Preparation method of disease-resistant biological bacterial fertilizer for increasing yield of field crops

文档序号：298014 发布日期：2021-11-26 浏览：41次中文

阅读说明：本技术 一种大田作物增产抗病生物菌肥的制备方法 (Preparation method of disease-resistant biological bacterial fertilizer for increasing yield of field crops ) 是由付刚锋李虎申谢荣芳苑学亮王子强李振娥罗秀英王洪峰商思森于 2021-09-07 设计创作，主要内容包括：本发明公开了一种大田作物增产抗病生物菌肥的制备方法,包括：步骤1,制备第一复合菌剂、第二复合菌剂；步骤2：从蜡蚧轮枝菌、白僵菌、绿僵菌的菌株库中筛选若干菌株,从中选取三种与步骤1中所得的第二复合菌剂拮抗最小的真菌菌株,进行扩大培养；步骤3,制得包衣剂；步骤4,发酵；步骤5,造粒、包衣；步骤6：真空低温烘干。本发明可解决现有技术中利用草木灰、糠醛渣、菌渣制备的大田生物菌肥,不具备多种不同功能活性微生物,微生物定殖存活性能低,不具备防病、防虫效果的问题。(The invention discloses a preparation method of a disease-resistant biological bacterial fertilizer for increasing yield of field crops, which comprises the following steps: step 1, preparing a first compound microbial inoculum and a second compound microbial inoculum; step 2: screening a plurality of strains from a strain library of verticillium lecanii, beauveria bassiana and metarhizium anisopliae, selecting three strains with the minimum antagonism with the second compound microbial inoculum obtained in the step 1, and carrying out expanded culture; step 3, preparing a coating agent; step 4, fermenting; step 5, granulating and coating; step 6: and (5) drying in vacuum at low temperature. The invention can solve the problems that the field biological bacterial fertilizer prepared by using plant ash, furfural residues and bacterial residues in the prior art does not have various microorganisms with different functional activities, has low microbial colonization survival performance and does not have the effects of disease prevention and insect prevention.)

1. A preparation method of disease-resistant biological bacterial fertilizer for increasing yield of field crops is characterized by comprising the following steps:

step 1: respectively activating and gradually amplifying protease producing bacteria, lipase producing bacteria, cellulase producing bacteria, amylase producing bacteria, potassium bacteria and phosphorus bacteria obtained from the common microorganism center of China Committee for culture Collection of microbial strains according to the provided strain specifications to prepare microbial inoculums, wherein the microbial inoculums prepared by international units are taken as metering units, and the microbial inoculums are prepared according to the ratio of (0.5-0.9) of protease to lipase to amylase to cellulase to (0.2-0.6) to (0.3-0.6) to (0.8-1.2) by taking FPA activity units as reference, and then adding 2-18% of a potassium bacterial agent and 5-15% of a phosphorus bacterial agent in volume ratio to prepare a first compound microbial inoculums for microbial active fermentation production; the number of viable bacteria contained in the first composite microbial inoculum is 6-8 multiplied by 109 cfu/ml;

respectively screening a plurality of strains from a strain library of bacillus thuringiensis, bacillus cereus and arthrobacter, selecting three strains with minimum antagonism with the first compound microbial agent from the strains, and respectively carrying out amplification culture, wherein the CFU ratio of the bacillus thuringiensis to the bacillus cereus to the arthrobacter is 10: 1-2: 1-2, compounding to form a second complex microbial inoculum; the number of viable bacteria contained in the second composite microbial inoculum is 6-8 multiplied by 109 cfu/ml;

step 2: screening a plurality of strains from a strain library of verticillium lecanii, beauveria bassiana and metarhizium anisopliae, selecting three strains with the minimum antagonism with the second composite microbial inoculum obtained in the step 1, carrying out amplification culture, and compounding in equal proportion to prepare a fungal microbial inoculum culture, wherein the microbial enzyme activity unit manufactured by international unit is taken as a measuring unit, and the fungal microbial inoculum culture contains 6-8 multiplied by 109cfu/ml of viable bacteria;

and step 3: and (3) mixing the fungus microbial inoculum culture obtained in the step (2) with starch, caramel pigment and water at the temperature of 30-40 ℃ according to the mass ratio of (0.5-1) to (3-6): (0.2-0.5): (20-45) uniformly mixing to prepare a coating agent;

and 4, step 4: crushing furfural residues and mushroom residues, mixing according to the weight ratio of 1:1.2-1.5, and adjusting the water content to 60-65% to obtain an organic matter to be fermented; piling organic matters to be fermented into a trapezoidal stack with the height of 1.5-1.8 m; inoculating a first compound microbial inoculum to the materials in the stack according to the inoculation amount of 0.3-0.8% of the dry weight of the materials in the stack to ensure that the microorganisms are fully contacted with the materials; aerobic fermentation is carried out at normal temperature, and the fermentation is carried out once a day for 5-7 days;

a plastic film is arranged on a top cover of the stack, so that the temperature of the materials in the stack is rapidly raised to 68-75 ℃, the duration is 3-5 days, and the materials in the stack are killed, the bacteria are killed, and the cellulose is degraded;

adding plant ash into the stack, adjusting the water content in the stack to 50-55%, and adjusting the pH value to 6.5-7.5;

adding the second composite microbial inoculum obtained in the step 1 according to 1-2% of the dry weight of the substances in the stack; mixing, and performing low-temperature oxygen-consuming stacking fermentation; turning and tamping the stacks once a day, keeping the temperature of the materials of 20-30 cm in the stacks at 28-35 ℃, and keeping the time for 8-10 days; turning and smashing until the temperature of the pile is the same as the outdoor temperature, and reducing the water content to 18-22% to obtain fermented organic matters;

and 5: putting the fermented organic matter into a granulator, and spraying a granulation auxiliary agent for granulation; drying the prepared particles in vacuum at low temperature; screening out particles meeting the requirements through a screening machine;

step 6: and (3) putting the prepared particles into a coating machine, uniformly scattering a chelating agent and a nitrification inhibitor on the surfaces of the particles, adding the coating agent obtained in the step (3) in a spray manner, and adhering the microbial inoculum to the surfaces of the organic particles to form a microbial inoculum coating to obtain a finished product.

2. The method for preparing the disease-resistant biological bacterial fertilizer for increasing yield of field crops according to claim 1, which is characterized in that: the granulation auxiliary agent is a byproduct slurry in amino acid production.

3. The method for preparing the disease-resistant biological bacterial fertilizer for increasing yield of field crops according to claim 1, which is characterized in that: the nitrification inhibitor is one of nitropyridine or thiosulfate.

4. The method for preparing the disease-resistant biological bacterial fertilizer for increasing yield of field crops according to claim 1, which is characterized in that: the chelating agent is an amino acid chelating agent.

5. The method for preparing the disease-resistant biological bacterial fertilizer for increasing yield of field crops according to claim 4, which is characterized in that: the amino acid chelating agent is one or more of methionine, lysine, glycine, cysteine and histidine.

6. The method for preparing the disease-resistant biological bacterial fertilizer for increasing yield of field crops according to claim 1, which is characterized in that: when the vacuum low-temperature drying is carried out, the temperature is 40-50 ℃, and the vacuum degree is-0.07 MPa to-0.08 MPa.

7. The method for preparing the disease-resistant biological bacterial fertilizer for increasing yield of field crops according to claim 1, which is characterized in that: the granulator is a disc granulator or a roller granulator.

8. The method for preparing the disease-resistant biological bacterial fertilizer for increasing yield of field crops according to claim 1, which is characterized in that: the grain diameter of the finished product is controlled to be 1.0-4.0 mm.

9. The method for preparing the disease-resistant biological bacterial fertilizer for increasing yield of field crops according to claim 1, which is characterized in that: the top width of the stack is 1-1.2 meters, the bottom width is 2-2.4 meters, the length of the stack is determined according to the field, and the volume of each stack is not less than 10m for carrying out the high-speed plantation.

Technical Field

The invention relates to the technical field of fertilizers, in particular to a preparation method of a disease-resistant biological bacterial fertilizer for increasing yield of field crops.

Background

The residue after burning of plants (herbs and woody plants) is called plant ash. Since plant ash is ash generated by burning plants, it is a mineral element contained in plants and almost all plant ash is contained therein. The content of potassium element is maximum, generally 6-12% of potassium is contained, more than 90% of potassium element is water-soluble and exists in the form of carbonate; secondly, phosphorus, which generally contains 1.5 to 3 percent; also contains calcium, magnesium, silicon, sulfur, and trace elements such as ferrum, manganese, copper, zinc, boron, molybdenum, etc. Therefore, the fertilizer is a natural fertilizer with wide source, low cost, complete nutrients and obvious fertilizer effect. However, the plant ash is alkaline and cannot be mixed with certain organic fertilizer raw materials, such as human excrement, stable manure, stack retting fertilizer or ammonium nitrogen fertilizer, and the like for application, otherwise, the volatilization loss of nitrogen is caused; the plant ash can not be mixed with the phosphate fertilizer for application, otherwise, phosphorus fixation can be caused, and the fertilizer efficiency of the phosphate fertilizer is reduced. If the plant ash is directly mixed with the raw materials of nitrogenous fertilizer and phosphate fertilizer, the fertilizer efficiency is reduced, and the environmental pollution is caused by the phenomena of volatilization of nitrogen, fixation of phosphorus and the like. In addition, the plant ash is light, so the plant ash is easy to move with wind when dry and easy to move with water when wet. When the plant ash is directly used as a fertilizer, a part of nutrients can not be absorbed and utilized by plants. Particularly, the plant ash has strong alkalinity and lower utilization rate in alkaline soil. A great deal of research shows that the plant effectiveness of the Ca, Fe, Zn and other medium and trace elements is extremely low under the alkaline condition of ash. Because the nutrient forms of plant ash burnt by different kinds of plants and at different temperatures are greatly different and are more ineffective, if the plant ash is directly applied without treatment, the fertilizer efficiency cannot be fully exerted on the one hand, and the adverse effect on crops can be caused on the other hand. These characteristics have greatly limited the use of plant ash for fertilizer production. The quantity of plant ash produced by burning plants in China every year is very large, taking a biological power plant as an example, the annual discharge amount of ash and smoke ash produced after power generation and combustion of straws used as crops is very large, if the ash and the smoke ash are not treated, atmospheric dust pollution is caused, and water pollution is caused when the ash and the smoke ash are discharged into water bodies such as rivers and lakes.

Chinese patent publication No. CN1189481A discloses a method for preparing high-potassium-calcium agricultural fertilizer by using plant ash, manure ash and the like. The fertilizer is prepared by stirring, grinding and sieving plant ash, manure ash, lime manure, industrial salt, potassium sulfate, potassium chloride and urine. The prepared fertilizer has the defects of high salinity and strong alkalinity, and most elements (such as P, Si, Fe, Zn, Cu, Mn, Mo and the like) in ash are fixed and are not easy to be absorbed and utilized by crops, so the fertilizer is not particularly suitable for alkaline soil.

Chinese patent CN200510095748.5 discloses a method for manufacturing plant straw ash multi-element fertilizer, which takes ash formed by burning, cracking and gasifying plant straws as main raw materials, adds inorganic waste acid solution, agricultural organic acid and/or organic solid wastes which can generate organic acid by composting reaction to reduce the pH value of the raw materials to 6.0-7.5, and then the raw materials are fermented for 1-10 days, and then the raw materials are dried and crushed into products or are added with 3-10% of adhesive after being crushed, and then the finished fertilizer is obtained by granulation. Although the pH value of the ash is reduced to be neutral, the method has the advantages that the beneficial microbial content in the fertilizer is low, and the defects of short fertilizer effect, easy loss, easy water pollution and the like of the original ash fertilizer are still not solved.

The organic fertilizer prepared from the furfural residues has the advantages that: firstly, increase farmland organic matter, improve soil structure. The furfural residues are rich in organic matters and humic acid, so that the number of soil aggregate structures can be increased, the soil permeability can be enhanced, and the physical and chemical properties of the soil can be improved after the application. And secondly, adjusting the pH value of the soil and improving the nutrient effectiveness. The furfural residue contains high free acid, can neutralize alkali root ions in alkaline soil such as saline-alkali soil, calcareous soil and rice soil, adjust the pH value of the soil, and increase the effectiveness of cations such as calcium, magnesium and zinc. Thirdly, the soil is activated, and the toxicity is reduced. The furfural residue contains high pentosan after decomposition, is a high-quality energy source of beneficial microorganisms in soil, can increase the activity of the microorganisms in the soil after application, and reduces the harm of heavy metal ions. The furfural residue is a byproduct of corncob decomposition, is discarded as an industrial waste at present, and pollutes the environment. The pH value of the furfural residue is about 1.90. The furfural residue is acidic, so that the furfural residue can be directly applied to the field as a fertilizer, can not be absorbed and utilized by crops, and the strong acidity of the furfural residue also influences the growth of the crops, so that the yield increase effect cannot be achieved, and the crop yield is influenced, even the crop is out of production. The furfural residue mainly comprises the following components: 76% of organic matter and 11.5% of humic acid.

The mushroom dregs are culture carrier for culturing edible mushroom, and the main components of the mushroom dregs are leftover of agricultural and sideline products, such as cottonseed shell, corncob, straw, coconut shell, wood dust, chicken manure and the like. In recent years, the cultivation of edible fungi in China is more and more generalized and scaled, and the result of the concentrated scale production is that the edible fungi residues after picking are largely discarded, which causes serious resource waste and environmental problems. Researches show that the waste mushroom dregs contain saccharides, organic acids, enzymes, bioactive substances, abundant proteins, and various beneficial components such as nitrogen, phosphorus, potassium, vitamins and the like, and have high utilization value. At present, the utilization of the mushroom dregs mainly focuses on the utilization of the mushroom dregs after treatment, feed for breeding industry, compost, culture medium and the like. The nutrient components and special physical properties contained in the mushroom dregs also attract the attention of many agricultural researchers at present.

Chinese patent CN201110372741.9 discloses a method for producing organic fertilizer by using plant ash and furfural residues of a biological power plant, which comprises the following steps: collecting furfural residues and plant ash of a biological power plant, and air-drying to make the water content of the furfural residues and the plant ash be 15-20%; mixing furfural residues and plant ash of a biological power plant according to the weight ratio of 2.7-3: 2.2-2.5, stirring, stacking and compacting, covering with a plastic film or canvas, standing for 24-36 hours, spreading, airing to remove water to make the water content be 8-10%, and crushing into powder with the granularity of 1-2 mm to obtain a mixture; and (5) spraying and granulating the mixture by using a granulator to obtain a finished product. The method can regulate soil structure, improve physical and chemical properties of soil, promote crop growth and increase yield by using organic fertilizer produced from plant ash and furfural residue of biological power plants. However, the fertilizer prepared by the method is still a single organic fertilizer, and the content of beneficial bacteria in the fertilizer is low.

The patent of publication No. CN104177138A discloses a biological bacterial fertilizer prepared by a solid fermentation technology and application thereof, the biological bacterial fertilizer is prepared by a fermentation raw material and a strain activating solution through the solid fermentation technology; the effective viable count of the strain activating solution is 4-30 hundred million/mL, and the volume-mass ratio of the addition amount of the strain activating solution to the fermentation raw material is 1-5L/100 kg; the fermentation raw materials comprise the following components in parts by mass: 30-50 parts of grape skin; 30-50 parts of mushroom dregs; 10-20 parts of furfural residues; 3-6 parts of plant ash; 0.2-0.8 part of inorganic salt, wherein the solid fermentation process comprises the mixing of fermentation raw materials, adding the strain activating solution into the mixed raw materials, uniformly stirring, flatly paving in a solid fermentation device for fermentation, and drying to obtain the biological bacterial fertilizer. The biological bacterial fertilizer prepared by the solid fermentation method has high effective viable count and strong strain competitiveness, and breaks through the problem of inactivation in the storage and use of strains of the traditional biological bacterial fertilizer. The disadvantage is that the consumption of plant ash is low.

The patent with the publication number of CN104003817A discloses a special disease-resistant compound microbial fertilizer for corn and also relates to a production method of the special disease-resistant compound microbial fertilizer for corn. The special disease-resistant compound microbial fertilizer for the corn comprises urea, a compound microbial agent, bran, chitosan, furfural residue, ammonium sulfate, potassium sulfate, monoammonium phosphate, rapeseed cake meal, manure, humic acid, sesame cake meal, trace elements, calcium superphosphate and plant ash. The bacterial fertilizer disclosed by the invention is balanced in nutrient elements and rich in trace element components, can effectively improve the soil productivity and fertility level, can obviously inhibit the commonly-occurring main corn diseases due to the addition of the compound microbial agent, is long in fertilizer efficiency, can effectively improve the corn yield by 25-30%, effectively prevents and controls the corn diseases, can effectively improve the soil after being used for a long time, and can play a role in the whole growth period of the corn. The disadvantage is that it is not resistant to insect pests.

Patent publication No. CN1724481B discloses a method for producing microbial active organic fertilizer by using waste mushroom dregs, which comprises the following steps: crushing the waste mushroom dregs, adding crushed crop straws or leftovers of a food processing factory, wetting with water to ensure that the water content of the materials is about 65-70%, inoculating 2-8% of an enzyme-producing compound microbial agent, placing the materials in a fermentation tank, sealing, fermenting for 5-15 days at a certain temperature, ventilating for 15 minutes every 24 hours in the fermentation process, and obtaining the microbial active organic fertilizer after the microbial biomass and the enzyme activity reach indexes. The method for producing the microbial active organic fertilizer by using the waste mushroom residues solves the problem of environmental pollution caused by conventional treatment of the waste mushroom residues, simultaneously recycles resources, and solves the problem of low-cost fertilizer sources for the production of green foods and organic foods. The method has the defects that the disease resistance problem is not solved, and in addition, after the mushrooms in the mushroom shed are harvested, mushroom residues contain a large amount of water, a large amount of mixed bacteria can be generated within one or two days after the mushrooms are grown out of the mushroom shed, and the preparation of the biological bacterial manure at the later stage is seriously influenced.

In the development process of agriculture in China, due to the fact that chemical fertilizers and pesticides are used excessively for a long time, a soil ecosystem is seriously damaged, vicious circle of soil conditions is caused, most prominently, soil is hardened, permeability is poor, soil fertility is lowered, plant diseases and insect pests are rampant, applied chemical fertilizers and crops can only absorb one third of the soil, the other two thirds of nitrogen, phosphorus and potassium and trace elements needed by the crops are solidified and lost by the soil, the crops cannot fully absorb needed nutrients, growth is inhibited, plants are small, diseases such as yellowing, lodging and rotting are prone to occurring, insect pest activities such as root-knot nematodes and stem nematodes are rampant, the crops are less harvested, and residual chemical fertilizers and pesticides on the crops threaten the health of people. At present, although there are various microbial agents which can resist insect pests, prevent diseases and promote the growth of crops, the effect is not ideal. The microbial fertilizer is a specific preparation containing living microorganisms, and the microbial organic fertilizer has the advantages of no damage to the soil structure, no toxicity and no harm to people, livestock and the like; the fertilizer efficiency is durable; can improve the crop yield and the crop quality and has the characteristics of low cost, but the microbial organic fertilizer has stronger selectivity and is easily restricted by factors such as soil, environment and the like.

Therefore, the biological bacterial fertilizer which can improve the soil fertility, improve the soil, protect the ecological environment, improve the crop yield and quality and can antagonize the soil-borne disease bacteria of the crops is developed, and the biological bacterial fertilizer has important significance for promoting the virtuous circle and the sustainable development of agriculture.

Disclosure of Invention

The invention aims to solve the problems that when biological bacterial manure prepared by using plant ash and the like in the prior art is used for planting field crops, nitrogen volatilization loss is easily caused when the plant ash content is high, and the bacterial manure bacterial content is low. Solves the problems that the organic fertilizer prepared by utilizing plant ash, furfural residues and mushroom residues in the prior art does not have various microorganisms with different functional activities, has low microorganism colonization survival performance and does not have the effects of disease prevention and insect prevention.

The technical scheme adopted by the invention is as follows.

A preparation method of disease-resistant biological bacterial fertilizer for increasing yield of field crops is characterized by comprising the following steps:

step 1: respectively activating and gradually expanding protease producing bacteria, lipase producing bacteria, cellulase producing bacteria, amylase producing bacteria, potassium bacteria and phosphorus bacteria obtained from China general microbiological culture Collection center (CGMCC) according to the provided strain specifications to prepare microbial inoculum, taking microbial enzyme activity unit of international unit system as a metering unit, wherein the activity unit of the cellulase is based on FPA activity unit, and adding 2-18 percent of protease, lipase, amylase and cellulase in a ratio of (0.5-0.9) to (0.2-0.6) to (0.3-0.6) to (0.8-1.2)Preparing a first compound microbial agent for fermentation production of the microbial active organic fertilizer by using a potassium bacterial agent in percentage by volume and a phosphorus bacterial agent in 5-15% by volume; the number of viable bacteria contained in the first composite microbial inoculum is 6-8 multiplied by 10⁹cfu/ml. The strains of the enzyme-producing compound microbial agent are all from China general microbiological culture Collection center (CGMCC), can be purchased through commercial approaches, and the culture and growth conditions of the strains are carried out according to the instructions provided by a strain preservation unit. The strains to be involved are mainly cellulase-producing strains (e.g., CGMCC No.: 3.316 or 3.2878, etc.), protease-producing strains (e.g., CGMCC No.: 1.230 or 1.265, etc.), lipase-producing strains (e.g., CGMCC No.: 2.1135 or 2.1405, etc.), amylase-producing strains (e.g., CGMCC No.: 1.803 or 1.836 or 1.831, etc.), phosphorus bacteria (e.g., organophosphorus bacteria or inorganic phosphorus bacteria, e.g., CGMCC No.: 1.217 or 1.223 or 1.220 or 1.823, etc.) and potassium bacteria (e.g., CGMCC No.: 1.910, etc.).

Respectively screening a plurality of strains from a strain library of bacillus thuringiensis, bacillus cereus and arthrobacter, selecting three strains with minimum antagonism with the first compound microbial agent from the strains, and respectively carrying out amplification culture, wherein the CFU ratio of the bacillus thuringiensis to the bacillus cereus to the arthrobacter is 10: 1-2: 1-2, compounding to form a second complex microbial inoculum; the second composite microbial inoculum contains viable bacteria with the number of 6-8 multiplied by 10⁹cfu/ml。

Step 2: screening a plurality of strains from a strain library of verticillium lecanii, beauveria bassiana and metarhizium anisopliae, selecting three strains with the minimum antagonism with the second composite microbial inoculum obtained in the step 1, carrying out amplification culture, and compounding in equal proportion to prepare a fungal microbial inoculum culture, wherein the microbial enzyme activity unit manufactured by international unit is taken as a measuring unit, and the fungal microbial inoculum culture contains 6-8 multiplied by 10 viable bacteria⁹cfu/ml。

And step 3: and (3) mixing the fungus microbial inoculum culture obtained in the step (2) with starch, caramel pigment and water at the temperature of 30-40 ℃ according to the mass ratio of (0.5-1) to (3-6): (0.2-0.5): (20-45) and mixing uniformly to prepare the coating agent.

And 4, step 4: crushing furfural residues and mushroom residues, mixing according to the weight ratio of 1:1.2-1.5, and adjusting the water content to 60-65% to obtain an organic matter to be fermented; piling organic matters to be fermented into a trapezoidal stack with the height of 1.5-1.8 m; inoculating a first compound microbial inoculum to the materials in the stack according to the inoculation amount of 0.3-0.8% of the dry weight of the materials in the stack to ensure that the microorganisms are fully contacted with the materials; aerobic fermentation is carried out at normal temperature, and the fermentation is carried out once a day for 5-7 days;

and (3) a plastic film is arranged on the top cover of the stack, so that the temperature of the materials in the stack is rapidly raised to 68-75 ℃, the duration is 3-5 days, and the materials in the stack are killed, the bacteria are killed, and the cellulose is degraded.

Adding plant ash into the stack, adjusting the water content in the stack to 50-55%, and adjusting the pH value to 6.5-7.5;

adding the second composite microbial inoculum obtained in the step 1 according to 1-2% of the dry weight of the substances in the stack; mixing, and performing low-temperature oxygen-consuming stacking fermentation; turning and tamping the stacks once a day, keeping the temperature of the materials of 20-30 cm in the stacks at 28-35 ℃, and keeping the time for 8-10 days; and turning and smashing until the temperature of the pile is the same as the outdoor temperature, and reducing the water content to 18-22% to obtain the fermented organic matter.

And 5: putting the fermented organic matter into a granulator, and spraying a granulation auxiliary agent for granulation; drying the prepared particles in vacuum at low temperature; screening out particles meeting the requirements through a screening machine;

As a preferred technical scheme, the granulation auxiliary agent is a slurry as a byproduct in the production of amino acid.

As a preferred technical scheme, the nitrification inhibitor is one of nitropyridine or thiosulfate.

Preferably, the chelating agent is an amino acid chelating agent.

As a preferred technical scheme, the amino acid chelating agent is one or more of methionine, lysine, glycine, cysteine and histidine.

As a preferred technical scheme, when the drying is carried out in vacuum at low temperature, the temperature is 40-50 ℃, and the vacuum degree is-0.07 MPa to-0.08 MPa.

As a preferred technical scheme, the granulator is a disc granulator or a roller granulator.

As a preferred technical scheme, the particle size of the finished product is controlled to be 1.0-4.0 mm.

As a preferred technical scheme, the top width of the stack is 1-1.2 meters, the bottom width of the stack is 2-2.4 meters, the length of the stack is determined according to the field, and the volume of each stack is not less than 10m for carrying out the year.

The invention has the beneficial effects that:

1. in the production process of the fertilizer, three times of fermentation are adopted. The first fermentation adopts protease producing bacteria, lipase producing bacteria, cellulase producing bacteria, amylase producing bacteria, potassium bacteria and phosphorus bacteria for normal temperature fermentation, and the furfural residue, cellulose, starch, saccharides and other substances in the residue are decomposed at low temperature. Then high-temperature stacking fermentation is adopted. As furfural residue and bacteria residue generate a large amount of acidic substances in the high-temperature fermentation process, various bacteria contained in the organic matter to be fermented are basically killed under the high-temperature acidity. A large amount of plant ash is added into the stack, and the pH value of substances in the stack can be adjusted by utilizing the advantage that the plant ash is alkaline. Is beneficial to the growth of bacillus thuringiensis, bacillus cereus and arthrobacterium during the third low-temperature stacking fermentation.

2. Bacillus thuringiensis is the bacterial insecticide with the largest production quantity and the most extensive application, can generate toxicity to more than 3000 kinds of insects and protozoa, and is researched by researchers particularly in killing mosquitoes and flies. The bacillus thuringiensis can produce a large amount of spores and toxic proteins mainly under the action of stomach toxicity, and after the spores are swallowed by insects, the spores can be propagated into a blood cavity in a large amount through insect intestinal tracts, so that insect septicemia is caused. The toxin produced by the thuringiensis can be divided into endotoxin and exotoxin, wherein the endotoxin is delta-endotoxin, also called parasporal crystal, and can decompose a large amount of small molecular toxin in insect intestinal tracts to kill insects, and the exotoxin is alpha-toxin and beta-toxin, wherein the alpha-toxin is insect intestinal tract destructive enzyme, and the beta-toxin is heat stable exotoxin and has specific pathogenic effect on mosquitoes and flies. However, Bacillus thuringiensis alone is not effective.

The bacillus cereus generates a plurality of active substances in the growth process and has obvious inhibition effect on pathogenic bacteria or conditional pathogenic bacteria; meanwhile, the bacillus cereus can rapidly consume free oxygen in the environment in the alimentary canal to form an intestinal hypoxia environment, is not beneficial to the growth and reproduction of intestinal aerobic organisms (coccidian), promotes the growth of beneficial anaerobic bacteria, generates organic acids such as lactic acid and the like, reduces the pH value of the intestinal canal, and indirectly inhibits the growth of other pathogenic bacteria. Strong bactericidal action. The bacillus thuringiensis and the bacillus cereus have good insecticidal effect when used in combination.

Arthrobacter is one of the most commonly isolated soil bacteria species, and is widely and almost ubiquitous on earth. Arthrobacter can be isolated from soil to plants, from mountains to seawater, from archaeological murals to clinical samples. Arthrobacter are widely distributed in part because of their ability to survive long periods of time under stress conditions caused by starvation, temperature changes, ionizing radiation, oxygen radicals, toxic compounds, and the like. For example, Arthrobacter can be isolated from soil in desert areas of Xinjiang; arthrobacter can be found in antarctic and arctic regions, glacier sludge, and ice dust and ice pits of alpine glaciers; arthrobacter is one of the most prevalent species among the bacteria isolated from leaks in the radionuclide reservoirs of the United states energy division of Washington. In recent years, the acquisition of genome sequences of multiple arthrobacter strains enables people to deeply understand the genomics basis of the wide environmental adaptability of the arthrobacter strains. The stress response related genes such as sigma factor, oxidative stress, osmotic stress, hunger stress, temperature stress and the like in the arthrobacter genome enable the arthrobacter genome to have outstanding capability of resisting environmental stress. The plasmids in arthrobacter often endow the strain with decomposition capability to more compounds and resistance to heavy metals, so that the strain has stronger environmental competitiveness. The above ability makes arthrobacter play an important role in the process of degrading environmental pollutants. In the prior art, the planting of vegetables such as eggplants and the like uses a large amount of pesticides and chemical fertilizers, causes serious environmental pollution of soil, and can play an important role in the process of degrading environmental pollutants by compounding arthrobacter to improve the fertilizer efficiency.

The above composite bacteria can form spores and parasporal crystals in the later growth stage, and can be matched with each other to improve the effect compared with single bacteria. The parasporal crystals are hydrolyzed by proteases into smaller units under specific conditions to produce toxicity. It is insoluble in water and organic solvents, but soluble in alkaline solutions. After the maggots are eaten into the preparation, the toxicity is released under the action of gastric juice, and the toxicity firstly acts on midgut cells of the maggots to paralyze the midgut. As the disease progresses, the adhesive substances of the intestinal wall cells are destroyed, the epithelial cells are peeled off and scattered in the intestine, the permeability of the middle intestine is out of control, the intestinal wall is perforated, and the intestinal contents, spores and bacteria invade the blood cavity in large quantity. The maggots die due to septicemia along with the mass propagation of bacteria. The main symptoms after the maggots are infected with the bacterial diseases are anorexia, food deprivation, slow action, vomiting and diarrhea and death after 1 to 2 days.

3. The verticillium lecanii is an entomopathogenic fungus with wider geographical distribution and host range, can parasitize various insects, and most commonly comprises homoptera aphids, whiteflies, scale insects and the like, thrips of thysanoptera, locusts of orthoptera, mirid bugs of hemiptera, leaf miners and aedes of diptera and the like. Metarhizium anisopliae belongs to Ascomycota, Hypocreales, Clavicipitaceae and Metarhizium, and is a broad-spectrum insecticidal fungus. The insect is mainly asexually propagated in nature, different insects are infected by active invasion of the body wall of the insect, yeast-like propagation can be carried out in the blood cavity of the insect to generate a large amount of insect thallus, and meanwhile, insecticidal toxins such as destituxin (non-ribosomal polypeptide toxin) and the like can be generated, so that the insect immunity is inhibited, and the insect disinfestation is accelerated. When the insect host does not exist, the metarhizium anisopliae can form rhizosphere symbiotic relationship with the root of the plant to carry out long-term storage. The two microorganisms can be infected through the digestive tract and the body wall of the maggots, conidia germinate to generate germ tubes when meeting proper temperature and humidity after contacting with the maggots to form hyphae, and simultaneously secrete chitinase and protein toxin to dissolve the maggot body wall to invade the maggots. In the body of the insect, hypha directly absorbs the body fluid nutrient of the insect, grows and continuously multiplies, so that the whole body of the insect is filled with the hypha, the circulation of blood is prevented, and metabolites of the hypha are greatly accumulated in the blood to cause the change of the physical and chemical properties of the blood, so that the maggots are metabolized and died. The maggots die 2-3 days after being infected with diseases, the dead maggots are soft, and then become dry and hard quickly due to the fact that moisture is captured by hyphae in a large amount. After the nutrients in the maggot bodies are absorbed completely, hypha extends out of the bodies along the valve gaps and internode membranes of the maggot bodies and produces conidia, and white fuzz is visible on the bodies.

Metarhizium anisopliae is an important biocontrol fungus, can parasitize various insects, is widely applied in production, and according to incomplete statistics, more than 200 insects can be infected and killed by the fungus all over the world. The bacterium has strong pathogenicity and good effect, is nontoxic to human, livestock and crops, and is one of the most studied and applied entomogenous fungi in the world at present. The research on the metarhizium anisopliae has been carried out for more than 100 years in the world, but the effect is unstable due to the restriction of a plurality of factors, and the large-scale production and application of the metarhizium anisopliae are greatly restricted. Until 1988, the emergence of pest control formulations for australian sugarcane and pasture had no major progress in their application. The invasion of the metarhizium anisopliae to the host is a comprehensive result of physiological and biochemical effects between the host and pathogenic bacteria, and the host can be infected through various ways such as a body wall, an air valve, an alimentary canal and the like, wherein the way of the body wall is a main way. The infection mode reflects the characteristics of the fungi. The pathogenic process can be generally divided into 9 stages: attaching conidia to insect epidermis; ② infectious units germinate on the epidermis, this process requires higher temperatures; (iii) penetration of the epidermis. The process is carried out by penetrating germ tube formed by germination of conidium into epidermis directly or forming attachment cell to attach on epidermis firmly, then generating slender invasion filament from attachment cell to penetrate into epidermis, under combined action of mechanical pressure generated by germination of germ tube and relevant enzyme secreted in growth process. The secreted enzyme can degrade protein, chitin and other substances in the body wall, so that the germ tube of spore germination can penetrate the body wall; mycelium grows in the body cavity; producing toxin. Many entomogenous fungi overcome the defense reaction of hosts before they invade the hosts extensively, and are considered to play an important role as toxins, which are known to be mostly small-molecule carboxypeptids and proteases; sixthly, the host dies; seventhly, the hyphae propagate in the host body in a large quantity and invade all organs of the host, and finally the worm body is rigid; allowing the hypha to penetrate out of the host body; ninthly, new infection units are generated and spread. The traditional view about the cause of the destructor of the metarhizium anisopliae is that hyphae rapidly proliferate in the polypide, and the host is finally killed after the nutrition of the host is absorbed by the mass consumption, but scholars think that the metarhizium anisopliae firstly secretes some substances in the process of invading the host, kills the host and then starts to breed the hyphae in large quantity.

In the invention, two fungi microbial agents are compounded. The microorganisms can be infected through the alimentary canal and the body wall of the maggots, conidia germinate to generate germ tubes when meeting proper temperature and humidity after contacting with the maggots to form hyphae, secrete chitinase and protein toxin at the same time, and dissolve the maggots body wall to invade the maggots. In the body of the insect, hypha directly absorbs the body fluid nutrient of the insect, grows and continuously multiplies, so that the whole body of the insect is filled with the hypha, the circulation of blood is prevented, and metabolites of the hypha are greatly accumulated in the blood to cause the change of the physical and chemical properties of the blood, so that the maggots are metabolized and died. The maggots die 2 to 3 days after being infected with diseases, the body of the dead maggots is soft, and then the maggots quickly become dry and hard because the hyphae take a large amount of moisture. After nutrients in the maggot bodies are absorbed completely, hypha extends out of the bodies along valve gaps and internode membranes of the maggot bodies and produces conidia, and white fuzz is visible on the bodies. The coating agent can be effectively prepared according to plant diseases and insect pests in the field. The granulation auxiliary agent is used for granulation, and is firstly decomposed and released in the using process of the bacterial manure, so that the mutual influence between fungi and other bacteria is reduced.

4. And the influence of a high-temperature environment on the activity of microorganisms is solved by adopting vacuum low-temperature drying, and the live microorganisms of the product can not be killed by cold air drying.

5. The invention makes full use of the furfural residues and the fungus residues to generate various amino acids and plant growth regulating substances in the decomposing process, adds the chelating agent, has the advantages that the nutrient components in the soil are not easy to be fixed, are easy to dissolve in water, are not dissociated, can be well absorbed and utilized by the plants, is mixed with other solid or liquid fertilizers for application without chemical reaction, and does not reduce the fertilizer efficiency of any fertilizer. The addition of the nitrification inhibitor can inhibit the biotransformation process of converting ammonium nitrogen into nitrate Nitrogen (NCT), and reduce the loss of nitrogen fertilizer in the form of nitrate nitrogen and the influence on the ecological environment by reducing the generation and accumulation of nitrate nitrogen in soil. But has no harm to people in the using range.

Drawings

FIG. 1 is a flow chart of the preparation method of the disease-resistant biological bacterial fertilizer for increasing yield of field crops.

Detailed Description

Example 1. A preparation method of disease-resistant biological bacterial fertilizer for increasing yield of field crops is characterized by comprising the following steps:

step 1: respectively activating and gradually amplifying protease producing bacteria, lipase producing bacteria, cellulase producing bacteria, amylase producing bacteria, potassium bacteria and phosphorus bacteria obtained from China general microbiological culture Collection center (CGMCC) according to provided strain specifications to prepare microbial inoculums, taking microbial inoculums prepared by international units as metering units, wherein the cellulase activity units take FPA activity units as reference, and adding 2% of potassium bacterial inoculums and 5% of phosphorus bacterial inoculums according to the ratio of 0.5: 0.2: 0.3: 0.8 of protease to lipase to cellulase to prepare a first compound microbial inoculums for fermentation production of the microbial active organic fertilizer; the first composite bacterial preparation has viable count of 6 × 10⁹cfu/ml。

Respectively screening 10 strains from a strain library of bacillus thuringiensis, bacillus cereus and arthrobacter, selecting three strains with minimum antagonism with a first compound microbial agent from the strains, and respectively carrying out amplification culture, wherein the CFU ratio of the bacillus thuringiensis to the bacillus cereus to the arthrobacter is 10: 1: 2, compounding to form a second complex microbial inoculum; the second composite bacterial preparation contains viable bacteria of 6 × 10⁹cfu/ml。

The selected strains are as follows: bacillus thuringiensis (CGMCC No. 4923), Arthrobacter CGMCC No. 7779; bacillus cereus (CGMCC No. 7069).

Step 2: screening 10 strains from a strain library of verticillium lecanii, beauveria bassiana and metarhizium anisopliae, selecting three strains with the minimum antagonism with the second compound microbial inoculum obtained in the step 1, carrying out amplification culture, and compounding in equal proportion to prepare a fungal microbial inoculum culture, wherein the microbial enzyme activity unit manufactured by international unit is taken as a measuring unit, and the fungal microbial inoculum culture contains viable count of 6 multiplied by 10⁹cfu/ml; during the antagonism test, the selected strain and the second complex microbial inoculum are mixed and inoculated on a culture medium, the culture is carried out at a proper temperature, a culture dish or a conical flask is taken out after a period of time, and the antagonism degree can be obtained by visual observation according to the characteristics of colony morphology or bacterial suspension and the like or microscopic examination.

The selected strains are as follows: beauveria bassiana (CGMCC No. 15866), Verticillium lecanii (CGMCC No. 8453), Metarhizium anisopliae (CGMCC No. 2375).

And step 3: and (3) mixing the fungal microbial inoculum culture obtained in the step (2) with starch, caramel pigment and water at the temperature of 30 ℃ according to the mass ratio of 0.5: 3: 0.2: 20 and mixing to obtain the coating agent.

And 4, step 4: crushing furfural residues and mushroom residues, mixing according to the weight ratio of 1:1.2, and adjusting the water content to 60% to obtain an organic matter to be fermented; piling organic matters to be fermented into a trapezoidal stack with the height of 1.5 meters; inoculating a first composite microbial inoculum to the materials in the stack according to the inoculation amount of 0.3 percent of the dry weight of the materials in the stack to ensure that the microorganisms are fully contacted with the materials; aerobic fermentation is carried out at normal temperature, and the fermentation is carried out once a day for 5 days. The fermentation temperature is 10-30 ℃.

And (3) a plastic film is arranged on a top cover of the stack, so that the temperature of the materials in the stack is rapidly raised to 68 ℃, the duration is 3 days, and the mixed bacteria of the materials in the stack are killed and the cellulose is degraded.

Adding plant ash into the stack, and adjusting the water content in the stack to 50%; the pH value is adjusted to 6.5.

Adding the second composite microbial inoculum obtained in the step 1 according to 1 percent of the dry weight of the substances in the stack; mixing, and performing low-temperature oxygen-consuming stacking fermentation; turning and tamping the stacks once a day, keeping the temperature of the 20 cm materials in the stacks at 28 ℃, and keeping the time for 8 days; and turning and smashing until the temperature of the pile is the same as the outdoor temperature and the moisture content is reduced to 18 percent, so as to obtain the fermented organic matter.

Step 6: and (3) putting the prepared particles into a coating machine, uniformly scattering a chelating agent and a nitrification inhibitor on the surfaces of the particles, adding the coating agent obtained in the step (3) in a spray manner, and adhering the microbial inoculum to the surfaces of the organic particles to form a microbial inoculum coating to obtain a finished product.

The granulation auxiliary agent is a byproduct slurry in amino acid production.

The weight ratio of the chelating agent to the nitrification inhibitor is 0.2 percent. The nitrification inhibitor is ammonium thiosulfate.

The chelating agent is an amino acid chelating agent. The amino acid chelating agent is glycine. When the mixture is dried in vacuum at low temperature, the temperature is 40 ℃, and the vacuum degree is-0.07 MPa.

The granulator is a disc granulator.

The grain diameter of the finished product is controlled to be 1.0 mm.

The top width of the stack is 1 meter, the bottom width is 2 meters, the stack length is determined according to the field, and the volume of each stack is not less than 10m for carrying out the cultivation.

Example 2. A preparation method of disease-resistant biological bacterial fertilizer for increasing yield of field crops is characterized by comprising the following steps:

step 1: respectively activating and gradually amplifying protease producing bacteria, lipase producing bacteria, cellulase producing bacteria, amylase producing bacteria, potassium bacteria and phosphorus bacteria obtained from China general microbiological culture Collection center (CGMCC) according to provided strain specifications to prepare microbial inoculums, taking microbial inoculums prepared by international units as metering units, wherein the microbial inoculums activity units of the international units are based on FPA activity units, and adding potassium bacteria microbial inoculums and phosphorus bacteria in a volume ratio of 15% and 8% according to a ratio of protease to lipase to amylase to cellulase of 0.7: 0.4: 0.5: 1.2 to prepare the microbial inoculums into a first compound microbial inoculums for fermentation production of the microbial active organic fertilizers; first composite bacteriumThe preparation contains viable bacteria of 6 × 10⁹cfu/ml。

Respectively screening a plurality of strains from a strain library of bacillus thuringiensis, bacillus cereus and arthrobacter, selecting three strains with minimum antagonism with the first compound microbial agent from the strains, and respectively carrying out amplification culture, wherein the CFU ratio of the bacillus thuringiensis to the bacillus cereus to the arthrobacter is 10: 2: 2, compounding to form a second complex microbial inoculum; the second composite bacterial preparation contains viable bacteria of 8 × 10⁹cfu/ml。

The selected strains are as follows: bacillus thuringiensis (CGMCC No. 4923), arthrobacterium (CGMCC No. 16246); bacillus cereus (CGMCC No. 7069).

Step 2: screening a plurality of strains from a strain library of verticillium lecanii, beauveria bassiana and metarhizium anisopliae, selecting three strains with the minimum antagonism with the second composite microbial inoculum obtained in the step 1, carrying out amplification culture, and carrying out equal proportion compounding to prepare a fungal microbial inoculum culture, wherein the microbial enzyme activity unit manufactured by international unit is taken as a measuring unit, and the fungal microbial inoculum culture contains viable count of 8 multiplied by 10⁹cfu/ml；

The selected strains are as follows: beauveria bassiana (CGMCC No. 15866), Verticillium lecanii (CGMCC No. 8453), Metarhizium anisopliae (CGMCC No. 8385).

And step 3: and (3) mixing the fungal microbial inoculum culture obtained in the step (2) with starch, caramel pigment and water at the temperature of 35 ℃ according to the mass ratio of 1: 5: 0.3: 35 and mixing to obtain the coating agent.

And 4, step 4: crushing furfural residues and mushroom residues, mixing according to the weight ratio of 1:1.3, and adjusting the water content to 65% to obtain an organic matter to be fermented; piling organic matters to be fermented into a trapezoidal stack with the height of 1.8 m; inoculating a first composite microbial inoculum to the materials in the stack according to the inoculation amount of 0.5 percent of the dry weight of the materials in the stack to ensure that the microorganisms are fully contacted with the materials; aerobic fermentation is carried out at normal temperature, and the fermentation is carried out once a day for 6 days;

a plastic film is arranged on a top cover of the stack, so that the temperature of the materials in the stack is quickly raised to 70 ℃, the duration is 3-5 days, and the materials in the stack are killed, the bacteria are killed, and the cellulose is degraded;

adding plant ash into the stack, and adjusting the water content in the stack to 50%; the pH value is adjusted to 7.5.

Adding the second composite microbial inoculum obtained in the step 1 according to 1-2% of the dry weight of the substances in the stack; mixing, and performing low-temperature oxygen-consuming stacking fermentation; turning and tamping the stacks once a day, keeping the temperature of the materials with the length of 30 cm in the stacks at 30 ℃, and keeping the time for 10 days; and turning and smashing until the temperature of the pile is the same as the outdoor temperature and the moisture content is reduced to 20%, and obtaining the fermented organic matter.

The granulation auxiliary agent is a byproduct slurry in amino acid production.

The weight ratio of the chelating agent to the nitrification inhibitor is 0.2 percent. The nitrification inhibitor is nitropyridine.

The chelating agent is an amino acid chelating agent. The amino acid chelating agent is methionine. When the drying is carried out in vacuum at low temperature, the temperature is 45 ℃, and the vacuum degree is-0.08 MPa.

A granulator drum granulator.

The grain diameter of the finished product is controlled to be 4.0 mm.

The top width of the stack is 1.2 meters, the bottom width is 2.4 meters, the length of the stack is determined according to the field, and the volume of each stack is not less than 10m for carrying out the high-speed cultivation.

Example 3. A preparation method of disease-resistant biological bacterial fertilizer for increasing yield of field crops is characterized by comprising the following steps:

step 1: respectively activating protease producing bacteria, lipase producing bacteria, cellulase producing bacteria, amylase producing bacteria, potassium bacteria and phosphorus bacteria obtained from China general microbiological culture Collection center according to provided strain specifications, performing amplification culture step by step to prepare into microbial inoculum, and taking microbial enzyme activity unit of International System of units as metering unit, whereinThe activity unit of the cellulase is based on the activity unit of FPA, and according to the proportion of 0.5: 0.6: 0.4: 1.2 of protease, lipase, amylase and cellulase, 18% of potassium bacterial agent and 15% of phosphorus bacterial agent in volume ratio are added to prepare a first compound microbial agent for fermentation production of the microbial active organic fertilizer; the first composite bacterial preparation contains viable bacteria with the number of 8 multiplied by 10⁹cfu/ml。

Respectively screening a plurality of strains from a strain library of bacillus thuringiensis, bacillus cereus and arthrobacter, selecting three strains with minimum antagonism with the first compound microbial agent from the strains, and respectively carrying out amplification culture, wherein the CFU ratio of the bacillus thuringiensis to the bacillus cereus to the arthrobacter is 10: 1-2: 1-2, compounding to form a second complex microbial inoculum; the second composite bacterial preparation contains viable bacteria of 8 × 10⁹cfu/ml。

The selected strain was the same as in example 1.

Step 2: screening a plurality of strains from a strain library of verticillium lecanii, beauveria bassiana and metarhizium anisopliae, selecting three strains with the minimum antagonism with the second composite microbial inoculum obtained in the step 1, carrying out amplification culture, and carrying out equal proportion compounding to prepare a fungal microbial inoculum culture, wherein the microbial enzyme activity unit manufactured by international unit is taken as a measuring unit, and the fungal microbial inoculum culture contains viable count of 8 multiplied by 10⁹cfu/ml；

The selected strain was the same as in example 1.

And step 3: and (3) mixing the fungus microbial inoculum culture obtained in the step (2) with starch, caramel pigment and water at the temperature of 30-40 ℃ according to the mass ratio of 1: 6: 0.5: 45 and mixing to obtain the coating agent.

And 4, step 4: crushing furfural residues and mushroom residues, mixing according to the weight ratio of 1:1.5, and adjusting the water content to 65% to obtain an organic matter to be fermented; piling organic matters to be fermented into a trapezoidal stack with the height of 1.8 m; inoculating a first composite microbial inoculum to the materials in the stack according to the inoculation amount of 0.8 percent of the dry weight of the materials in the stack to ensure that the microorganisms are fully contacted with the materials; aerobic fermentation is carried out at normal temperature, and the fermentation is carried out once a day for 7 days;

a plastic film is arranged on a top cover of the stack, so that the temperature of the materials in the stack is rapidly raised to 75 ℃, the duration is 5 days, and the mixed bacteria of the materials in the stack are killed and the cellulose is degraded;

adding plant ash into the stack, and adjusting the water content in the stack to 55%; the pH value is adjusted to 7.

Adding the second composite microbial inoculum obtained in the step 1 according to 2% of the dry weight of the substances in the stack; mixing, and performing low-temperature oxygen-consuming stacking fermentation; turning and tamping the stacks once a day, keeping the temperature of the materials of 30 cm in the stacks at 35 ℃, and keeping the time for 10 days; and turning and smashing until the temperature of the pile is the same as the outdoor temperature, and reducing the water content to 22% to obtain the fermented organic matter.

The granulation auxiliary agent is a byproduct slurry in amino acid production.

The weight ratio of the chelating agent to the nitrification inhibitor is 0.2 percent. The nitrification inhibitor is thiosulfate.

The chelating agent is an amino acid chelating agent. The amino acid chelating agent is cysteine. When the drying is carried out in vacuum at low temperature, the temperature is 50 ℃, and the vacuum degree is-0.08 MPa.

A granulator drum granulator.

The grain diameter of the finished product is controlled to be 3.0 mm.

Example 4. A preparation method of disease-resistant biological bacterial fertilizer for increasing yield of field crops is characterized by comprising the following steps:

step 1: respectively taking protease producing bacteria, lipase producing bacteria, cellulase producing bacteria, amylase producing bacteria, potassium bacteria and phosphorus bacteria obtained from China general microbiological culture Collection center (CGMCC) according to the provided strainsPerforming activation and stepwise amplification culture on the microorganism to prepare a microbial inoculum, taking a microbial enzyme activity unit prepared by international units as a metering unit, wherein the cellulase activity unit takes an FPA activity unit as a standard, and adding 16% of a potassium bacterial inoculum and 12% of a phosphorus bacterial inoculum in volume ratio according to the ratio of protease, lipase, amylase and cellulase being 0.6: 0.5: 0.6: 1.1; the first composite bacterial preparation contains viable bacteria with the number of 8 multiplied by 10⁹cfu/ml。

Respectively screening a plurality of strains from a strain library of bacillus thuringiensis, bacillus cereus and arthrobacter, selecting three strains with minimum antagonism with the first compound microbial agent from the strains, and respectively carrying out amplification culture, wherein the CFU ratio of the bacillus thuringiensis to the bacillus cereus to the arthrobacter is 10: 1: 2, compounding to form a second complex microbial inoculum; the second composite bacterial preparation contains viable bacteria of 8 × 10⁹cfu/ml。

The selected strain was the same as in example 2.

Step 2: screening a plurality of strains from a strain library of verticillium lecanii, beauveria bassiana and metarhizium anisopliae, selecting three strains with the minimum antagonism with the second composite microbial inoculum obtained in the step 1, carrying out amplification culture, and carrying out equal proportion compounding to prepare a fungal microbial inoculum culture, wherein the microbial enzyme activity unit manufactured by international unit is taken as a measuring unit, and the fungal microbial inoculum culture contains viable count of 8 multiplied by 10⁹cfu/ml；

The selected strain was the same as in example 2.

And step 3: and (3) mixing the fungal microbial inoculum culture obtained in the step (2) with starch, caramel pigment and water at the temperature of 30-40 ℃ according to the mass ratio of 0.7: 5: 0.3: 41 mixing to obtain coating agent.

And 4, step 4: crushing furfural residues and mushroom residues, mixing according to the weight ratio of 1:1.5, and adjusting the water content to 62% to obtain an organic matter to be fermented; piling organic matters to be fermented into a trapezoidal stack with the height of 1.8 m; inoculating a first composite microbial inoculum to the materials in the stack according to the inoculation amount of 0.8 percent of the dry weight of the materials in the stack to ensure that the microorganisms are fully contacted with the materials; aerobic fermentation is carried out at normal temperature, and the fermentation is carried out once a day for 7 days;

adding plant ash into the stack, and adjusting the water content in the stack to 55%; the pH value is adjusted to 7.5.

Adding the second composite microbial inoculum obtained in the step 1 according to 1.5 percent of the dry weight of the substances in the stack; mixing, and performing low-temperature oxygen-consuming stacking fermentation; turning and tamping the stacks once a day, keeping the temperature of 30 cm materials in the stacks at 32 ℃, and keeping the time for 10 days; and turning and smashing until the temperature of the pile is the same as the outdoor temperature and the moisture content is reduced to 20%, and obtaining the fermented organic matter.

The granulation auxiliary agent is a byproduct slurry in amino acid production.

The weight ratio of the chelating agent to the nitrification inhibitor is 0.2 percent. The nitrification inhibitor is nitropyridine.

The chelating agent is an amino acid chelating agent. The amino acid chelating agent is histidine. When the drying is carried out in vacuum at low temperature, the temperature is 40 ℃, and the vacuum degree is-0.08 MPa.

A granulator drum granulator.

The grain diameter of the finished product is controlled to be 3.0 mm.

Example 5. A preparation method of disease-resistant biological bacterial fertilizer for increasing yield of field crops is characterized by comprising the following steps:

step 1: separately producing proteases obtained from China general microbiological culture Collection center (CGMCC)The method comprises the following steps of (1) activating and carrying out amplification culture on bacteria, lipase producing bacteria, cellulase producing bacteria, amylase producing bacteria, potassium bacteria and phosphorus bacteria according to a provided strain specification to prepare a microbial inoculum, wherein the microbial enzyme activity unit prepared by international unit is taken as a metering unit, the activity unit of cellulase is based on the activity unit of FPA, and the potassium bacterial inoculum and the phosphorus bacterial inoculum are added according to the ratio of 0.6: 0.5: 0.6: 1.2 of protease to lipase to amylase to cellulase to prepare a first compound microbial inoculum for fermentation production of the microbial activity; the first composite bacterial preparation has viable count of 7 × 10⁹cfu/ml。

Respectively screening a plurality of strains from a strain library of bacillus thuringiensis, bacillus cereus and arthrobacter, selecting three strains with minimum antagonism with the first compound microbial agent from the strains, and respectively carrying out amplification culture, wherein the CFU ratio of the bacillus thuringiensis to the bacillus cereus to the arthrobacter is 10: 1:1, compounding to form a second complex microbial inoculum; the second composite bacterial preparation contains viable bacteria of 7 × 10⁹cfu/ml。

The selected strain was the same as in example 1.

Step 2: screening a plurality of strains from a strain library of verticillium lecanii, beauveria bassiana and metarhizium anisopliae, selecting three strains with the minimum antagonism with the second composite microbial inoculum obtained in the step 1, carrying out amplification culture, and carrying out equal proportion compounding to prepare a fungal microbial inoculum culture, wherein the microbial enzyme activity unit manufactured by international unit is taken as a measuring unit, and the fungal microbial inoculum culture contains viable count of 7 multiplied by 10⁹cfu/ml；

The selected strain was the same as in example 1.

And 4, step 4: crushing furfural residues and mushroom residues, mixing according to the weight ratio of 1:1.5, and adjusting the water content to 62% to obtain an organic matter to be fermented; piling organic matters to be fermented into a trapezoidal stack with the height of 1.7 m; inoculating a first composite microbial inoculum to the materials in the stack according to the inoculation amount of 0.8 percent of the dry weight of the materials in the stack to ensure that the microorganisms are fully contacted with the materials; aerobic fermentation is carried out at normal temperature, and the fermentation is carried out once a day for 6 days;

a plastic film is arranged on a top cover of the stack, so that the temperature of the materials in the stack is quickly raised to 70 ℃, the duration is 4 days, and the mixed bacteria of the materials in the stack are killed and the cellulose is degraded;

adding plant ash into the stack, and adjusting the water content in the stack to 55%; the pH value is adjusted to 7.2.

Adding the second composite microbial inoculum obtained in the step 1 according to 1.5 percent of the dry weight of the substances in the stack; mixing, and performing low-temperature oxygen-consuming stacking fermentation; turning and tamping the stacks once a day, keeping the temperature of the materials with the length of 30 cm in the stacks at 35 ℃, and keeping the time for 8 days; and turning and smashing until the temperature of the pile is the same as the outdoor temperature and the moisture content is reduced to 20%, and obtaining the fermented organic matter.

The granulation auxiliary agent is a byproduct slurry in amino acid production.

The weight ratio of the chelating agent to the nitrification inhibitor is 0.2 percent. The nitrification inhibitor is nitropyridine.

The chelating agent is an amino acid chelating agent. The amino acid chelating agent is histidine. When the mixture is dried in vacuum at low temperature, the temperature is 40 ℃, and the vacuum degree is-0.07 MPaMPa.

A granulator drum granulator.

The grain diameter of the finished product is controlled to be 3.0 mm.

Example 6. A preparation method of disease-resistant biological bacterial fertilizer for increasing yield of field crops is characterized by comprising the following steps.

Step 1: respectively activating and gradually amplifying protease producing bacteria, lipase producing bacteria, cellulase producing bacteria, amylase producing bacteria, potassium bacteria and phosphorus bacteria obtained from the China Committee for culture Collection of microorganisms according to the provided strain specification to prepare microbial inoculum, taking the microbial enzyme activity unit of International Unit (International Unit) as a metering unit, wherein the cellulase activity unit takes FPA activity unit as the standard, and adding 10% by volume of potassium bacterial inoculum and 8% by volume of phosphorus bacterial inoculum according to the ratio of protease to lipase to amylase to cellulase being 0.5: 0.2: 0.6: 0.8 to prepare a first compound microbial inoculum for fermentation production of the microbial activity organic fertilizer; the first composite bacterial preparation contains viable bacteria with the number of 8 multiplied by 10⁹cfu/ml。

Respectively screening a plurality of strains from a strain library of bacillus thuringiensis, bacillus cereus and arthrobacter, selecting three strains with minimum antagonism with the first compound microbial agent from the strains, and respectively carrying out amplification culture, wherein the CFU ratio of the bacillus thuringiensis to the bacillus cereus to the arthrobacter is 10: 1: 22, compounding to form a second complex microbial inoculum; the second composite bacterial preparation contains viable bacteria of 8 × 10⁹cfu/ml; the strain selection was the same as in example 2.

Step 2: screening a plurality of strains from a strain library of verticillium lecanii, beauveria bassiana and metarhizium anisopliae, selecting three strains with the minimum antagonism with the second composite microbial inoculum obtained in the step 1, carrying out amplification culture, and carrying out equal proportion compounding to prepare a fungal microbial inoculum culture, wherein the microbial enzyme activity unit manufactured by international unit is taken as a measuring unit, and the fungal microbial inoculum culture contains viable count of 8 multiplied by 10⁹cfu/ml; the strain selection was the same as in example 2.

and 4, step 4: crushing furfural residues and mushroom residues, mixing according to the weight ratio of 1:1.2, and adjusting the water content to 60-65% to obtain organic matters to be fermented; piling organic matters to be fermented into a trapezoidal stack with the height of 1.5 meters; inoculating a first compound microbial inoculum to the materials in the stack according to the inoculation amount of 0.3-0.8% of the dry weight of the materials in the stack to ensure that the microorganisms are fully contacted with the materials; aerobic fermentation is carried out at normal temperature, and the fermentation is carried out once a day for 5-7 days;

a plastic film is arranged on a top cover of the stack, so that the temperature of the materials in the stack is rapidly raised to 68 ℃, the duration is 5 days, and the mixed bacteria of the materials in the stack are killed and the cellulose is degraded;

adding plant ash into the stack, and adjusting the water content in the stack to 50%; the pH value is adjusted to 6.8.

Adding the second composite microbial inoculum obtained in the step 1 according to 1 percent of the dry weight of the substances in the stack; mixing, and performing low-temperature oxygen-consuming stacking fermentation; turning and tamping the stacks once a day, keeping the temperature of 20 cm materials in the stacks at 35 ℃, and keeping the time for 10 days; turning and smashing until the temperature of the pile is the same as the outdoor temperature and the moisture content is reduced to 18 percent, and obtaining fermented organic matters;

The granulation auxiliary agent is a byproduct slurry in amino acid production.

The weight ratio of the chelating agent to the nitrification inhibitor is 0.2 percent. The nitrification inhibitor is nitropyridine.

The chelating agent is an amino acid chelating agent.

The amino acid chelating agent is methionine. When the mixture is dried in vacuum at low temperature, the temperature is 40 ℃, and the vacuum degree is-0.07 MPaMPa.

The microbial agent developed by the company can promote the growth and development of crops, improve the yield structure and the product quality and increase the yield by carrying out field tests on the crops such as eggplants, corns and the like.

A granulator drum granulator.

The grain diameter of the finished product is controlled to be 3.0 mm.

Test example 1

1. Test site: shandong chat the village of the town Luzhuang in the east Changfu district of the city, and the village Lu is good at the land parcel.

2. Experiment time, planting in 1 month and 17 days in 2021, and harvesting in 5 months and 7 days in 2021.

3.1 test soil, moisture soil, typical moisture soil, light loam, flat terrain, better irrigation and drainage conditions, moderate fertility, and soil conditions, see table 1.

TABLE 1 test soil conditions

Organic matter (g/kg)	Basic hydrolyzed nitrogen (mg/kg)	Quick-acting phosphorus (mg/kg)	Quick-acting potassium (mg/kg)	pH
					15.9	90	46	210	7.9

3.2 eggplant, round miscellaneous No. 2.

3.3 test fertilizers: the fertilizer product prepared by the method in the embodiment 3 has the following technical indexes: the effective viable count (cfu) is more than or equal to 10.0 hundred million/g, the organic matter is more than or equal to 40 percent, and the extracellular polysaccharide is more than or equal to 1.0 mg/g.

3.4 test methods.

And (3) experimental design: three treatments of experimental design are carried out, each interval is repeated for three times, 9 experimental cells are counted, the area of each cell is 30 square meters, all the cells are arranged randomly, and a protection row is arranged.

K1, applying the test fertilizer and top dressing.

CK, inactivation matrix + topdressing (control).

K2, conventional topdressing.

3.5 fertilizing method.

Conventional fertilization: before sowing, applying base fertilizer, and applying additional fertilizer when the Menga bud appears. The nitrogen-potassium compound fertilizer is applied once every 10 days, and 10kg of the nitrogen-potassium compound fertilizer is applied once per mu.

Base application of the fertilizer to be tested: 40kg of the test fertilizer is applied to each mu of land, is applied together with other base fertilizer, and is uniformly mixed with the ploughed soil.

And thirdly, inactivating the matrix, wherein the time method is consistent with that of the fertilizer for test. The field management is consistent for the three treatments.

4 results and analysis.

4.1 Effect of different treatments on the biological traits of eggplant.

Tests show that the root system of the eggplant is strong, the plant grows vigorously, and the fruits are purplish red and bright in the plot where the fertilizer for the tests is applied. According to the records of field observation of the test field, the average single fruit diameters of K1 and K2 are respectively 13.5cm and 12.2cm, and are respectively increased by 1.0 cm and reduced by 0.3cm compared with the control CK; the average plant heights of K1 and K2 are 78.9cm and 76.2cm respectively, and are increased by 2.1cm and reduced by 0.6cm respectively compared with that of a control CK; the number of results of K1 and K2 single plants is not greatly different from that of the control CK; k1 shows no diseased plants, diseased leaves and pests, and CK and K2 show a small amount of diseased plants, diseased leaves and pests.

4.2 Effect of fertilizer application for test on eggplant yield, see Table 2

TABLE 2 Effect of application of fertilizers for testing on eggplant yield

4.3 analysis of variance of the experimental data statistics, as in Table 3.

TABLE 3 analysis of variance of test data statistics

4.4 the difference between treatments was significant and compared by PLSD method as shown in Table 4.

TABLE 4 multiple comparative analysis

The data analysis shows that the difference between K1 and CK reaches an extremely significant level, and the difference between K2 and CK is not significant.

4.5 benefit situation analysis

The benefits of eggplant application with fertilizer for the test are shown in Table 5

TABLE 5 benefit of eggplant application for test fertilizers

Experiments on eggplants show that the fertilizer product in the embodiment 3 of the invention has obvious yield increasing effect on the eggplants and can play a more obvious role in preventing diseases and resisting insects.

Test example 2. 1. Test site: shandong province, chat about Shenwei village and Weizhuang village in Shenzhou county.

2. Experiment time, planting in 1 month and 15 days in 2021, and harvesting in 5 months and 6 days in 2021.

3.1 test soil, moisture soil, light loam, flat topography, better irrigation and drainage conditions, moderate fertility, soil condition, see table 6.

TABLE 6 test soil conditions

3.2 eggplant, Luqie No. 1.

3.3 test fertilizers: example 4 the fertilizer product produced by the process.

3.4 test methods.

And (3) experimental design: three intervals are designed in the test, each interval is repeated for three times, 9 test cells are counted, the area of each cell is 30 square meters, all the cells are randomly arranged, and protection rows are arranged.

K1, applying the test fertilizer and top dressing.

CK, basal inactivated substrate + topdressing (control).

K2, conventional topdressing.

3.5 fertilizing method.

Conventional fertilization: preparing soil before sowing as base fertilizer, and applying additional fertilizer when the Menga bud appears. Sequentially applying nitrogen-potassium compound fertilizer every 10 days, 10kg per mu.

Basal application of the test fertilizer: 40kg of test organic fertilizer is applied to each mu, and is applied together with other basal fertilizer, and is uniformly mixed with the ploughed soil.

The inactivated substrate was applied basal for a time method consistent with the fertilizer tested. The field management is consistent for the three treatments.

4 results and analysis.

4.1 Effect of different treatments on eggplant biological shape.

Tests show that the root system of the eggplant is strong, the plant grows vigorously, and the fruits are purplish red and bright in the plot where the fertilizer for the tests is applied. According to the records of field observation of the test field, the average single fruit diameters of K1 and K2 are respectively 26.5cm and 25.1cm, and are respectively increased by 1.2cm and reduced by 0.2cm compared with the control CK; the average plant heights of K1 and K2 are 75.6cm and 73.3cm respectively, and are increased by 1.9cm and reduced by 0.4cm respectively compared with that of a control CK; the number of results of K1 and K2 single plants is not greatly different from that of the control CK; k1 shows no diseased plants, diseased leaves and pests, and CK and K2 show a small amount of diseased plants, diseased leaves and pests.

4.2 Effect of fertilizer application for test on eggplant yield, see Table 7

TABLE 7 Effect of application of fertilizers for testing on eggplant yield

4.3 analysis of variance of the experimental data statistics, as in Table 8.

TABLE 8 analysis of variance factors for test data statistics

4.4 the differences between treatments were significant and compared by PLSD method, as shown in Table 9.

TABLE 9 multiple comparative analysis

The data analysis shows that the difference between K1 and CK reaches an extremely significant level, and the difference between K2 and CK is not significant.

4.5 benefits of eggplant application for the Fertilizer tested, as shown in Table 5

TABLE 5 benefit of eggplant application for test fertilizers

The fertilizer product of the embodiment 4 of the invention has obvious yield increasing effect on eggplants and can play a more obvious disease-resistant effect.

Test example 3. The application effect on corn is achieved.

In 2020, the test was carried out in autumn corn field in Shandong province in Houying Luzhuang village in east Changfu district of chat city. The product obtained in the embodiment 5 of the invention is applied (test technical indexes that the effective viable count (cfu) is more than or equal to 10.0 hundred million/g, the organic matter is more than or equal to 40 percent, and the extracellular polysaccharide is more than or equal to 1.0 mg/g).

Physiological manifestation of the treatment: test observation on 7/25/2020: compared with the plot without the bacterial manure, the plot with the bacterial manure has the advantages that the capillary roots of the corn are more, the third layer of aerial roots is larger than the nodal roots, the leaves are green and more, and the straws are high.

When the corns are harvested in 10/2/2020, compared with the plots without the bacterial manure, the plots with the product obtained by applying the invention are stronger in straws, have no bacterial wilt, short bald tips and more grains per ear, and have obvious disease prevention effect, the grain number per ear is increased by 18.87, the thousand seed weight is increased by 4.04g, and the increase rate is 1.41%; the yield is increased by 29.2 kg/mu, the yield is increased by 5.05 percent, and the yield increasing effect is obvious.

Test example 4. The test was conducted on the corn of the village of the towns in the beam village in the high shop county in 1.2020.

2 test content and methods

2.1 basic conditions of the test field

The test is carried out in the land plots of the farmers of the Yangchuncun village and the vanderby in the Beam villages in Gangtang county from 13 days 6 and 10 and 2 days 2020, the soil type is saline-alkali soil, and the fertility level is low in the middle.

TABLE 6 soil nutrient and content table

2.2 test field climate conditions

The average day number of the frost-free period in Gaogang county is 201 days, the effective accumulated temperature is 4445.2C, the perennial rainfall is 541.6mm, and the rainfall condition from the sowing period to the harvesting period of the corn in 2020: 29.6mm in beam village and town 6 month, 19 days, 9 days, 6 months, 20 days, 7 days, 39.5mm in average in county, 3.7mm in beam village and town 7.7 mm in 16 days, 7 months, 2 days, 06 mm in beam village and town 4.8mm in county, 7 months, 3 days, 4 days, 16 days, 7 months, 4 days, 16 mm in beam village and town 7.2mm in county, 7.8mm in average in county, 45.5mm in 17 days, 7 months, 14 days, 7 months, 15 days, 07 mm in 7 months, 25.3mm in beam village and town; 21 days 21 months-22 days 7 months at 06 days 32.5mm in beam village and town, 22.5mm in average county, 2.0mm in beam village and town, 8 months 30 days 18 to 8 months 31 days 5, and 13.7mm in average county; the beam villages and towns are 24.5mm at 9 months, 2 days, 15 hours to 23 hours, and the average size in the whole county is 28.1 mm; 43.2mm in beam villages and towns from 08 hours at 9 months and 14 days to 16 hours at 9 months and 17 days, and 56.9mm in average in county

2.2 test materials

The tested crop is corn, the variety is denuded in sea 605, the seedling emerges to mature for 101 days, the plant height is 259 cm, the ear position is 99 cm, the number of adult plant leaves is 19-20, the seedling leaf sheath is purple, the leaf is green, the leaf margin is purple, the anther is yellow green, and the glume is light purple. The planting density is 4000-; the previous crop is wheat, the variety is 22 Jimai, the plant height is 75 cm, and the annual output is 500 kg/mu.

3 protocol and results of the test

3.1 test design and method

The experiment was run with 2 treatments and no replicates. The plots are in the north-south direction, and each treatment area is 1 mu;

treatment 1: the total nutrient of nitrogen, phosphorus and potassium of the applied controlled release fertilizer is more than or equal to 45 percent, the proportion is (27-9-9), and the dosage is 40 kg/mu;

and (3) treatment 2: the total nutrient of nitrogen, phosphorus and potassium of the applied controlled release fertilizer is more than or equal to 45 percent, the mixture ratio is (27-9-9), and the dosage is 40 kg/mu plus 40 kg/mu of the fertilizer product of the invention example 5 (the test fertilizer is applied as seed fertilizer at one time).

3.2 assay biological Property analysis

The treatment with the microbial agent showed:

test observation on 7/25/2020: the third layer of aerial roots with more capillary roots and larger node roots appear early, the leaves are green and many, and the straws are high.

The corns are all ripe and have aphids when being harvested in 10, 2 and 10 months in 2020, and the straws are strong, the bacterial wilt is less, the bald tip is short and the grain number per spike is large when the bacterial manure of the embodiment 3 is applied.

3.3 test results

And (3) measuring yield in 10-month-2 days in 2020, selecting 3 points for each treatment, checking 20 ears for each point, selecting 6 ears for each point, totaling 18 ears, bringing the ears back to the room for air drying, weighing thousand grain weight, and calculating theoretical yield. The row spacing is 0.65 m, and the plant spacing is 5m, and 45 plants are arranged in two rows.

TABLE 2 statistics of maize yields

Treatment of	Mu plant number (plant)	Grain number of spike (number)	Thousand Kernel weight (g)	Yield (kg/mu)
					Process 1	4515	526.73	286.14	578.4
Treatment 2	4515	545.60	290.18	607.6

As can be seen from table 2: the grain number of the applied controlled release fertilizer and the bacterial fertilizer in the example 3 is increased by 18.87 compared with the grain number of the applied controlled release fertilizer, the thousand-grain weight is increased by 4.04g, and the increase rate is 1.41%: the yield is increased by 29.2 kg/mu, the yield is increased by 5.05 percent, and the yield increasing effect is obvious.

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