Preparation method of a microbial inoculum and an organic fertilizer

Abstract

The present invention belongs to the field of microbial technology and discloses a preparation method of a microbial inoculant and an organic fertilizer. The microbial inoculant comprises the following raw materials in parts by weight: 10-20 parts of a compound bacterium agent, 5-10 parts of a promoter, and 5-10 parts of a nutrient substance; the compound bacterium agent comprises Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus; the promoter comprises modified saponins. The present invention utilizes the synergistic prevention and control effect on disease prevention and control when Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus are used in combination, greatly reduces diseases, and realizes increased production and income. At the same time, the surface activity of the 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 nutrient components in the organic fertilizer to reduce the loss of nutrients; trehalose is added as a nutrient component for the rapid proliferation of the microbial flora.

Description

Technical Field

[0001] The present invention relates to the technical field of microorganisms, and particularly to a preparation method of a microbial inoculant and organic fertilizer. Background Art

[0002] With the advancement of agricultural modernization, the demand for efficient and environmentally friendly fertilizers is increasing day by day. Although traditional chemical fertilizers can quickly provide the nutrient elements required by plants, long-term use will lead to problems such as soil acidification, hardening, and nutrient loss, and at the same time cause 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 inoculants and organic fertilizers has gradually received attention. Microbial inoculants can decompose organic matter, release nutrients, and inhibit pathogenic bacteria through the metabolic activities of microorganisms, thereby improving soil fertility and plant disease resistance. Organic fertilizers are rich in organic matter, which 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 of the current microbial inoculants and organic fertilizers on the market have the following problems: insufficient microbial activity: The microbial inoculants with single strains have limited effects in actual 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 ability: The prevention and control effects of traditional organic fertilizers on soil-borne diseases are not obvious, and it is difficult to effectively inhibit the growth of pathogenic bacteria. Complex preparation process: The existing preparation methods often require complex equipment and processes, increasing production costs. Summary of the Invention

[0005] The purpose of the present invention is to provide a new preparation method of a microbial inoculant and organic fertilizer. By optimizing the formula and preparation process of the microbial inoculant and combining the characteristics of organic fertilizers, 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 invention purpose, the present invention provides the following technical solutions:

[0007] The present invention provides a microbial inoculant, comprising the following raw materials in parts by weight:

[0008] Compound bacterium agent 10 - 20 parts, promoter 5 - 10 parts, nutrient substance 5 - 10 parts;

[0009] The compound bacterium agent includes Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus;

[0010] The promoter includes modified saponins.

[0011] Preferably, in the above microbial inoculant, the mass ratio of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus in the composite inoculant is 2-3:1-2:1-2;

[0012] The preservation number of Bacillus velezensis is CGMCC 28606;

[0013] The preservation number of Pseudomonas monteilii is CICC 25156;

[0014] The preservation number of Lactobacillus pentosus is ATCC 8041;

[0015] The nutrient substances include zinc amino acid, calcium amino acid, and fulvic acid; the mass ratio of zinc amino acid, calcium amino acid, and fulvic acid is 1-3:1-3:4-5.

[0016] Preferably, in the above microbial inoculant, the preparation method of the modified saponins includes the following steps:

[0017] Mix saponins, maleic anhydride, a catalyst, and a solvent, and carry out a reaction to obtain modified saponins;

[0018] The mass ratio of the saponins to maleic anhydride is 1:0.5-2;

[0019] The mass ratio of the saponins to the catalyst is 1:0.02-0.1;

[0020] The dosage ratio of the saponins to the solvent is 1 g:10-20 mL;

[0021] The reaction conditions are: temperature is 60-100 °C, time is 4-12 h, and the atmosphere is nitrogen or argon.

[0022] The present invention also provides a preparation method of a microbial inoculant, including the following steps:

[0023] Respectively inoculate Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus strains into LB medium for the first culture to obtain the seed solutions of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus respectively;

[0024] Respectively inoculate the seed solutions of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus into a new LB medium for the second culture to obtain the fermentation broths of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus respectively;

[0025] Mix the fermentation broths of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus to obtain a composite inoculant;

[0026] Mix the compound microbial agent, promoter and nutrients to obtain the microbial inoculum.

[0027] Preferably, in the above method for preparing the microbial inoculum, the temperature of the first culture is 25 - 30 °C, and the time of the first culture is 20 - 24 h;

[0028] The temperature of the second culture is 25 - 30 °C, and the time of the second culture is 40 - 48 h.

[0029] The present invention also provides an organic fertilizer, comprising the following raw materials in parts by weight:

[0030] 50 - 100 parts of crop waste, 40 - 50 parts of feces, 20 - 30 parts of microbial inoculum, 10 - 20 parts of humic acid, 1 - 5 parts of trehalose, 5 - 10 parts of potassium chloride, and 5 - 10 parts of diammonium phosphate;

[0031] The microbial inoculum is the above-mentioned microbial inoculum or the microbial inoculum prepared by the above-mentioned method for preparing the microbial inoculum.

[0032] The present invention also provides a method for preparing an organic fertilizer, comprising the following steps:

[0033] Mix part of the crop waste and part of the humic acid, and conduct calcination to obtain an intermediate;

[0034] Mix the intermediate, the remaining crop waste, feces and water to obtain a mixture; then add the microbial inoculum for composting fermentation to obtain a fermentation product;

[0035] Mix the fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate, and conduct granulation to obtain the organic fertilizer.

[0036] Preferably, in the above method for preparing the organic fertilizer, the temperature of the calcination is 300 - 500 °C, and the time of the calcination is 2 - 5 h.

[0037] Preferably, in the above method for preparing the organic fertilizer, the mass ratio of the part of the crop waste to the remaining crop waste is 1:1 - 2;

[0038] The mass ratio of the part of the humic acid to the remaining humic acid is 1:1 - 2.

[0039] Preferably, in the above method for preparing the organic fertilizer, the water content of the mixture is 50 - 60 wt%;

[0040] The temperature of the fermentation is 50 - 60 °C, and the time of the fermentation is 15 - 20 d.

[0041] It can be seen from the above technical solutions that, compared with the prior art, the present invention has the following beneficial effects:

[0042] (1) The microbial inoculant provided by the present invention includes a compound inoculant, a promoter, and nutrients. The compound inoculant includes Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus. The antibacterial substances produced by Bacillus velezensis and the antibiotics secreted by Pseudomonas monteilii act together to more effectively inhibit the growth of pathogenic bacteria. At the same time, Pseudomonas monteilii decomposes complex organic substances to provide more nutrients for Bacillus velezensis, promoting its growth and metabolism. Lactobacillus pentosus decomposes complex carbohydrates such as cellulose to release simple sugars, providing a carbon source for Bacillus velezensis and promoting its growth. Moreover, Lactobacillus pentosus produces lactic acid through fermentation to regulate the acid-base balance of the soil, and metabolites such as lactic acid can inhibit the growth of pathogenic bacteria. That is, by utilizing the synergistic control effect of the combined use of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus on disease prevention and control, the limitation problem of a single strain is solved, the disease is significantly reduced, and the increase in production and income is achieved. At the same time, the surface activity of the modified saponin can promote the attachment of microorganisms to the surface of soil particles and plant roots, improving the activity and utilization efficiency of microorganisms. It can also form a stable complex with the nutrient components in the organic fertilizer, reducing nutrient loss. Trehalose is added as a nutrient component for the rapid proliferation of the microbial flora.

[0043] (2) The organic fertilizer provided by the present invention includes crop waste, manure, microbial inoculant, humic acid, trehalose, potassium chloride, and diammonium phosphate. The raw materials in the organic fertilizer act synergistically to improve the fertilizer efficiency and stability of the organic fertilizer and enhance its promoting effect on plant growth.

[0044] (3) In the preparation method of the organic fertilizer provided by the present invention, part of the crop waste and part of the humic acid are first mixed and calcined, and then fermented and granulated. 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, which can increase the soil aggregation, improve the air permeability and water retention of the 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 the organic fertilizer, but also realizes the resource utilization of agricultural waste, which has important economic and environmental significance. Detailed implementation mode

[0045] The present invention provides a microbial inoculant, which includes the following raw materials in parts by weight:

[0046] Compound inoculant 10 - 20 parts, promoter 5 - 10 parts, nutrients 5 - 10 parts.

[0047] In the present invention, the weight parts of the compound inoculant in the microbial inoculant can be 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 parts.

[0048] In the present invention, the weight portion of the promoter in the microbial inoculant can be 5, 6, 7, 8, 9 or 10 portions.

[0049] In the present invention, the weight portion of the nutrient in the microbial inoculant can be 5, 6, 7, 8, 9 or 10 portions.

[0050] In the present invention, the compound inoculant includes Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus.

[0051] In the present invention, the mass ratio of Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus in the compound inoculant is 2-3:1-2:1-2. As an example, the mass ratio of Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus in the compound inoculant can be 2:1:1, 2:1:2, 2:2:1, 3:1:1, 3:2:1, 3:1:2 or 3:2:2.

[0052] In the present invention, the preservation number of Bacillus velezensis is CGMCC 28606; the preservation number of Pseudomonas monteilii is CICC 25156; the preservation number of Lactobacillus pentosus is ATCC 8041.

[0053] In the present invention, the promoter includes modified saponins.

[0054] In the present invention, the preparation method of the modified saponins includes the following steps:

[0055] Mix saponins, maleic anhydride, a catalyst and a solvent, and carry out a reaction to obtain modified saponins.

[0056] By grafting maleic anhydride onto saponins, it can promote the attachment of microorganisms to the surface of soil particles and plant roots, improve the activity and utilization efficiency of microorganisms; the modified saponins can form a stable complex with the nutrient components in the microbial inoculant, reduce the loss of nutrients, and extend the validity period of the fertilizer.

[0057] In the present invention, the mass ratio of saponins to maleic anhydride is 1:0.5-2. As an example, the mass ratio of saponins to maleic anhydride can be 1:0.5, 1:0.8, 1:1, 1:1.2, 1:15, 1:1.8 or 1:2.

[0058] In the present invention, the mass ratio of saponins to the catalyst is 1:0.02-0.1. As an example, the mass ratio of saponins 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.

[0059] In the present invention, the dosage ratio of the saponin to the solvent is 1 g: 10 - 20 mL. As an example, the dosage ratio of the saponin to the solvent can 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.

[0060] In the present invention, the catalyst includes at least one of p-toluenesulfonic acid, sulfuric acid, and 4-dimethylaminopyridine.

[0061] In the present invention, the solvent includes at least one of N,N-dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, and acetone.

[0062] In the present invention, the conditions of the reaction are as follows: the temperature is 60 - 100 °C, the time is 4 - 12 h, and the atmosphere is 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 4 h, 5 h, 6 h, 7 h, 8 h, 9 h, 10 h, 11 h, or 12 h.

[0063] In the present invention, the nutrient includes zinc amino acid, calcium amino acid, and fulvic acid.

[0064] In the present invention, the mass ratio of zinc amino acid, calcium amino acid, and fulvic acid is 1 - 3: 1 - 3: 4 - 5. As an example, the masses of zinc amino acid, calcium amino acid, and fulvic acid can be 1: 1: 4, 1: 2: 4, 2: 3: 5, 3: 3: 4, or 3: 3: 5.

[0065] The present invention also provides a method for preparing a microbial inoculant, comprising the following steps:

[0066] Inoculate Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus strains into LB medium for the first culture respectively to obtain the seed solutions of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus;

[0067] Inoculate the seed solutions of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus into fresh LB medium for the second culture respectively to obtain the fermentation broths of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus;

[0068] Mix the fermentation broths of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus to obtain a compound inoculant;

[0069] Mix the compound inoculant, promoter, and nutrient to obtain the microbial inoculant.

[0070] In the present invention, the temperature of the first cultivation is 25 - 30 °C, and the time of the first cultivation is 20 - 24 h. As an example, the temperature of the first cultivation can be 25 °C, 26 °C, 27 °C, 28 °C, 29 °C or 30 °C, and the time of the first cultivation can be 20 h, 21 h, 22 h, 26 h or 24 h.

[0071] In the present invention, the temperature of the second cultivation is 25 - 30 °C, and the time of the second cultivation is 40 - 48 h. As an example, the temperature of the second cultivation can be 25 °C, 26 °C, 27 °C, 28 °C, 29 °C or 30 °C, and the time of the second cultivation can be 40 h, 41 h, 42 h, 43 h, 44 h, 45 h, 46 h, 47 h or 48 h.

[0072] The present invention also provides an organic fertilizer, comprising the following raw materials in parts by weight:

[0073] 50 - 100 parts of crop waste, 40 - 50 parts of feces, 20 - 30 parts of microbial inoculum, 10 - 20 parts of humic acid, 1 - 5 parts of trehalose, 5 - 10 parts of potassium chloride, and 5 - 10 parts of diammonium phosphate;

[0074] The microbial inoculum is the above - mentioned microbial inoculum or the microbial inoculum prepared by the above - mentioned preparation method of the microbial inoculum.

[0075] In the present invention, the parts by weight of crop waste in the organic fertilizer can be 50, 55, 60, 65, 70, 75, 78, 85, 90, 95 or 100 parts.

[0076] In the present invention, the parts by weight of feces in the organic fertilizer can be 40, 41, 42, 43, 44, 45, 46, 48, 49 or 50 parts.

[0077] In the present invention, the parts by weight of microbial inoculum in the organic fertilizer can be 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 parts.

[0078] In the present invention, the parts by weight of humic acid in the organic fertilizer can be 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 parts.

[0079] In the present invention, the parts by weight of trehalose in the organic fertilizer can be 1, 2, 3, 4 or 5 parts.

[0080] In the present invention, the parts by weight of potassium chloride in the organic fertilizer can be 5, 6, 7, 8, 9 or 10 parts.

[0081] In the present invention, the parts by weight of diammonium phosphate in the organic fertilizer can be 5, 6, 7, 8, 9 or 10 parts.

[0082] The present invention also provides a method for preparing organic fertilizer, comprising the following steps:

[0083] Mix a part of crop waste and a part of humic acid, and conduct calcination to obtain an intermediate;

[0084] Mix the intermediate, the remaining crop waste, feces and water to obtain a mixture; then add a microbial inoculant for compost fermentation to obtain a fermentation product;

[0085] Mix the fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate, and conduct granulation to obtain organic fertilizer.

[0086] In the present invention, the temperature of the calcination is 300 - 500 °C, and the time of the calcination is 2 - 5 h. As an example, the temperature of the calcination 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 time of the calcination can be 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h or 5 h.

[0087] In the present invention, the mass ratio of the part of crop waste to the remaining crop waste is 1:1 - 2.

[0088] In the present invention, the mass ratio of the part of humic acid to the remaining humic acid is 1:1 - 2.

[0089] In the present invention, the water content of the mixture is 50 - 60 wt%. As an example, the water content of the mixture can be 50 wt%, 52 wt%, 54 wt%, 55 wt%, 56 wt%, 58 wt% or 60 wt%.

[0090] In the present invention, the temperature of the fermentation is 50 - 60 °C, and the time of the fermentation is 15 - 20 d. As an example, the temperature of the fermentation can be 50 °C, 52 °C, 54 °C, 55 °C, 56 °C, 57 °C, 58 °C or 60 °C, and the time of the fermentation can be 15 d, 16 d, 17 d, 18 d, 19 d or 20 d.

[0091] Next, the technical solutions in the embodiments of the present invention will be described clearly and completely. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts shall fall within the protection scope of the present invention.

[0092] Example 1

[0093] An organic fertilizer, comprising the following raw materials in parts by weight:

[0094] 65 parts of crop waste, 40 parts of feces, 20 parts of microbial inoculant, 10 parts of humic acid, 4 parts of trehalose, 6 parts of potassium chloride, and 8 parts of diammonium phosphate;

[0095] The microbial inoculant comprises raw materials in the following parts by weight:

[0096] 13 parts of compound bacteria agent, 8 parts of promoter, and 8 parts of nutrient substance;

[0097] The compound bacteria agent comprises Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus in a mass ratio of 2:1:2. Its preparation method includes:

[0098] Inoculate Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus strains into LB medium respectively and culture at 30 °C for 24 h to obtain the seed solutions of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus respectively. Among them, the composition of LB medium is: 10 g of glucose, 5 g of beef extract, 1.5 g of sodium chloride, 0.05 g of FeSO4·7H2O, 0.15 g of K2HPO4, 0.5 g of ZnSO4·7H2O, 1000 mL of distilled water, pH 7.0, sterilization conditions: 121 °C, 30 min;

[0099] Inoculate the seed solutions of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus into fresh LB medium respectively and culture at 30 °C for 48 h to obtain the fermentation broths of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus respectively. Among them, the composition of LB medium is: 10 g of glucose, 5 g of beef extract, 1.5 g of sodium chloride, 0.05 g of FeSO4·7H2O, 0.15 g of K2HPO4, 0.5 g of ZnSO4·7H2O, 1000 mL of distilled water, pH 7.0, sterilization conditions: 121 °C, 30 min;

[0100] Mix the fermentation broths of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus to obtain the compound bacteria agent;

[0101] Mix the compound bacteria agent, promoter, and nutrient substance to obtain the microbial inoculant.

[0102] The promoter is modified saponin. The preparation method of modified saponin includes the following steps:

[0103] Mix saponin, maleic anhydride, catalyst 4-dimethylaminopyridine, and N,N-dimethylformamide, and react at 80 °C for 8 h under a nitrogen atmosphere to obtain modified saponin. Among them, the mass ratio of saponin to maleic anhydride is 1:1.5, the mass ratio of saponin to catalyst 4-dimethylaminopyridine is 1:0.08, and the dosage ratio of saponin to N,N-dimethylformamide is 1 g:20 mL;

[0104] The nutrients include zinc amino acid, calcium amino acid and fulvic acid with a mass ratio of 1:1:4;

[0105] A method for preparing organic fertilizer comprises the following steps:

[0106] Mix half of the crop waste and half of the humic acid, and calcine at 300 °C for 2 h to obtain an intermediate;

[0107] Mix the intermediate, the remaining crop waste, manure and water to obtain a mixture with a water content of 50 wt%; then add a microbial inoculum and carry out composting fermentation at 55 °C for 20 d to obtain a fermentation product;

[0108] Mix the fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate, and granulate to obtain the organic fertilizer.

[0109] Example 2

[0110] An organic fertilizer, comprising the following raw materials in parts by weight:

[0111] 70 parts of crop waste, 50 parts of manure, 30 parts of microbial inoculum, 15 parts of humic acid, 3 parts of trehalose, 10 parts of potassium chloride and 10 parts of diammonium phosphate;

[0112] The microbial inoculum comprises the following raw materials in parts by weight:

[0113] 16 parts of compound bacterium agent, 6 parts of promoter, 10 parts of nutrients;

[0114] The compound bacterium agent comprises Paenibacillus bellii, Pseudomonas monteilii and Lactobacillus pentosus with a mass ratio of 2:2:1, and its preparation method comprises:

[0115] Inoculate Paenibacillus bellii, Pseudomonas monteilii and Lactobacillus pentosus strains into LB medium respectively and culture at 30 °C for 24 h to obtain seed solutions of Paenibacillus bellii, Pseudomonas monteilii and Lactobacillus pentosus respectively; wherein, the composition of LB medium is: 10 g of glucose, 5 g of beef extract, 1.5 g of sodium chloride, 0.05 g of FeSO4·7H2O, 0.15 g of K2HPO4, 0.5 g of ZnSO4·7H2O, 1000 mL of distilled water, pH 7.0, sterilization condition: 121 °C, 30 min;

[0116] The seed solutions of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus were respectively inoculated into fresh LB medium and cultured at 30 °C for 48 h to obtain the fermentation broths of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus. Among them, the composition of the LB medium was: 10 g of glucose, 5 g of beef extract, 1.5 g of sodium chloride, 0.05 g of FeSO4·7H2O, 0.15 g of K2HPO4, 0.5 g of ZnSO4·7H2O, 1000 mL of distilled water, pH 7.0, sterilization conditions: 121 °C, 30 min;

[0117] The fermentation broths of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus were mixed to obtain a compound microbial agent;

[0118] The compound microbial agent, promoter, and nutrient were mixed to obtain a microbial inoculum.

[0119] The promoter was modified saponin, and the preparation method of the modified saponin included the following steps:

[0120] Saponin, maleic anhydride, catalyst 4-dimethylaminopyridine, and N,N-dimethylformamide were mixed and reacted at 70 °C for 10 h under a nitrogen atmosphere to obtain modified saponin. Among them, 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 dosage ratio of saponin to N,N-dimethylformamide was 1 g:20 mL;

[0121] The nutrient included zinc amino acid, calcium amino acid, and fulvic acid with a mass ratio of 2:1:4;

[0122] The preparation method of the organic fertilizer included the following steps:

[0123] 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;

[0124] The intermediate, the remaining crop waste, manure, and water were mixed to obtain a mixture with a water content of 55 wt%. Then, the microbial inoculum was added and composted at 55 °C for 20 d to obtain a fermentation product;

[0125] The fermentation product, the remaining humic acid, trehalose, potassium chloride, and diammonium phosphate were mixed and granulated to obtain the organic fertilizer.

[0126] Example 3

[0127] An organic fertilizer comprising the following raw materials in parts by weight:

[0128] 90 parts of crop waste, 45 parts of manure, 25 parts of microbial inoculum, 18 parts of humic acid, 5 parts of trehalose, 8 parts of potassium chloride, and 7 parts of diammonium phosphate;

[0129] The microbial inoculant includes raw materials in the following parts by weight:

[0130] 15 parts of compound bacterium agent, 9 parts of promoter, and 7 parts of nutrient substance;

[0131] The compound bacterium agent includes Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus with a mass ratio of 3:2:1. Its preparation method includes:

[0132] Inoculate Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus strains into LB medium respectively and culture them at 30 °C for 24 h to obtain the seed solutions of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus respectively; among them, the composition of LB medium is: 10 g of glucose, 5 g of beef extract, 1.5 g of sodium chloride, 0.05 g of FeSO4·7H2O, 0.15 g of K2HPO4, 0.5 g of ZnSO4·7H2O, 1000 mL of distilled water, pH 7.0, sterilization conditions: 121 °C, 30 min;

[0133] Inoculate the seed solutions of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus into fresh LB medium respectively and culture them at 30 °C for 48 h to obtain the fermentation broths of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus respectively; among them, the composition of LB medium is: 10 g of glucose, 5 g of beef extract, 1.5 g of sodium chloride, 0.05 g of FeSO4·7H2O, 0.15 g of K2HPO4, 0.5 g of ZnSO4·7H2O, 1000 mL of distilled water, pH 7.0, sterilization conditions: 121 °C, 30 min;

[0134] Mix the fermentation broths of Bacillus velezensis, Pseudomonas monteilii, and Lactobacillus pentosus to obtain the compound bacterium agent;

[0135] Mix the compound bacterium agent, promoter, and nutrient substance to obtain the microbial inoculant.

[0136] The promoter is modified saponin. The preparation method of modified saponin includes the following steps:

[0137] Mix saponin, maleic anhydride, catalyst 4-dimethylaminopyridine, and N,N-dimethylformamide, and react them at 75 °C for 8 h under a nitrogen atmosphere to obtain modified saponin; among them, the mass ratio of saponin to maleic anhydride is 1:1, the mass ratio of saponin to catalyst 4-dimethylaminopyridine is 1:0.05, and the dosage ratio of saponin to N,N-dimethylformamide is 1 g:15 mL;

[0138] The nutrient substance includes zinc amino acid, calcium amino acid, and fulvic acid with a mass ratio of 3:1:5;

[0139] The preparation method of the organic fertilizer includes the following steps:

[0140] Mix half of the crop waste and half of the humic acid, and calcine at 300 °C for 3 h to obtain an intermediate product.

[0141] Mix the intermediate product, the remaining crop waste, manure and water to obtain a mixture with a water content of 55 wt%; then add a microbial inoculant and carry out composting fermentation at 55 °C for 20 d to obtain a fermentation product.

[0142] Mix the fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate, and granulate to obtain an organic fertilizer.

[0143] Comparative Example 1

[0144] An organic fertilizer, which is the same as that in Example 1, except that: the compound bacteria in the microbial inoculant include Bacillus velezensis and Pseudomonas monteilii with a mass ratio of 2:1.

[0145] Comparative Example 2

[0146] An organic fertilizer, which is the same as that in Example 1, except that: the compound bacteria in the microbial inoculant include Bacillus velezensis and Lactobacillus pentosus with a mass ratio of 2:2.

[0147] Comparative Example 3

[0148] An organic fertilizer, which is the same as that in Example 1, except that: the compound bacteria in the microbial inoculant include Pseudomonas monteilii and Lactobacillus pentosus with a mass ratio of 1:2.

[0149] Comparative Example 4

[0150] An organic fertilizer, which is the same as that in Example 1, except that: the microbial inoculant does not contain a promoter.

[0151] Comparative Example 5

[0152] An organic fertilizer, which is the same as that in Example 1, except that: the promoter in the microbial inoculant is saponin.

[0153] Comparative Example 6

[0154] An organic fertilizer, which is the same as that in Example 1, except that:

[0155] The preparation method of the organic fertilizer includes the following steps:

[0156] Mix the crop waste, manure and water to obtain a mixture with a water content of 50 wt%; then add a microbial inoculant and carry out composting fermentation at 55 °C for 20 d to obtain a fermentation product.

[0157] Mix the fermentation product, humic acid, trehalose, potassium chloride and diammonium phosphate, and granulate to obtain an organic fertilizer.

[0158] Experimental Example 1: Experiment on the Promotion of Plant Growth by Organic Fertilizer

[0159] 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 (100 kg / mu), Comparative Example 5 group (100 kg / mu), and Comparative Example 6 group (100 kg / mu) were set up in the experiment. Each treatment group was set with 3 replicates, the experimental plot area was 25 square meters, and the initial soil pH was 4.6.

[0160] The organic fertilizers in Examples 1 - 3 and Comparative Examples 1 - 6 were applied to the test fields respectively, and then rotary tillage was carried out with a rotary tiller to mix evenly, and then sowing was carried out.

[0161] All treatment groups were fertilized uniformly, with a one - time basal application and no top - dressing in the later stage.

[0162] After the corn matured, the yield (kg / mu), yield increase rate (%), and root dry weight (g / plant) of the corn in each group were recorded respectively.

[0163] The calculation formula for the yield increase rate is as follows:

[0164] Yield increase rate (%) = (yield of the experimental group - yield of the blank control group) / yield of the blank control group × 100%; The specific experimental data are shown in Table 1.

[0165] Table 1

[0166]

[0167] Experimental Example 2: Experiment on the Promotion of Cucumber Growth

[0168] Cucumber seedlings with consistent growth were selected for transplanting, and divided into a blank control group, Examples 1 - 3 groups, and Comparative Examples 1 - 6. The blank control group was not fertilized, and the other treatment groups used the organic fertilizers in Examples 1 - 3 and Comparative Examples 1 - 6 respectively. At the time of transplanting, 10 g of organic fertilizer was used for each plant root, evenly spread on the soil surface of the breeding tray and covered with soil. There were 100 seedlings in each treatment, with 3 replicates. After sufficient watering, the plant height (cm), root - shoot ratio, and dry weight (g) were measured after 18 d. The specific experimental data are shown in Table 2.

[0169] Table 2

[0170]

[0171] As can be seen from Table 2, the organic fertilizers prepared in Examples 1 to 3 of the present invention have an obvious promoting effect on the growth of cucumbers. The plant height and dry weight are significantly increased compared with the blank control group and the comparative examples. 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 cucumbers.

[0172] Test Example 3 Determination of Indoor Control Effect on Cucumbers

[0173] The active spores of Pythium aphanidermatum, Rhizoctonia solani and Fusarium oxysporum were respectively inoculated into sorghum grain medium and cultured at 28 °C until the hyphae covered the surface of each sorghum grain. After evenly mixing the 3 kinds of fungus-infected sorghum grains, they were evenly spread on the surface soil 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, and 10 g / m 2 After mixing with the soil, it was evenly spread on the soil surface of the breeding tray, cucumber seeds were evenly sown and covered with soil. There were 100 seedlings in each treatment, with 3 replicates, and they were cultured at 20 °C. After emergence, the disease incidence was observed, and the disease incidence rate and control effect were calculated. No fungicide was applied in the blank control group. The calculation formulas for the disease incidence rate and control effect are as follows:

[0174] Disease incidence rate = number of diseased plants / total number of plants × 100%;

[0175] Control effect (%) = [(control disease incidence rate - treatment disease incidence rate) / control disease incidence rate] × 100%; The specific experimental data are shown in Table 3.

[0176] Table 3

[0177]

[0178] As can be seen from Table 3, the organic fertilizers prepared in Examples 1 to 3 of the present invention have an obvious inhibitory effect on the disease incidence rate of cucumbers, far superior to the blank control group and the comparative examples. That is, the synergistic control effect on disease prevention and control when using Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus in combination solves the limitation problem of single strains, greatly reduces diseases, and realizes increased production and income.

[0179] The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A microbial inoculum, characterized in that, It comprises the following raw materials in parts by weight: Compound bacterium agent: 10 - 20 parts, promoter: 5 - 10 parts, nutrient substance: 5 - 10 parts; The compound bacterium agent is composed of Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus; The promoter includes modified saponin; The mass ratio of Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus in the compound bacterium agent is 2 - 3:1 - 2:1 - 2; The preservation number of Bacillus velezensis is CGMCC NO.28606; The preservation number of Pseudomonas monteilii is CICC 25156; The preservation number of Lactobacillus pentosus is ATCC 8041; The nutrient substance includes zinc amino acid, calcium amino acid and fulvic acid; the mass ratio of zinc amino acid, calcium amino acid and fulvic acid is 1 - 3:1 - 3:4 - 5; The preparation method of the modified saponin includes the following steps: Mix saponin, maleic anhydride, catalyst and solvent, and carry out reaction to obtain modified saponin; The mass ratio of saponin to maleic anhydride is 1:0.5 - 2; The mass ratio of saponin to catalyst is 1:0.02 - 0.1; The dosage ratio of saponin to solvent is 1g:10 - 20mL; The reaction conditions: temperature is 60 - 100°C, time is 4 - 12h, and the atmosphere is nitrogen or argon.

2. The preparation method of the microbial inoculum according to claim 1, characterized in that, It includes the following steps: Respectively inoculate Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus strains into LB medium for the first culture to obtain the seed solutions of Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus respectively; Respectively inoculate the seed solutions of Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus into a new LB medium for the second culture to obtain the fermentation broths of Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus respectively; Mix the fermentation broths of Bacillus velezensis, Pseudomonas monteilii and Lactobacillus pentosus to obtain a compound bacterium agent; Mix the compound bacterium agent, promoter and nutrient substance to obtain a microbial bacterium agent.

3. The preparation method of the microbial inoculum according to claim 2, characterized in that, The temperature of the first culture is 25 - 30°C, and the time of the first culture is 20 - 24h; The temperature of the second culture is 25 - 30°C, and the time of the second culture is 40 - 48h.

4. An organic fertilizer, characterized in that, It comprises the following raw materials in parts by weight: Agricultural crop waste: 50 - 100 parts, feces: 40 - 50 parts, microbial bacterium agent: 20 - 30 parts, humic acid: 10 - 20 parts, trehalose: 1 - 5 parts, potassium chloride: 5 - 10 parts, and diammonium phosphate: 5 - 10 parts; The microbial bacterium agent is the microbial bacterium agent described in claim 1 or the microbial bacterium agent prepared by the preparation method of the microbial bacterium agent described in claim 2 or 3.

5. The preparation method of the organic fertilizer according to claim 4, characterized in that, It includes the following steps: Mix part of the agricultural crop waste and part of the humic acid, and carry out calcination to obtain an intermediate; Mix the intermediate, the remaining agricultural crop waste, feces and water to obtain a mixture; then add the microbial bacterium agent and carry out compost fermentation to obtain a fermentation product; Mix the fermentation product, the remaining humic acid, trehalose, potassium chloride and diammonium phosphate, and carry out granulation to obtain organic fertilizer.

6. The preparation method of the organic fertilizer according to claim 5, characterized in that The temperature of the calcination is 300 - 500°C, and the time of the calcination is 2 - 5h.

7. The preparation method of the organic fertilizer according to claim 5, characterized in that The mass ratio of the partial crop waste to the remaining crop waste is 1:1 to 2; The mass ratio of the partial humic acid to the remaining humic acid is 1:1 to 2.

8. The preparation method of the organic fertilizer according to claim 5, characterized in that, The water content of the mixture is 50 to 60 wt%; The fermentation temperature is 50 to 60 °C, and the fermentation time is 15 to 20 d.

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