CN113185354A - Microbial agent capable of effectively promoting crop rooting and preparation method thereof - Google Patents
Microbial agent capable of effectively promoting crop rooting and preparation method thereof Download PDFInfo
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- CN113185354A CN113185354A CN202110485693.8A CN202110485693A CN113185354A CN 113185354 A CN113185354 A CN 113185354A CN 202110485693 A CN202110485693 A CN 202110485693A CN 113185354 A CN113185354 A CN 113185354A
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- 230000001737 promoting effect Effects 0.000 title claims abstract description 16
- 238000000855 fermentation Methods 0.000 claims abstract description 111
- 230000004151 fermentation Effects 0.000 claims abstract description 111
- 241000194032 Enterococcus faecalis Species 0.000 claims abstract description 45
- 229940032049 enterococcus faecalis Drugs 0.000 claims abstract description 44
- 239000003337 fertilizer Substances 0.000 claims abstract description 31
- 239000007788 liquid Substances 0.000 claims abstract description 31
- 241000194107 Bacillus megaterium Species 0.000 claims abstract description 29
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- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 25
- 230000001580 bacterial effect Effects 0.000 claims abstract description 21
- 238000001035 drying Methods 0.000 claims abstract description 17
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- 239000003795 chemical substances by application Substances 0.000 claims abstract description 12
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- 238000002156 mixing Methods 0.000 claims abstract description 11
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- 229910052700 potassium Inorganic materials 0.000 claims abstract description 11
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- 238000000034 method Methods 0.000 claims abstract description 8
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- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 9
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 9
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 8
- 239000002054 inoculum Substances 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims 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 description 7
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- 230000001954 sterilising effect Effects 0.000 claims description 7
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- XQGPKZUNMMFTAL-UHFFFAOYSA-L dipotassium;hydrogen phosphate;trihydrate Chemical compound O.O.O.[K+].[K+].OP([O-])([O-])=O XQGPKZUNMMFTAL-UHFFFAOYSA-L 0.000 claims description 5
- XNCMOUSLNOHBKY-UHFFFAOYSA-H iron(3+);trisulfate;heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+3].[Fe+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O XNCMOUSLNOHBKY-UHFFFAOYSA-H 0.000 claims description 5
- 239000011777 magnesium Substances 0.000 claims description 5
- 229910052749 magnesium Inorganic materials 0.000 claims description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 5
- 229910052757 nitrogen Inorganic materials 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 239000001509 sodium citrate Substances 0.000 claims description 5
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims description 5
- 229960004793 sucrose Drugs 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 4
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 4
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 4
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- 239000007858 starting material Substances 0.000 claims description 2
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- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical class C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/02—Other organic fertilisers from peat, brown coal, and similar vegetable deposits
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
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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
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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
- C05G3/80—Soil conditioners
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- 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
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Abstract
The invention discloses a microbial agent capable of effectively promoting crop rooting and a preparation method thereof, and mainly relates to the field of biological bacterial fertilizers. The method comprises the following steps: fermenting enterococcus faecalis, performing secondary fermentation on saccharomycetes, introducing the saccharomycetes into the enterococcus faecalis for mixed fermentation, adding bacillus subtilis liquid and bacillus megaterium liquid into mixed fermentation liquid, wherein the proportion of the bacillus subtilis liquid to the bacillus megaterium liquid is 15-25 parts by volume: 15-25 parts of bacillus megatherium: mixing 50-70 parts of fermentation liquor, uniformly mixing, and adding 5-10% of potassium fulvate in mass proportion of the mixed liquor to obtain basic mother liquor; and (4) boiling and drying the basic mother liquor to obtain a finished bacterial fertilizer product. The invention has the beneficial effects that: it can effectively promote the rooting of plants and the absorption of nutrient elements, thereby promoting the growth of plants.
Description
Technical Field
The invention relates to the field of biological bacterial fertilizers, in particular to a microbial agent capable of effectively promoting crop rooting and a preparation method thereof.
Background
In recent decades, agricultural planting in China uses a great amount of chemical fertilizers and pesticides, so that a series of problems such as soil hardening, soil acidification, soil-borne diseases and the like occur successively. The problems of long-term excessive use of chemical fertilizers and pesticides, high multiple cropping index and soil continuous productivity obstacle caused by continuous cropping cause the problems of soil quality health and agricultural product quality safety. The soil environment of China is not optimistic overall, 20.27 hundred million acres of cultivated land exist in China, and 19.4% of cultivated land pollutants exceed the standard. Soil conditioners are needed to solve the problems occurring in the soil. The microbial fertilizer plays a role in converting soil organic matters into humus, can increase the granular structure of soil, improve the water and fertilizer retention capacity, activate nutrients solidified by the soil, improve the utilization rate of a fertilizer, and simultaneously, beneficial microorganisms secrete various beneficial substances and various enzymes to the soil, so that the conversion of the nutrients is effectively promoted, the occurrence of soil-borne diseases is reduced, and the polluted soil is repaired.
The existing microorganism is in line with the defect of slow effect when the bacterial manure is specifically applied, and a blank window period which cannot effectively play a role exists in the bacterial planting period, so that the effect period is long, and the application effect of the product is influenced.
Disclosure of Invention
The invention aims to provide a microbial agent capable of effectively promoting crop rooting and a preparation method thereof, and the microbial agent can effectively promote plant rooting and nutrient element absorption so as to promote plant growth.
In order to achieve the purpose, the invention is realized by the following technical scheme:
a microbial agent capable of effectively promoting crop rooting comprises the following steps:
fermenting enterococcus faecalis to obtain a primary enterococcus faecalis strain;
secondary fermentation is carried out on the saccharomycetes to obtain primary saccharomycetes;
introducing the primary yeast strain into the primary enterococcus faecalis strain for mixed fermentation to obtain mixed fermentation liquor;
adding bacillus subtilis liquid and bacillus megatherium liquid into the mixed fermentation liquid, wherein the ratio of the bacillus subtilis liquid to the bacillus megatherium liquid is 15-25 parts by volume: 15-25 parts of bacillus megatherium: mixing 50-70 parts of fermentation liquor, uniformly mixing, and adding 5-10% of potassium fulvate in mass proportion of the mixed liquor to obtain basic mother liquor;
and (4) boiling and drying the basic mother liquor to obtain a finished bacterial fertilizer product.
Further, in the enterococcus faecalis primary species, not less than 15 single bacteria are observed in each visual field under a microscope with a magnification of 16 times and 100 times;
the content of the yeast in the primary yeast strain is more than 0.5 hundred million/ml.
Further, the mixed fermentation comprises the steps of introducing primary yeast strains into primary enterococcus faecalis strains, wherein the total amount is 7-9 tons, stirring is set to be 180-220rpm/min, the air volume is 220-250 cubic per hour, the temperature is set to be 35-37 ℃, mixed fermentation is started, observation is carried out after 8-12 hours of fermentation, more than 20 enterococcus faecalis in a 16 x 100-time amplified visual field, the count of saccharomycetes by a haemocytometer is more than 1 hundred million/ml, ammonia water is fed again to keep the pH value between 5.2-5.5, and the mixed fermentation is kept for 10-15 hours, so that the feeding is stopped when the feeding amount of the ammonia water is more than 0.5 ton, and the mixed fermentation is kept for 5-6 hours, thus obtaining the yeast enterococcus faecalis obtained.
Further, the enterococcus faecalis fermentation comprises the following steps:
inoculating enterococcus faecalis in the culture medium with the inoculation amount of 5-15%, controlling the fermentation temperature between 30-40 deg.C, standing and fermenting;
introducing air at the same interval in the fermentation process, setting the air flow to be 150 cubic/h at the time of 15 seconds every time for 3 hours, culturing the mixture to 12 to 16 hours, observing the mixture under a microscope of 16 times to 100 times, adding ammonia water when the air flow is not less than 5 single bacteria in each visual field, adjusting the pH to 5.8 to 6.2, continuing fermenting the mixture to 24 to 30 hours, adding ammonia water to keep the pH between 5.0 and 6.0, observing the mixture under a microscope of 16 times to 100 times, stopping the feeding when the air flow is not less than 15 single bacteria in each visual field, and obtaining the enterococcus faecalis primary strain.
Further, the yeast fermentation comprises:
inoculating yeast into a 37 ℃ yeast culture medium, wherein the inoculation amount is 3-7%, the culture temperature is set to be 37 ℃, the rotating speed is 100rpm/min, the rotating speed is increased by 20rpm every 1 hour until 220rpm/min is kept unchanged, the ventilation volume is 120 cubic meters per hour initially, the ventilation volume is increased to 300 cubic meters per hour after 8 hours, and after 12-18 hours of culture, not less than 3-5 yeasts are in a microscopic view field to obtain a first-grade yeast strain;
transferring all the primary yeast strains into a secondary fermentation tank, adding a yeast culture medium, and metering the volume of the secondary fermentation tank to 3.5-4 tons at 10 tons; culturing in secondary fermentation tank for 7-15 hr until the content of yeast is greater than 0.5 hundred million/ml to obtain primary yeast strain.
Further, the raw materials of the culture medium of the enterococcus faecalis comprise: 6% of molasses, 0.4% of yeast powder, 0.4% of peptone, 0.03% of dipotassium hydrogen phosphate and 5% of brown sugar;
the raw materials of the culture medium of the yeast comprise: 1% of yeast powder, 0.5% of peptone and 0.5% of sodium chloride.
Further, the bacillus subtilis liquid is prepared by the following method: 20g/L of sucrose, 5g/L of ammonium sulfate, 45g/L of bran, 2.0g/L of sodium citrate, 0.4g/L of dipotassium phosphate trihydrate, 0.5g/L of magnesium sulfate heptahydrate and 0.03g/L of ferric sulfate heptahydrate, wherein the pH value is natural, the mixture is fermented for 24-38 hours at 37 ℃ according to the inoculum size of 7%, the viable count reaches 200 hundred million/ml, and the spore formation rate is more than 98%, so that the composition is obtained;
the bacillus megaterium liquid is prepared by the following method: 18g/L of corn paste, 55g/L of molasses, 0.5g/L of calcium chloride, 3g/L of dipotassium phosphate and 0.4g/L of magnesium sulfate, wherein the pH value of the fermentation start is 6.8-7.2; fermenting at 37 deg.C for 24-30 hr to obtain fermented product with viable count of 5 hundred million/ml and spore rate of over 90%.
Furthermore, the bacterial fertilizer has the moisture content of less than 15 percent and is granular, the content of humic acid in the bacterial fertilizer is 5-7 percent, the content of magnesium is 3-7 percent, the content of nitrogen is 10-17 percent, and the content of bacillus subtilis and bacillus megatherium is 2-15 hundred million/g.
The preparation method of the compound microbial fertilizer is another aspect of the invention.
Compared with the prior art, the invention has the beneficial effects that:
the microbial agent produced by the method contains metabolites (microbial derivatives) produced by complex fermentation of various microorganisms, including organic acids, various enzymes, auxin analogs and the like, and can effectively promote rooting of plants and absorption of nutrient elements, thereby promoting growth of the plants.
The traditional microbial agent needs an adaptation period of 3-5 days after being applied to cultivated land, then gradually propagates in soil, the population quantity starts to increase, secondary metabolites generated in the propagation process can promote plant growth and prevent diseases, the whole period starts to gradually show effects at least 7-10 days later, the product enriches the metabolites in the fermentation process, can play a role in promoting plant growth 3-5 days after being applied to cultivated land, and greatly shortens the period of effectiveness.
Therefore, the product effectively overcomes the defect of slow effect of the microbial agent, and a plurality of water-soluble metabolites begin to act during the field planting of strains, thereby greatly shortening the effect taking period of the microbial fertilizer.
Drawings
FIG. 1 is a flow chart of the present invention.
FIG. 2 is a photograph showing the results of the verification test in example 2 of the present invention.
FIG. 3 is a photograph showing the results of a verification experiment in example 3 of the present invention.
Figure 4 is a photograph of a product of the invention.
FIG. 5 is a photograph of an experimental sample of examples 2 and 3 of the present invention.
FIG. 6 is a report of microbial detection of the product obtained in example 1 of the present invention
FIG. 7 is a report of the detection of Bacillus megaterium in the product of example 1 of the present invention.
FIG. 8 is a report of Bacillus subtilis detection in the product of example 1 of the present invention.
FIG. 9 shows the microscopic examination results of example 1 of the present invention (the left picture shows primary yeast species (16X 100) and the right picture shows primary enterococcus faecalis species (16X 100))
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and these equivalents also fall within the scope of the present application.
The instruments, reagents, materials and the like used in the following examples are conventional instruments, reagents, materials and the like in the prior art and are commercially available in a normal manner unless otherwise specified. Unless otherwise specified, the experimental methods, detection methods, and the like described in the following examples are conventional experimental methods, detection methods, and the like in the prior art.
Example 1: preparation of composite microbial fertilizer
1. Enterococcus faecalis fermentation
1.1 preparation of culture medium for enterococcus faecalis fermentation
The formula comprises the following components: 6% of molasses, 0.4% of yeast powder, 0.4% of peptone, 0.03% of dipotassium hydrogen phosphate, 5% of brown sugar and the balance of softened water;
preparing according to the formula, keeping the pH value natural, metering to 3.8 tons in a 10-ton fermentation tank, sterilizing at 116 deg.C under 0.8-0.9Mpa for 30min, and cooling to room temperature to obtain enterococcus faecalis culture medium for use.
1.2 enterococcus faecalis fermentation
Inoculating enterococcus faecalis into the culture medium in the step 1), controlling the fermentation temperature to be 38-40 ℃, and standing for fermentation, wherein the inoculation amount is 10%.
Air is introduced into the fermentation process at the same time interval, the time is set to be 3 hours, 15 seconds are carried out each time, and the air volume is 120 cubic meters per hour.
Culturing for 13.5 hours, observing under a microscope of 16X 100 times, if 7 single bacteria exist in each visual field, adding ammonia water, adjusting the pH to 5.8-6.2, continuing to ferment for 28 hours, constantly adding ammonia water to maintain the pH between 5.0-6.0, observing under a microscope of 16X 100 times, if 20 single bacteria exist in each visual field, stopping adding, and obtaining the enterococcus faecalis primary strain. The introduction of the yeast primary species can begin.
2. Fermentation with yeast
2.1 preparation of Yeast culture Medium
The formula is as follows: 1% of yeast powder, 0.5% of peptone, 0.5% of sodium chloride and the balance of softened water.
Adjusting pH to 5.55, adding into 200L fermentation tank, diluting to 130L, sterilizing at 121 deg.C and 0.9-1.0Mpa for 20min, and cooling to 37 deg.C to obtain yeast culture medium;
2.2 Yeast inoculation and Primary fermentation
Inoculating yeast into a 37 ℃ yeast culture medium, wherein the inoculation amount is 5%, the culture temperature is set to be 37 ℃, the rotating speed is 100rpm/min, the rotating speed is increased by 20rpm every 1 hour until the rotating speed is 220rpm/min, the rotating speed is kept still, the ventilation volume is 120 cubic meters per hour initially, and the ventilation volume is increased to 300 cubic meters per hour after 8 hours. After 14.5 hours of culture, 5 yeasts are in the visual field of microscopic examination, and a primary yeast strain is obtained.
2.3 Secondary fermentation of Yeast
Transferring the primary yeast strains into a secondary fermentation tank, using 2.1 yeast culture medium, and metering the volume of the secondary fermentation tank to 3.5-4 tons for 10 tons.
Culturing in secondary fermentation tank for 11 hr until the content of yeast is greater than 0.68 hundred million/ml, and completing secondary fermentation to obtain primary yeast strain.
3. Preparation of Bacillus subtilis liquid
The preparation method comprises the following steps: the culture medium comprises 20g/L of cane sugar, 5g/L of ammonium sulfate, 45g/L of bran, 2.0g/L of sodium citrate, 0.4g/L of dipotassium phosphate trihydrate, 0.5g/L of magnesium sulfate heptahydrate and 0.03g/L of ferric sulfate heptahydrate, the pH value is natural, the inoculation amount is 7%, after fermentation is carried out for 28 hours at 37 ℃, the viable count reaches 200 hundred million/ml, the spore formation rate is more than 98%, and the culture is stopped for later use.
4. Preparation of Bacillus megaterium (TMQJ05) bacterial liquid
The preparation method comprises the following steps: in the culture medium, 18g/L of corn paste, 55g/L of molasses, 3g/L of calcium chloride and 0.4g/L of magnesium sulfate are used, and the pH of the fermentation starting material is 6.8-7.2; after fermenting for 27 hours at 37 ℃, the number of viable bacteria can reach 5 hundred million/ml, the spore rate is more than 90 percent, and the culture is stopped for standby.
5. Mixed fermentation
Introducing primary yeast strains into primary enterococcus faecalis strains, wherein the total amount is 8.5 tons, stirring is set to be 220rpm/min, the air volume is 230 cubic per hour, the temperature is set to be 37 ℃, mixed fermentation is started, microscopic examination is performed every two hours to ensure that normal fermentation is not infected with infectious microbes, observation is performed after 10 hours of mixed fermentation, 23 enterococcus faecalis exist in a 16 x 100-time enlarged visual field, the count of yeast by a haemocytometer exceeds 1 hundred million/ml, ammonia water is fed again to keep the pH value between 5.2 and 5.5, the pH value is kept for 12.3 hours, the feeding is stopped when the feeding amount of the ammonia water is more than 0.55 tons, and the mixed fermentation is kept for 5.5 hours, so that mixed fermentation liquid is obtained.
6. Composite fermentation
Adding the fermented bacillus subtilis and bacillus megatherium into the mixed fermentation liquor, wherein the ratio is as follows: 22 parts of bacillus subtilis: 17 parts of bacillus megatherium: and mixing 66 parts of mixed fermentation liquor, fully stirring, and adding 9% of potassium fulvate (requiring that the potassium fulvate is 65% of humic acid and the potassium content is more than or equal to 18.2%, and is fully water-soluble) in mass proportion of the mixed liquor to obtain the basic mother liquor.
7. Fluidized drying
And (4) carrying out boiling drying on the basic mother liquor.
The carrier, namely the boiling drying carrier, is one of magnesium sulfate heptahydrate and ammonium sulfate or the arbitrary proportion of the two, and the mesh number of the magnesium sulfate heptahydrate and the ammonium sulfate is required to be 10-20 meshes.
Proportioning-the proportion of boiling drying carrier and mother liquor is 9 parts: 10 portions, the air inlet temperature in the drying process is 102 ℃, the air outlet temperature is 53 ℃, and the material temperature is 62 ℃.
The final product, namely the compound microbial fertilizer, is obtained by boiling and drying, wherein the water content of the bacterial fertilizer is less than 8.5 percent, the bacterial fertilizer is granulated, the content of humic acid is 6.8 percent, the content of magnesium is 4.0 percent, the content of nitrogen is 11.4 percent, and the content of bacillus subtilis and bacillus megatherium is 14.3 hundred million/g. Example 2: preparation of composite microbial fertilizer
1. Enterococcus faecalis fermentation
1.1 preparation of culture medium for enterococcus faecalis fermentation
The formula comprises the following components: 6% of molasses, 0.4% of yeast powder, 0.4% of peptone, 0.03% of dipotassium hydrogen phosphate, 5% of brown sugar and the balance of softened water;
preparing according to the formula, keeping the pH value natural, metering to 4 tons in a 10-ton fermentation tank, sterilizing at 116 deg.C under 0.8-0.9Mpa for 30min, and cooling to room temperature to obtain enterococcus faecalis culture medium for use.
1.2 enterococcus faecalis fermentation
Inoculating enterococcus faecalis into the culture medium in the step 1), controlling the fermentation temperature to be 38-40 ℃, and standing for fermentation, wherein the inoculation amount is 6%.
Air is introduced at the same time interval in the fermentation process, the time is set to be 3 hours, 15 seconds are carried out each time, and the air volume is 130 cubic meters per hour.
Culturing for 15.8 hours, observing under a microscope of 16 times 100 times, if 8 single bacteria exist in each visual field, adding ammonia water, adjusting the pH to 5.8-6.2, continuing to ferment for 26.5 hours, constantly adding ammonia water to maintain the pH between 5.0-6.0, observing under a microscope of 16 times 100 times, if 16 single bacteria exist in each visual field, stopping adding, and obtaining an initial strain of enterococcus faecalis obtained. Introduction of yeast may begin.
2. Fermentation with yeast
2.1 preparation of Yeast culture Medium
The formula is as follows: 1% of yeast powder, 0.5% of peptone, 0.5% of sodium chloride and the balance of softened water.
Adjusting pH to 5.5, adding into 200L fermentation tank, diluting to 130L, sterilizing at 121 deg.C and 0.9-1.0Mpa for 20min, and cooling to 37 deg.C to obtain yeast culture medium;
2.2 Yeast inoculation and Primary fermentation
Inoculating yeast into a 37 ℃ yeast culture medium, wherein the inoculation amount is 4%, the culture temperature is set to be 37 ℃, the rotating speed is 100rpm/min, the rotating speed is increased by 20rpm every 1 hour until the rotating speed is 220rpm/min, the rotating speed is kept still, the ventilation volume is 120 cubic meters per hour initially, and the ventilation volume is increased to 300 cubic meters per hour after 8 hours. After culturing for 16 hours, 5 yeasts exist in the visual field of microscopic examination, and a primary yeast strain is obtained.
2.3 Secondary fermentation of Yeast
Transferring all the primary yeast strains into a secondary fermentation tank, and using 2.1 yeast culture medium, wherein the volume of the secondary fermentation tank is 10 tons and is 3.5-4 tons.
Culturing in a secondary fermentation tank for 11.5 hours until the content of yeast is 0.68 hundred million/ml, and completing secondary fermentation to obtain primary secondary fermentation yeast seeds.
3. Preparation of Bacillus subtilis liquid
The preparation method comprises the following steps: the culture medium comprises 20g/L of cane sugar, 5g/L of ammonium sulfate, 45g/L of bran, 2.0g/L of sodium citrate, 0.4g/L of dipotassium phosphate trihydrate, 0.5g/L of magnesium sulfate heptahydrate and 0.03g/L of ferric sulfate heptahydrate, the pH value is natural, the inoculation amount is 7%, the fermentation is carried out for 27 hours at the temperature of 37 ℃, the viable count reaches 233 hundred million/ml, and the spore formation rate is more than 98% for later use.
4. Preparation of Bacillus megaterium (TMQJ05) bacterial liquid
The preparation method comprises the following steps: the culture medium is 18g/L of corn paste, 55g/L of molasses, 3g/L of calcium chloride, 0.5g/L of dipotassium phosphate and 0.4g/L of magnesium sulfate, and the pH value of the fermentation starting is 6.8-7.2; fermenting at 37 deg.C for 29 hr until viable count reaches 6.2 hundred million/ml and spore rate is above 90%.
5. Mixed fermentation
Introducing the secondary fermentation liquor of the yeast into the fermentation liquor of enterococcus faecalis, wherein the total quantity is 7-9 tons, the stirring speed is 210rpm/min, the air quantity is 235 cubic per hour, the temperature is set to be 35-37 ℃, mixed fermentation is started, microscopic examination is carried out every two hours to ensure that the normal fermentation is not infected with infectious microbes, observation is carried out after 10.5 hours of mixed fermentation, more than 20 enterococcus faecalis carried out in a 16 x 100-time amplified visual field, a blood count plate counts the yeast and exceeds 1.6 hundred million/ml, ammonia water is fed again to keep the pH value to be 5.2-5.5, the operation is kept for 13 hours, and the feeding is stopped and the mixed fermentation is kept for 6 hours when the feeding amount of the ammonia water is more than 0.52 tons, so that the mixed fermentation liquor is obtained.
6. Composite fermentation
Adding the fermented bacillus subtilis and bacillus megatherium into the mixed fermentation liquor, wherein the ratio is as follows: 16 parts of bacillus subtilis: 23 parts of bacillus megatherium: and mixing 55 parts of mixed fermentation liquor, fully stirring, and adding 6% of potassium fulvate (required to be fully water-soluble, namely potassium fulvate: 68% of humic acid and 17.8% of potassium content) in mass proportion of the mixed liquor to obtain the basic mother liquor.
7. Fluidized drying
And (4) carrying out boiling drying on the basic mother liquor.
The carrier, namely the boiling drying carrier, is one of magnesium sulfate heptahydrate and ammonium sulfate or the arbitrary proportion of the two, and the mesh number of the magnesium sulfate heptahydrate and the ammonium sulfate is required to be 10.
Proportioning-the proportion of boiling drying carrier and mother liquor is 5 parts: 5 parts, wherein the air inlet temperature in the drying process is 100 ℃, the air outlet temperature is 52 ℃, and the material temperature is 63 ℃.
The final product, namely the compound microbial fertilizer, is obtained by boiling and drying, wherein the water content of the bacterial fertilizer is lower than 9.2 percent and the bacterial fertilizer is granulated, the content of humic acid is 6.8 percent, the content of magnesium is 6.8 percent, the content of nitrogen is 10.3 percent, and the content of bacillus subtilis and bacillus megatherium is 12.8 hundred million/g.
Example 3: application example experiment using the microbial Fertilizer product obtained in example 1
Designing a water culture experiment on the produced product, selecting wheat seeds with full and consistent grains, accelerating germination in a germination accelerating disc, selecting the wheat seeds with consistent growth vigor when the bud length is 0.8-1.2cm, carefully transplanting the wheat seeds into planting cotton, and placing the wheat seeds into a basic culture solution for culture and observation.
Designing a blank: a basic nutrient solution.
Experimental groups: the produced products are respectively an experimental group 1, an experimental group 2 and an experimental group 3 according to the dilution times of 10 ten thousand times, 5 ten thousand times and 1 ten thousand times.
And observing after 8 days of culture, and finding that the root system added with the finished product is obviously superior to the basic nutrient solution, and the capillary root is vigorous at 1 ten thousand times, so that the absorption area of the fertilizer is increased, and the main root is increased by more than 10%.
The experimental results are shown in figure 2.
Example 4: application example experiment using the microbial Fertilizer product obtained in example 1
Designing a pot experiment on the finished product, selecting corn seeds with full and consistent grains, planting the corn seeds in a flowerpot with the diameter of 20cm, watering 5 seeds in each pot, culturing in a dark place until the seeds break the soil and germinate, performing illumination culture, starting treatment when the height of the seedlings is about 2cm, applying the finished product with water, watering once every two days, treating once every 8 days, treating three times in total, and culturing for 30 days.
Designing a blank: a basic nutrient solution.
Experimental groups: the produced products are respectively an experimental group 1 and an experimental group 2 according to the dilution times of 5 ten thousand times and 1 ten thousand times.
And observing after 8 days of culture, and finding that the root system added with the finished product is obviously superior to the basic nutrient solution.
The results of the experiment are shown in FIG. 3.
The main roots are obviously increased, the growth rate is more than 25%, the capillary roots are more dense, the dry ploughing weight is increased by more than 20% compared with the blank, and the promotion effect is obvious.
Claims (10)
1. A microbial agent capable of effectively promoting crop rooting is characterized by comprising the following steps:
fermenting enterococcus faecalis to obtain a primary enterococcus faecalis strain;
secondary fermentation is carried out on the saccharomycetes to obtain primary saccharomycetes;
introducing the primary yeast strain into the primary enterococcus faecalis strain for mixed fermentation to obtain mixed fermentation liquor;
adding bacillus subtilis liquid and bacillus megatherium liquid into the mixed fermentation liquid, wherein the ratio of the bacillus subtilis liquid to the bacillus megatherium liquid is 15-25 parts by volume: 15-25 parts of bacillus megatherium: mixing 50-70 parts of fermentation liquor, uniformly mixing, and adding 5-10% of potassium fulvate in mass proportion of the mixed liquor to obtain basic mother liquor;
and (4) boiling and drying the basic mother liquor to obtain a finished bacterial fertilizer product.
2. The microbial inoculant being effective for promoting rooting in crops as claimed in claim 1, wherein said enterococcus faecalis is of a primary species, wherein no less than 15 individual microbes are observed under a microscope of 16 x 100 times;
the content of the yeast in the primary yeast strain is more than 0.5 hundred million/ml.
3. The microbial inoculant for promoting rooting of crops as claimed in claim 1, wherein the mixed fermentation comprises introducing primary yeast into primary enterococcus faecalis, the total amount is 7-9 tons, stirring is set at 220rpm/min, air flow is 220-.
4. The microbial inoculant according to claim 1, wherein the enterococcus faecalis fermentation process comprises:
inoculating enterococcus faecalis in the culture medium with the inoculation amount of 5-15%, controlling the fermentation temperature between 30-40 deg.C, standing and fermenting;
introducing air at the same interval in the fermentation process, setting the air flow to be 150 cubic/h at the time of 15 seconds every time for 3 hours, culturing the mixture to 12 to 16 hours, observing the mixture under a microscope of 16 times to 100 times, adding ammonia water when the air flow is not less than 5 single bacteria in each visual field, adjusting the pH to 5.8 to 6.2, continuing fermenting the mixture to 24 to 30 hours, adding ammonia water to keep the pH between 5.0 and 6.0, observing the mixture under a microscope of 16 times to 100 times, stopping the feeding when the air flow is not less than 15 single bacteria in each visual field, and obtaining the enterococcus faecalis primary strain.
5. The microbial inoculant according to claim 1, wherein the yeast fermentation comprises:
inoculating yeast into a 37 ℃ yeast culture medium, wherein the inoculation amount is 3-7%, the culture temperature is set to be 37 ℃, the rotating speed is 100rpm/min, the rotating speed is increased by 20rpm every 1 hour until 220rpm/min is kept unchanged, the ventilation volume is 120 cubic meters per hour initially, the ventilation volume is increased to 300 cubic meters per hour after 8 hours, and after 12-18 hours of culture, not less than 3-5 yeasts are in a microscopic view field to obtain a first-grade yeast strain;
transferring all the primary yeast strains into a secondary fermentation tank, adding a yeast culture medium, and metering the volume of the secondary fermentation tank to 3.5-4 tons at 10 tons; culturing in secondary fermentation tank for 7-15 hr until the content of yeast is greater than 0.5 hundred million/ml to obtain primary yeast strain.
6. The microbial inoculant effective in promoting rooting in crops of claim 1, 4 or 5, wherein the culture medium of enterococcus faecalis comprises: 6% of molasses, 0.4% of yeast powder, 0.4% of peptone, 0.03% of dipotassium hydrogen phosphate, 5% of brown sugar and the balance of softened water;
the raw materials of the culture medium of the yeast comprise: 1% of yeast powder, 0.5% of peptone, 0.5% of sodium chloride and the balance of softened water.
7. The microbial inoculant of claim 1, wherein the microbial inoculant is effective to promote rooting of a crop,
the bacillus subtilis liquid is prepared by the following method: the culture medium is 20g/L of cane sugar, 5g/L of ammonium sulfate, 45g/L of bran, 2.0g/L of sodium citrate, 0.4g/L of dipotassium phosphate trihydrate, 0.5g/L of magnesium sulfate heptahydrate and 0.03g/L of ferric sulfate heptahydrate, the pH value is natural, the mixture is fermented for 24 to 38 hours at the temperature of 37 ℃ according to the inoculation amount of 7 percent, the viable count reaches 200 hundred million/ml, and the spore formation rate is more than 98 percent, so that the biological enzyme preparation is obtained;
the bacillus megaterium liquid is prepared by the following method: in the culture medium, 18g/L of corn paste, 55g/L of molasses, 3g/L of calcium chloride and 0.4g/L of magnesium sulfate are used, and the pH of the fermentation starting material is 6.8-7.2; fermenting at 37 deg.C for 24-30 hr until viable count reaches 5 hundred million/ml and spore rate is above 90%.
8. The microbial agent capable of effectively promoting crop rooting according to claim 1, wherein the bacterial fertilizer has a moisture content of less than 15% and is granular, the bacterial fertilizer has a humic acid content of 5-7%, a magnesium content of 3-7%, a nitrogen content of 10-17%, and a total content of bacillus subtilis and bacillus megaterium of 2-15 hundred million/g.
9. A preparation method of a microbial agent capable of effectively promoting crop rooting is characterized by comprising the following steps: fermenting enterococcus faecalis to obtain a primary enterococcus faecalis strain;
secondary fermentation is carried out on the saccharomycetes to obtain primary saccharomycetes;
introducing the primary yeast strain into the primary enterococcus faecalis strain for mixed fermentation to obtain mixed fermentation liquor;
adding bacillus subtilis liquid and bacillus megatherium liquid into the mixed fermentation liquid, wherein the ratio of the bacillus subtilis liquid to the bacillus megatherium liquid is 15-25 parts by volume: 15-25 parts of bacillus megatherium: mixing 50-70 parts of fermentation liquor, uniformly mixing, and adding 5-10% of potassium fulvate in mass proportion of the mixed liquor to obtain basic mother liquor;
and (4) boiling and drying the basic mother liquor to obtain a finished bacterial fertilizer product.
10. The method for preparing the microbial inoculum for effectively promoting the rooting of the crops according to claim 9, which is characterized by comprising the following steps:
6 percent of molasses, 0.4 percent of yeast powder, 0.4 percent of peptone, 0.03 percent of dipotassium phosphate, 5 percent of brown sugar and the balance of softened water are added, the constant volume is 3.5 to 4 tons in a 10-ton fermentation tank, sterilization is carried out for 30min at 116 ℃ and 0.8 to 0.9Mpa, the temperature is reduced to room temperature after the sterilization is finished, an enterococcus faecalis inoculated in the culture medium, the inoculation amount is 5 to 15 percent, the fermentation temperature is controlled to be between 30 and 40 ℃, standing fermentation is carried out, air is introduced at the same time interval in the fermentation process, the time is set to be 3 hours, the air volume is 150 cubic meters per hour per 15 seconds per time, the culture is carried out for 12 to 16 hours, the observation is carried out under a microscope of 16 times 100 times, when not less than 5 single bacteria exist in each visual field, ammonia water is added, the PH is adjusted to be 5.8 to 6.2, the fermentation is continued to 24 to 30 hours, and the ammonia water is added at the moment to keep the PH value between 5.0 and 6.0 to 6.0, observing under a microscope with the magnification of 16 times 100 times, stopping feeding no less than 15 single bacteria in each visual field, and obtaining the primary strain of enterococcus faecalis. Yeast can be introduced;
preparing 1% of yeast powder, 0.5% of peptone and 0.5% of sodium chloride, supplementing the rest with softened water, fixing the volume to 130L in a 200L fermentation tank, adjusting the pH to 5.3-5.8, 121 ℃, sterilizing at 0.9-1.0Mpa for 20min, cooling to 37 ℃ to obtain a yeast culture medium, inoculating yeast into the 37 ℃ yeast culture medium, wherein the inoculation amount is 3-7%, the culture temperature is set to 37 ℃, the rotating speed is 100rpm/min, increasing the rotating speed to 20rpm every 1 hour until 220rpm/min keeps still, the ventilation volume is initial 120 cubic meters per hour, increasing the ventilation volume to 300 cubic meters per hour after 8 hours, culturing for 12-18 hours, not less than 3-5 yeasts in a microscopic examination field to obtain a primary yeast, transferring the primary yeast into a secondary constant volume fermentation tank, using the yeast culture medium, and enabling the secondary fermentation tank to be 10 tons to 3.5-4 tons, culturing in secondary fermentation tank for 7-15 hr until the content of yeast is greater than 0.5 hundred million/ml, and completing secondary fermentation to obtain primary yeast strain;
in a culture medium which comprises 20g/L of cane sugar, 5g/L of ammonium sulfate, 45g/L of bran, 2.0g/L of sodium citrate, 0.4g/L of dipotassium hydrogen phosphate trihydrate, 0.5g/L of magnesium sulfate heptahydrate, 0.03g/L of ferric sulfate heptahydrate and the balance of softened water, fermenting for 24-38 hours at 37 ℃ according to the inoculation amount of 7 percent, wherein the number of viable bacteria reaches 200 hundred million/ml, and the spore formation rate is more than 98 percent, thus obtaining the bacillus subtilis liquid;
the pH of the initial fermentation is 6.8-7.2 in a culture medium which comprises 18g/L of corn paste, 55g/L of molasses, 3g/L of calcium chloride, 0.5g/L of dipotassium phosphate, 0.4g/L of magnesium sulfate and the balance of softened water; fermenting at 37 deg.C for 24-30 hr until viable count reaches 5 hundred million/ml and spore rate is above 90% to obtain Bacillus megaterium (TMQJ05) bacterial liquid;
introducing primary yeast strains into primary enterococcus faecalis strains, wherein the total amount is 7-9 tons, stirring is set to be 180-;
adding the fermented bacillus subtilis and bacillus megatherium into the mixed fermentation liquor, wherein the ratio is as follows: 15-25 parts of bacillus subtilis: 15-25 parts of bacillus megatherium: mixing 50-70 parts of mixed fermentation liquor, fully stirring, and adding 5-10% of potassium fulvate in mass proportion of the mixed liquor to obtain basic mother liquor;
and (3) obtaining a finished product of the compound microbial fertilizer through boiling and drying, wherein the water content of the bacterial fertilizer is lower than 15%, the bacterial fertilizer is granular, the humic acid content is 5-7%, the magnesium content is 3-7%, the nitrogen content is 10-17%, and the content of bacillus subtilis and bacillus megatherium is 2-15 hundred million/g.
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