Bio-organic fertilizer for preventing and treating root rot of codonopsis pilosula as well as preparation method and application of bio-organic fertilizer

Abstract

The invention relates to the technical field of bio-fertilizers, and particularly discloses a bio-organic fertilizer for preventing and treating root rot of codonopsis pilosula as well as a preparation method and application of the bio-organic fertilizer. The bio-organic fertilizer for preventing and treating root rot of codonopsis pilosula provided by the invention comprises a kitchen fermentation base material and a complex microbial inoculant in a weight ratio of 100: (1-10), the complex microbial inoculant comprises one or more of bacillus atrophaeus, bacillus coagulans, bacillus licheniformis, bacillus velezensis and brevibacillus laterosporus; the invention further provides a preparation method of the bio-organic fertilizer and application of the bio-organic fertilizer in prevention and treatment of root rot of codonopsis pilosula. The bio-organic fertilizer provided by the invention can degrade glyphosate in soil on one hand, and also can reduce the occurrence of root rot of codonopsis pilosula on the other hand, so that the growth of codonopsis pilosula and the accumulation of active value components of codonopsis pilosula can be promoted.

Description

Technical Field

[0001] The present application relates to the technical field of biological fertilizers, and in particular to a biological organic fertilizer for preventing and treating Codonopsis pilosula root rot, and a preparation method and application thereof. Background Art

[0003] In order to solve the problem caused by excessive use of chemical pesticides and fertilizers in the cultivation of Codonopsis pilosula, two methods are currently commonly used in the industry. The first method is to directly use physical or chemical methods to reduce pesticide residues in the soil, such as using adsorbents to adsorb herbicides such as glyphosate, but this method has limited effect and high cost; the second method is to improve the soil by adding microorganisms, using the adaptability of microorganisms to the environment and antagonism to pathogens to reduce the incidence of root rot, thereby improving the health of the soil and the number of Codonopsis pilosula seedlings, but this method is expensive, and a single microbial preparation has an insufficient antibacterial spectrum and cannot effectively prevent and control a variety of pathogens, making it difficult to meet the diverse needs in practical applications.

[0004] Therefore, the development of a broad-spectrum bio-organic fertilizer that can degrade chemical pesticide residues and effectively prevent and treat Codonopsis root rot is of great significance to the healthy and sustainable development of the Codonopsis industry. Summary of the invention

[0005] In order to degrade chemical pesticide residues in Codonopsis pilosula cultivation and achieve effective prevention and treatment of Codonopsis pilosula root rot, the present application provides a biological organic fertilizer for preventing and treating Codonopsis pilosula root rot, and a preparation method and application thereof.

[0006] In the first aspect, the present application provides a biological organic fertilizer for preventing and treating Codonopsis pilosula root rot, using the following technical solution: A biological organic fertilizer for preventing and treating root rot of Codonopsis pilosula comprises a kitchen fermentation base material and a composite bacterial agent in a weight ratio of 100:(1-10); the composite bacterial agent comprises one or more of Bacillus atrophaeus, Bacillus coagulans, Bacillus licheniformis, Bacillus velezii and Brevibacillus laterosporus.

[0007] The present application uses a food waste fermentation base material and a composite bacterial agent to prepare a biological organic fertilizer, wherein the food waste fermentation base material is a base material prepared by high-temperature aerobic fermentation of food waste, which contains a variety of humic acids, macroelements such as nitrogen, phosphorus, potassium, trace elements such as zinc, iron, manganese, and active substances. The food waste fermentation base material can participate in the degradation process of glyphosate, especially in the presence of microorganisms, humic acid can promote the biodegradation of glyphosate and reduce pesticide residues in the soil; in addition, the food waste fermentation base material can provide sufficient nutrients for the composite bacterial agent, and can significantly improve the soil structure, increase soil activity, and promote the growth and development of the root system of Codonopsis pilosula. The composite bacterial agent has excellent antifungal activity, and the secondary metabolites it produces can effectively inhibit the reproduction of pathogenic bacteria of Codonopsis pilosula root rot, destroy its hyphae growth, and promote the biodegradation of glyphosate and reduce chemical pesticide residues. Specifically, the secondary metabolites of Bacillus Velezii, Bacillus laterosporus, etc. contain a variety of proteases and lipopeptide antibiotics, which can significantly inhibit the mycelial growth and spore germination of the pathogenic bacteria of Codonopsis root rot, increase the permeability of the pathogenic bacteria cell membrane, aggravate the release of intracellular nucleic acid contents, and then cause cell rupture to death, thereby effectively preventing and controlling the occurrence of Codonopsis root rot. In summary, the bio-organic fertilizer provided by the present application can not only effectively degrade the glyphosate content in the soil through the combined action of the kitchen fermentation base material and the composite bacterial agent, but also significantly reduce the occurrence of Codonopsis root rot, promote the healthy growth of Codonopsis and the accumulation of medicinal active ingredients, and has a good prospect for use.

[0008] In the present application, glyphosate will form aminomethylphosphine, sarcosine, phosphoric acid, glycine and carbon dioxide after degradation, and the above substances can be used as phosphorus sources, carbon sources or nitrogen sources for the growth of composite bacterial agents, promote the growth and reproduction of microorganisms, and further achieve the prevention and control of Codonopsis pilosula root rot.

[0009] Optionally, the weight ratio of the kitchen fermentation base material to the composite bacterial agent is 100:(3-7).

[0010] In the present application, the weight ratio of the kitchen fermentation base material and the compound bacterial agent will affect the control effect of Codonopsis pilosula diseased soil and the growth of Codonopsis pilosula. Through experimental research, it was found that by further controlling the weight ratio of the kitchen fermentation base material and the compound bacterial agent within the above range, the obtained biological organic fertilizer has a better growth-promoting effect on Codonopsis pilosula and a better control effect on diseased soil, and can make the dry weight of Codonopsis pilosula reach more than 12.0g / grain, and the control effect on diseased soil can reach more than 75%.

[0011] In some embodiments, the weight ratio of the kitchen fermentation base and the composite bacterial agent can be 100:(1-3), 100:(1-5), 100:(1-7), 100:(1-10), 100:(3-5), 100:(3-7), 100:(3-10), 100:(5-7), 100:(5-10) or 100:(7-10).

[0012] In a specific embodiment, the weight ratio of the kitchen fermentation base and the composite bacterial agent can also be 100:1, 100:3, 100:5, 100:7 or 100:10.

[0013] Optionally, the composite bacterial agent contains Bacillus Velez subtilis, Bacillus atrophaeus and Brevibacillus laterosporus, and the weight ratio of Bacillus Velez subtilis, Bacillus atrophaeus and Brevibacillus laterosporus is (1-5):(1-2):(1-3).

[0014] Optionally, the weight ratio of Bacillus Velez, Bacillus atrophaeus and Brevibacillus laterosporus is (2-4): (1.3-1.6): (1.5-2.5).

[0015] Through experimental research, the present application found that by further using Bacillus velez, Bacillus atrophaeus and Bacillus laterosporus to prepare a composite bacterial agent, and controlling the weight ratio of the three bacteria within the range of (2-4): (1.3-1.6): (1.5-2.5), the obtained composite bacterial agent has a better comprehensive inhibitory effect on the three pathogens of Fusarium oxysporum, Fusarium acuminate and Fusarium trilineatum, and a better prevention and control effect on Codonopsis pilosula root rot.

[0016] In some embodiments, the weight ratio of the Bacillus Velezii, Bacillus atrophaeus and Bacillus laterosporus can be (1-2): 0.6:2, (1-3): 0.6:2, (1-4): 0.6:2, (1-5): 0.6:2, (2-3): 0.6:2, (2-4): 0.6:2, (2-5): 0.6:2, (3-4): 0.6:2, (3-5): 0.6:2, (4-5): 0.6:2, (1-3): 0.6:2, (1-4): 0.6:2, (1-5): 0.6:2, (2-3): 0.6:2, (2-4): 0.6:2, (2-5): 0.6:2, (3-4): 0.6:2, (3-5): 0.6:2, (4-5): 0.6:2, (1-3): 0.6:2, (1-3): 0.6:2, (1-4): 0.6:2. 3:(1-2):2, 3:(1.3-1.6):2, 3:(1.3-2):2, 3:(1.6-2):2, 3:1.6:(1-1.5), 3:1.6:(1-2), 3:1.6:(1-2.5), 3:1.6:(1-3), 3:1.6:(1.5-2), 3:1.6:(1.5-2.5), 3:1.6:(1.5-3), 3:1.6:(2-2.5), 3:1.6:(2-3) or 3:1.6:(2.5-3).

[0017] In a specific embodiment, the weight ratio of the Bacillus Velez, Bacillus atrophaeus and Bacillus laterosporus can also be 1:1.6:2, 2:1.6:2, 3:1.6:2, 4:1.6:2, 5:1.6:2, 3:1:2, 3:1.3:2, 3:2:2, 3:1.6:1, 3:1.6:1.5, 3:1.6:2..5 or 3:1.6:3.

[0018] Optionally, the Bacillus Velez is Bacillus Velez BGB-89R, the Bacillus atrophaeus is Bacillus atrophaeus BGB-98R, and the Brevibacillus laterosporus is Brevibacillus laterosporus BGB-173R.

[0019] Optionally, the composite bacterial agent further comprises a protective agent, which is a mixture of trehalose, egg white powder, cephalin and maltitol.

[0020] Optionally, the total humic acid content of the kitchen fermentation base material is 38-42% by mass and the water content is ≤10%.

[0021] In a second aspect, the present application provides a method for preparing a bio-organic fertilizer, comprising the following steps: preparing a kitchen kitchen fermentation base material, a composite bacterial agent, and mixing materials.

[0022] In the third aspect, the application provides an application of the biological organic fertilizer in the prevention and treatment of Codonopsis pilosula root rot.

[0023] Optionally, the pathogen of the root rot of Codonopsis pilosula is one or more of Fusarium oxysporum, Columnar spore fungus, Fusarium acuminate, Fusarium solani, Fusarium equisetum, Fusarium avenae, Fusarium odoratum, Fusarium pseudoverticillium, Fusarium lanuginosa, Fusarium trilineata, Fusarium solani and Echinococcus robustus.

[0024] In summary, this application has the following beneficial effects: 1. The present application uses kitchen fermentation base material and composite bacterial agent as raw materials to prepare biological organic fertilizer. By controlling the weight ratio of the kitchen fermentation base material and the composite bacterial agent within the range of 100:(1-10), the obtained biological organic fertilizer can degrade the glyphosate content in the soil on the one hand, and reduce the occurrence of Codonopsis pilosula root rot on the other hand, thereby promoting the growth of Codonopsis pilosula and the accumulation of active ingredients with medicinal value.

[0025] 2. The present application further adopts a weight ratio of (2-4): (1.3-1.6): (1.5-2.5) of Bacillus Velezii, Bacillus atrophaeus and Bacillus laterosporus to prepare a composite bacterial agent. The obtained composite bacterial agent has a better comprehensive inhibitory effect on the three pathogens of Fusarium oxysporum, Fusarium acuminate and Fusarium trilineatum. When it is mixed with a kitchen fermentation base material, the obtained biological organic fertilizer has a better effect on promoting the growth of Codonopsis pilosula and preventing and controlling diseased soil. DETAILED DESCRIPTION

[0026] The present application provides a biological organic fertilizer for preventing and treating Codonopsis pilosula root rot, comprising a kitchen fermentation base material and a composite bacterial agent in a weight ratio of 100:(1-10); further, the kitchen fermentation base material and the composite bacterial agent in a weight ratio of 100:(3-7).

[0027] The composite bacterial agent contains a protective agent and one or more of Bacillus atrophaeus, Bacillus coagulans, Bacillus licheniformis, Bacillus velezii and Bacillus laterosporus, and the protective agent is a mixture of trehalose, egg white powder, cephalin and maltitol in a weight ratio of 1:0.5:0.02:0.2. Further, the composite bacterial agent contains Bacillus velezii, Bacillus atrophaeus and Bacillus laterosporus, and the weight ratio of Bacillus velezii, Bacillus atrophaeus and Bacillus laterosporus is (1-5): (1-2): (1-3); further, the weight ratio of Bacillus velezii, Bacillus atrophaeus and Bacillus laterosporus is (2-4): (1.3-1.6): (1.5-2.5).

[0028] The kitchen fermentation base material is prepared by the method provided in ZL 2010102693567; the mass content of total humic acid in the kitchen fermentation base material is 38-42%, the mass content of free humic acid is 35-40%, the mass content of water-soluble humic acid is 14-16%, and the water content of the kitchen fermentation base material is ≤10%.

[0029] The preparation method of the composite bacterial agent provided in the present application comprises the following steps: (1) Fermentation: Fermentation medium is loaded into a fermentation device, and the ventilation volume of the fermentation device is adjusted to 0.8-1.2vvm, the temperature is 28-35°C, and the rotation speed is 150-220rpm. Bacillus seed liquid is inoculated into the fermentation medium according to a volume percentage of 0.1-1%, and fermentation begins. After 16h-20h, the dissolved oxygen is controlled at 30-50%. After 24-48h of fermentation growth, a sterile glucose solution with a mass percentage of 20-35% is added at a flow rate of 80-150g / h, and the total sugar supplement concentration is about 30-60g / L. The pH value and dissolved oxygen content reach the lowest point, and the fermentation continues. After about 45-65h, microscopic examination is performed. When the sporulation amount reaches more than 90%, the fermentation is completed and the tank is put into storage.

[0030] (2) centrifuging the fermentation liquid in a vertical centrifuge at a speed of 5000-8000 rpm to obtain a Bacillus concentrate and a Bacillus supernatant, storing the Bacillus supernatant at 4° C. for later use, detecting the bacterial content of the Bacillus concentrate, and cold drying at 4° C.-8° C. to obtain a Bacillus powder with a bacterial content of 50-200 billion CFU / g.

[0031] (3) preparing different types of Bacillus powder respectively according to the method of step (1-2); mixing different types of Bacillus powder in a certain proportion, and then adding a protective agent and mixing evenly to obtain a composite bacterial agent.

[0032] The strains used in the examples of the present application are all strains disclosed in the prior art. The Bacillus coagulans is Bacillus coagulans BC2000, with a deposit number of CGMCC No. 21621; the Bacillus Velez is Bacillus Velez BGB-89R, with a deposit number of CGMCC No. 28824; the Bacillus licheniformis is Bacillus licheniformis BGB-83R, with a deposit number of CGMCC NO. 24182; the Bacillus atrophaeus is Bacillus atrophaeus BGB-98R, with a deposit number of CGMCC No. 27752; the Bacillus laterosporus is Bacillus laterosporus BGB-173R, with a deposit number of CGMCC No. 28825; the raw materials, reagents, solvents, etc. used in the present application can all be obtained commercially.

[0033] The present application is further described in detail below in combination with preparation examples, embodiments and performance testing experiments.

[0034] Preparation Example 1-14 Preparation Examples 1-14 each provide a composite bacterial agent.

[0035] The difference between the above preparation examples is that the types and ratios of the strains in the composite bacterial agent are specifically shown in Table 1 below.

[0036] The preparation method of the composite bacterial agent provided in Preparation Example 1-14 comprises the following steps: (1) Fermentation: Fermentation medium is loaded into a fermentation tank, and the ventilation volume, temperature and rotation speed of the fermentation tank are adjusted to 0.8 vvm, 30°C and 180 rpm, and the Bacillus seed liquid is inoculated into the fermentation medium at a volume percentage of 1%, and fermentation is started. After 16 hours, the dissolved oxygen is controlled at 40%. After 24 hours of fermentation growth, a sterile glucose solution with a mass percentage of 30% is added at a flow rate of 100 g / h, and the total sugar supplement concentration is about 50 g / L. The pH value and dissolved oxygen content reach the lowest point, and the fermentation continues. After about 48 hours of microscopic examination, when the sporulation amount reaches more than 90%, the fermentation is completed and the tank is removed.

[0037] (2) The fermentation liquid was centrifuged in a vertical centrifuge at a speed of 6000 rpm to obtain a Bacillus concentrate and a Bacillus supernatant. The Bacillus supernatant was stored at 4° C. for later use. The bacterial content of the Bacillus concentrate was tested and the concentrate was lyophilized at 4° C. to obtain a Bacillus powder with a bacterial content of 50-200 billion CFU / g.

[0038] (3) According to the method of step (1-2), different types of Bacillus powders are prepared respectively (the bacterial content of Bacillus Velez powder is 200 billion CFU / g; the bacterial content of Bacillus atrophaeus powder is 200 billion CFU / g; the bacterial content of Bacillus laterosporus powder is 200 billion CFU / g; the bacterial content of Bacillus coagulans powder is 5 billion CFU / g; the bacterial content of Bacillus licheniformis powder is 200 billion CFU / g); the Bacillus powders are mixed evenly according to the types and proportions shown in Table 1 to obtain composite powders; then the composite powders are mixed evenly with a protective agent at a weight ratio of 20:1 to obtain a composite bacterial agent. The protective agent is a mixture of trehalose, egg white powder, cephalin and maltitol in a weight ratio of 1:0.5:0.02:0.2.

[0039] Table 1 Types and ratios of bacterial powders in the composite bacterial agents provided in Preparation Examples 1-14 Comparative Preparation Example 1 Comparative Preparation Example 1 provides a composite bacterial agent.

[0040] The difference between the above-mentioned comparative preparation example and Preparation Example 3 is that the type and ratio of the bacterial powder in the composite bacterial agent are: Bacillus Velez, Bacillus subtilis (Bacillus subtilis BJ-1, deposit number CCTCCNO: M2017443, the bacterial content of Bacillus subtilis powder is 200 billion CFU / g) and Bacillus laterosporus in a weight ratio of 2:1.6:2.

[0041] Comparative Preparation Example 2 Comparative Preparation Example 2 provides a composite bacterial agent.

[0042] The difference between the above comparative preparation example and preparation example 3 is that the type and ratio of the bacterial powder in the composite bacterial agent are: Bacillus Velez, Bacillus atrophaeus and Bacillus megaterium (Bacillus megaterium ZLP-21, the preservation number is CGMCC No. 25030, and the bacterial content of Bacillus megaterium powder is 200 billion CFU / g) in a weight ratio of 2:1.6:2.

[0043] Pathogen inhibition effect The inhibitory effects of the composite bacterial agents provided in Preparation Examples 1-14 and Comparative Preparation Examples 1-2 on the three pathogenic bacteria, namely, Fusarium oxysporum strains, Fusarium acuminate strains, and Fusarium trilineatum strains, as well as the inhibitory effects of the corresponding composite supernatants on the spore germination of the three pathogenic bacteria, namely, Fusarium oxysporum strains, Fusarium acuminate strains, and Fusarium trilineatum strains.

[0044] (I) Spore inhibition test: (1) Preparation of test samples: The supernatants of each Bacillus (containing the metabolites of each bacterium) were filtered using a 0.45 μm sterile filter membrane, and then the supernatants of each Bacillus were mixed according to the weight ratio of the corresponding bacterial powders of Preparation Examples 1-14 and Comparative Preparation Examples 1-2 (as shown in Table 1) to obtain composite supernatants A1-A14 and B1-B2; the composite supernatants A1-A14 and B1-B2 were used as test samples for spore inhibition effect detection.

[0045] (2) Preparation of pathogens: Inoculate the bacterial cakes (5 mm in diameter) of the pathogenic bacteria Fusarium oxysporum, Fusarium acuminate, and Fusarium trilineatum into the center of the PDA culture plate and culture them in the dark at 25°C for 7 days for reproduction. Take 5 bacterial cakes (5 mm in diameter) from the edge of the colony and inoculate them into PDA liquid culture medium. Culture them in the dark at 25°C and 180 rpm / min for 48 hours to obtain the fermentation broth of the three pathogens. Determine the number of spores in the fermentation broth of each pathogen and adjust it to a bacterial content of 1×10 5 CFU / mL.

[0046] (3) Spore inhibition test: The composite supernatants A1-A14 and B1-B2 were mixed with the pathogen fermentation broth at a volume ratio of 1:1, and 2 mL was added to the strains in a 24-well plate and cultured at 25°C for 24 h; they were respectively treated groups 1-14 and control groups 1-2; blank PDA liquid culture medium was used instead of the composite supernatant as the blank control group. Each group was repeated 3 times. The spore germination of the composite was observed under an optical microscope, and the spore germination rate and spore germination inhibition rate were calculated, and the average value of the three groups was taken. The results are shown in Table 2 below.

[0047] The calculation formula of spore germination rate is: spore germination rate (%) = number of germinated spores / total number of spores × 100%; The calculation formula of spore germination inhibition rate is: spore germination inhibition rate (%) = [(C 1 -C 2 ) / C 1 ]×100%; where C 1 is the spore germination rate of the blank control group, C 2 is the spore germination inhibition rate of the treatment group or the control group.

[0048] (II) Pathogen inhibition test: Three kinds of pathogens (5mm cake) were inoculated to the center of the PDA plate, and the composite bacterial powders prepared in Preparation Examples 1-14 and Comparative Preparation Examples 1-2 were streaked with a sterile inoculation loop at a position about 1 cm away from the edge of the plate on both sides of the pathogenic bacterial cake. The plate inoculated with only the pathogenic bacterial cake was used as the blank control group, and each group was repeated 3 times. All plates were placed at 28°C for dark culture until the pathogenic mycelium of the blank control group grew to the edge of the plate. The pathogen inhibition rate of each composite bacterial powder was calculated, and the results are shown in Table 2 below.

[0049] The calculation formula of pathogen inhibition rate is: inhibition rate (%) = [(R 1 -R 2 ) / R 1 ]×100%; where R 1 is the radius of the pathogen colony in the blank control group, R 2 is the radius of the pathogen colony in the treatment group or control group.

[0050] Table 2 Results of spore germination inhibition rate and pathogen inhibition rate test According to the detection results in Table 2, the composite supernatant A1-A14 (including any three of Bacillus atrophaeus, Bacillus coagulans, Bacillus licheniformis, Bacillus velezii and Brevibacillus laterosporus) can achieve a spore germination inhibition rate of more than 90% on the three pathogens of Bacillus oxysporum, Fusarium spp. and Fusarium trilineatum; and the composite bacterial agent prepared by any three bacterial powders of Bacillus atrophaeus, Bacillus coagulans, Bacillus licheniformis, Bacillus velezii and Brevibacillus laterosporus in Preparation Examples 1-14 can achieve a pathogen inhibition rate of more than 50% on the three pathogens of Bacillus oxysporum, Fusarium spp. and Fusarium trilineatum; and the composite supernatant B1 (including Bacillus velezii, Bacillus subtilis and Brevibacillus laterosporus) can achieve a spore germination inhibition rate of more than 50% on Fusarium spp. and Fusarium trilineatum. The spore germination inhibition rate of the two pathogens, Fusarium, is only 77.66-82.84%, and the pathogen inhibition rate of the composite bacterial agent prepared by Bacillus Velez, Bacillus subtilis and Brevibacillus laterosporus in Comparative Preparation Example 1 on Fusarium acutum and Fusarium trilineum is only 40.9-46.9%; while the spore germination inhibition rate of the composite supernatant B2 (containing Bacillus Velez, Bacillus atrophaeus and Bacillus megaterium) on Fusarium oxysporum and Fusarium trilineum is only 86.03-89.11%, and the pathogen inhibition rate of the composite bacterial agent prepared by Bacillus Velez, Bacillus subtilis and Brevibacillus laterosporus in Comparative Preparation Example 1 on Fusarium oxysporum and Fusarium trilineum is only 43.7-49.6%. Therefore, it is shown that the composite bacterial agent prepared by using any three bacterial powders of Bacillus atrophaeus, Bacillus coagulans, Bacillus licheniformis, Bacillus velez and Bacillus laterosporus in the present application has a good inhibitory effect on Fusarium oxysporum, Fusarium acuminate and Fusarium trilineatum, and can achieve effective prevention and treatment of Codonopsis pilosula root rot.

[0051] Further comparison found that the composite supernatants A2-A4, A7, and A10-11 had a spore germination inhibition rate of 94.02-94.61% (≥94%) for Spore germination of Fusarium oxysporum, a spore germination inhibition rate of 95.08-95.61% (≥95%) for Fusarium acuminate, and a spore germination inhibition rate of 92.01-92.39% (≥92%) for Fusarium trilineatum; and the composite bacterial agents provided in Preparation Examples 2-4, Preparation Examples 7, and Preparation Examples 10-11 had a pathogen inhibition rate of 60.5-65.4% (≥60%) for Spore oxysporum, a pathogen inhibition rate of 70.8-73.5% (≥70%) for Fusarium acuminate, and a pathogen inhibition rate of 55.2-58.9% (≥55%) for Fusarium trilineatum. Therefore, it is explained that the present application further uses Bacillus velezensis, Bacillus atrophaeus and Brevibacillus laterosporus to prepare the composite bacterial agent, and controls the weight ratio of the three bacteria within the range of (2-4): (1.3-1.6): (1.5-2.5). The obtained composite bacterial agent has a better comprehensive inhibitory effect on the three pathogens, namely, Fusarium oxysporum, Fusarium acuminate and Fusarium trilineatum, and has a better prevention and control effect on Codonopsis pilosula root rot.

[0052] Examples 1-14 Embodiments 1-14 respectively provide a kind of biological organic fertilizer.

[0053] The difference between the above embodiments is that the composite bacterial agent used in the biological organic fertilizer is derived from Preparation Examples 1-14 respectively.

[0054] The preparation method of the bio-organic fertilizer provided in Examples 1-14 comprises the following steps: (1) Preparation of fermented kitchen material: The fermented kitchen material was prepared according to the method provided in ZL 2010102693567. The mass content of total humic acid, the mass content of free humic acid, and the mass content of water-soluble humic acid in the obtained fermented kitchen material was 41.6%, 37.3%, and 14.8%, respectively. The water content of the fermented kitchen material was 8.5%.

[0055] (2) Preparation of composite bacterial agent: proceed according to Preparation Example 1-14.

[0056] (3) The kitchen kitchen fermentation base material and the composite bacterial agent are uniformly mixed in a weight ratio of 100:5 to obtain a biological organic fertilizer.

[0057] Examples 15-18 Embodiments 15-18 respectively provide a kind of biological organic fertilizer.

[0058] The difference between the above embodiment and embodiment 3 is that the weight ratio of the kitchen fermentation base material to the composite bacterial agent is specifically shown in Table 3 below.

[0059] Table 3 Weight ratio of the fermented base material of the food and the composite bacterial agent in the bio-organic fertilizer provided in Examples 15-18 Example Weight ratio of kitchen fermentation base material and compound bacterial agent 3 100：5 15 100：1 16 100：3 17 100：7 18 100：10 Comparative Example 1-2 Comparative Examples 1-2 respectively provide a kind of biological organic fertilizer.

[0060] The difference between the above comparative example and Example 3 is that the composite bacterial agent used in the biological organic fertilizer is derived from comparative preparation examples 1-2 respectively.

[0061] Comparative Example 3 Comparative Example 3 provides a bio-organic fertilizer, which is a kitchen kitchen fermentation base material.

[0062] Effect of controlling root rot of Codonopsis pilosula The control effect of the biological organic fertilizer provided by Examples 1-18 and Comparative Examples 1-3 on Codonopsis pilosula root rot was investigated, and the results are shown in Table 4 below.

[0063] (1) Test method: The soil in which 41% glyphosate isopropylamine salt solution (dosage of 2150 g active ingredient / hectare) was used in the last season of planting Codonopsis pilosula was used as a test site, and weeding was carried out. The test site was divided into 22 areas (each area was 6m×3m), of which 18 areas were treated with the biological organic fertilizer provided by Examples 1-18 (test areas 1-18), 3 areas were treated with the biological organic fertilizer provided by Comparative Examples 1-3 (control areas 1-3); and 1 area was not treated at all (blank control area).

[0064] Before planting Codonopsis pilosula, apply 100kg / mu of bio-organic fertilizer to the soil, plough and mix thoroughly, and till to a depth of 35cm; apply 10kg / mu of urea as topdressing. Then plant Codonopsis pilosula seedlings (seedling height about 23cm, stem diameter about 3.3mm, single seedling fresh weight about 2.2g, seedling size uniform and disease-free) at a rate of 10 plants / m 2 Planting density should be kept at a reasonable level. During the planting period, pests and diseases should be monitored and controlled in a timely manner to avoid the use of chemical pesticides that are harmful to Codonopsis pilosula. Glyphosate degradation rate survey, yield measurement and root rot control effect survey should be conducted 18 months after planting.

[0065] (2) Glyphosate degradation rate survey method: A 1m×1m sample plot was set up in each area of ​​the test site. After removing dead branches and leaves from each sample plot, a circular knife was used to take samples, and then 500g of 0-20cm surface soil mixed sample was collected in an S-shaped manner. The fresh soil was sieved through a 2mm sieve and stored in a 4℃ refrigerator for testing. The glyphosate residue in the fresh soil was determined by high performance liquid chromatography. The specific method can be referred to "Determination of glyphosate residues in soil by pre-column derivatization-solid phase extraction-high performance liquid chromatography fluorescence method" (Wang Yunru, Qin Yuyan, Deng Youzhan, et al. Pre-column derivatization-solid phase extraction-high performance liquid chromatography fluorescence method [J]. Agricultural Research and Application, 2019, 32(Z1): 28-33.1.4).

[0066] (3) Yield measurement: Fifteen plants were randomly sampled from each area, their roots were taken and dried, the dry weight of Codonopsis pilosula was weighed, and the average dry weight of Codonopsis pilosula in each area was calculated.

[0067] (4) Investigation on the effect of root rot prevention and control: Five sampling points were selected in each plot using the five-point sampling method, and 1.0 m 2 Conduct a diseased plant survey. Calculate the disease index and control effect. The calculation formula is as follows: Disease index = 100 × Σ (number of diseased plants at each level × disease level) / (total number of plants surveyed × highest disease level) Control effect = (disease index of blank control area - disease index of treated area) / disease index of blank control area × 100% Note: The disease grading standard is level 0: no disease; level 1: the diseased area accounts for less than 5% of the whole plant; level 3: the diseased area accounts for 6% to 25% of the whole plant; level 5: the diseased area accounts for 26% to 50% of the whole plant; level 7: the diseased area accounts for 51% to 75% of the whole plant; level 9: the diseased area accounts for more than 76% of the whole plant.

[0068] Table 4 Test results of the control effect of Codonopsis root rot According to the test results of Table 4, the bio-organic fertilizer provided by Examples 1-18 of the present application is applied to the diseased soil of Codonopsis pilosula planted with pesticides, and the glyphosate content in the soil can be degraded to 0.37-0.46 mg / kg (≤0.50 mg / kg), so that the average dry weight of Codonopsis pilosula reaches 10.7-12.8 g / grain (≥10 g / grain), and the control effect on the diseased soil can reach 72.2-77.3% (≥70%); while the bio-organic fertilizer provided by Comparative Examples 1-3 is applied to the diseased soil of Codonopsis pilosula planted with pesticides, and the glyphosate content in the soil can only be degraded to 0.79-0.96 mg / kg, so that the average dry weight of Codonopsis pilosula is only 8.53-9.28 g, and the control effect on the diseased soil is only 48.7-65.8%. Therefore, it is shown that the biological organic fertilizer made from the food fermentation base material and the composite bacterial agent provided in the present application can not only degrade glyphosate in the soil, but also reduce the occurrence of Codonopsis pilosula root rot and promote the growth and development of Codonopsis pilosula roots, thereby obtaining a high yield of Codonopsis pilosula.

[0069] The test results of test area 1-14 show that after being treated with the biological organic fertilizer provided by Example 1, Example 5-6, Example 8-9, and Example 12, the average dry weight of Codonopsis pilosula is 11.2-11.8g / grain, and the control effect on diseased soil is 75.1-76.0%; and after being treated with the biological organic fertilizer provided by Example 2-4, Example 7, and Example 10-11, the average dry weight of Codonopsis pilosula can reach 12.0-12.8g / grain (≥12.0g / grain), and the control effect on diseased soil can reach 76.2-77.3% (≥70%). Therefore, it is explained that the present application adopts a weight ratio of (2-4): (1.3-1.6): (1.5-2.5) of Bacillus velezensis, Bacillus atrophaeus and Bacillus laterosporus to prepare a composite bacterial agent and mix it with a kitchen fermentation base material, and the obtained biological organic fertilizer has a better effect on the growth promotion of Codonopsis pilosula and the control effect of diseased soil.

[0070] The test results of test area 3 and test area 15-17 show that after test area 15 and test area 18 were treated with the biological organic fertilizer provided by Example 15 and Example 18, the average dry weight of Codonopsis pilosula was 10.7-11.3 g / grain, and the control effect on diseased soil was 72.2-74.7%; and after test area 3 and test area 16-17 were treated with the biological organic fertilizer provided by Example 3 and Example 16-17, the average dry weight of Codonopsis pilosula could reach 12.0-12.8 g / grain (≥12.0 g / grain), and the control effect on diseased soil could reach 75.4-77.3% (≥75%). Therefore, it is explained that the present application further controls the weight ratio of the kitchen fermentation base material to the composite bacterial agent within the range of 100: (3-7), and the obtained biological organic fertilizer has a better growth-promoting effect on Codonopsis pilosula and a better control effect on diseased soil.

[0071] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A biological organic fertilizer for preventing and treating Codonopsis pilosula root rot, characterized in that: The invention comprises a kitchen fermentation base material and a composite bacterial agent in a weight ratio of 100:(1-10); the composite bacterial agent comprises one or more of Bacillus atrophaeus, Bacillus coagulans, Bacillus licheniformis, Bacillus velezii and Bacillus laterosporus.

2. The bio-organic fertilizer according to claim 1, characterized in that The weight ratio of the kitchen kitchen fermentation base material to the composite bacterial agent is 100:(3-7).

3. The bio-organic fertilizer according to claim 1, characterized in that The composite bacterial agent contains Bacillus Velez, Bacillus atrophaeus and Brevibacillus laterosporus, and the weight ratio of Bacillus Velez, Bacillus atrophaeus and Brevibacillus laterosporus is (1-5): (1-2): (1-3).

4. The bio-organic fertilizer according to claim 3, characterized in that The weight ratio of the Bacillus Velez, Bacillus atrophaeus and Bacillus laterosporus is (2-4): (1.3-1.6): (1.5-2.5).

5. The bio-organic fertilizer according to claim 3, characterized in that The Bacillus Velez is Bacillus Velez BGB-89R, the Bacillus atrophaeus is Bacillus atrophaeus BGB-98R, and the Brevibacillus laterosporus is Brevibacillus laterosporus BGB-173R.

6. The bio-organic fertilizer according to claim 1, characterized in that The composite bacterial agent also contains a protective agent, which is a mixture of trehalose, egg white powder, cephalin and maltitol.

7. The bio-organic fertilizer according to claim 1, characterized in that The total humic acid content of the kitchen fermentation base material is 38-42% by mass and the water content is ≤10%.

8. The preparation method of the bio-organic fertilizer as described in any one of claims 1-7, characterized in that, The following steps are involved: Prepare kitchen fermentation base materials, compound bacterial agents and mixed materials.

9. Application of the bio-organic fertilizer as described in any one of claims 1 to 7 in the prevention and treatment of Codonopsis pilosula root rot.

10. The use according to claim 9, characterized in that: The pathogens of the root rot of Codonopsis pilosula are one or more of Fusarium oxysporum, Columnar spore fungi, Fusarium acuminate, Fusarium solani, Fusarium equisetum, Fusarium avenae, Fusarium odoratum, Fusarium pseudoverticillium, Fusarium lanuginosa, Fusarium trilineata, Fusarium solani and Echinops robustus.

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