite bacterial fertilizer for promoting the colonization of bacillus; wherein the mass ratio of the mixture II to the mixture I is 1:10000.
Example 3
The embodiment provides a preparation method of a compound bacterial fertilizer for promoting bacillus colonization, which comprises the following steps:
s1, activating bacillus amyloliquefaciens on a solid LB (LB) culture medium by adopting a plate streaking method, picking a single colony after a single colony grows out, inoculating the single colony into a liquid LB culture medium, carrying out shake culture for 8 hours at a shaking table 40 ℃, then inoculating the single colony into a seed culture medium according to an inoculum size of 3%, and carrying out shake culture for 8 hours at the shaking table 40 ℃ to obtain bacillus amyloliquefaciens seed liquid; wherein the seed culture medium consists of corn starch, bean pulp, calcium carbonate and ammonium sulfate according to the mass ratio of 50:30:3:3; preparing bacillus megatherium seed liquid by the same method; uniformly mixing bacillus amyloliquefaciens seed liquid and bacillus megatherium seed liquid to obtain mixed bacillus seed liquid;
s2, uniformly mixing peptone and ferric chloride to obtain a mixture I, uniformly mixing rapeseed meal and the mixture I according to the mass ratio of 6000:1, then adding water according to the mass ratio of the rapeseed meal to the water of 1:0.5, uniformly mixing, piling to obtain a mixed material pile, and mixing bacillus seed liquid obtained in the step S1 according to the mass ratio of 1L/m 3 Adding the mixture into the mixture pile, and uniformly mixing to obtain a pile body, wherein the volume of the pile body is 10m 3 The height of the pile body is 0.9m;
composting the pile for 10d, wherein the temperature of the pile is 45 ℃, the pile temperature is reduced after composting is finished, the materials are loose, and no odor exists, so that the initial bacterial manure is obtained;
s3, mixing palmitic acid and gamma-aminobutyric acid to obtain a mixture II, adding the mixture II into the initial bacterial fertilizer obtained in the step S2, and uniformly mixing to obtain the composite bacterial fertilizer for promoting bacillus colonization; wherein the mass ratio of the palmitic acid to the mixture I is 1:1000.
Comparative example 1
This comparative example is substantially the same as example 1 except that stearic acid is not used, i.e., the bacterial manure of this comparative example is obtained after composting is completed.
Comparative example 2
The comparative example is basically the same as example 1, except that the prepared compound bacterial fertilizer for promoting bacillus colonization is placed at a high temperature of 120 ℃ and a high pressure of 1MPa for 1 hour, and the bacillus amyloliquefaciens in the finally obtained compound bacterial fertilizer is inactivated at a high temperature and a high pressure.
Fertilizer efficiency test
The compound bacterial fertilizers prepared in examples 1-3 and comparative examples 1-2 are subjected to neutral protease enzyme activity detection, easily oxidized organic matter content detection and water-soluble amino acid content detection.
Neutral proteinase is the main enzyme needed by bacillus to decompose protein macromolecules in materials such as rapeseed meal, bean pulp, cottonseed meal and the like, and the neutral proteinase decomposes the protein into substances such as amino acid, polypeptide and the like of small molecules so as to help promote the absorption of nitrogen elements by crops, thereby improving fertilizer efficiency.
The easily oxidized organic matter is organic carbon matter which is easily oxidized and has high activity, and is a main source for obtaining organic nutrition by plants; the water-soluble amino acid is a micromolecular amino acid substance formed after the cake which is difficult to dissolve in water is hydrolyzed by neutral proteinase secreted by bacillus, etc., and the substance has small molecular weight, is easy to dissolve in water and is easy to be absorbed by plant root systems; the concentration of the easily oxidized organic matters and the content of the water-soluble amino acids are main indexes for reacting the fertilizer efficiency of the organic fertilizer.
The specific detection method comprises the following steps:
(1) Detection of enzyme activity of neutral protease in compound bacterial fertilizer
Weighing 1g of casein, weighing 200 mu L of 2% sodium hydroxide solution, adding the casein and the sodium hydroxide solution into a boric acid buffer solution with the pH of 100 mLpH=7.4, stirring for 5min in a boiling water bath until no precipitate exists, continuing to perform water bath for 15min, cooling, adjusting the pH=7.4, and preserving to obtain casein solution;
weighing 4.24g of sodium carbonate, and dissolving in 100mL of distilled water to prepare sodium carbonate solution;
in the composting process of example 1, 10g of the enzyme sample is dissolved in 30mL of water at the 5d, 10d, 15d and 20d of composting, and the enzyme samples of each fermentation period of example 1 are obtained after stirring for 10 min; examples 2 to 3 and comparative examples 1 to 2 enzyme samples of each fermentation period of examples 2 to 3 and comparative examples 1 to 2 were obtained by the same method as described above;
then placing the casein solution into a 40 ℃ constant temperature water bath kettle to preheat for 5min, simultaneously taking 1mL of enzyme samples of examples 1-3 and comparative examples 1-2, respectively, adding the enzyme samples into a test tube, placing the test tube into a 40 ℃ water bath for 2min, then adding the preheated casein solution, uniformly mixing, placing the test tube into a 40 ℃ water bath for 10min, adding 2mL of trichloroacetic acid, uniformly mixing, taking out, standing for 10min, centrifuging for 2min at 12000rpm, taking 1mL of supernatant, mixing with 5mL of sodium carbonate solution and 1mL of Fulin reagent, placing the mixture into a 40 ℃ water bath for 20min, and measuring absorbance at 680 nm. In addition, the detection is also provided with a blank sample group without adding any enzyme sample, the blank sample group is directly added into an empty test tube, trichloroacetic acid is added into the test tube, then casein solution is added into the test tube, and the rest steps are the same as the enzyme sample. The measurement results are shown in Table 1.
TABLE 1 results of neutral protease enzyme Activity assay
As can be seen from Table 1, the neutral protease activities of examples 1 to 3 are maintained at higher levels in composting of 5d, 10d, 15d and 20d, which means that the compound bacterial fertilizer of the invention contains high-density bacillus and can well meet the material decomposition requirements.
The neutral protease activity of comparative example 1 was not substantially changed compared to example 1, because the main material and the auxiliary material were used although the rhizosphere colonization inducer was not used in comparative example 1, that is, the main material and the auxiliary material normally exert a synergistic effect during composting, and the remaining conditions were substantially the same as example 1, so that the enzyme sample test result of comparative example 1 was not greatly different from example 1, which also means that the synergistic effect of the main material and the auxiliary material has a remarkable effect on the fertilizer efficiency of the compound bacterial fertilizer.
In comparison with example 1, no enzyme activity was detected in both comparative example 2 and the blank because bacillus was inactivated at high temperature and high pressure in comparative example 2, and no bacillus was present in the blank, and thus no neutral protease enzyme activity was detected in both comparative example 2 and the blank.
(2) Content detection of easily-oxidized organic matters
In the composting process of the embodiment 1, 0.1g is taken at the 5d, 10d, 15d and 20d of composting respectively, the composting process is placed in a 50mL conical flask, 10mL of potassium permanganate solution with the concentration of 10g/L is added, the stirring and the mixing are carried out for 1h, the stirring and the mixing are carried out at 10000rpm for 5min, 1mL of supernatant fluid is taken and diluted with 5mL of water, the light absorption value is measured at 565nm, and the proportion of the easily oxidized organic matters in the total organic matters in each fermentation period of the embodiment 1 is calculated; examples 2 to 3 and comparative examples 1 to 2 the same method as described above was used to obtain the ratio of the readily oxidizable organic substance to the total organic substance in each fermentation period of examples 2 to 3 and comparative examples 1 to 2. The measurement results are shown in Table 2.
TABLE 2 detection results of easily oxidizable organic substance content
As can be seen from Table 2, the ratio of the easily-oxidizable organic matters in examples 1-3 in the total organic matters is obviously improved in the composting process of 5d, 10d, 15d and 20d, which shows that the content of the easily-oxidizable organic matters in the composting fermentation process is gradually increased along with the extension of the fermentation time, so that the fertilizer efficiency of the compound bacterial fertilizer is also gradually enhanced.
Compared with example 1, the ratio of the easily oxidized organic matters in the total organic matters in the compound bacterial manure of comparative example 1 is almost unchanged, because the steps and conditions before the end of composting in comparative example 1 are basically the same as those in example 1, the detection results are not greatly different, and the synergistic effect of the main materials and the auxiliary materials has obvious influence on the fertilizer efficiency of the compound bacterial manure.
The ratio of the easily oxidizable organic substance in comparative example 2 to the total organic substance is significantly lower than in example 1 because the bacillus in comparative example 2 has been inactivated, and thus, the ratio of the easily oxidizable organic substance in comparative example 2 to the total organic substance is not significantly improved with the increase of the fermentation time without containing active bacillus.
(3) Water-soluble amino acid content detection
0.028g glycine is weighed and dissolved in 100mL distilled water to prepare glycine standard solution; weighing 0.2g of ninhydrin, and dissolving in 10mL of ethanol to prepare a ninhydrin color solution; taking 0mL,0.2mL,0.4mL,0.6mL,0.8mL and 1mL of glycine standard solution respectively, putting the glycine standard solution in a test tube, adding distilled water to 1mL, adding 4mL of phosphate buffer solution with pH=5.5 respectively, adding 0.2mL of ninhydrin color development solution, uniformly mixing, sealing a test tube mouth, boiling water bath for 15min, taking 1mL of ninhydrin color development solution, adding 5mL of water for dilution, measuring a light absorption value at a position of 572nm, and manufacturing a standard curve.
In the composting process of the example 1, 1g of the water is taken to be dissolved in 10mL of water and fully vibrated and uniformly mixed for 20min respectively at the 5d, 10d, 15d and 20d of composting, the ninhydrin color development method is used for developing color, the light absorption value is measured at 572nm, and the water-soluble amino acid content is calculated according to the standard curve; examples 2 to 3 and comparative examples 1 to 2 the water-soluble amino acid contents in examples 2 to 3 and comparative examples 1 to 2 were obtained in the same manner as described above. The results are shown in Table 3.
TABLE 3 detection results of the content of Water-soluble amino acids
As can be seen from Table 3, the water-soluble amino acid contents of examples 1 to 3 are obviously improved in composting of 5d, 10d, 15d and 20d, which shows that the water-soluble amino acid content in the composting fermentation process is gradually increased along with the extension of the fermentation time, namely the fertilizer efficiency of the compound bacterial fertilizer is also gradually enhanced.
The water-soluble amino acid content of comparative example 1 was not substantially changed as compared with example 1, since the steps and conditions before the end of composting in comparative example 1 were substantially the same as those of example 1, and thus the detection results were not much different, which also indicates that the synergistic effect of the main material and the auxiliary material had a remarkable effect on the fertilizer efficiency of the compound bacterial fertilizer.
The water-soluble amino acid content in comparative example 2 was significantly lower than that in example 1, because the bacillus in comparative example 2 had been inactivated, and thus, in the case of not containing active bacillus, the water-soluble amino acid contents in comparative example 2 and comparative example 3 were not significantly increased with the increase of fermentation time.
Field test
The field test effect verification is carried out on the compound bacterial fertilizers prepared in the examples 1-3 and the comparative examples 1-2:
1. test time and place
Time: 2021 month 3 to 2021 month 10
Location: east-west lake area test field in Wuhan city of Hubei province
2. Test materials
Test fertilizer: the compound bacterial fertilizers prepared in examples 1-3 and comparative examples 1-2; commercial Bacillus-free organic fertilizer (as control group)
Test crop: tobacco leaf
3. Test protocol
The field application method of the fertilizer comprises the following steps: the compound bacterial fertilizers prepared in examples 1-3 and comparative examples 1-2 and the commercial organic fertilizer are applied in a hole application mode, the application amount is 50 kg/mu, and crop seedlings are transplanted after application; after the organic fertilizer of the control group was applied, the same number of bacillus as in example 1 was also applied to the roots of each tobacco plant by root irrigation.
And (3) field management: soil preparation, fertilizer base fertilizer application, deep ploughing, ridging, hole digging, artificial hole sowing, compound bacterial fertilizer application, tobacco seedling transplanting, planting density of 1000 plants/mu and watering and irrigation according to soil moisture content. The field conditions and the management conditions of examples 1 to 3 and comparative examples 1 to 2 and the control group were all kept the same.
4. Detection method of soil colonisation bacillus
And taking soil with depth of 3-20 cm from 0-30 cm around crops in 10, 30, 50 and 70d after transplanting tobacco seedlings, and detecting the density of bacillus by adopting a dilution coating method. The results are shown in Table 4.
TABLE 4 Bacillus rhizosphere colonization density
As can be seen from Table 4, the Bacillus density of examples 1 to 3 was increased by more than 10 times compared with the control group, which demonstrates that the Bacillus can be efficiently colonized at the tobacco root after the application of the Bacillus colonization promoting compound bacterial fertilizer of the present invention.
The bacillus density of comparative examples 1-2 was significantly reduced compared to example 1, especially in the soil of comparative example 2, with almost no bacillus, probably because the bacillus in the bacterial manure was affected by biological and non-biological factors in the soil due to the fact that no rhizosphere colonization inducer was used in comparative example 1, and the bacillus density in the soil was low; in comparative example 2, after the composite bacterial manure is subjected to high-temperature and high-pressure treatment, bacillus with activity hardly exists, rhizosphere colonization cannot be performed in soil, so that bacillus cannot be detected in soil, and the fact that natural bacillus does not exist in soil influences the detection result is also indicated;
the very low bacillus density of the control group compared to example 1 suggests that common commercial organic fertilizers without bacillus do not promote efficient rhizosphere colonization of bacillus, and thus the density of bacillus detected in the soil is very low.
5. Detection of growth promoting effect of bacterial manure on tobacco
After the tobacco enters the maturity stage, 50 pieces of tobacco are randomly selected, the tobacco leaf area of the largest tobacco leaf of each piece of tobacco is measured, and the average value is calculated, so that the largest tobacco leaf areas of the examples 1-3, the comparative examples 1-2 and the control group are obtained, and the results are shown in Table 5.
Wherein the tobacco leaf area (cm) 2 ) =length (cm) ×width (cm) × 0.6345.
Table 5 maximum tobacco leaf area
As can be seen from Table 5, the maximum tobacco leaf areas of examples 1 to 3 are each 830cm, as compared with comparative example 1 2 About, the compound bacterial fertilizer provided by the invention can effectively promote the fertilizer efficiency of organic fertilizer and promote the growth of crops.
The maximum tobacco leaf area of comparative example 1 was reduced as compared with example 1, because the tobacco growth in comparative example 1 was poor because the rhizosphere colonization inducer was not used in comparative example 1 and bacillus was not allowed to colonize around the tobacco root, thereby promoting tobacco growth.
The maximum tobacco leaf area of comparative example 2 was reduced as compared with example 1, because bacillus in comparative example 2 was inactive and had no growth promoting effect on tobacco, and thus the tobacco in comparative example 2 grew slightly worse.
In comparison with example 1, although bacillus was applied in the control group, the general organic fertilizer failed to promote the high-density growth and efficient colonization of bacillus, so that the bacillus in the control group was not ideal for the tobacco promotion effect.
The above detailed description describes in detail the practice of the invention, but the invention is not limited to the specific details of the above embodiments. Many simple modifications and variations of the technical solution of the present invention are possible within the scope of the claims and technical idea of the present invention, which simple modifications are all within the scope of the present invention.
Claims (10)
1. A rhizosphere colonization inducer comprising one or more of stearic acid, palmitic acid, malic acid, and gamma-aminobutyric acid.
2. A composite bacterial fertilizer for promoting bacillus colonization, comprising the rhizosphere colonization inducer of claim 1.
3. The compound bacterial fertilizer for promoting the colonization of bacillus according to claim 2, wherein the compound bacterial fertilizer further comprises bacillus, a main material and an auxiliary material; the bacillus comprises one or more of bacillus amyloliquefaciens, bacillus megaterium, bacillus licheniformis, bacillus bailii and bacillus mucilaginosus.
4. The compound bacterial fertilizer for promoting the colonization of bacillus according to claim 2, wherein the total viable count in the compound bacterial fertilizer is more than or equal to 1.0 multiplied by 10 10 CFU/g。
5. The compound bacterial fertilizer for promoting bacillus colonization according to claim 3, wherein the main material comprises one or more of rapeseed meal, soybean meal, cottonseed meal, bran, edible fungus residue, chaff, crop straw, wood chips, molasses; the mass ratio of the main material to the rhizosphere colonization inducer is (1000-10000): 1.
6. The compound bacterial fertilizer for promoting bacillus colonization according to claim 3, wherein the auxiliary materials comprise one or more of peptone, yeast powder, ammonium sulfate, manganese sulfate, ferric chloride and sodium chloride; the mass ratio of the main material to the auxiliary material is (100-10000): 1.
7. The method for preparing a compound bacterial fertilizer for promoting the colonization of bacillus according to any one of claims 1 to 6, which is characterized by comprising the following steps:
inoculating, activating and culturing bacillus strains, and inoculating the bacillus strains into a seed culture medium for culturing to obtain bacillus seed liquid; uniformly mixing the main materials, the auxiliary materials and water, piling to obtain a mixed material pile, adding the bacillus seed liquid into the mixed material pile, uniformly mixing, and composting to obtain an initial bacterial fertilizer;
adding the rhizosphere colonization inducer into the initial bacterial fertilizer, and uniformly mixing to obtain the composite bacterial fertilizer for promoting the colonization of bacillus.
8. The method according to claim 7, wherein the seed medium is inoculated with the activated and cultured strain of Bacillus in an inoculum size of 1 to 3% by volume of the seed medium.
9. The method according to claim 7, wherein the seed medium comprises corn starch, soybean meal, calcium carbonate and ammonium sulfate in a mass ratio of (6.5-50): 10-70): 1-6): 1.
10. The method according to claim 7, wherein the Bacillus seed solution is added to the mixed material pile in an amount of (1-3) L/m 3 。
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