CN112225595A - Water-soluble compound microbial fertilizer and preparation method thereof - Google Patents
Water-soluble compound microbial fertilizer and preparation method thereof Download PDFInfo
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- CN112225595A CN112225595A CN202011134954.3A CN202011134954A CN112225595A CN 112225595 A CN112225595 A CN 112225595A CN 202011134954 A CN202011134954 A CN 202011134954A CN 112225595 A CN112225595 A CN 112225595A
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- C05—FERTILISERS; MANUFACTURE THEREOF
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- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
Abstract
The invention belongs to the technical field of microbial fertilizers, and particularly relates to a water-soluble compound microbial fertilizer and a preparation method thereof. The water-soluble composite microbial fertilizer provided by the invention comprises yeast fermentation waste liquid dry powder, inorganic water-soluble fertilizer and functional bacteria powder in a mass ratio of (68-85) to (15-30) to (0.3-3.5). The water-soluble composite microbial fertilizer provided by the invention takes the water-soluble yeast fermentation waste liquid dry powder as an organic carrier, makes up the defect that the traditional composite microbial fertilizer is not water-soluble, expands the application mode of the composite microbial organic fertilizer, avoids the damage of the traditional organic fertilizer to crops, and can effectively improve the soil fertility by compounding the water-soluble composite microbial fertilizer with the functional bacteria powder and the inorganic water-soluble fertilizer according to a certain proportion. Experimental results of the embodiment show that the water-insoluble content of the water-soluble compound microbial fertilizer provided by the invention is 0.08-0.25%, and the requirement that the water-insoluble content of the water-soluble compound microbial fertilizer is less than or equal to 5% in the standard of common water-soluble fertilizers is met.
Description
Technical Field
The invention belongs to the technical field of microbial fertilizers, and particularly relates to a water-soluble compound microbial fertilizer and a preparation method thereof.
Background
The compound microbial fertilizer is a ternary compound fertilizer formed by organic fertilizer, inorganic fertilizer and functional bacteria, embodies the comprehensive effect of three effects in one, and has the effects of activating soil, improving soil fertility, inhibiting diseases, strengthening soil microbial function, increasing yield and income and the like.
The organic raw materials commonly used at present comprise straws, livestock and poultry manure, cake fertilizer, algae fertilizer, mushroom residue, industrial byproducts and the like. However, the organic raw materials are often not thoroughly decomposed, the phenomena of seedling burning and seedling death can occur after the organic raw materials are used, harmful bacteria carried by the organic raw materials can cause secondary pollution to soil, the activity of adsorbed effective functional bacteria is also inhibited, and the yield and the quality of crops can be influenced; in addition, the compound microbial fertilizer produced by the organic raw materials is generally used as a base fertilizer, is rarely used for topdressing, and cannot exert the advantages of a 'ternary compound fertilizer' in the middle and later periods of faster crop growth and more nutrient absorption.
Disclosure of Invention
The invention aims to provide a water-soluble compound microbial fertilizer and a preparation method thereof. The water-soluble microbial fertilizer provided by the invention has low content of water-insoluble substances, and can be applied to fertilization modes such as broadcasting application, hole application, flushing application, drip irrigation, spraying application and the like.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention provides a water-soluble composite microbial fertilizer, which comprises yeast fermentation waste liquid dry powder, inorganic water-soluble fertilizer and functional bacteria powder; the mass ratio of the yeast fermentation waste liquid dry powder to the inorganic water-soluble fertilizer to the functional bacteria powder is (68-85) to (15-30) to (0.3-3.5).
Preferably, the mass content of water-soluble organic matters in the yeast fermentation waste liquid dry powder is not less than 60%, the mass content of water-soluble fulvic acid is not less than 50%, the mass content of nitrogen, phosphorus and potassium is not less than 10%, and the mass content of crude protein is 18-22%.
Preferably, the yeast fermentation waste liquid dry powder also comprises B vitamins, vitamin C, inositol, polysaccharide, trace elements, mycoprotein, nucleic acid, surfactant and growth promoting factors.
Preferably, the pH value of the yeast fermentation waste liquid dry powder is 5-6.
Preferably, the inorganic water-soluble fertilizer is a nitrogen-phosphorus-potassium compound fertilizer.
Preferably, the inorganic water soluble fertilizer comprises two or more of urea, ammonium chloride, ammonium sulfate, monopotassium phosphate, monoammonium phosphate, potassium sulfate and potassium nitrate.
Preferably, the functional bacteria powder comprises one or more of gel-like bacillus powder, bacillus subtilis powder and bacillus licheniformis powder.
Preferably, the preparation of each bacterial powder in the functional bacterial powder comprises the following steps:
(1) sequentially performing strain activation, primary fermentation, secondary fermentation and tertiary fermentation on the strain to obtain a zymocyte liquid;
(2) carrying out membrane filtration concentration on the zymocyte liquid obtained in the step (1) to obtain concentrated bacteria liquid;
(3) and (3) carrying out spray drying on the concentrated bacterial liquid obtained in the step (2) to obtain bacterial powder.
Preferably, the effective viable count of the water-soluble compound microbial fertilizer is more than or equal to 0.2 hundred million/g, and the organic matter content is more than or equal to 20 percent, calculated as N, P2O5And K2The total nutrient content of O is 8.0-25.0%, the rate of mixed bacteria is less than or equal to 30%, the water content is less than or equal to 30%, and the pH value is 5.5-8.5.
The invention also provides a preparation method of the water-soluble compound microbial fertilizer in the technical scheme, which comprises the following steps: and mixing the yeast fermentation waste liquid dry powder, the inorganic water-soluble fertilizer and the functional bacteria powder to obtain the water-soluble composite microbial fertilizer.
The invention provides a water-soluble composite microbial fertilizer, which comprises yeast fermentation waste liquid dry powder, inorganic water-soluble fertilizer and functional bacteria powder; the mass ratio of the yeast fermentation waste liquid dry powder to the inorganic water-soluble fertilizer to the functional bacteria powder is (68-85) to (15-30) to (0.3-3.5). The water-soluble composite microbial fertilizer provided by the invention takes the water-soluble yeast fermentation waste liquid dry powder as an organic carrier, makes up the defect that the traditional composite microbial fertilizer is not water-soluble, expands the application mode of the composite microbial organic fertilizer, avoids the damage of the traditional organic fertilizer to crops, and can effectively improve the soil fertility by compounding the water-soluble composite microbial fertilizer with the functional bacteria powder and the inorganic water-soluble fertilizer according to a certain proportion. Experimental results of the embodiment show that the water-insoluble content of the water-soluble compound microbial fertilizer provided by the invention is 0.08-0.25%, and the requirement that the water-insoluble content of the water-soluble compound microbial fertilizer is less than or equal to 5% in the standard of common water-soluble fertilizers is met.
Biological preservation Instructions
The Bacillus mucilaginosus (Bacillus licheiformis) is preserved at the common microorganism center of China Committee for culture Collection of microorganisms, the specific address is the microorganism research institute of China academy of sciences No.3, West Lu No.1 Hospital, North Kyoho, Beijing, the preservation number is CGMCC No.3995, and the preservation time is 7 months and 2 days in 2010;
bacillus subtilis (Bacillus subtilis) with a preservation place of China general microbiological culture Collection center, specifically, with a microorganism research institute of China academy of sciences No.3, Xilu No.1, Beijing, the morning area, the preservation number of CGMCC NO.10248 and a preservation time of 2014, 12 months and 29 days;
bacillus licheniformis (Bacillus licheniformis), the preservation place is China general microbiological culture Collection center, the concrete address is the microbiological research institute of China academy of sciences No.3, Xilu No.1, Beijing, the south China, the storage number is CGMCC No.10741, the preservation date is 2015, 4 months and 24 days.
Detailed Description
The invention provides a water-soluble composite microbial fertilizer, which comprises yeast fermentation waste liquid dry powder, inorganic water-soluble fertilizer and functional bacteria powder; the mass ratio of the yeast fermentation waste liquid dry powder to the inorganic water-soluble fertilizer to the functional bacteria powder is (68-85) to (15-30) to (0.3-3.5).
The water-soluble compound microbial fertilizer provided by the invention comprises yeast fermentation waste liquid dry powder. In the invention, the yeast fermentation waste liquid dry powder is a water-soluble organic raw material, so that the defect that the traditional compound microbial fertilizer is not water-soluble is overcome, the application mode of the compound microbial organic fertilizer is expanded, and the damage of the traditional organic fertilizer to crops is avoided; in addition, the utilization of the yeast fermentation waste liquid dry powder provides a way for resource utilization of waste, and has environmental protection benefits.
In the invention, the mass content of the water-soluble organic matters in the yeast fermentation waste liquid dry powder is preferably not less than 60%, and more preferably 65-75%; the mass content of the water-soluble fulvic acid in the yeast fermentation waste liquid dry powder is preferably not less than 50%, and more preferably 55-65%; the mass content of nitrogen, phosphorus and potassium in the yeast fermentation waste liquid dry powder is preferably not less than 10%, and more preferably 12-15%; the mass content of the crude protein in the yeast fermentation waste liquid dry powder is preferably 18-22%, and more preferably 20%. In the invention, the main fulvic acid is an organic matter of a plant source, is a biological stimulant for promoting the growth of crops, and simultaneously increases the colloid content of soil, promotes the formation of a granular structure, improves the soil porosity, and improves the cation exchange capacity and the fertilizer retention capacity of the soil.
In the invention, the yeast fermentation waste liquid dry powder preferably further comprises B vitamins, vitamin C, inositol, polysaccharide, trace elements, mycoprotein, nucleic acid, surfactant and growth promoting factors.
In the invention, the pH value of the yeast fermentation waste liquid dry powder is preferably 5-6.
The source of the yeast fermentation waste liquid dry powder is not particularly limited in the present invention, and commercially available products known to those skilled in the art may be used. In the present invention, the yeast fermentation waste liquid dried powder is preferably derived from Angel Yeast Co.
The water-soluble compound microbial fertilizer provided by the invention comprises an inorganic water-soluble fertilizer. In the present invention, the inorganic water-soluble fertilizer is preferably a nitrogen-phosphorus-potassium compound fertilizer, more preferably comprises two or more of urea, ammonium chloride, ammonium sulfate, monopotassium phosphate, monoammonium phosphate, potassium sulfate and potassium nitrate, and most preferably a combination of urea, monopotassium phosphate and potassium nitrate, or a combination of ammonium chloride, monoammonium phosphate and potassium dihydrogen phosphate, or a combination of urea, monoammonium phosphate and potassium sulfate, or a combination of ammonium sulfate, monoammonium phosphate and potassium sulfate.
The invention has no special limit on the dosage of each component in the inorganic water-soluble fertilizer, and the nutrient proportion required by crops can be met. The sources of the components in the inorganic water-soluble fertilizer are not particularly limited, and the inorganic water-soluble fertilizer can be prepared from commercially available products which are well known to those skilled in the art. In the invention, the inorganic water-soluble fertilizer is a water-soluble fertilizer and provides nitrogen, phosphorus and potassium required by the growth of crops.
The water-soluble compound microbial fertilizer provided by the invention comprises functional bacterial powder. In the present invention, the functional bacteria powder preferably includes one, two or three of a gel-like bacillus powder, a bacillus subtilis powder and a bacillus licheniformis powder, and more preferably a combination of the gel-like bacillus powder and the bacillus subtilis powder, or a combination of the gel-like bacillus powder and the bacillus licheniformis powder. In the invention, the microorganisms contained in the functional bacteria powder have the effects of dissolving phosphorus, dissolving potassium and fixing nitrogen, so that the utilization rate of the nitrogen, phosphorus and potassium fertilizer is improved; meanwhile, the microbial inoculum is used as a beneficial bacterium, occupies ecological niches, reduces the infection of pathogenic bacteria, has the effect of preventing and treating soil-borne diseases, supplements soil microorganisms and improves the number and activity of floras.
In the present invention, the preparation of each of the functional bacterial powders preferably comprises the steps of:
(1) sequentially performing strain activation, primary fermentation, secondary fermentation and tertiary fermentation on the strain to obtain a zymocyte liquid;
(2) carrying out membrane filtration concentration on the zymocyte liquid obtained in the step (1) to obtain concentrated bacteria liquid;
(3) and (3) carrying out spray drying on the concentrated bacterial liquid obtained in the step (2) to obtain bacterial powder.
The invention preferably carries out strain activation, primary fermentation, secondary fermentation and tertiary fermentation on strains in sequence to obtain zymogen liquid.
In the invention, the strain used for preparing the gel-like Bacillus powder is preferably the gel-like Bacillus (Bacillus licheiformis), the preservation place is the common microorganism center of China Committee for culture Collection of microorganisms, the specific address is the institute of microorganisms of China academy of sciences No.3, West Lu No.1, North Cheng, Yang ward, Beijing, the preservation number is CGMCC No.3995, and the preservation time is 2010, 7 months and 2 days. In the invention, the paenibacillus mucilaginosus has the effects of dissolving phosphorus, potassium and nitrogen, releasing medium and trace elements held by soil, inhibiting germs, promoting growth and increasing yield.
In the invention, the strain used for preparing the Bacillus subtilis powder is preferably Bacillus subtilis (Bacillus subtilis), the storage place is the common microorganism center of China Committee for culture Collection of microorganisms, the specific address is the microorganism research institute of China academy of sciences No.3, West Lu No.1, North Chen West province, in Beijing, the storage number is CGMCC NO.10248, and the storage time is 29 days 12 months 2014. In the invention, the bacillus subtilis can be rapidly and massively propagated and colonized at the rhizosphere, body surface or in vivo of a plant and in soil, performs nutrition and space site competition, and has the effects of inhibiting bacteria, sterilizing, improving soil fertilizer efficiency and promoting crop growth.
In the invention, the strain used for preparing the Bacillus licheniformis powder is preferably Bacillus licheniformis (Bacillus licheniformis), the preservation place is the common microorganism center of China Committee for culture Collection of microorganisms, the specific address is the microorganism research institute of China academy of sciences No.3, West Lu No.1, North Chen West province, in the Korean area, in Beijing, the preservation number is CGMCC No.10741, and the preservation date is 2015, 4 months and 24 days. In the invention, the bacillus licheniformis can improve the disease resistance, cold resistance and drought resistance of crops; increase soil nutrients, improve soil structure and improve the utilization rate of chemical fertilizers; promoting organic matters in the soil to be decomposed into humus and stimulating the growth of crops; promoting the growth of crops and increasing the yield; has certain effects of fixing nitrogen, dissolving phosphorus and potassium.
In the present invention, the strain activation preferably includes: inoculating the strain into an activation culture medium for activation to obtain an activated strain. In the invention, the activation temperature is preferably 28-30 ℃, and more preferably 29 ℃; the activation time is preferably 24-28 h, and more preferably 26 h.
In the present invention, the strain activation medium of paenibacillus mucilaginosus is preferably a solid medium, and the composition of the solid medium preferably includes: 10-15 g/L of glucose, 0.3-0.5 g/L of dipotassium hydrogen phosphate, 0.2-0.4 g/L of magnesium sulfate, 0.2-0.3 g/L of sodium chloride, 1-1.5 g/L of calcium carbonate and 20-25 g/L of agar powder. In the invention, the initial pH value of the activation-grade culture medium of the Paenibacillus mucilaginosus is preferably 7.0-7.5.
In the present invention, the activating medium for bacillus subtilis preferably includes: 10-15 g/L of glucose, 8-10 g/L of peptone, 5-10 g/L of sodium chloride and 0.5-1 g/L of calcium carbonate. In the invention, the initial pH value of the activation medium of the bacillus subtilis is preferably 7.0-7.5.
In the present invention, the activating medium for bacillus licheniformis preferably comprises: 8-10 g/L of peptone, 2-3 g/L of beef extract, 3-5 g/L of sodium chloride and 5-18 g/L of agar. In the invention, the initial pH value of the activation medium of the bacillus licheniformis is preferably 7.0-7.4.
In the present invention, the primary fermentation preferably comprises: inoculating the activated strain into a primary culture medium for culturing to obtain primary fermentation liquid. In the invention, the temperature of the primary fermentation is preferably 28-30 ℃, and more preferably 29 ℃; the time of the primary fermentation is preferably 45-50 h, and more preferably 48 h; the first-stage fermentation is preferably shake flask fermentation.
In the present invention, the first-order culture medium of paenibacillus mucilaginosus is preferably a solid culture medium, and the composition of the solid culture medium preferably comprises: 12-16 g/L of sucrose, 0.4-0.8 g/L of dipotassium phosphate, 0.3-0.5 g/L of magnesium sulfate, 0.2-0.3 g/L of sodium chloride, 2.0-3.0 g/L of calcium carbonate and 20-25 g/L of agar powder. In the invention, the initial pH value of the primary culture medium of the paenibacillus mucilaginosus is preferably 7.0-7.5.
In the present invention, the primary culture medium of Bacillus subtilis preferably comprises: 12-18 g/L of sucrose, 8-10 g/L of peptone, 5-10 g/L of sodium chloride and 0.5-1 g/L of calcium carbonate. In the invention, the initial pH value of the primary culture medium of the bacillus subtilis is preferably 7.0-7.5.
In the present invention, the primary culture medium of bacillus licheniformis preferably comprises: 8-10 g/L of peptone, 3-4 g/L of beef extract, 3-5 g/L of sodium chloride and 5-18 g/L of agar. In the invention, the initial pH value of the primary culture medium of the bacillus licheniformis is preferably 7.0-7.5.
In the present invention, the secondary fermentation preferably comprises: inoculating the primary fermentation liquid into a secondary culture medium for culturing to obtain secondary fermentation liquid. In the invention, the temperature of the secondary fermentation is preferably 28-30 ℃, and more preferably 29 ℃; the secondary fermentation time is preferably 48-72 h, and more preferably 54-66 h; the secondary fermentation is preferably a seed tank fermentation.
In the present invention, the secondary medium of Paenibacillus mucilaginosus preferably comprises: 5.5-9 g/L of starch, 1.6-2.0 g/L of yeast powder, 6-8 g/L of soybean meal, 1-1.4 g/L of magnesium sulfate, 2-3 g/L of dipotassium phosphate, 5.5-8.5 g/L of calcium carbonate and 0.2-0.3 g/L of sodium chloride. In the invention, the initial pH value of the secondary culture medium of the paenibacillus mucilaginosus is preferably 7.0-7.5.
In the present invention, the secondary medium of Bacillus subtilis preferably comprises: 7-12 g/L of brown sugar, 2-2.5 g/L of starch, 10-12 g/L of soybean meal, 2-3 g/L of monopotassium phosphate, 1-2 g/L of dipotassium phosphate, 0.5-1 g/L of calcium carbonate, 0.5-1 g/L of magnesium sulfate, 0.5-1 g/L of calcium chloride, 0.2-0.5 g/L of manganese sulfate and 0.01-0.05 g/L of ferric chloride. In the invention, the initial pH value of the secondary culture medium of the bacillus subtilis is preferably 7.0-7.5.
In the present invention, the secondary culture medium of bacillus licheniformis preferably comprises: 10-15 g/L of cane sugar, 10-12 g/L of soybean meal, 1-2 g/L of monopotassium phosphate and 0.1-0.3 g/L of zinc sulfate. In the invention, the initial pH value of the secondary culture medium of the bacillus licheniformis is preferably 6.5-7.0.
In the present invention, the tertiary fermentation preferably comprises: and inoculating the secondary fermentation liquid into a tertiary culture medium for fermentation to obtain a zymogen liquid. In the invention, the inoculation amount of the secondary fermentation liquid is preferably 4-6% of the mass of the tertiary culture medium, and more preferably 5%; the third stage fermentation is preferably a fermentor fermentation.
In the present invention, the tertiary medium of each species is preferably the same as the secondary medium of the corresponding species in composition.
In the invention, the temperature of the third-stage fermentation is preferably 30-35 ℃, and more preferably 32 ℃; the time of the tertiary fermentation is preferably 3-5 days, and more preferably 4 days. In the present invention, the process of the three-stage fermentation is preferably aerated, and the aeration is preferably performed by: the ventilation volume is 10-20 m within the first 24h of fermentation3H; after fermenting for 24 hours, the ventilation volume is 15-24 m3H is used as the reference value. In the present invention, the three-stage fermentation is preferably performed under stirring; the stirring frequency is preferably 1-2 times/h, and the stirring time is preferably 10-20 min.
In the invention, the number of strains in the zymocyte liquid is preferably 5-20 hundred million/mL.
After the zymophyte liquid is obtained, the invention preferably performs membrane filtration and concentration on the zymophyte liquid to obtain concentrated bacteria liquid. In the present invention, the membrane filtration concentration is preferably performed using a membrane filter. In the invention, the membrane filtration concentration is preferably 5 to 10 times.
In the invention, the number of strains in the concentrated bacterial liquid is preferably 20-200 hundred million/mL.
After the concentrated bacterial liquid is obtained, the concentrated bacterial liquid is preferably subjected to spray drying to obtain bacterial powder. In the invention, the temperature of the air inlet of the spray drying is preferably 70-100 ℃, more preferably 80-90 ℃, and the temperature of the air outlet is preferably 40-60 ℃, more preferably 45-55 ℃; the feeding speed of the spray drying is preferably 8-15L/min, and more preferably 10-12L/min. In the invention, the spray drying preferably adopts a compound of skimmed milk powder and sodium glutamate which are sterilized at 121 ℃ as a protective agent; the ratio of the skimmed milk powder to the sodium glutamate is preferably (1-3) to 1, and more preferably 2 to 1. In the invention, the dosage of the protective agent is preferably 5-10% of the mass of the concentrated bacterial liquid. In the invention, the spray drying can improve the resistance of spores generated by the functional bacteria to adverse factors such as high temperature, high pressure, drying, oxidation, acid and alkali, toxic substances and the like within the parameter range, and the activity of the functional bacteria is maintained in the drying process.
In the invention, the effective viable count of the water-soluble compound microbial fertilizer is preferably more than or equal to 0.2 hundred million/g, and the organic matter is preferably more than or equal to 20 percent, calculated as N, P2O5And K2The total nutrient content of the O is preferably 8.0-25.0% by mass, the mixed bacteria rate is preferably less than or equal to 30%, the water content is preferably less than or equal to 30%, and the pH value is preferably 5.5-8.5.
In the invention, the mass ratio of the yeast fermentation waste liquid dry powder, the inorganic water-soluble fertilizer and the functional bacteria powder is (68-85): 15-30): 0.3-3.5, preferably (70-82): 17-28): 0.5-3.0, more preferably (73-80): 19-26): 0.5-2.0.
The water-soluble composite microbial fertilizer provided by the invention takes the water-soluble yeast fermentation waste liquid dry powder as an organic carrier, so that the defect that the traditional composite microbial fertilizer is not water-soluble is overcome, the application mode of the composite microbial organic fertilizer is expanded, the damage of the traditional organic fertilizer to crops is avoided, and the water-soluble composite microbial fertilizer is compounded with the functional bacteria powder and the inorganic water-soluble fertilizer according to a certain proportion, so that the soil fertility is effectively improved; in addition, the water-soluble compound microbial fertilizer provided by the invention has stable bacterial count, and can ensure the stability of the product.
The invention also provides a preparation method of the microbial fertilizer in the technical scheme, which comprises the following steps: and mixing the yeast fermentation waste liquid dry powder, the inorganic water-soluble fertilizer and the functional bacteria powder to obtain the water-soluble composite microbial fertilizer.
The operation of the mixing is not particularly limited in the present invention, and the technical scheme of material mixing known to those skilled in the art can be adopted. In the present invention, the mixing is preferably performed under stirring conditions; the stirring time is preferably 20-30 min.
The preparation method provided by the invention is simple and easy to operate, and is convenient for industrial production.
The following examples are provided to illustrate the water-soluble compound microbial fertilizer and the preparation method thereof in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
Water-soluble composite microbial fertilizer 1: 80% of yeast fermentation waste liquid dry powder, 8% of urea, 4.5% of monopotassium phosphate, 6% of potassium nitrate, 1.0% of paenibacillus mucilaginosus powder and 0.5% of bacillus subtilis powder; the mass ratio of the yeast fermentation waste liquid dry powder to the inorganic water-soluble fertilizer to the functional bacteria powder is 80:18.5: 1.5.
In the embodiment, the water-soluble compound microbial fertilizer 1 contains 53% of organic matters, 24.1% of NPK, 1.2 hundred million/g of effective bacteria, 0.2% of mixed bacteria, 0.55% of water and 5.7 of pH value.
The yeast fermentation waste liquid dry powder is from Angel yeast GmbH, and has fulvic acid content of 60%, nitrogen phosphorus potassium content of 13%, crude protein content of 20%, and pH value of 6.0.
The preparation method of the jelly-like paenibacillus strain powder comprises the following steps:
(1) activating strains: inoculating the strain of the Paenibacillus mucilaginosus into an activation culture medium for activation, wherein the activation temperature is 29 ℃, and the activation time is 26 hours, so as to obtain the activated strain. Activation medium composition: 12g/L of glucose, 0.3g/L of dipotassium phosphate, 0.3g/L of magnesium sulfate, 0.2g/L of sodium chloride, 1.5g/L of calcium carbonate and 20g/L of agar powder, and the initial pH value is 7.0.
(2) Primary fermentation: inoculating the activated strain into a primary culture medium for culturing to obtain primary fermentation liquid. The temperature of the primary fermentation is 29 ℃, and the time of the primary fermentation is 48 hours. The first-stage fermentation medium comprises the following components: 14g/L of sucrose, 0.6g/L of dipotassium phosphate, 0.4g/L of magnesium sulfate, 0.3g/L of sodium chloride, 2.5g/L of calcium carbonate and 20g/L of agar powder, and the initial pH value is 7.0.
(2) Secondary fermentation: inoculating the primary fermentation liquid into a secondary culture medium for culturing, wherein the fermentation temperature is 29 ℃, and the fermentation time is 60 hours, so as to obtain the secondary fermentation liquid. The composition of the secondary culture medium is as follows: 7.5g/L of starch, 1.8g/L of yeast powder, 7g/L of soybean meal, 1.2g/L of magnesium sulfate, 2.8g/L of dipotassium phosphate, 6.5g/L of calcium carbonate and 0.3g/L of sodium chloride, and the initial pH value is 7.5.
(3) And (3) three-stage fermentation: inoculating the secondary fermentation liquid into a tertiary culture medium for fermentation, wherein the inoculation amount is 5% of the mass of the tertiary culture medium, the fermentation temperature is 32 ℃, and the ventilation volume in the first 24h of fermentation is 15m3H; the ventilation capacity after 24h fermentation is 18m3And h, stirring frequency is 2 times per hour, stirring is carried out for 15min each time, and fermentation time is 4 days, so as to obtain the zymophyte liquid. The composition of the tertiary medium is the same as that of the secondary medium.
(4) Concentration: after the zymophyte liquid is obtained, the zymophyte liquid is concentrated by 10 times by adopting a membrane filter to obtain concentrated bacteria liquid.
(5) And (3) drying: drying the concentrated bacterial liquid by adopting a spray drying process, wherein the feeding speed is 12 mL/min, the temperature of a dried air inlet is 85 ℃, the temperature of a spray air outlet is 50 ℃, and a compound of skimmed milk powder and sodium glutamate sterilized at 121 ℃ is used as a protective agent to obtain bacterial powder, wherein the ratio of the skimmed milk powder to the sodium glutamate is 2:1, and the dosage of the protective agent is 10% of the mass of the concentrated bacterial liquid.
The preparation method of the bacillus subtilis powder comprises the following steps:
the technological process and technological parameters of the bacillus subtilis powder refer to a 'preparation method of the bacillus subtilis powder in jelly shape', and the two methods are mainly different in culture medium components. The components of the culture medium of the bacillus subtilis are as follows:
activating a culture medium: 15g/L glucose, 10g/L peptone, 8g/L sodium chloride and 0.7 g/L calcium carbonate, and the initial pH value is 7.0.
Primary culture medium: 16g/L of sucrose, 10g/L of peptone, 9g/L of sodium chloride and 0.8g/L of calcium carbonate, and the initial pH value is 7.0.
Secondary culture medium, tertiary culture medium: 10g/L of brown sugar, 2g/L of starch, 11g/L of soybean meal, 2g/L of monopotassium phosphate, 1g/L of dipotassium phosphate, 1g/L of calcium carbonate, 0.9g/L of magnesium sulfate, 0.8g/L of calcium chloride, 0.4g/L of manganese sulfate and 0.02g/L of ferric chloride, and the initial pH value is 7.5.
And putting the prepared paenibacillus jelly powder, the prepared bacillus subtilis powder, the dried yeast fermentation waste liquid powder, the urea, the monopotassium phosphate and the potassium nitrate into a stirring tank according to the proportion, stirring for 25min, fully mixing uniformly, taking out of the tank and packaging.
Example 2
Water-soluble compound microbial fertilizer 2: 75.5% of yeast fermentation waste liquid dry powder, 9% of ammonium chloride, 9% of monoammonium phosphate, 4.5% of monopotassium phosphate, 1.0% of paenibacillus jelly powder and 1.0% of bacillus licheniformis powder; the mass ratio of the yeast fermentation waste liquid dry powder to the inorganic water-soluble fertilizer to the functional bacteria powder is 75.5:22.5: 2.
In the embodiment, the water-soluble compound microbial fertilizer 2 contains 50% of organic matters, 24.9% of NPK, 0.7 hundred million/g of effective bacteria, 0.1% of mixed bacteria rate, 0.6% of water and 5.7 of pH value.
The preparation method of the bacillus licheniformis powder comprises the following steps:
the technological process and technological parameters of the bacillus licheniformis powder refer to the 'preparation method of the paenibacillus jelly-like powder' in the example 1, and the two are mainly different in culture medium components. The components of the culture medium of the bacillus licheniformis are as follows:
activating a culture medium: 9g/L of peptone, 3g/L of beef extract, 4g/L of NaCl and 12g/L of agar, and the initial pH value is 7.2.
Primary culture medium: 10g/L of peptone, 4g/L of beef extract, 4g/L of sodium chloride and 10g/L of agar, and the initial pH value is 7.2.
Secondary culture medium, tertiary culture medium: 12g/L of sucrose, 12g/L of soybean meal, 1.5g/L of monopotassium phosphate and 0.2g/L of zinc sulfate, and the initial pH value is 7.0.
In this example, the sources of the yeast fermentation waste liquid dry powder and the paenibacillus jelly powder were the same as in example 1, and the preparation method was the same as in example 1.
Example 3
Water-soluble composite microbial fertilizer 3: 77.5 percent of yeast fermentation waste liquid dry powder, 5 percent of urea, 5 percent of monoammonium phosphate, 12 percent of potassium sulfate and 0.5 percent of bacillus subtilis powder; the mass ratio of the yeast fermentation waste liquid dry powder to the inorganic water-soluble fertilizer to the functional bacteria powder is 77.5:22: 0.5.
In the embodiment, the water-soluble compound microbial fertilizer 3 contains 51.3 percent of organic matters, 24.6 percent of NPK, 1.0 hundred million/g of effective bacteria, 0.12 percent of mixed bacteria, 0.33 percent of water and 5.8 of pH value.
In this example, the sources of the yeast fermentation waste liquid dry powder and the bacillus subtilis powder are the same as those in example 1, and the preparation method is the same as that in example 1.
Example 4
Water-soluble composite microbial fertilizer 4: 73% of yeast fermentation waste liquid dry powder, 10% of ammonium sulfate, 5% of monoammonium phosphate, 10% of potassium sulfate and 2.0% of bacillus licheniformis powder; the mass ratio of the yeast fermentation waste liquid dry powder to the inorganic water-soluble fertilizer to the functional bacteria powder is 73:25: 2.
The water-soluble compound microbial fertilizer 4 prepared by the embodiment contains 48.3% of organic matters, 22.6% of NPK, 1.0 hundred million/g of effective bacteria, 0.08% of mixed bacteria rate, 0.43% of water and 5.9 of pH value.
In this example, the sources of the yeast fermentation waste liquid dry powder and the Bacillus licheniformis powder are the same as those in example 2, and the preparation method is the same as that in example 2.
Performance testing
1. Solubility analysis of composite microbial fertilizer
The solubility of the water-soluble composite microbial fertilizers of examples 1 to 4 was verified by referring to a method of "determination of water-insoluble content and pH of water-soluble fertilizer" (NY/T1973-2010).
1g of a sample (accurate 0.001g) was weighed, placed in a beaker, added to 250mL of water and stirred well for 3 min. Filtering with a glass crucible type filter which is dried in a drying oven at (110 +/-2) DEG C to constant weight in advance, transferring all residues into the filter with water as little as possible, transferring the filter into the drying oven at 110 ℃, drying for 1h, taking out the filter, transferring the filter into a dryer, cooling to room temperature, and weighing. Three replicates of each example gave the average values shown in table 1.
TABLE 1 detection results of water-insoluble substances
As can be seen from Table 1, the average water-insoluble content of the water-soluble compound microbial fertilizer in the examples 1 to 4 is 0.08 to 0.25 percent, the water-insoluble content required by the standard of general water-soluble fertilizers is less than or equal to 5 percent, and the water-soluble compound microbial fertilizer provided by the invention has lower water-insoluble content and better water solubility and can meet the requirements of water-soluble fertilizers.
2. Functional bacteria stability analysis of composite microbial fertilizer
In each example, 3 bags are sealed by 1kg of aluminum foil bags and stored in a room-temperature dry place, the number of effective viable bacteria is detected every three months, and a sample is taken from the unopened packaging bag every time until 6 months of storage. The effective viable count detection method is carried out according to a standard method of microbial fertilizer product detection regulations (NY/T2321-2013).
TABLE 2 effective viable count variation for different products
| Examples | Initial number of bacteria (hundred million/g) | 3 months of bacteria (hundred million/g) | 6 months bacterium number (hundred million/g) |
| Example 1 | 1.2 | 1.18 | 1.09 |
| Example 2 | 0.7 | 0.65 | 0.67 |
| Example 3 | 1.0 | 0.99 | 0.92 |
| Example 4 | 1.0 | 0.95 | 0.90 |
As can be seen from Table 2, the effective viable count of each example does not change much during the storage process for up to six months, and is still within the range that the effective viable count required by the composite microbial fertilizer (NY/T798-2015) is more than or equal to 0.2 hundred million/g, which indicates that the water-soluble composite microbial fertilizer provided by the invention has stable bacterial count and can ensure the stability of the product.
As can be seen from the comparative examples and the examples, the water-soluble compound microbial fertilizer provided by the invention has the advantages of low content of water-insoluble substances, good water solubility, stable bacterial count and capability of ensuring the stability of the product, and can meet the requirements of water-soluble fertilizers.
The foregoing is merely a preferred embodiment of the invention and is not intended to limit the invention in any manner. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be construed as the protection scope of the present invention.
Claims (10)
1. A water-soluble composite microbial fertilizer comprises yeast fermentation waste liquid dry powder, inorganic water-soluble fertilizer and functional bacteria powder; the mass ratio of the yeast fermentation waste liquid dry powder to the inorganic water-soluble fertilizer to the functional bacteria powder is (68-85) to (15-30) to (0.3-3.5).
2. The water-soluble composite microbial fertilizer as claimed in claim 1, wherein the mass content of water-soluble organic matters in the yeast fermentation waste liquid dried powder is not less than 60%, the mass content of water-soluble fulvic acid is not less than 50%, the mass content of nitrogen, phosphorus and potassium is not less than 10%, and the mass content of crude protein is 18-22%.
3. The water-soluble composite microbial fertilizer according to claim 1 or 2, wherein the yeast fermentation waste liquid dry powder further comprises B vitamins, vitamin C, inositol, polysaccharide, trace elements, mycoprotein, nucleic acid, surfactant and growth promoting factor.
4. The water-soluble composite microbial fertilizer according to claim 1, wherein the pH value of the yeast fermentation waste liquid dried powder is 5-6.
5. The water-soluble composite microbial fertilizer according to claim 1, wherein the inorganic water-soluble fertilizer is a nitrogen-phosphorus-potassium compound fertilizer.
6. The water-soluble composite microbial fertilizer according to claim 5, wherein the inorganic water-soluble fertilizer comprises two or more of urea, ammonium chloride, ammonium sulfate, monopotassium phosphate, monoammonium phosphate, potassium sulfate and potassium nitrate.
7. The water-soluble composite microbial fertilizer according to claim 1, wherein the functional bacterial powder comprises one or more of paenibacillus mucilaginosus powder, bacillus subtilis powder and bacillus licheniformis powder.
8. The water-soluble composite microbial fertilizer according to claim 7, wherein the preparation of each of the functional bacterial powders comprises the following steps:
(1) sequentially performing strain activation, primary fermentation, secondary fermentation and tertiary fermentation on the strain to obtain a zymocyte liquid;
(2) carrying out membrane filtration concentration on the zymocyte liquid obtained in the step (1) to obtain concentrated bacteria liquid;
(3) and (3) carrying out spray drying on the concentrated bacterial liquid obtained in the step (2) to obtain bacterial powder.
9. The water-soluble composite microbial fertilizer as claimed in claim 1, wherein the effective viable count of the water-soluble composite microbial fertilizer is more than or equal to 0.2 hundred million/g, the organic matter content of the water-soluble composite microbial fertilizer is more than or equal to 20 percent, and N, P percent is calculated2O5And K2The total nutrient content of O is 8.0-25.0%, the rate of mixed bacteria is less than or equal to 30%, the water content is less than or equal to 30%, and the pH value is 5.5-8.5.
10. The preparation method of the water-soluble composite microbial fertilizer of any one of claims 1 to 9, comprising the following steps: and mixing the yeast fermentation waste liquid dry powder, the inorganic water-soluble fertilizer and the functional bacteria powder to obtain the water-soluble composite microbial fertilizer.
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