A selenium-rich functional bio-organic fertilizer, a preparation method and application thereof

Abstract

The application provides a selenium-rich functional bio-organic fertilizer, a preparation method and application thereof, and belongs to the technical field of bio-fertilizers.The selenium-rich functional bio-organic fertilizer comprises a substrate and a fermentation inoculum; the substrate comprises the following components in a weight fraction: 300-450 parts of selenium-rich chicken manure, 75-150 parts of bacterial rods, 50-100 parts of corn husks and 25-50 parts of humic acid; the amount of the fermentation inoculum is 1‰-3‰ of the mass of the substrate; and the fermentation inoculum comprises bacillus and rhodopseudomonas.The selenium-rich functional bio-organic fertilizer provides an efficient, safe and practical method for the production of selenium-rich vegetable agricultural products and the prevention of plant fungal diseases, has low cost, simple preparation process, is convenient for industrialization, and has a wide market prospect.

Description

Technical Field

[0001] This invention belongs to the field of bio-fertilizer technology, and particularly relates to a selenium-enriched functional bio-organic fertilizer, its preparation method, and its application. Background Technology

[0002] With the large-scale and intensive development of livestock and poultry farming, livestock and poultry manure, as a byproduct of farming, is constantly increasing. Improper treatment not only breeds a large number of harmful bacteria but also poses a serious threat to the sustainable development of the ecological environment, becoming a major source of agricultural non-point source pollution. Compared with other treatment methods, aerobic composting is simple to operate, requires fewer technical standards, and is economical. It can not only remove harmful substances from compost but also allow compost products to be used in agricultural planting, turning waste into treasure and achieving the goal of agricultural planting and breeding cycles. This has significant practical implications for reducing environmental pollution and carbon emissions.

[0003] The key to producing organic fertilizer from livestock and poultry manure lies in the organic fertilizer fermentation agent and the composting process. These factors not only affect the composting effect of the organic fertilizer but also directly impact the product's function and efficacy. In existing technologies, the composting of bio-organic fertilizers suffers from slow temperature rise and a long composting cycle. The products are primarily used to enhance soil fertility, with limited effectiveness in controlling plant diseases. Summary of the Invention

[0004] In view of this, the purpose of this invention is to provide a selenium-enriched functional bio-organic fertilizer, its preparation method, and its application, solving the technical problems of long composting cycles and limited product functions in existing bio-organic fertilizers. This invention selects dominant microbial strains with composting and antibacterial effects as fermentation agents and uses chicken manure from egg-laying chicken farms that produce selenium-enriched eggs as the main material to produce selenium-enriched functional organic fertilizer. This process not only results in rapid temperature rise and a long duration of high-temperature phase, leading to thorough composting, but also promotes plant growth, improves the nutritional value of fruits and vegetables, improves the soil microbial community structure, and effectively inhibits the occurrence of pathogens.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a selenium-enriched bio-organic fertilizer, comprising a substrate and a fermentation agent; the substrate comprises the following components by weight fraction: 300-450 parts selenium-enriched chicken manure, 75-150 parts mushroom sticks, 50-100 parts corn husks, and 25-50 parts humic acid; the amount of the fermentation agent is 1‰ to 3‰ of the substrate weight.

[0006] Preferably, the fermentation agent comprises Bacillus and Rhodopseudomonas; the mass ratio of Bacillus to Rhodopseudomonas is 3:2; The Bacillus species include Bacillus subtilis, Bacillus belye, or Bacillus amyloliquefaciens.

[0007] This invention also provides a method for preparing the aforementioned selenium-enriched bio-organic fertilizer, comprising the following steps: Selenium-enriched chicken manure, mushroom sticks, corn husks, and humic acid are mixed, inoculated with fermentation agents, piled up, fermented, decomposed, and aged to obtain the selenium-enriched bio-organic fertilizer.

[0008] Preferably, the height of the stack is 1.2~1.5m, the width of the stack is 1.5~3.0m, and the spacing between the stacks is 0.8~1.2m.

[0009] Preferably, during the heating and high-temperature stages of the fermentation process, the pile is turned over once every 2-3 days, and during the cooling stage when the pile temperature is below 50°C, the pile is turned over once every 7-10 days.

[0010] Preferably, the composting time is 20-30 days, and the aging time is 25-35 days.

[0011] This invention also provides the application of the aforementioned selenium-enriched bio-organic fertilizer in the prevention and control of plant pathogens.

[0012] Preferably, the plant pathogens include one or more of Fusarium oxysporum, Botrytis cinerea, Alternaria alternata, and Fusarium solani.

[0013] This invention also provides the application of the aforementioned selenium-enriched bio-organic fertilizer in vegetable cultivation.

[0014] Preferably, the vegetable includes chives.

[0015] Compared with the prior art, the present invention has the following beneficial effects: 1. The fermentation process is simple and easy to promote: The fermentation process of the selenium-enriched functional bio-organic fertilizer provided by this invention adopts windrow fermentation, which can realize the large-scale production of organic fertilizer with just a turning machine. The process is simple and easy to promote.

[0016] 2. Rapid temperature rise and thorough decomposition: The selected fermentation agent can raise the pile temperature to above 50℃ within 2-4 days, and the high temperature can reach above 70℃. The high temperature stage is maintained for more than 15 days, ensuring thorough decomposition and effectively killing insect eggs and pathogenic microorganisms in the material.

[0017] 3. Multifunctional properties including disease resistance and growth promotion: The selenium-enriched functional bio-organic fertilizer produced on a large scale according to this invention has a selenium content of 0.912 mg / kg and an effective live bacteria count of 2.6 × 10⁻⁶. 8 CFU / mL, with both growth-promoting and antibacterial effects.

[0018] 4. Broad application prospects: This selenium-enriched functional bio-organic fertilizer can not only prevent and control plant diseases, but also produce selenium-enriched agricultural products, achieving the effects of increasing yield and improving quality. Attached Figure Description

[0019] Figure 1 The inhibition rates of different Bacillus strains against plant pathogens in Example 1; Figure 2 Example 2 shows the changes in fermentation temperature for different material combinations; Figure 3 Example 3: Temperature variation during large-scale fermentation of selenium-enriched functional bio-organic fertilizer; Figure 4 Example 4 illustrates the inhibitory effect of selenium-enriched functional organic fertilizer dilution on plant pathogens; Figure 5 Example 5 illustrates the inhibitory effect of organic fertilizers fermented with different combinations of microbial agents on plant pathogens. Figure 6 This is the effect of selenium-enriched organic fertilizer on the growth promotion of chives in Example 5. Detailed Implementation

[0020] This invention provides a selenium-enriched bio-organic fertilizer, comprising a substrate and a fermentation agent; the substrate comprises the following components by weight fraction: 300-450 parts selenium-enriched chicken manure, 75-150 parts mushroom sticks, 50-100 parts corn husks, and 25-50 parts humic acid; the amount of the fermentation agent is 1‰ to 3‰ of the substrate weight.

[0021] In this invention, the preferred amount of selenium-enriched chicken manure is 400 parts; the preferred amount of the mushroom stick is 100 parts; the preferred amount of corn husk is 70 parts; the preferred amount of humic acid is 30 parts; the preferred amount of the fermentation agent is 2‰~2.5‰; the fermentation agent includes Bacillus and Rhodopseudomonas; the mass ratio of Bacillus to Rhodopseudomonas is 3:2; the Bacillus includes Bacillus subtilis, Bacillus belyceae, or Bacillus amyloliquefaciens.

[0022] This invention also provides a method for preparing the aforementioned selenium-enriched bio-organic fertilizer, comprising the following steps: Selenium-enriched chicken manure, mushroom sticks, corn husks, and humic acid are mixed, inoculated with fermentation agents, piled up, fermented, decomposed, and aged to obtain the selenium-enriched bio-organic fertilizer.

[0023] In this invention, the height of the stack is 1.2~1.5m, the width of the stack is 1.5~3.0m, and the spacing between the stacks is 0.8~1.2m; during the fermentation process, in the heating and high-temperature stages, the stack is turned over once every 2~3 days; in the cooling stage, when the stack temperature is below 50℃, the stack is turned over once every 7~10 days; the maturation time is 20~30 days, preferably 25 days; when the material turns dark brown, has no ammonia odor, and the temperature naturally drops to ambient temperature ±5℃, it indicates that the maturation is complete; the aging time is preferably 25~35 days, more preferably 30 days.

[0024] This invention also provides the application of the aforementioned selenium-enriched bio-organic fertilizer in the prevention and control of plant pathogens. In this invention, the plant pathogens include one or more of Fusarium oxysporum, Botrytis cinerea, Alternaria alternata, and Fusarium solani.

[0025] This invention also provides the application of the aforementioned selenium-enriched bio-organic fertilizer in vegetable cultivation. In this invention, the vegetable includes chives.

[0026] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0027] Example 1

[0028] Screening of antibacterial Bacillus

[0029] 1. Preparation of pathogens

[0030] Prepare PDA medium (1 L): 200.0 g peeled potatoes, 20.0 g glucose, 18-20 g agar, and distilled water to a final volume of 1 L. Sterilize at 115℃ for 20 min; cool to approximately 50℃, pour into 85 mm sterile petri dishes to make PDA plates, inoculate with plant pathogen mycelia, and incubate at 25℃ for 5 days until the mycelia have completely covered the plates. The pathogen used in the experiment was obtained from the Agricultural Microbial Germplasm Resource Bank of Tianjin Academy of Agricultural Sciences.

[0031] 2. Preparation of Bacillus fermentation broth

[0032] Prepare a liquid culture medium consisting of 1% peptone, 1% soluble starch, 0.3% sodium chloride, 0.1% dipotassium hydrogen sulfate, and 0.1% magnesium sulfate, bringing the volume to 1 L. Dispense 100 mL into 250 mL Erlenmeyer flasks and sterilize at 121°C for 20 min. Inoculate different Bacillus strains into the liquid culture medium cooled to 30°C and incubate at 30°C for 2 days. Determine the bacterial count when it reaches 10⁻⁶. 9 The probiotic fermentation broth can be obtained at CFU / mL.

[0033] 3. In vitro antibacterial test – Oxford cup method

[0034] Antagonistic bacteria were initially screened using the plate confrontation method. Holes were punched at the edge of the plant pathogen using a 6mm diameter punch, and the bacterial blocks were inoculated into the center of a PDA plate. Probiotic fermentation broth (Oxford cups) was symmetrically inoculated 2cm away from the pathogen. Petri dishes without probiotic fermentation broth served as controls. Each treatment had three replicates, and all were incubated in the dark at 25℃. When the diameter of the pathogen in the control group reached 3 / 4 of the petri dish diameter, the diameter of the pathogen colony was measured using the cross-sectional method.

[0035] Experimental results: such as Figure 1As shown. Of the nine Bacillus strains selected, eight showed significant inhibitory effects against the tested pathogens. Specifically, K8, S1, YC1, and HYS showed outstanding inhibitory effects against cucumber wilt; H4, K8, S1, and YC1 showed outstanding inhibitory effects against cucumber gray mold; S1, YC1, and HF showed outstanding inhibitory effects against tomato early blight; and S1, YC1, HYS, K8, SX, and HF showed outstanding inhibitory effects against tomato root rot. Finally, K8 was selected. Bacillus subtilis H4 Bacillus amyloliquefaciens ) and S1 ( Bacillus velezensis It is used as a fermentation Bacillus.

[0036] Example 2

[0037] Screening of optimal combinations of fermentation inoculants

[0038] The bacterial strains used in this invention include Bacillus K8 (see CN112011486A - A strain of Bacillus subtilis K8 that simultaneously nitrifies and denitrifies and its application), H4 (see CN101792723A - A strain of Bacillus amyloliquefaciens and its application in aquaculture), and S1 (see literature: Fengfeng Zhang, Fengxing Xie, Ke Zhou, et al. Nitrogen removal performance of novel isolated Bacillus sp. capable of simultaneous heterotrophic nitrification and aerobic denitrification. Applied Biochemistry and Biotechnology, 2022, 194: 3196–3211), and Rhodopseudomonas (photosynthetic bacteria PSB) (see literature: Zhang Fengfeng, Zhou Ke, Zhao Yujie, et al. Study on culture conditions of photosynthetic bacteria. Tianjin Agricultural Sciences, 22000099, 1155(2): 99-111.).

[0039] 1. Material ratio: Combination 1: 3 tons of selenium-enriched chicken manure, 1.5 tons of mushroom sticks, 1.0 ton of corn husks, and 0.5 tons of humic acid; Combination 2: 4 tons of selenium-enriched chicken manure, 1 ton of mushroom sticks, 0.7 tons of corn husks, and 0.3 tons of humic acid; Combination 3: 4.5 tons of selenium-enriched chicken manure, 0.75 tons of mushroom sticks, 0.5 tons of corn husks, and 0.25 tons of humic acid. 2. Inoculum formulation ratio: Combination 1: Bacillus subtilis: Rhodopseudomonas aeruginosa = 3:2; Combination 2: Bacillus belyssus: Rhodopseudomonas aeruginosa = 3:2; Combination 3: Bacillus amyloliquefaciens: Rhodopseudomonas aeruginosa = 3:2; 3. Add materials 1-3 in proportion and mix evenly. Add 15kg of fermentation inoculant for each combination (the inoculant amount is 2.5‰ of the material weight). The control group is the one without fermentation inoculant. There are a total of 10 treatments. Mix the materials and inoculant evenly. The combinations are shown in Table 1.

[0040] Table 1. Dosage of each treatment material and microbial agent

[0041] 4. Stack fermentation: The optimal combination of microbial agents is selected by analyzing the temperature changes during the fermentation process and the post-fermentation moisture, pH, organic matter, nutrients, selenium content, effective viable bacteria count, and heavy metal content.

[0042] Since the inoculation temperatures of different bacterial agent combinations using the same material combination are relatively close, the same material combination...

[0043] Three different inoculation treatments were combined into one treatment, and the average value was taken. Temperature changes of different material combinations were compared, and the results are as follows: Figure 2 As shown, the inoculation treatment had a higher temperature than the control and a faster temperature rise, reaching 55℃ 4 days earlier than the uninoculated control. The high temperature above 55℃ lasted for more than 15 days, while the control only lasted 11 days. Comparing the differences between the different treatments, it was found that material combination 2 had a higher temperature throughout the fermentation process, with the highest temperature reaching 72.3℃.

[0044] Table 2 Indicators for each treatment

[0045] Table 2 shows that the moisture content and pH value of the bacterial treatment were lower than those of the control; the organic matter content, nitrogen content, selenium content, and number of viable bacteria were higher than those of the control; the mortality rate of ascarids was 100%, and the number of fecal coliforms was significantly lower than that of the control, indicating that the bacterial treatment is beneficial to the decomposition of organic fertilizer and the improvement of product quality. Comparison of different bacterial inoculation combinations revealed that the bacterial agent combination 2 treatment had the highest number of viable bacteria.

[0046] Table 3 Heavy metal content of each treatment

[0047] As shown in Table 3, the heavy metal content of all treatments, including the control, was lower than the NY884-2012 standard.

[0048] Example 3

[0049] Large-scale production of selenium-enriched functional bio-organic fertilizer

[0050] Based on the results of Example 2, the selected material ratio is combination 2, and the selected microbial agent ratio is combination 2.

[0051] 1. Scale up combination 2 in Example 2 by a factor of 230. Mix 920 tons of selenium-enriched chicken manure, 230 tons of mushroom sticks, 161 tons of corn husks, and 69 tons of humic acid using a turner.

[0052] 2. Inoculate the fermentation agent of combination 2 at a rate of 2‰ (2.76 tons) and spray it onto the surface of the material with a high-pressure water gun. Turn the material over once with a turning machine to ensure that the agent and the material are evenly mixed.

[0053] 3. The materials with added microbial agents are stacked in the organic fertilizer fermentation workshop. The stacks are 3m wide, 1.5m high, and 75m long, with a 1.2m interval between the stacks, forming 4 stacks.

[0054] 4. Fermentation: During the heating and high-temperature stages, turn the pile once every 2-3 days. During the cooling stage, when the pile temperature is below 50℃, turn the pile once every 7-10 days. After 25 days of composting, when the material turns dark brown, has no ammonia odor, and the temperature naturally drops to within ±5℃ of the ambient temperature, it indicates that composting is complete. After aging for 30 days, you will obtain selenium-enriched functional bio-organic fertilizer.

[0055] Depend on Figure 3 It can be seen that, due to the large amount of fermentation material, it was divided into 4 piles. The temperature of the material in the large-scale fermentation reached above 55℃ on the 3rd and 4th days of stacking, and this high temperature persisted for more than 18 days, indicating that the material was relatively thoroughly decomposed. After decomposition, the effective viable bacteria count reached 2.6 × 10⁻⁶. 7 The concentration of CFU / mL and the selenium content reached 0.912 mg / kg.

[0056] Example 4

[0057] Selenium-enriched functional bio-organic fertilizer has antibacterial effects against plant pathogens.

[0058] 1. Add 1g of the decomposed selenium-enriched functional bio-organic fertilizer prepared in Example 3 to 3mL of sterile water and incubate at 30℃ for 1h to obtain organic fertilizer leachate. 2. A comparative experiment was conducted using the plate confrontation method to determine the antibacterial effect of selenium-enriched functional bio-organic fertilizer on plant pathogens.

[0059] Prepare PDA medium (1L): 200.0g peeled potato, 20.0g glucose, 18-20g agar, distilled water to a final volume of 1L, sterilize at 115℃ for 20min; cool to about 50℃, pour into 85mm sterile petri dishes to make PDA plates, inoculate with plant pathogen mycelia, incubate at 25℃ for 5 days, and use when the mycelia have covered the petri dishes.

[0060] Plate confrontation method: Using a 6mm diameter punch, holes were made at the edge of the plant pathogen plate. The inoculum was inoculated into the center of a PDA plate. Oxford cups were placed symmetrically 2cm away from the pathogen, and 0.1mL of organic fertilizer leachate was added. A petri dish without organic fertilizer leachate served as a control. Each treatment was repeated in triplicate and incubated in the dark at 25℃. When the diameter of the pathogen in the control group reached 3 / 4 of the petri dish diameter, the diameter of the pathogen colony was measured using the cross-sectional method.

[0061] Experimental results: Figure 4 It can be seen that selenium-enriched functional bio-organic fertilizer has a significant inhibitory effect on four pathogens: cucumber gray mold, cucumber wilt, tomato early blight and tomato root rot, with inhibition rates of 53.6%, 66.13%, 2.85% and 42.86%, respectively.

[0062] Example 5

[0063] The antibacterial effect of selenium-enriched functional bio-organic fertilizer on plant leaves

[0064] 1. Take 2g of the selenium-enriched functional bio-organic fertilizer prepared in Examples 2-1, 1-2, 1-3 and Example 3 respectively, add it to 20mL of sterile water, and incubate at 30℃ for 1h to obtain organic fertilizer leachate; 2. Soak the leaves in the extract for 1 hour and then air dry them. Use sterile water treatment as a control and make wounds on the leaves with a sterile needle.

[0065] 3. Use a punch to make holes in the pathogen plate, pick up the fungal cake and place it on the wound of the leaf; After being placed in a 25℃ incubator for 2 days, the size and length of leaf lesions were observed, and the inhibition effect was calculated using a formula.

[0066] Inhibition effect (%) = (Length of control lesion - Length of treated lesion) / (Length of control lesion - Diameter of mycelial cake) × 100

[0067] Experimental results: such as Figure 5 As shown, the selenium-enriched functional bio-organic fertilizer of the present invention has a good antibacterial effect on the inoculation of tomato early blight strain on detached leaves. The length of lesions on detached leaves after treatment with organic fertilizer leachate is significantly different from that of the control. The inhibitory effects relative to the control are 48.48%, 54.55%, 53.03%, and 40.9%, respectively. Among them, 1-2 in Example 2 have the strongest antibacterial effect.

[0068] Example 6

[0069] The growth-promoting effect of selenium-enriched functional bio-organic fertilizer on chives

[0070] 1. The selenium-enriched functional bio-organic fertilizer prepared in Example 3 was applied as a base fertilizer in holes at a rate of 1500 kg / mu; no fertilizer was applied to the control group.

[0071] 2. Chives are transplanted by root, with a plant spacing of 15cm and a row spacing of 30cm; 3. Routine management involves only watering appropriately, and no fertilization is required during this period.

[0072] Experimental results: Figure 5 It can be seen that the average plant height of the treatment with selenium-enriched functional bio-organic fertilizer reached over 30cm, while the plant height of the control was only about 16cm. The plant height of the treatment was significantly higher than that of the control, and the number of tillers was also significantly higher than that of the control. This indicates that the organic fertilizer of this invention has a significant promoting effect on the growth of chives.

[0073] As can be seen from the above embodiments, the present invention provides a selenium-enriched functional organic fertilizer, its preparation method, and its application. This method features a simple production process and low cost, and the produced organic fertilizer has multiple effects, including promoting growth, preventing disease, and improving soil structure. The promotion and application of this technology will have a positive impact on soil structure improvement, plant disease control, and the production of green vegetables.

[0074] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A selenium-enriched bio-organic fertilizer, characterized in that, It includes a substrate and a fermentation agent; the substrate comprises the following components by weight fraction: 300-450 parts selenium-enriched chicken manure, 75-150 parts mushroom sticks, 50-100 parts corn husks, and 25-50 parts humic acid; the amount of the fermentation agent is 1‰ to 3‰ of the substrate weight.

2. The selenium-enriched bio-organic fertilizer according to claim 1, characterized in that, The fermentation agent includes Bacillus and Rhodopseudomonas; the mass ratio of Bacillus to Rhodopseudomonas is 3:

2. The Bacillus species include Bacillus subtilis, Bacillus belye, or Bacillus amyloliquefaciens.

3. The method for preparing selenium-enriched bio-organic fertilizer according to claim 1 or 2, characterized in that, Includes the following steps: Selenium-enriched chicken manure, mushroom sticks, corn husks, and humic acid are mixed, inoculated with fermentation agents, piled up, fermented, decomposed, and aged to obtain the selenium-enriched bio-organic fertilizer.

4. The preparation method according to claim 3, characterized in that, The height of the stack is 1.2~1.5m, the width of the stack is 1.5~3.0m, and the spacing between the stacks is 0.8~1.2m.

5. The preparation method according to claim 3, characterized in that, During the fermentation process, in the heating and high-temperature stages, the pile is turned over once every 2-3 days; in the cooling stage, when the pile temperature is below 50℃, the pile is turned over once every 7-10 days.

6. The preparation method according to claim 3, characterized in that, The fermentation time is 20-30 days, and the aging time is 25-35 days.

7. The application of the selenium-enriched bio-organic fertilizer according to claim 1 or 2 in the prevention and control of plant pathogens.

8. The application according to claim 7, characterized in that, The plant pathogens include one or more of Fusarium oxysporum, Botrytis cinerea, Alternaria alternata, and Fusarium solani.

9. The application of the selenium-enriched bio-organic fertilizer as described in claim 1 or 2 in vegetable cultivation.

10. The application according to claim 9, characterized in that, The vegetables mentioned include chives.

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