CN116694526B - Composite microbial agent for promoting rooting and preparation method thereof - Google Patents
Composite microbial agent for promoting rooting and preparation method thereof Download PDFInfo
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- CN116694526B CN116694526B CN202310712363.7A CN202310712363A CN116694526B CN 116694526 B CN116694526 B CN 116694526B CN 202310712363 A CN202310712363 A CN 202310712363A CN 116694526 B CN116694526 B CN 116694526B
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- azotobacter vinelandii
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 113
- 238000002360 preparation method Methods 0.000 title claims abstract description 31
- 230000001737 promoting effect Effects 0.000 title claims abstract description 20
- 239000002131 composite material Substances 0.000 title claims description 8
- 238000000855 fermentation Methods 0.000 claims abstract description 153
- 230000004151 fermentation Effects 0.000 claims abstract description 153
- 241000589149 Azotobacter vinelandii Species 0.000 claims abstract description 45
- 241000588914 Enterobacter Species 0.000 claims abstract description 37
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 37
- 229920001661 Chitosan Polymers 0.000 claims abstract description 17
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000004202 carbamide Substances 0.000 claims abstract description 17
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 13
- 150000001875 compounds Chemical class 0.000 claims abstract description 12
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 8
- QJZYHAIUNVAGQP-UHFFFAOYSA-N 3-nitrobicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid Chemical compound C1C2C=CC1C(C(=O)O)C2(C(O)=O)[N+]([O-])=O QJZYHAIUNVAGQP-UHFFFAOYSA-N 0.000 claims abstract description 7
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims abstract description 7
- 229960000892 attapulgite Drugs 0.000 claims abstract description 7
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- 238000000034 method Methods 0.000 claims description 16
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- 238000003199 nucleic acid amplification method Methods 0.000 claims description 11
- DAJSVUQLFFJUSX-UHFFFAOYSA-M sodium;dodecane-1-sulfonate Chemical compound [Na+].CCCCCCCCCCCCS([O-])(=O)=O DAJSVUQLFFJUSX-UHFFFAOYSA-M 0.000 claims description 10
- 244000063299 Bacillus subtilis Species 0.000 claims description 9
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 5
- 239000011259 mixed solution Substances 0.000 claims description 5
- 239000002068 microbial inoculum Substances 0.000 abstract description 23
- 230000012010 growth Effects 0.000 abstract description 8
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- 238000009629 microbiological culture Methods 0.000 description 24
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- 238000011282 treatment Methods 0.000 description 13
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- 235000005822 corn Nutrition 0.000 description 9
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 8
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 8
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- 239000000843 powder Substances 0.000 description 8
- 239000011780 sodium chloride Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 239000005720 sucrose Substances 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 239000012137 tryptone Substances 0.000 description 8
- 239000012138 yeast extract Substances 0.000 description 8
- 239000001963 growth medium Substances 0.000 description 5
- 229910052757 nitrogen Inorganic materials 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- 241000589151 Azotobacter Species 0.000 description 4
- 241001322378 Bacillus paralicheniformis Species 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 4
- 241000982938 Enterobacter cancerogenus Species 0.000 description 4
- 239000012880 LB liquid culture medium Substances 0.000 description 4
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- 239000008272 agar Substances 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 229910052698 phosphorus Inorganic materials 0.000 description 4
- 230000008635 plant growth Effects 0.000 description 4
- 235000010958 polyglycerol polyricinoleate Nutrition 0.000 description 4
- 201000009030 Carcinoma Diseases 0.000 description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 3
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- 229910052700 potassium Inorganic materials 0.000 description 3
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- 239000000575 pesticide Substances 0.000 description 2
- 239000003375 plant hormone Substances 0.000 description 2
- 239000011591 potassium Substances 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 229930192334 Auxin Natural products 0.000 description 1
- 201000004569 Blindness Diseases 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 241001057636 Dracaena deremensis Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000005977 Ethylene Substances 0.000 description 1
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- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 241000881860 Paenibacillus mucilaginosus Species 0.000 description 1
- 241000235342 Saccharomycetes Species 0.000 description 1
- 239000000589 Siderophore Substances 0.000 description 1
- 241000223260 Trichoderma harzianum Species 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
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- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 1
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- 239000013543 active substance Substances 0.000 description 1
- 238000005904 alkaline hydrolysis reaction Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
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- 244000052616 bacterial pathogen Species 0.000 description 1
- FFBHFFJDDLITSX-UHFFFAOYSA-N benzyl N-[2-hydroxy-4-(3-oxomorpholin-4-yl)phenyl]carbamate Chemical compound OC1=C(NC(=O)OCC2=CC=CC=C2)C=CC(=C1)N1CCOCC1=O FFBHFFJDDLITSX-UHFFFAOYSA-N 0.000 description 1
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- 239000002274 desiccant Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000005069 ears Anatomy 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 244000037666 field crops Species 0.000 description 1
- 230000005669 field effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000007952 growth promoter Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000003617 indole-3-acetic acid Substances 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 235000009973 maize Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
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- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000002786 root growth Effects 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
- C05D9/00—Other inorganic fertilisers
-
- 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
-
- 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
- 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
- C05G5/00—Fertilisers characterised by their form
- C05G5/10—Solid or semi-solid fertilisers, e.g. powders
- C05G5/12—Granules or flakes
-
- 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
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/065—Azotobacter
-
- 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
- C12R2001/38—Pseudomonas
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The invention discloses a compound microbial agent for promoting rooting and a preparation method thereof, belonging to the technical field of microorganisms. The microbial agent comprises microbial fermentation broth and a microbial carrier, wherein the volume mass ratio of the microbial fermentation broth to the microbial carrier is 1L:1kg. The invention is compounded with three functional strains of enterobacter canescens, azotobacter vinelandii and bacillus paratyphenius to form a core microbial inoculum, and the three strains are mixed in equal proportion, so that the promoting effect of the microbial inoculum on roots is better than that of a single strain, the three strains are compounded for use, and the three strains promote each other, so that the microbial inoculum has stronger adaptability to complex environment of rhizosphere in practical application. Meanwhile, diatomite, attapulgite and the like are used as carriers, chitosan and sodium dodecyl sulfate are added to fully adsorb and disperse the microbial inoculum, so that the acting time and the acting stability of the microbial inoculum are prolonged, and urea humic acid and the like provide nutrient substances for crops and promote the growth of the crops.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a compound microbial agent for promoting rooting and a preparation method thereof.
Background
Promoting yield increase is the primary task of modern efficient agriculture. In order to achieve the purpose, technicians in the planting industry often blindly overuse fertilizers, especially fertilizers, which not only increase the planting cost and waste resources, but also cause the problems of soil hardening and acidification of cultivated lands, quality degradation of agricultural products and the like, and the problems are accompanied by non-point source pollution which seriously affects the quality of surface water and underground water. Therefore, the control of the fertilizer dosage is a realistic requirement for promoting cost saving, efficiency improvement, energy saving and emission reduction, and has very important significance for guaranteeing national grain safety, agricultural product quality safety and agricultural ecological safety. The research and development of the functional microbial fertilizer and the screening and compounding of the agricultural probiotics are one of the important means for realizing fertilizer reduction.
Rhizosphere growth promoting bacteria (PGPR) are beneficial bacteria which can freely live in the soil surrounding the root or attach to the surface and inside the root, can promote plant growth and the absorption and utilization of mineral nutrition, and can inhibit harmful organisms. Rhizosphere growth promoters may promote plant growth, either directly or indirectly, through one or more mechanisms. Through the actions of nitrogen fixation, phosphorus decomposition, potassium decomposition, plant hormone production, ethylene reduction by enzymolysis and the like, soil nutrients are increased, the absorption and utilization of root systems to mineral substances are promoted, and the plant growth is directly stimulated and regulated; on the other hand, by producing antibiotics and secreting siderophores, HCN and the like, it can inhibit harmful pathogenic microorganisms in soil, indirectly protect and promote plant growth. Inoculating rhizosphere growth-promoting bacteria can not only reduce the use of chemical fertilizers and pesticides, but also reduce the environmental pollution and food safety problems caused by using a large amount of chemical fertilizers and pesticides.
The development, development and application of the novel PGPR composition and production process composite microbial inoculum are the directions of one hot spot of the current microbial fertilizer. The aim of the complexation is to achieve complementation of the pro-active effect and amplification of the efficacy between strains including PGPR, and the combination of PGPR with other beneficial flora.
At present, although a plurality of composite microbial agents are combined, the existing microbial agents still have the defects of low survival rate, low infection rate and short shelf life, so that the effective viable count is greatly reduced during field application, and the field effect is also reduced to different degrees. For example, patent application CN201910314496.2 discloses a compound microbial fertilizer comprising a microbial agent and a microbial agent carrier; the microbial inoculum is prepared by mixing and activating bacillus subtilis, trichoderma harzianum, bacillus licheniformis and saccharomycetes; the microbial inoculum carrier comprises fermented sludge, fermented straw, nitrification inhibitor, fermented cow dung, water-retaining agent and the like. Patent application CN201811264239.4 discloses a composite microbial agent, which is formed by mixing a composite microbial agent, trace elements, diatomite and a drying agent; the composite microbial inoculum consists of bacillus subtilis, bacillus licheniformis, bacillus mucilaginosus and lactobacillus plantarum.
In addition, the existing microbial agents are still not deeply studied in efficacy, and most of the microbial agents are only applied to the field as beneficial microorganisms, so that the research on the influence on the growth and development of crops after application is lacking, and the blindness and the lack of pertinency of the fertilization of the existing microbial fertilizers are caused.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a novel microbial agent, a specific strain combination is screened as a functional strain to promote rooting, and meanwhile, an efficient carrier is used for loading the microbial agent, so that the stability and the durability of microbial action are improved.
In order to achieve the technical purpose, the invention adopts the following technical scheme:
the compound microbial agent for promoting rooting comprises microbial fermentation broth and a microbial carrier, wherein the volume mass ratio of the microbial fermentation broth to the microbial carrier is 1L:1kg.
Further, the microbial fermentation broth comprises an enterobacter oncogenic fermentation broth with a preservation number of CGMCC1.14969, a Azotobacter vinelandii fermentation broth with a preservation number of CGMCC1.7741 and a Bacillus licheniformis fermentation broth with a preservation number of CGMCC1.15832.
The enterobacter canceratus (Enterobacter cancerogenus) is purchased from China general microbiological culture collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 11 months and 20 days 2014, deposit number: CGMCC1.14969.
The invention relates to a Violet azotobacter (Azotobacter vinelandii), which is purchased from China general microbiological culture Collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 24 days of 2008, 7 months, deposit number: CGMCC1.7741.
The bacillus licheniformis (Bacillus paralicheniformis) of the invention is purchased from China general microbiological culture collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 20 days of 9 months of 2015, deposit number: CGMCC1.15832.
Furthermore, the volume ratio of the enterobacter canescens fermentation liquor, the Weinilan azotobacter fermentation liquor and the paraBacillus licheniformis fermentation liquor is 1:1:1.
Further, the preparation methods of the enterobacter canescens fermentation liquor, the Weinilan azotobacter fermentation liquor and the parabacillus licheniformis fermentation liquor are as follows: respectively activating enterobacter, azotobacter vinelandii and Bacillus paratlicheniformis, performing amplification culture and industrial fermentation to obtain effective viable bacteria with concentration of 1×10 4 -1×10 6 And mixing the three fermentation broths according to the volume ratio of 1:1:1 to obtain the microbial fermentation broth.
The method for activating, expanding and culturing the enterobacter canescens and the azotobacter vinelandii and the side bacillus licheniformis and carrying out industrial fermentation comprises the following steps:
A. respectively inoculating three strains into LB solid culture medium for activation, culturing at 28-30deg.C for 24h, respectively inoculating the activated three strains into LB liquid culture medium filled with 100mL, shake culturing at 28-30deg.C at 260r/min for 12h, and taking out as seed solution;
B. seed solutions of three strains are respectively mixed according to the volume ratio of 1:10 access fermenter containing 1LLB liquid MediumIndustrial fermentation is carried out in the presence of 1X 10 active bacteria 4 -1×10 6 And (3) completing fermentation at cfu/mL to obtain fermentation liquor of three bacteria.
The formula of the LB solid medium comprises the following components: 10g of tryptone, 10g of NaCl, 8g of yeast extract powder, 8g of sucrose, 15g of agar and 1000mL of distilled water, and controlling the pH value of the LB solid medium to be 7-7.5.
The formula of the LB liquid medium comprises the following components: 10g of tryptone, 10g of NaCl, 8g of yeast extract powder, 8g of sucrose and 1000ml of distilled water, and controlling the pH value of the LB liquid medium to be 7-7.5.
Further, the microbial carrier is diatomite, attapulgite and humic acid according to the mass ratio of 5:1: (1-3) and mixing.
Further, the microbial agent also comprises urea, chitosan and sodium dodecyl sulfonate, and the solid-to-liquid ratio of the microbial fermentation broth to the urea, the chitosan and the sodium dodecyl sulfonate is 1L: (1-5 g): (0.5-1 g): (0.1-0.5 g).
A preparation method of a compound microbial agent for promoting rooting comprises the following steps:
(1) Preparing a microbial fermentation broth: respectively activating enterobacter, azotobacter vinelandii and Bacillus paratlicheniformis, performing amplification culture and industrial fermentation to obtain effective viable bacteria with concentration of 1×10 4 -1×10 6 Mixing the three fermentation broths according to the volume ratio of 1:1:1 to obtain a microbial fermentation broth;
(2) Adding urea, chitosan and sodium dodecyl sulfate into the microbial fermentation broth, dispersing a microbial carrier in the mixed solution, enabling the carrier to fully adsorb the microbial fermentation broth, drying, and finally granulating to obtain the microbial agent.
The microbial agent is applied to soil before sowing, the application amount is 10-15 kg/mu, and the microbial agent is suitable for rooting of various crops, especially suitable for field crops such as corn, wheat and the like, has the effects of root increasing, rooting, root strengthening and the like, has the effects of disease resistance and growth promotion, and realizes the yield increase of crops.
Advantageous effects
The invention is compounded with three functional strains to form a core microbial inoculum, wherein the enterobacter canescens has the capabilities of dissolving phosphorus, producing indoleacetic acid and secreting auxin, has beneficial growth promoting characteristics, but is singly used, has single function, and the generated active substances are insufficient to stably act on soil for a long time. Therefore, the invention uses the Azotobacter vinelandii and the Bacillus paratyphenius simultaneously. The Weiniland azotobacter has strong nitrogen fixation effect, enriches the nutrient element composition in soil, can secrete a large amount of polysaccharide, enzyme and other bioactive substances, stimulates and promotes the activity of other microorganisms, and can generate plant hormone to promote the growth of crops. The side lichen bacillus can produce various antioxidant substances, remove the antioxidant substances, eliminate putrefaction, inhibit pathogenic bacteria and have obvious disease-resistant and growth-promoting effects. In particular, after the three bacteria are mixed in equal proportion, the root growth promoting effect of the strain is better than that of a single strain, the three bacteria are compounded for use and promote each other, and in practical application, the strain has stronger adaptability to the complex environment of the rhizosphere. Meanwhile, diatomite, attapulgite and the like are used as carriers, chitosan and sodium dodecyl sulfate are added to fully adsorb and disperse the microbial inoculum, so that the acting time and the acting stability of the microbial inoculum are prolonged, and urea humic acid and the like provide nutrient substances for crops and promote the growth of the crops.
Detailed Description
The technical scheme of the present invention is further described below with reference to specific examples, but is not limited thereto.
Example 1
The compound microbial agent for promoting rooting comprises microbial fermentation broth and a microbial carrier, wherein the volume mass ratio of the microbial fermentation broth to the microbial carrier is 1L:1kg.
The microbial fermentation broth comprises a bacillus subtilis fermentation broth with a preservation number of CGMCC1.14969, a azotobacter vinelandii fermentation broth with a preservation number of CGMCC1.7741 and a bacillus licheniformis fermentation broth with a preservation number of CGMCC1.15832.
Enterobacter carcinomatosis (Enterobacter cancerogenus), purchased from China general microbiological culture Collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 11 months and 20 days 2014, deposit number: CGMCC1.14969.
Azotobacter vinelandii (Azotobacter vinelandii), available from China general microbiological culture Collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 24 days of 2008, 7 months, deposit number: CGMCC1.7741.
Bacillus licheniformis (Bacillus paralicheniformis), purchased from China general microbiological culture collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 20 days of 9 months of 2015, deposit number: CGMCC1.15832.
The volume ratio of the enterobacter oncogenic fermentation liquor to the Azotobacter vinelandii fermentation liquor to the Bacillus licheniformis fermentation liquor is 1:1:1.
The preparation method of the enterobacter oncogenic fermentation broth, the Violet azotobacter vinelandii fermentation broth and the Bacillus licheniformis fermentation broth comprises the following steps: respectively activating enterobacter, azotobacter vinelandii and Bacillus paratlicheniformis, performing amplification culture and industrial fermentation to obtain effective viable bacteria with concentration of 1×10 4 -1×10 6 And mixing the three fermentation broths according to the volume ratio of 1:1:1 to obtain the microbial fermentation broth.
The method for activating, expanding and culturing the enterobacter canescens and the azotobacter vinelandii and the side bacillus licheniformis and carrying out industrial fermentation comprises the following steps:
A. respectively inoculating three strains into LB solid culture medium for activation, culturing at 28-30deg.C for 24h, respectively inoculating the activated three strains into LB liquid culture medium filled with 100mL, shake culturing at 28-30deg.C at 260r/min for 12h, and taking out as seed solution;
B. seed solutions of three strains are respectively mixed according to the volume ratio of 1:10 access to 1LLB liquidIndustrial fermentation is carried out in a fermenter of a bulk culture medium, and the effective viable bacteria concentration of each fermenter reaches 1X 10 4 -1×10 6 And (3) completing fermentation at cfu/mL to obtain fermentation liquor of three bacteria.
The formula of the LB solid medium comprises the following components: 10g of tryptone, 10g of NaCl, 8g of yeast extract powder, 8g of sucrose, 15g of agar and 1000mL of distilled water, and controlling the pH value of the LB solid medium to be 7-7.5.
The formula of the LB liquid medium comprises the following components: 10g of tryptone, 10g of NaCl, 8g of yeast extract powder, 8g of sucrose and 1000ml of distilled water, and controlling the pH value of the LB liquid medium to be 7-7.5.
The microbial carrier is diatomite, attapulgite and humic acid according to the mass ratio of 5:1:1, and mixing.
The microbial agent also comprises urea, chitosan and sodium dodecyl sulfonate, and the solid-liquid ratio of the microbial fermentation broth to the urea, the chitosan and the sodium dodecyl sulfonate is 1L:1g:0.5g:0.1g.
A preparation method of a compound microbial agent for promoting rooting comprises the following steps:
(1) Preparing a microbial fermentation broth: respectively activating enterobacter, azotobacter vinelandii and Bacillus paratlicheniformis, performing amplification culture and industrial fermentation to obtain effective viable bacteria with concentration of 1×10 4 -1×10 6 Mixing the three fermentation broths according to the volume ratio of 1:1:1 to obtain a microbial fermentation broth;
(2) Adding urea, chitosan and sodium dodecyl sulfate into the microbial fermentation broth, dispersing a microbial carrier in the mixed solution, enabling the carrier to fully adsorb the microbial fermentation broth, drying, and finally granulating to obtain the microbial agent.
Example 2
The compound microbial agent for promoting rooting comprises microbial fermentation broth and a microbial carrier, wherein the volume mass ratio of the microbial fermentation broth to the microbial carrier is 1L:1kg.
The microbial fermentation broth comprises a bacillus subtilis fermentation broth with a preservation number of CGMCC1.14969, a azotobacter vinelandii fermentation broth with a preservation number of CGMCC1.7741 and a bacillus licheniformis fermentation broth with a preservation number of CGMCC1.15832.
Enterobacter carcinomatosis (Enterobacter cancerogenus), purchased from China general microbiological culture Collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 11 months and 20 days 2014, deposit number: CGMCC1.14969.
Azotobacter vinelandii (Azotobacter vinelandii), available from China general microbiological culture Collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 24 days of 2008, 7 months, deposit number: CGMCC1.7741.
Bacillus licheniformis (Bacillus paralicheniformis), purchased from China general microbiological culture collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 20 days of 9 months of 2015, deposit number: CGMCC1.15832.
The volume ratio of the enterobacter oncogenic fermentation liquor to the Azotobacter vinelandii fermentation liquor to the Bacillus licheniformis fermentation liquor is 1:1:1.
The preparation method of the enterobacter oncogenic fermentation broth, the Violet azotobacter vinelandii fermentation broth and the Bacillus licheniformis fermentation broth comprises the following steps: respectively activating enterobacter, azotobacter vinelandii and Bacillus paratlicheniformis, performing amplification culture and industrial fermentation to obtain effective viable bacteria with concentration of 1×10 4 -1×10 6 And mixing the three fermentation broths according to the volume ratio of 1:1:1 to obtain the microbial fermentation broth.
The method for activating, expanding and culturing the enterobacter canescens and the azotobacter vinelandii and the side bacillus licheniformis and carrying out industrial fermentation comprises the following steps:
A. respectively inoculating three strains into LB solid culture medium for activation, culturing at 28-30deg.C for 24h, respectively inoculating the activated three strains into LB liquid culture medium filled with 100mL, shake culturing at 28-30deg.C at 260r/min for 12h, and taking out as seed solution;
B. seed solutions of three strains are respectively mixed according to the volume ratio of 1:10 are put into a fermentation tank filled with 1LLB liquid medium for industrial fermentation, and the effective viable bacteria concentration of each fermentation tank reaches 1 multiplied by 10 4 -1×10 6 And (3) completing fermentation at cfu/mL to obtain fermentation liquor of three bacteria.
The formula of the LB solid medium comprises the following components: 10g of tryptone, 10g of NaCl, 8g of yeast extract powder, 8g of sucrose, 15g of agar and 1000mL of distilled water, and controlling the pH value of the LB solid medium to be 7-7.5.
The formula of the LB liquid medium comprises the following components: 10g of tryptone, 10g of NaCl, 8g of yeast extract powder, 8g of sucrose and 1000ml of distilled water, and controlling the pH value of the LB liquid medium to be 7-7.5.
The microbial carrier is diatomite, attapulgite and humic acid according to the mass ratio of 5:1:2, mixing.
The microbial agent also comprises urea, chitosan and sodium dodecyl sulfonate, and the solid-liquid ratio of the microbial fermentation broth to the urea, the chitosan and the sodium dodecyl sulfonate is 1L:3g:0.8g:0.3g.
A preparation method of a compound microbial agent for promoting rooting comprises the following steps:
(1) Preparing a microbial fermentation broth: respectively activating enterobacter, azotobacter vinelandii and Bacillus paratlicheniformis, performing amplification culture and industrial fermentation to obtain effective viable bacteria with concentration of 1×10 4 -1×10 6 Mixing the three fermentation broths according to the volume ratio of 1:1:1 to obtain a microbial fermentation broth;
(2) Adding urea, chitosan and sodium dodecyl sulfate into the microbial fermentation broth, dispersing a microbial carrier in the mixed solution, enabling the carrier to fully adsorb the microbial fermentation broth, drying, and finally granulating to obtain the microbial agent.
Example 3
The compound microbial agent for promoting rooting comprises microbial fermentation broth and a microbial carrier, wherein the volume mass ratio of the microbial fermentation broth to the microbial carrier is 1L:1kg.
The microbial fermentation broth comprises a bacillus subtilis fermentation broth with a preservation number of CGMCC1.14969, a azotobacter vinelandii fermentation broth with a preservation number of CGMCC1.7741 and a bacillus licheniformis fermentation broth with a preservation number of CGMCC1.15832.
Enterobacter carcinomatosis (Enterobacter cancerogenus), purchased from China general microbiological culture Collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 11 months and 20 days 2014, deposit number: CGMCC1.14969.
Azotobacter vinelandii (Azotobacter vinelandii), available from China general microbiological culture Collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 24 days of 2008, 7 months, deposit number: CGMCC1.7741.
Bacillus licheniformis (Bacillus paralicheniformis), purchased from China general microbiological culture collection center (China General Microbiological Culture CollectionCenter, CGMCC), address: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 20 days of 9 months of 2015, deposit number: CGMCC1.15832.
The volume ratio of the enterobacter oncogenic fermentation liquor to the Azotobacter vinelandii fermentation liquor to the Bacillus licheniformis fermentation liquor is 1:1:1.
The preparation method of the enterobacter oncogenic fermentation broth, the Violet azotobacter vinelandii fermentation broth and the Bacillus licheniformis fermentation broth comprises the following steps: respectively activating enterobacter, azotobacter vinelandii and Bacillus paratlicheniformis, performing amplification culture and industrial fermentation to obtain effective viable bacteria with concentration of 1×10 4 -1×10 6 And mixing the three fermentation broths according to the volume ratio of 1:1:1 to obtain the microbial fermentation broth.
The method for activating, expanding and culturing the enterobacter canescens and the azotobacter vinelandii and the side bacillus licheniformis and carrying out industrial fermentation comprises the following steps:
A. respectively inoculating three strains into LB solid culture medium for activation, culturing at 28-30deg.C for 24h, respectively inoculating the activated three strains into LB liquid culture medium filled with 100mL, shake culturing at 28-30deg.C at 260r/min for 12h, and taking out as seed solution;
B. seed solutions of three strains are respectively mixed according to the volume ratio of 1:10 are put into a fermentation tank filled with 1LLB liquid medium for industrial fermentation, and the effective viable bacteria concentration of each fermentation tank reaches 1 multiplied by 10 4 -1×10 6 And (3) completing fermentation at cfu/mL to obtain fermentation liquor of three bacteria.
The formula of the LB solid medium comprises the following components: 10g of tryptone, 10g of NaCl, 8g of yeast extract powder, 8g of sucrose, 15g of agar and 1000mL of distilled water, and controlling the pH value of the LB solid medium to be 7-7.5.
The formula of the LB liquid medium comprises the following components: 10g of tryptone, 10g of NaCl, 8g of yeast extract powder, 8g of sucrose and 1000ml of distilled water, and controlling the pH value of the LB liquid medium to be 7-7.5.
The microbial carrier is diatomite, attapulgite and humic acid according to the mass ratio of 5:1:3, mixing.
The microbial agent also comprises urea, chitosan and sodium dodecyl sulfonate, and the solid-liquid ratio of the microbial fermentation broth to the urea, the chitosan and the sodium dodecyl sulfonate is 1L:5g:1g:0.5g.
A preparation method of a compound microbial agent for promoting rooting comprises the following steps:
(1) Preparing a microbial fermentation broth: respectively activating enterobacter, azotobacter vinelandii and Bacillus paratlicheniformis, performing amplification culture and industrial fermentation to obtain effective viable bacteria with concentration of 1×10 4 -1×10 6 Mixing the three fermentation broths according to the volume ratio of 1:1:1 to obtain a microbial fermentation broth;
(2) Adding urea, chitosan and sodium dodecyl sulfate into the microbial fermentation broth, dispersing a microbial carrier in the mixed solution, enabling the carrier to fully adsorb the microbial fermentation broth, drying, and finally granulating to obtain the microbial agent.
Comparative example 1
The kinds and proportions of functional bacteria were changed, and the other preparation methods were the same as in example 3, and comparative examples were set. The volume ratio of the enterobacter oncogenic fermentation broth, the azotobacter vinelandii fermentation broth and the Bacillus licheniformis fermentation broth is set to be 1:2:1, and the rest raw materials and the preparation process are the same as in example 3.
Comparative example 2
The kinds and proportions of functional bacteria were changed, and the other preparation methods were the same as in example 3, and comparative examples were set. The volume ratio of the enterobacter oncogenic fermentation broth, the azotobacter vinelandii fermentation broth and the Bacillus licheniformis fermentation broth is set to be 1:1:2, and the rest raw materials and the preparation process are the same as in example 3.
Comparative example 3
The kinds and proportions of functional bacteria were changed, and the other preparation methods were the same as in example 3, and comparative examples were set. The volume ratio of the enterobacter oncogenic fermentation broth, the azotobacter vinelandii fermentation broth and the Bacillus licheniformis fermentation broth is set to be 2:1:1, and the rest raw materials and the preparation process are the same as in example 3.
Comparative example 4
The kinds and proportions of functional bacteria were changed, and the other preparation methods were the same as in example 3, and comparative examples were set. Namely, the volume ratio of the intestinal bacillus subtilis fermentation liquor, the Azotobacter vinelandii fermentation liquor and the Bacillus licheniformis fermentation liquor is set to be 1:1:0, namely, only two bacteria fermentation liquor is used, and the rest raw materials and the preparation process are the same as those of the example 3.
Comparative example 5
The kinds and proportions of functional bacteria were changed, and the other preparation methods were the same as in example 3, and comparative examples were set. Namely, the volume ratio of the intestinal bacillus subtilis fermentation liquor, the Azotobacter vinelandii fermentation liquor and the Bacillus licheniformis fermentation liquor is set to be 1:0:1, namely, only two bacteria fermentation liquor is used, and the rest raw materials and the preparation process are the same as those of the example 3.
Comparative example 6
The kinds and proportions of functional bacteria were changed, and the other preparation methods were the same as in example 3, and comparative examples were set. Namely, the volume ratio of the intestinal bacillus subtilis fermentation liquor, the Azotobacter vinelandii fermentation liquor and the Bacillus licheniformis fermentation liquor is set to be 0:1:1, namely, only two bacteria fermentation liquor is used, and the rest raw materials and the preparation process are the same as those of the example 3.
Comparative example 7
The kinds and proportions of functional bacteria were changed, and the other preparation methods were the same as in example 3, and comparative examples were set. Namely, the volume ratio of the enterobacter oncogenic fermentation broth, the azotobacter vinelandii fermentation broth and the Bacillus licheniformis fermentation broth is set to be 1:0:0, namely, only one bacterial fermentation broth is used, and the rest raw materials and the preparation process are the same as those in example 3.
Comparative example 8
The kinds and proportions of functional bacteria were changed, and the other preparation methods were the same as in example 3, and comparative examples were set. Namely, the volume ratio of the enterobacter oncogenic fermentation broth, the azotobacter vinelandii fermentation broth and the Bacillus licheniformis fermentation broth is set to be 0:1:0, namely, only one bacterial fermentation broth is used, and the rest raw materials and the preparation process are the same as those in example 3.
Comparative example 9
The kinds and proportions of functional bacteria were changed, and the other preparation methods were the same as in example 3, and comparative examples were set. Namely, the volume ratio of the enterobacter oncogenic fermentation broth, the azotobacter vinelandii fermentation broth and the Bacillus licheniformis fermentation broth is set to be 0:0:1, namely, only one bacterial fermentation broth is used, and the rest raw materials and the preparation process are the same as those in example 3.
Firstly, the positive effect of microbial fermentation bacteria liquid on promoting rooting is examined, sterile triangular flasks (100 ml) are taken and divided into 13 groups, 3 are arranged in each triangular flask, 5 waxy corn seeds are placed in each triangular flask, and the microbial fermentation bacteria liquid obtained in examples 1-3 and comparative examples 1-9 are respectively added into the corresponding triangular flasks so that the fermentation liquid just penetrates half of the seeds. While group 13 was added sterile water as a blank. After 15 days of culture, the CI600 plant root system growth monitoring system counts the average total root length, the average root number and the average root diameter of the corn. The experimental results are shown in table 1:
TABLE 1 rooting of maize
| Average total root length of individual plant cm | Average number of individual plants | Average root diameter mm | |
| Example 1 | 121.65 | 62.36 | 3.69 |
| Example 2 | 120.69 | 61.89 | 3.66 |
| Example 3 | 121.18 | 62.78 | 3.71 |
| Comparative example 1 | 112.69 | 56.12 | 3.12 |
| Comparative example 2 | 113.77 | 56.05 | 3.12 |
| Comparative example 3 | 114.98 | 56.99 | 3.11 |
| Comparative example 4 | 101.36 | 43.21 | 3.05 |
| Comparative example 5 | 100.26 | 41.22 | 3.03 |
| Comparative example 6 | 96.59 | 40.36 | 3.01 |
| Comparative example 7 | 65.37 | 36.96 | 3.00 |
| Comparative example 8 | 62.78 | 36.10 | 2.91 |
| Comparative example 9 | 61.23 | 35.87 | 2.85 |
| Sterile water control group | 11.20 | 8.12 | 1.20 |
As can be seen from the data in Table 1, the fermentation broths of the three strains of the invention have strong rooting effect on corn, while the synergistic balance effect among the three strains is broken and the rooting effect is slightly weakened in comparative examples 1-9 with the composition and the proportion of the microbial inoculum changed. In order to further verify the practical application effect, a planting test is performed.
Planting test
Crop: corn, zhongnuo No. one
Basic conditions of test field: the test is carried out in a test field of the agricultural academy of sciences in Yi city in 2021, the soil is sandy soil, and the soil nutrition condition is as follows: 110.6mg/kg of alkaline hydrolysis nitrogen, 129.0mg/kg of total nitrogen, 15.9mg/kg of quick-acting phosphorus, 140mg/kg of quick-acting potassium and 132mg/kg of organic matters.
The test was performed in 10 treatments, each repeated 3 times, and the results were averaged. The base fertilizer is applied to each treatment group conventionally, and the application amount of each mu of N, P, K is respectively 10 kg/mu, 6 kg/mu and 7 kg/mu.
Treatment 1: conventional fertilization + 10 kg/mu of example 3 microbial inoculum;
treatment 2: conventional fertilization and 10 kg/mu of microbial inoculum of comparative example 1;
treatment 3: conventional fertilization and 10 kg/mu of microbial inoculum of comparative example 2;
treatment 4: conventional fertilization and comparative example 3 microbial inoculum 10 kg/mu;
treatment 5: conventional fertilization and comparative example 4 microbial inoculum 10 kg/mu;
treatment 6: conventional fertilization and 10 kg/mu of the microbial inoculum of comparative example 5;
treatment 7: conventional fertilization and comparative example 6 microbial inoculum 10 kg/mu;
treatment 8: conventional fertilization and comparative example 7 microbial inoculum 10 kg/mu;
treatment 9: conventional fertilization and 10 kg/mu of the microbial inoculum of comparative example 8;
treatment 10: conventional fertilization and comparative example 9 microbial inoculum 10 kg/mu;
treatment 11: and (5) conventional fertilization.
Test measurement index and method
Measuring yield and checking species: 10 ears of corn are randomly selected for seed examination in the mature period, and main factors such as yield and yield increase are measured. The yield per cell was calculated as 2 lines per cell and the yield per unit area was converted to 14% water content.
Dry root system: when corn grows to 7 leaf period, 3 representative corn root systems are collected in each district, the root systems are washed clean by clean water, then the surface moisture is absorbed by filter paper, and the corn is dried to constant weight at 80 ℃, and the average root system dry weight of each treated corn plant is calculated.
TABLE 2 planting test results
| Root system dry weight (g/plant) | Yield (kg/hm) 2 ） | Yield increase (%) | Thousand grain weight/g | |
| Process 1 | 2.12 | 11789.6 | 12.03 | 351.69 |
| Process 2 | 2.01 | 11674.5 | 10.94 | 346.22 |
| Process 3 | 2.01 | 11602.3 | 10.25 | 345.01 |
| Process 4 | 2.03 | 11623.9 | 10.46 | 345.55 |
| Process 5 | 1.91 | 11321.3 | 7.58 | 332.15 |
| Process 6 | 1.88 | 11210.1 | 6.53 | 330.47 |
| Process 7 | 1.85 | 11212.9 | 6.55 | 330.55 |
| Process 8 | 1.52 | 10693.1 | 1.61 | 323.69 |
| Process 9 | 1.52 | 10699.9 | 1.68 | 324.78 |
| Process 10 | 1.49 | 10702.9 | 1.71 | 325.87 |
| Process 11 | 1.12 | 10523.3 | - | 319.56 |
It should be noted that the above-mentioned embodiments are merely some, but not all embodiments of the preferred mode of carrying out the invention. It is evident that all other embodiments obtained by a person skilled in the art without making any inventive effort, based on the above-described embodiments of the invention, shall fall within the scope of protection of the invention.
Claims (2)
1. The compound microbial agent for promoting rooting is characterized by comprising microbial fermentation broth and a microbial carrier, wherein the volume mass ratio of the microbial fermentation broth to the microbial carrier is 1L:1kg;
the microbial fermentation broth comprises a bacillus subtilis fermentation broth with a preservation number of CGMCC1.14969, a azotobacter vinelandii fermentation broth with a preservation number of CGMCC1.7741 and a bacillus licheniformis fermentation broth with a preservation number of CGMCC 1.15832;
the volume ratio of the enterobacter oncogenic fermentation liquor to the Violet azotobacter vinelandii fermentation liquor to the auxiliary Bacillus licheniformis fermentation liquor is 1:1:1;
the preparation method of the enterobacter oncogenic fermentation broth, the Violet azotobacter vinelandii fermentation broth and the Bacillus licheniformis fermentation broth comprises the following steps: respectively activating enterobacter, azotobacter vinelandii and Bacillus paratlicheniformis, performing amplification culture and industrial fermentation to obtain effective viable bacteria with concentration of 1×10 4 -1×10 6 Mixing the three fermentation broths according to the volume ratio of 1:1:1 to obtain a microbial fermentation broth;
the microbial carrier is diatomite, attapulgite and humic acid according to the mass ratio of 5:1: (1-3) mixing;
the microbial agent also comprises urea, chitosan and sodium dodecyl sulfonate, and the solid-liquid ratio of the microbial fermentation broth to the urea, the chitosan and the sodium dodecyl sulfonate is 1L: (1-5 g): (0.5-1 g): (0.1-0.5 g).
2. A method for preparing the rooting-promoting composite microbial agent according to claim 1, which is characterized by comprising the following steps:
(1) Preparing a microbial fermentation broth: respectively activating enterobacter, azotobacter vinelandii and Bacillus paratlicheniformis, performing amplification culture and industrial fermentation to obtain effective viable bacteria with concentration of 1×10 4 -1×10 6 Mixing the three fermentation broths according to the volume ratio of 1:1:1 to obtain a microbial fermentation broth;
(2) Adding urea, chitosan and sodium dodecyl sulfate into the microbial fermentation broth, dispersing a microbial carrier in the mixed solution, enabling the carrier to fully adsorb the microbial fermentation broth, drying, and finally granulating to obtain the microbial agent.
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