Preparation method of microbial agent and organic fertilizer

Abstract

The invention belongs to the technical field of microorganisms, and discloses a preparation method of a microbial agent and an organic fertilizer. The microbial agent comprises the following raw materials in parts by weight: 10-20 parts of a complex microbial agent, 5-10 parts of an accelerant and 5-10 parts of nutrient substances, the complex microbial inoculants comprise bacillus velezensis, pseudomonas monteilii and lactobacillus pentosus; the accelerant comprises modified saponin. The synergistic prevention and control effect on disease prevention and control when the bacillus velezensis, the pseudomonas monteilii and the lactobacillus pentosus are used in a composite mode is utilized, diseases are greatly reduced, and yield and income are increased. Meanwhile, the surface activity of the modified saponin can promote attachment of microorganisms on the surfaces of soil particles and plant roots, and the activity and utilization efficiency of the microorganisms are improved; the fertilizer can also form a stable compound with nutritional ingredients in the organic fertilizer, so that the loss of nutrients is reduced; trehalose is added to serve as a nutritional ingredient for rapid proliferation of microbial flora.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a method for preparing a microbial agent and an organic fertilizer. Background Art

[0002] With the advancement of agricultural modernization, the demand for efficient and environmentally friendly fertilizers is increasing. Although traditional chemical fertilizers can quickly provide the nutrients required by plants, long-term use will lead to problems such as soil acidification, compaction, nutrient loss, and environmental pollution. Therefore, the development of a new type of fertilizer that can both improve soil fertility and reduce environmental pollution has become a research hotspot in the agricultural field.

[0003] In recent years, the combination of microbial agents and organic fertilizers has gradually attracted attention. Microbial agents can decompose organic matter, release nutrients, and inhibit pathogens through the metabolic activities of microorganisms, thereby improving soil fertility and plant disease resistance. Organic fertilizers are rich in organic matter and can improve soil structure, increase soil water retention and aeration. Combining the two can not only improve the utilization rate of fertilizers, but also realize the resource utilization of agricultural waste and reduce environmental pollution.

[0004] However, most microbial agents and organic fertilizers on the market currently have the following problems: Insufficient microbial activity: Microbial agents of a single strain have limited effects in practical applications and are difficult to meet the needs of complex soil environments. Poor fertilizer stability: Organic fertilizers are easily decomposed during storage and use, resulting in nutrient loss and reduced fertilizer efficiency. Weak disease prevention and control capabilities: Traditional organic fertilizers have little effect on the prevention and control of soil-borne diseases and are difficult to effectively inhibit the growth of pathogens. Complex preparation process: Existing preparation methods often require complex equipment and processes, which increases production costs. Summary of the invention

[0005] The purpose of the present invention is to provide a novel method for preparing a microbial agent and an organic fertilizer. By optimizing the formula and preparation process of the microbial agent and combining the characteristics of the organic fertilizer, a new type of efficient, stable and environmentally friendly fertilizer is developed, aiming to improve soil fertility, promote plant growth, reduce environmental pollution, and effectively prevent and control soil-borne diseases.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: The present invention provides a microbial agent, comprising the following raw materials in parts by weight: 10-20 parts of compound bacterial agent, 5-10 parts of promoter, 5-10 parts of nutrients; The composite bacterial agent includes Bacillus Velezii, Pseudomonas monteri and Lactobacillus pentosus; The accelerator includes modified saponin.

[0007] Preferably, in the above-mentioned microbial agent, the mass ratio of Bacillus Velezii, Pseudomonas montersii and Lactobacillus pentosus in the composite agent is 2-3:1-2:1-2; The deposit number of the Bacillus Velez is CGMCC 28606; The deposit number of the Pseudomonas monteri is CICC 25156; The deposit number of the Lactobacillus pentosus is ATCC 8041; The nutrients include amino acid zinc, amino acid calcium and fulvic acid; the mass ratio of the amino acid zinc, amino acid calcium and fulvic acid is 1-3:1-3:4-5.

[0008] Preferably, in the above-mentioned microbial agent, the preparation method of the modified saponin comprises the following steps: mixing saponin, maleic anhydride, a catalyst and a solvent, and reacting them to obtain a modified saponin; The mass ratio of the saponin to maleic anhydride is 1:0.5-2; The mass ratio of the saponin to the catalyst is 1:0.02-0.1; The dosage ratio of the saponin and the solvent is 1 g: 10-20 mL; The reaction conditions are as follows: temperature of 60-100° C., reaction time of 4-12 h, and atmosphere of nitrogen or argon.

[0009] The present invention also provides a method for preparing a microbial agent, comprising the following steps: Inoculating Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus strains into LB medium for first culture respectively, to obtain seed liquids of Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus respectively; Inoculating the seed liquid of Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus into new LB medium for second culture respectively, and obtaining fermentation liquid of Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus respectively; The fermentation broths of Bacillus Velezii, Pseudomonas monteri and Lactobacillus pentosus are mixed to obtain a composite bacterial agent; The composite bacterial agent, the promoter and the nutrient substances are mixed to obtain the microbial agent.

[0010] Preferably, in the above-mentioned method for preparing the microbial agent, the temperature of the first culture is 25-30° C., and the time of the first culture is 20-24 hours; The temperature of the second culture is 25-30° C., and the time of the second culture is 40-48 hours.

[0011] The present invention also provides an organic fertilizer, comprising the following raw materials in parts by weight: 50-100 parts of crop waste, 40-50 parts of feces, 20-30 parts of microbial agents, 10-20 parts of humic acid, 1-5 parts of trehalose, 5-10 parts of potassium chloride and 5-10 parts of diammonium phosphate; The microbial agent is the above-mentioned microbial agent or a microbial agent prepared by the above-mentioned preparation method of the microbial agent.

[0012] The present invention also provides a method for preparing an organic fertilizer, comprising the following steps: Mixing part of the crop waste and part of the humic acid, and calcining the mixture to obtain an intermediate; The intermediate, the remaining crop waste, feces and water are mixed to obtain a mixture; then a microbial agent is added to perform composting fermentation to obtain a fermentation product; The fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate are mixed and granulated to obtain an organic fertilizer.

[0013] Preferably, in the above-mentioned method for preparing organic fertilizer, the calcination temperature is 300-500° C., and the calcination time is 2-5 hours.

[0014] Preferably, in the above-mentioned method for preparing organic fertilizer, the mass ratio of the part of the crop waste to the remaining crop waste is 1:1-2; The mass ratio of the partial humic acid to the remaining humic acid is 1:1-2.

[0015] Preferably, in the above method for preparing organic fertilizer, the water content of the mixture is 50-60wt%; The fermentation temperature is 50-60°C, and the fermentation time is 15-20 days.

[0016] It can be seen from the above technical solution that compared with the prior art, the present invention has the following beneficial effects: (1) The microbial agent provided by the present invention includes a composite agent, a promoter and nutrients, wherein the composite agent includes Bacillus velez, Pseudomonas monterey and Lactobacillus pentosus. The antibacterial substances produced by Bacillus velez and the antibiotics secreted by Pseudomonas monterey work together to more effectively inhibit the growth of pathogens; at the same time, Pseudomonas monterey decomposes complex organic matter, provides more nutrients for Bacillus velez, and promotes its growth and metabolism; Lactobacillus pentosus decomposes complex carbohydrates such as cellulose, releases simple sugars, provides a carbon source for Bacillus velez, and promotes its growth. Lactobacillus pentosus produces lactic acid through fermentation to adjust the acid-base balance of the soil, and metabolites such as lactic acid can inhibit the growth of pathogens. That is, the synergistic control effect of Bacillus velez, Pseudomonas monterey and Lactobacillus pentosus on disease control when used in combination solves the limitation of a single strain, greatly reduces diseases, and achieves increased production and income. At the same time, the surface activity of modified saponins can promote the attachment of microorganisms to the surface of soil particles and plant roots, improve the activity and utilization efficiency of microorganisms; it can also form a stable complex with the nutrients in organic fertilizers to reduce nutrient loss; trehalose is added as a nutrient for the rapid proliferation of microbial flora.

[0017] (2) The organic fertilizer provided by the present invention comprises crop waste, feces, microbial agents, humic acid, trehalose, potassium chloride and diammonium phosphate. The synergistic effect between the raw materials in the organic fertilizer improves the fertilizer efficiency and stability of the organic fertilizer, and enhances its promoting effect on plant growth.

[0018] (3) In the preparation method of organic fertilizer provided by the present invention, part of crop waste and part of humic acid are first mixed, calcined, and then fermented and mixed to obtain biochar. After calcination, the crop waste and humic acid form biochar and part of the organic-inorganic composite material. The product has high adsorption and pore structure, can increase the agglomeration of soil, improve the air permeability and water retention of soil; the porous structure provides a good habitat for soil microorganisms, which is conducive to the growth and reproduction of microorganisms; it not only improves the quality and fertilizer efficiency of organic fertilizer, but also realizes the resource utilization of agricultural waste, which has important economic and environmental significance. DETAILED DESCRIPTION

[0019] The present invention provides a microbial agent, comprising the following raw materials in parts by weight: 10~20 parts of compound bacterial agent, 5~10 parts of promoter, and 5~10 parts of nutrients.

[0020] In the present invention, the weight parts of the composite microbial agent in the microbial agent can be 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 parts.

[0021] In the present invention, the weight portion of the promoter in the microbial agent can be 5, 6, 7, 8, 9 or 10 parts.

[0022] In the present invention, the weight proportion of the nutrients in the microbial agent may be 5, 6, 7, 8, 9 or 10 parts.

[0023] In the present invention, the composite bacterial agent comprises Bacillus Velezii, Pseudomonas montersii and Lactobacillus pentosus.

[0024] In the present invention, the mass ratio of Bacillus velez, Pseudomonas montenosus and Lactobacillus pentosus in the composite bacterial agent is 2-3:1-2:1-2. As an example, the mass ratio of Bacillus velez, Pseudomonas montenosus and Lactobacillus pentosus in the composite bacterial agent can be 2:1:1, 2:1:2, 2:2:1, 3:1:1, 3:2:1, 3:1:2 or 3:2:2.

[0025] In the present invention, the deposit number of the Bacillus velez is CGMCC 28606; the deposit number of the Pseudomonas monteridium is CICC 25156; and the deposit number of the Lactobacillus pentosus is ATCC 8041.

[0026] In the present invention, the accelerator comprises modified saponin.

[0027] In the present invention, the preparation method of the modified saponin comprises the following steps: Saponin, maleic anhydride, a catalyst and a solvent are mixed and reacted to obtain a modified saponin.

[0028] By using maleic anhydride to graft saponin, the attachment of microorganisms to the surface of soil particles and plant roots can be promoted, thereby improving the activity and utilization efficiency of microorganisms; modified saponin can form a stable complex with the nutrients in microbial agents, reduce nutrient loss, and extend the shelf life of fertilizers.

[0029] In the present invention, the mass ratio of the saponin to maleic anhydride is 1:0.5 to 2. As an example, the mass ratio of the saponin to maleic anhydride can be 1:0.5, 1:0.8, 1:1, 1:1.2, 1:15, 1:1.8 or 1:2.

[0030] In the present invention, the mass ratio of the saponin to the catalyst is 1:0.02 to 0.1. As an example, the mass ratio of the saponin to the catalyst can be 1:0.02, 1:0.03, 1:0.04, 1:0.05, 1:0.06, 1:0.07, 1:0.08, 1:0.09 or 0.1.

[0031] In the present invention, the usage ratio of the saponin to the solvent is 1 g: 10-20 mL. As an example, the usage ratio of the saponin to the solvent may be 1 g: 10 mL, 1 g: 11 mL, 1 g: 12 mL, 1 g: 14 mL, 1 g: 15 mL, 1 g: 16 mL, 1 g: 17 mL, 1 g: 18 mL or 1 g: 20 mL.

[0032] In the present invention, the catalyst includes at least one of p-toluenesulfonic acid, sulfuric acid and 4-dimethylaminopyridine.

[0033] In the present invention, the solvent includes at least one of N,N-dimethylformamide, dimethyl sulfoxide, tetrahydrofuran and acetone.

[0034] In the present invention, the reaction conditions are: temperature of 60-100°C, time of 4-12h, and atmosphere of nitrogen or argon. As an example, the reaction temperature can be 60°C, 65°C, 70°C, 75°C, 80°C, 85°C, 90°C, 95°C or 100°C; the reaction time can be 4h, 5h, 6h, 7h, 8h, 9h, 10h, 11h or 12h.

[0035] In the present invention, the nutrients include amino acid zinc, amino acid calcium and fulvic acid.

[0036] In the present invention, the mass ratio of amino acid zinc, amino acid calcium and fulvic acid is 1-3:1-3:4-5. As an example, the mass ratio of amino acid zinc, amino acid calcium and fulvic acid can be 1:1:4, 1:2:4, 2:3:5, 3:3:4 or 3:3:5.

[0037] The present invention also provides a method for preparing a microbial agent, comprising the following steps: Inoculating Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus strains into LB medium for first culture respectively, to obtain seed liquids of Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus respectively; Inoculating the seed liquid of Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus into new LB medium for second culture respectively, and obtaining fermentation liquid of Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus respectively; The fermentation broths of Bacillus Velezii, Pseudomonas monteri and Lactobacillus pentosus are mixed to obtain a composite bacterial agent; The composite bacterial agent, the promoter and the nutrient substances are mixed to obtain the microbial agent.

[0038] In the present invention, the temperature of the first culture is 25-30°C, and the time of the first culture is 20-24h. As an example, the temperature of the first culture can be 25°C, 26°C, 27°C, 28°C, 29°C or 30°C, and the time of the first culture can be 20h, 21h, 22h, 26h or 24h.

[0039] In the present invention, the temperature of the second culture is 25-30°C, and the time of the second culture is 40-48 hours. As an example, the temperature of the second culture can be 25°C, 26°C, 27°C, 28°C, 29°C or 30°C, and the time of the second culture can be 40h, 41h, 42h, 43h, 44h, 45h, 46h, 47h or 48h.

[0040] The present invention also provides an organic fertilizer, comprising the following raw materials in parts by weight: 50-100 parts of crop waste, 40-50 parts of feces, 20-30 parts of microbial agents, 10-20 parts of humic acid, 1-5 parts of trehalose, 5-10 parts of potassium chloride and 5-10 parts of diammonium phosphate; The microbial agent is the above-mentioned microbial agent or a microbial agent prepared by the above-mentioned preparation method of the microbial agent.

[0041] In the present invention, the weight parts of the crop waste in the organic fertilizer can be 50, 55, 60, 65, 70, 75, 78, 85, 90, 95 or 100 parts.

[0042] In the present invention, the weight parts of feces in the organic fertilizer can be 40, 41, 42, 43, 44, 45, 46, 48, 49 or 50 parts.

[0043] In the present invention, the weight parts of the microbial agent in the organic fertilizer can be 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 parts.

[0044] In the present invention, the weight parts of humic acid in the organic fertilizer can be 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 parts.

[0045] In the present invention, the weight portion of trehalose in the organic fertilizer can be 1, 2, 3, 4 or 5 parts.

[0046] In the present invention, the weight parts of potassium chloride in the organic fertilizer can be 5, 6, 7, 8, 9 or 10 parts.

[0047] In the present invention, the weight parts of diammonium phosphate in the organic fertilizer can be 5, 6, 7, 8, 9 or 10 parts.

[0048] The present invention also provides a method for preparing an organic fertilizer, comprising the following steps: Mixing part of the crop waste and part of the humic acid, and calcining the mixture to obtain an intermediate; The intermediate, the remaining crop waste, feces and water are mixed to obtain a mixture; then a microbial agent is added to perform composting fermentation to obtain a fermentation product; The fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate are mixed and granulated to obtain an organic fertilizer.

[0049] In the present invention, the calcination temperature is 300-500°C, and the calcination time is 2-5h. As an example, the calcination temperature can be 300°C, 320°C, 340°C, 350°C, 360°C, 370°C, 400°C, 420°C, 450°C, 460°C, 480°C or 500°C, and the calcination time can be 2h, 2.5h, 3h, 3.5h, 4h, 4.5h or 5h.

[0050] In the present invention, the mass ratio of the part of the crop waste to the remaining crop waste is 1:1-2.

[0051] In the present invention, the mass ratio of the partial humic acid to the remaining humic acid is 1:1-2.

[0052] In the present invention, the water content of the mixture is 50-60wt%. As an example, the water content of the mixture can be 50wt%, 52wt%, 54wt%, 55wt%, 56wt%, 58wt% or 60wt%.

[0053] In the present invention, the fermentation temperature is 50-60°C, and the fermentation time is 15-20 days. As an example, the fermentation temperature can be 50°C, 52°C, 54°C, 55°C, 56°C, 57°C, 58°C or 60°C, and the fermentation time can be 15 days, 16 days, 17 days, 18 days, 19 days or 20 days.

[0054] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0055] Example 1

[0056] An organic fertilizer comprises the following raw materials in parts by weight: 65 parts of crop waste, 40 parts of feces, 20 parts of microbial agents, 10 parts of humic acid, 4 parts of trehalose, 6 parts of potassium chloride and 8 parts of diammonium phosphate; The microbial agent comprises the following raw materials in parts by weight: 13 parts of compound bacterial agent, 8 parts of promoter, 8 parts of nutrients; The composite bacterial agent comprises Bacillus velez, Pseudomonas monteri and Lactobacillus pentosus in a mass ratio of 2:1:2, and the preparation method thereof comprises: Bacillus velez, Pseudomonas montenosus and Lactobacillus pentosus strains were inoculated into LB medium and cultured at 30°C for 24 hours to obtain seed solutions of Bacillus velez, Pseudomonas montenosus and Lactobacillus pentosus respectively; wherein the composition of LB medium is: 10g glucose, 5g beef extract, 1.5g sodium chloride, 0.05g FeSO4·7H2O, 0.15g K2HPO4, 0.5g ZnSO4·7H2O, 1000mL distilled water, pH 7.0, sterilization conditions: 121°C, 30 min; The seed liquid of Bacillus Velez, Pseudomonas montenosus and Lactobacillus pentosus were inoculated into a new LB medium and cultured at 30°C for 48 hours to obtain fermentation liquids of Bacillus Velez, Pseudomonas montenosus and Lactobacillus pentosus respectively; wherein the composition of the LB medium is: 10g glucose, 5g beef extract, 1.5g sodium chloride, 0.05g FeSO4·7H2O, 0.15g K2HPO4, 0.5g ZnSO4·7H2O, 1000mL distilled water, pH 7.0, sterilization conditions: 121°C, 30 min; The fermentation broths of Bacillus Velezii, Pseudomonas monteri and Lactobacillus pentosus are mixed to obtain a composite bacterial agent; The composite bacterial agent, the promoter and the nutrient substances are mixed to obtain the microbial agent.

[0057] The accelerator is modified saponin, and the preparation method of the modified saponin comprises the following steps: Saponin, maleic anhydride, catalyst 4-dimethylaminopyridine and N,N-dimethylformamide were mixed, and reacted at 80°C for 8 hours under a nitrogen atmosphere to obtain a modified saponin; wherein the mass ratio of saponin to maleic anhydride was 1:1.5, the mass ratio of saponin to catalyst 4-dimethylaminopyridine was 1:0.08, and the amount ratio of saponin to N,N-dimethylformamide was 1 g:20 mL; The nutrients include amino acid zinc, amino acid calcium and fulvic acid in a mass ratio of 1:1:4; The preparation method of organic fertilizer comprises the following steps: Half of the crop waste and half of the humic acid were mixed and calcined at 300 °C for 2 h to obtain an intermediate; The intermediate, the remaining crop waste, the feces and water are mixed to obtain a mixture with a water content of 50 wt %; then a microbial agent is added to carry out composting fermentation at 55° C. for 20 days to obtain a fermentation product; The fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate are mixed and granulated to obtain an organic fertilizer.

[0058] Example 2

[0059] An organic fertilizer comprises the following raw materials in parts by weight: 70 parts of crop waste, 50 parts of feces, 30 parts of microbial agents, 15 parts of humic acid, 3 parts of trehalose, 10 parts of potassium chloride and 10 parts of diammonium phosphate; The microbial agent comprises the following raw materials in parts by weight: 16 parts of compound bacterial agent, 6 parts of promoter, 10 parts of nutrients; The composite bacterial agent comprises Bacillus velez, Pseudomonas monteri and Lactobacillus pentosus in a mass ratio of 2:2:1, and the preparation method thereof comprises: Bacillus velez, Pseudomonas montenosus and Lactobacillus pentosus strains were inoculated into LB medium and cultured at 30°C for 24 hours to obtain seed solutions of Bacillus velez, Pseudomonas montenosus and Lactobacillus pentosus respectively; wherein the composition of LB medium is: 10g glucose, 5g beef extract, 1.5g sodium chloride, 0.05g FeSO4·7H2O, 0.15g K2HPO4, 0.5g ZnSO4·7H2O, 1000mL distilled water, pH 7.0, sterilization conditions: 121°C, 30 min; The seed liquid of Bacillus Velez, Pseudomonas montenosus and Lactobacillus pentosus were inoculated into a new LB medium and cultured at 30°C for 48 hours to obtain fermentation liquids of Bacillus Velez, Pseudomonas montenosus and Lactobacillus pentosus respectively; wherein the composition of the LB medium is: 10g glucose, 5g beef extract, 1.5g sodium chloride, 0.05g FeSO4·7H2O, 0.15g K2HPO4, 0.5g ZnSO4·7H2O, 1000mL distilled water, pH 7.0, sterilization conditions: 121°C, 30 min; The fermentation broths of Bacillus Velezii, Pseudomonas monteri and Lactobacillus pentosus are mixed to obtain a composite bacterial agent; The composite bacterial agent, the promoter and the nutrient substances are mixed to obtain the microbial agent.

[0060] The accelerator is modified saponin, and the preparation method of the modified saponin comprises the following steps: Saponin, maleic anhydride, catalyst 4-dimethylaminopyridine and N,N-dimethylformamide were mixed, and reacted at 70°C for 10 hours under a nitrogen atmosphere to obtain a modified saponin; wherein the mass ratio of saponin to maleic anhydride was 1:1.5, the mass ratio of saponin to catalyst 4-dimethylaminopyridine was 1:0.06, and the amount ratio of saponin to N,N-dimethylformamide was 1 g:20 mL; The nutrients include amino acid zinc, amino acid calcium and fulvic acid in a mass ratio of 2:1:4; The preparation method of organic fertilizer comprises the following steps: Half of the crop waste and half of the humic acid were mixed and calcined at 300 °C for 3 h to obtain an intermediate; The intermediate, the remaining crop waste, the feces and water are mixed to obtain a mixture with a water content of 55 wt %; then a microbial agent is added to carry out composting fermentation at 55° C. for 20 days to obtain a fermentation product; The fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate are mixed and granulated to obtain an organic fertilizer.

[0061] Example 3

[0062] An organic fertilizer comprises the following raw materials in parts by weight: 90 parts of crop waste, 45 parts of feces, 25 parts of microbial agents, 18 parts of humic acid, 5 parts of trehalose, 8 parts of potassium chloride and 7 parts of diammonium phosphate; The microbial agent comprises the following raw materials in parts by weight: 15 parts of compound bacterial agent, 9 parts of promoter, 7 parts of nutrients; The composite bacterial agent comprises Bacillus velez, Pseudomonas monteri and Lactobacillus pentosus in a mass ratio of 3:2:1, and the preparation method thereof comprises: Bacillus velez, Pseudomonas montenosus and Lactobacillus pentosus strains were inoculated into LB medium and cultured at 30°C for 24 hours to obtain seed solutions of Bacillus velez, Pseudomonas montenosus and Lactobacillus pentosus respectively; wherein the composition of LB medium is: 10g glucose, 5g beef extract, 1.5g sodium chloride, 0.05g FeSO4·7H2O, 0.15g K2HPO4, 0.5g ZnSO4·7H2O, 1000mL distilled water, pH 7.0, sterilization conditions: 121°C, 30 min; The seed liquid of Bacillus Velez, Pseudomonas montenosus and Lactobacillus pentosus were inoculated into a new LB medium and cultured at 30°C for 48 hours to obtain fermentation liquids of Bacillus Velez, Pseudomonas montenosus and Lactobacillus pentosus respectively; wherein the composition of the LB medium is: 10g glucose, 5g beef extract, 1.5g sodium chloride, 0.05g FeSO4·7H2O, 0.15g K2HPO4, 0.5g ZnSO4·7H2O, 1000mL distilled water, pH 7.0, sterilization conditions: 121°C, 30 min; The fermentation broths of Bacillus Velezii, Pseudomonas monteri and Lactobacillus pentosus are mixed to obtain a composite bacterial agent; The composite bacterial agent, the promoter and the nutrient substances are mixed to obtain the microbial agent.

[0063] The accelerator is modified saponin, and the preparation method of the modified saponin comprises the following steps: Saponin, maleic anhydride, catalyst 4-dimethylaminopyridine and N,N-dimethylformamide were mixed, and reacted at 75°C for 8 hours under a nitrogen atmosphere to obtain a modified saponin; wherein the mass ratio of saponin to maleic anhydride was 1:1, the mass ratio of saponin to catalyst 4-dimethylaminopyridine was 1:0.05, and the amount ratio of saponin to N,N-dimethylformamide was 1 g:15 mL; The nutrients include amino acid zinc, amino acid calcium and fulvic acid in a mass ratio of 3:1:5; The preparation method of organic fertilizer comprises the following steps: Half of the crop waste and half of the humic acid were mixed and calcined at 300 °C for 3 h to obtain an intermediate; The intermediate, the remaining crop waste, the feces and water are mixed to obtain a mixture with a water content of 55 wt %; then a microbial agent is added to carry out composting fermentation at 55° C. for 20 days to obtain a fermentation product; The fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate are mixed and granulated to obtain an organic fertilizer.

[0064] Comparative Example 1 An organic fertilizer is consistent with that in Example 1, except that the composite microbial agent in the microbial agent includes Bacillus Velezii and Pseudomonas monteriodii in a mass ratio of 2:1.

[0065] Comparative Example 2 An organic fertilizer is consistent with that in Example 1, except that the composite microbial agent in the microbial agent includes Bacillus Velez and Lactobacillus pentosus in a mass ratio of 2:2.

[0066] Comparative Example 3 An organic fertilizer is consistent with that in Example 1, except that the composite microbial agent in the microbial agent includes Pseudomonas montosus and Lactobacillus pentosus in a mass ratio of 1:2.

[0067] Comparative Example 4 An organic fertilizer, which is the same as that in Example 1, except that the microbial agent does not contain a promoter.

[0068] Comparative Example 5 An organic fertilizer, which is consistent with that in Example 1, except that the promoter in the microbial agent is saponin.

[0069] Comparative Example 6 An organic fertilizer, which is consistent with that in Example 1, except that: The preparation method of organic fertilizer comprises the following steps: The crop waste, feces and water are mixed to obtain a mixture with a water content of 50wt%; then a microbial agent is added to carry out composting fermentation at 55°C for 20 days to obtain a fermentation product; The fermentation product, humic acid, trehalose, potassium chloride and diammonium phosphate are mixed and granulated to obtain organic fertilizer.

[0070] Test Example 1: Experiment on promoting plant growth with organic fertilizer The experiment set up a blank control group (CK), Example 1 group (100 kg / mu), Example 2 group (100 kg / mu), Example 3 group (100 kg / mu), Comparative Example 1 group (100 kg / mu), Comparative Example 2 group (100 kg / mu), Comparative Example 3 group (100 kg / mu), Comparative Example 4 group (100kg / mu), Comparative Example 5 group (100 kg / mu) and Comparative Example 6 group (100 kg / mu). Each treatment group was set up for 3 repetitions, the experimental plot area was 25 square meters, and the initial soil pH was 4.6.

[0071] The organic fertilizers in Examples 1 to 3 and Comparative Examples 1 to 6 were respectively applied to the test fields, tilled and mixed by a rotary tiller, and then sown.

[0072] All treatment groups applied fertilizers in the same way, with a one-time basal application and no topdressing in the later period.

[0073] When the corn matured, the corn yield (kg / mu), yield increase rate (%) and root dry weight (g / plant) of each group were recorded.

[0074] The calculation formula of the yield increase rate is as follows: Yield increase rate (%) = (yield of experimental group - yield of blank control group) / yield of blank control group × 100%; specific experimental data are shown in Table 1.

[0075] Table 1

[0076] Test Example 2: Experiment on promoting cucumber growth Cucumber seedlings with consistent growth were selected for transplanting and divided into a blank control group, Example 1-3 groups, and Comparative Examples 1-6. The blank control group was not fertilized, and the other treatment groups were respectively treated with the organic fertilizers in Examples 1-3 and Comparative Examples 1-6. During transplanting, 10 g of organic fertilizer was used for each root, evenly spread on the soil surface of the breeding tray and covered with soil. 100 seedlings were treated for each treatment, repeated 3 times, and watered sufficiently. After 18 days, the plant height (cm), root-to-crown ratio, and dry weight (g) were measured. The specific experimental data are shown in Table 2.

[0077] Table 2

[0078] As can be seen from Table 2, the organic fertilizers prepared in Examples 1 to 3 of the present invention have a significant promoting effect on the growth of cucumber, and the plant height and dry weight are significantly increased compared with the blank control group and the comparative example. It can be seen that the organic fertilizers prepared in Examples 1 to 3 of the present invention have a certain promoting effect on the growth of cucumber.

[0079] Test Example 3: Indoor protection effect determination on cucumber Active spores of Pythium aphanidermatum, Rhizoctonia solani and Fusarium oxysporum were inoculated into sorghum grain culture medium respectively and cultured at 28°C until mycelium covered the surface of each sorghum grain. Equal amounts of the three kinds of infected sorghum grains were mixed and evenly spread on the topsoil of the breeding tray (20 g / m2), covered with 1 cm of soil, and then the organic fertilizers in Examples 1 to 3 and Comparative Examples 1 to 6 were taken, each taking 10 g / m2. 2 After mixing the soil, evenly spread it on the soil surface of the breeding tray, evenly sow cucumber seeds and cover with soil, 100 seedlings per treatment, 3 replicates, culture at 20°C, observe the disease after emergence, calculate the incidence rate and control effect. No bacterial agent was applied to the blank control group. The calculation formulas for the incidence rate and control effect are as follows: Incidence rate = number of diseased plants / total number of plants × 100%; Control effect (%) = [(control incidence-treatment incidence) / control incidence] × 100%; specific experimental data are shown in Table 3.

[0080] Table 3

[0081] As shown in Table 3, the organic fertilizers prepared in Examples 1 to 3 of the present invention have a clear inhibitory effect on the incidence of cucumber, which is far superior to the blank control group and the comparative example. That is, the synergistic control effect of disease control when Bacillus velez, Pseudomonas monteri and Lactobacillus pentosus are used in combination to solve the limitation problem of a single strain, greatly reduce diseases, and achieve increased production and income. The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A microbial agent, characterized in that: The invention comprises the following raw materials in parts by weight: 10-20 parts of compound bacterial agent, 5-10 parts of promoter, 5-10 parts of nutrients; The composite bacterial agent includes Bacillus Velezii, Pseudomonas monteri and Lactobacillus pentosus; The accelerator includes modified saponin.

2. The microbial agent according to claim 1, characterized in that The mass ratio of Bacillus velez, Pseudomonas monteri and Lactobacillus pentosus in the composite bacterial agent is 2-3:1-2:1-2; The deposit number of the Bacillus Velez is CGMCC 28606; The deposit number of the Pseudomonas monteri is CICC 25156; The deposit number of the Lactobacillus pentosus is ATCC 8041; The nutrients include amino acid zinc, amino acid calcium and fulvic acid; the mass ratio of the amino acid zinc, amino acid calcium and fulvic acid is 1-3:1-3:4-5.

3. The microbial agent according to claim 1, characterized in that: The preparation method of the modified saponin comprises the following steps: mixing saponin, maleic anhydride, a catalyst and a solvent, and reacting them to obtain a modified saponin; The mass ratio of the saponin to maleic anhydride is 1:0.5-2; The mass ratio of the saponin to the catalyst is 1:0.02-0.1; The dosage ratio of the saponin and the solvent is 1 g: 10-20 mL; The reaction conditions are as follows: temperature of 60-100° C., reaction time of 4-12 h, and atmosphere of nitrogen or argon.

4. The method for preparing the microbial agent according to any one of claims 1 to 3, characterized in that: The following steps are involved: Inoculating Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus strains into LB medium for first culture respectively, to obtain seed liquids of Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus respectively; Inoculating the seed liquid of Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus into new LB medium for second culture respectively, and obtaining fermentation liquid of Bacillus Velezii, Pseudomonas montenosus and Lactobacillus pentosus respectively; The fermentation broths of Bacillus Velezii, Pseudomonas monteri and Lactobacillus pentosus are mixed to obtain a composite bacterial agent; The composite bacterial agent, the promoter and the nutrient substances are mixed to obtain the microbial agent.

5. The method for preparing the microbial agent according to claim 4, characterized in that: The temperature of the first culture is 25-30° C., and the time of the first culture is 20-24 hours; The temperature of the second culture is 25-30° C., and the time of the second culture is 40-48 hours.

6. An organic fertilizer, characterized in that: The invention comprises the following raw materials in parts by weight: 50-100 parts of crop waste, 40-50 parts of feces, 20-30 parts of microbial agents, 10-20 parts of humic acid, 1-5 parts of trehalose, 5-10 parts of potassium chloride and 5-10 parts of diammonium phosphate; The microbial agent is the microbial agent described in any one of claims 1 to 3 or a microbial agent prepared by the preparation method of the microbial agent described in claim 4 or 5.

7. The method for preparing the organic fertilizer according to claim 6, wherein The following steps are involved: Mixing part of the crop waste and part of the humic acid, and calcining the mixture to obtain an intermediate; The intermediate, the remaining crop waste, feces and water are mixed to obtain a mixture; then a microbial agent is added to perform composting fermentation to obtain a fermentation product; The fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate are mixed and granulated to obtain an organic fertilizer.

8. The method for preparing organic fertilizer according to claim 7, wherein The calcination temperature is 300-500° C., and the calcination time is 2-5 hours.

9. The method for preparing organic fertilizer according to claim 7, wherein The mass ratio of the part of the crop waste to the remaining crop waste is 1:1-2; The mass ratio of the partial humic acid to the remaining humic acid is 1:1-2.

10. The method for preparing organic fertilizer according to claim 7, characterized in that, The water content of the mixture is 50-60wt%; The fermentation temperature is 50-60°C, and the fermentation time is 15-20 days.

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