Method for preparing fermentation preparation for biological fertilizer
By hydrolyzing corn cobs and fermenting them with Rhodotorula glutinis and Candida tropicalis, combined with Bacillus licheniformis, bio-fertilizers are prepared, solving the problems of high cost and unstable fertilizer effect of bio-fertilizers, and achieving efficient promotion of crop growth and disease control.
Patent Information
- Application Number
- CN202511045715.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-29
- Publication Date
- 2025-11-07
AI Technical Summary
Existing bio-fertilizers have high production costs, long fermentation cycles, and unstable fertilizer effects. The application technology of corn cobs in bio-fertilizers is insufficient, and traditional processing methods cause resource waste and environmental pollution.
Corn cob is crushed and hydrolyzed, then fermented using activated liquids of Rhodotorula glutinis and Candida tropicalis, combined with Bacillus licheniformis and Bacillus mucilaginosus to prepare a fermentation agent. This agent is then mixed with basic fertilizer to form a compound microbial agent, allowing for direct fermentation and avoiding the detoxification step.
It can improve crop growth, reduce diseases, inhibit the accumulation of cadmium, increase the content of the trace element selenium, promote crop growth, and enhance fertilizer efficiency.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of biological fertilizer, in particular to a method for preparing a fermentation preparation for biological fertilizer. BACKGROUND
[0002] The treatment of agricultural waste has been an important issue for global agricultural sustainable development. Among them, corn cob, as a byproduct after corn planting, has the characteristics of large yield, rich in cellulose, hemicellulose and lignin. However, the traditional treatment methods of corn cob (such as incineration, landfill, etc.) not only cause resource waste, but also cause serious pollution to the environment. In recent years, with the popularization of the concept of resource recycling, the conversion of corn cob into high value-added products has become a feasible solution.
[0003] Biological fertilizer, as a kind of green and environmentally friendly fertilizer product, has gradually been favored by the market because it can improve soil structure, increase soil fertility, reduce the use of chemical fertilizers, and reduce the incidence of plant diseases. The core of biological fertilizer is to use the metabolic action of microorganisms to decompose organic matter into nutrients that plants can absorb, while promoting the reproduction of beneficial microorganisms in the soil and inhibiting the growth of pathogenic bacteria. However, most of the biological fertilizers on the market currently have problems such as high preparation cost, long fermentation period, and unstable fertilizer efficiency, which limits their large-scale application.
[0004] At present, the technology of applying corn cob to biological fertilizer still needs to be developed. SUMMARY
[0005] (I) Technical problems solved
[0006] In view of the deficiencies of the prior art, the present application provides a method for preparing a fermentation preparation for biological fertilizer.
[0007] (II) Technical solutions
[0008] In order to achieve the above-mentioned purpose, the present application provides the following technical solutions: a method for preparing a fermentation preparation for biological fertilizer, comprising the following steps:
[0009] S1, after the corn cob is crushed, hydrolysis is carried out to obtain a hydrolysate;
[0010] S2, inoculating Rhodotorula glutinis activation liquid and Candida tropicalis activation liquid into the hydrolysate for fermentation to obtain a fermentation liquid;
[0011] S3, centrifuging the fermentation liquid, collecting the precipitate, and spray drying to obtain a fermentation preparation.
[0012] Further, the hydrolysis treatment comprises: mixing the corncob powder with an acid solution in a mass-volume ratio of 1:2-4 to obtain a mixture; and subjecting the mixture to acid hydrolysis treatment at a temperature of 120-140 DEG C for 0.5-2 hours to obtain a hydrolysis solution; the acid solution is 0.5-2% sulfuric acid and 2-4% phosphoric acid in volume fraction.
[0013] Further, the inoculation volume ratio of the Rhodotorula mucilaginosa activation solution and the Candida tropicalis activation solution is 1:2-4.
[0014] Further, a composite microbial agent comprises: a fermentation preparation, Bacillus licheniformis and Bacillus mycoides.
[0015] Further, the mass ratio of the fermentation preparation, Bacillus licheniformis and Bacillus mycoides is 3-5:1:2-3.
[0016] Further, a bio-fertilizer comprises: the composite microbial agent and a base fertilizer.
[0017] Further, the base fertilizer comprises: 12-15 parts of urea, 20-24 parts of phosphate fertilizer, 20-26 parts of potassium fertilizer, 1-5 parts of nitro-humic acid, 1-5 parts of ammonium nitrate and 1-6 parts of ferrous sulfate.
[0018] Further, the addition amount of the composite microbial agent is 1-3% of the base fertilizer.
[0019] (Three) beneficial technical effects
[0020] The present application utilizes Candida tropicalis and Rhodotorula mucilaginosa to ferment and culture the hydrolyzed corncob, and the obtained microbial bodies can effectively promote the growth of crops, increase the yield, reduce the disease, inhibit the enrichment of metal cadmium in crops, and simultaneously increase the content of trace element selenium in crops. Moreover, the hydrolysis solution is directly subjected to fermentation treatment without detoxification treatment, so that some active ingredients beneficial to the growth of crops can be avoided from being lost in the detoxification process.
[0021] The bio-fertilizer of the present application can be used for rice, wheat, tomato, potato, cucumber, corn and other crops to increase the yield of crops, inhibit the disease of crops, inhibit the enrichment of metal cadmium in crops, and simultaneously increase the content of trace element selenium in crops. DETAILED DESCRIPTION
[0022] The technical solutions in the embodiments of the present application will be clearly and completely described below in combination with the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.
[0023] The components of the seed culture medium formula of the present application are all commercially available unless otherwise specified.
[0024] Example 1
[0025] 1. 100 g of crushed corncob was mixed with 300 mL of acid solution (1% v / v sulfuric acid and 2% v / v phosphoric acid), and heated at 130°C for 1 h to obtain a hydrolysis liquor. The hydrolysis liquor was subjected to suction filtration, and after the first suction filtration, the filter residue was washed with 180 mL of water for secondary suction filtration. The two filtrates were combined to obtain a corncob hydrolysate.
[0026] 2. Candida tropicalis was cultured in a seed culture medium (3% w / v malt extract powder) for 24 h to obtain a Candida tropicalis activation solution.
[0027] Rhodotorula mucilaginosa was cultured in a seed culture medium (glucose: 15 g / L, (NH4)2SO4: 2 g / L, yeast powder: 1 g / L, KH2PO4: 7 g / L, Na2HPO4: 5 g / L, MgSO4: 1.5 g / L) for 24 h to obtain a Rhodotorula mucilaginosa activation solution.
[0028] 3. The Rhodotorula mucilaginosa activation solution and the Candida tropicalis activation solution were inoculated into the corncob hydrolysate at a volume ratio of 1:3 and a total inoculation amount of 4%, and cultured at 160 rpm and 30°C for 24 h to obtain a fermentation product. The fermentation product was centrifuged at 5000 rpm for 5 min, and the precipitate was collected, washed with deionized water, and spray dried to obtain a fermentation preparation.
[0029] 4. The fermentation preparation, Bacillus licheniformis, and Bacillus mycoides were mixed at a mass ratio of 4:1:2 to obtain a composite microbial agent.
[0030] 5. The composite microbial agent was mixed with a basic fertilizer (urea 21 kg, superphosphate 24 kg, potassium sulfate 25 kg, nitro humic acid 2 kg, ammonium nitrate 5 kg, ferrous sulfate 6 kg) at a proportion of 2% to obtain a bio-fertilizer.
[0031] Comparative Example 1: Rhodotorula mucilaginosa as a fermentation strain
[0032] 1. 100 g of crushed corncob was mixed with 300 mL of acid solution (1% v / v sulfuric acid and 2% v / v phosphoric acid), and heated at 130°C for 1 h to obtain a hydrolysis liquor. The hydrolysis liquor was subjected to suction filtration, and after the first suction filtration, the filter residue was washed with 180 mL of water for secondary suction filtration. The two filtrates were combined to obtain a corncob hydrolysate.
[0033] 2. Rhodotorula mucilaginosa was cultured in a seed medium (glucose: 15 g / L, (NH4)2SO4: 2 g / L, yeast powder: 1 g / L, KH2PO4: 7 g / L, Na2HPO4: 5 g / L, MgSO4: 1.5 g / L) for 24 h to obtain a Rhodotorula mucilaginosa activation solution.
[0034] 3. The Rhodotorula mucilaginosa activation solution was inoculated into the corncob hydrolysate at an inoculation amount of 4%, and cultured at 160 rpm and 30°C for 24 h to obtain a fermentation product. The fermentation product was centrifuged at 5000 rpm for 5 min, and the precipitate was collected, washed with deionized water, and spray-dried to obtain a fermentation preparation.
[0035] 4. The fermentation preparation, Bacillus licheniformis, and Bacillus mycoides were mixed at a mass ratio of 4:1:2 to obtain a composite microbial agent.
[0036] 5. The composite microbial agent was mixed with a base fertilizer (urea 21 kg, superphosphate 24 kg, potassium sulfate 25 kg, nitro humic acid 2 kg, ammonium nitrate 5 kg, ferrous sulfate 6 kg) at a proportion of 2% to obtain a biofertilizer.
[0037] Comparative Example 2: Candida tropicalis as a fermentation strain
[0038] 1. 100 g of crushed corncob was mixed with 300 mL of acid solution (1% v / v sulfuric acid and 2% v / v phosphoric acid), and heated at 130°C for 1 h to obtain a hydrolysis solution. The hydrolysis solution was suction-filtered, and after the first suction filtration, the filter residue was washed with 180 mL of water, and the second suction filtration was performed. The two filtrates were combined to obtain a corncob hydrolysate.
[0039] 2. Candida tropicalis was cultured in a seed medium (3% w / v malt extract powder) for 24 h to obtain a Candida tropicalis activation solution.
[0040] 3. The Candida tropicalis activation solution was inoculated into the corncob hydrolysate at an inoculation amount of 4%, and cultured at 160 rpm and 30°C for 24 h to obtain a fermentation product. The fermentation product was centrifuged at 5000 rpm for 5 min, and the precipitate was collected, washed with deionized water, and spray-dried to obtain a fermentation preparation.
[0041] 4. The fermentation preparation, Bacillus licheniformis, and Bacillus mycoides were mixed at a mass ratio of 4:1:2 to obtain a composite microbial agent.
[0042] 5. The composite microbial agent was mixed with a base fertilizer (urea 21 kg, superphosphate 24 kg, potassium sulfate 25 kg, nitro humic acid 2 kg, ammonium nitrate 5 kg, ferrous sulfate 6 kg) at a proportion of 2% to obtain a biofertilizer.
[0043] Comparative Example 3: addition of an activated carbon detoxification step
[0044] 1. 100 g of crushed corncob was mixed with 300 mL of acid solution (1% v / v sulfuric acid and 2% v / v phosphoric acid), and heated at 130°C for 1 h to obtain a hydrolysis liquor. The hydrolysis liquor was filtered, and after the first filtration, the residue was washed with 180 mL of water, and the second filtration was performed. The two filtrates were combined to obtain the corncob hydrolysis liquor.
[0045] 2. The pH of the corncob hydrolysis liquor was adjusted to 4.0, and then 1% activated carbon was added. After stirring at 30°C for 1 h, the mixture was filtered, and the supernatant was collected to obtain a detoxified hydrolysis liquor.
[0046] 3. Candida tropicalis was cultured in a seed medium (3% w / v malt extract powder) for 24 h to obtain a Candida tropicalis activation solution.
[0047] Rhodotorula mucilaginosa was cultured in a seed medium (glucose: 15 g / L, (NH4)2SO4: 2 g / L, yeast powder: 1 g / L, KH2PO4: 7 g / L, Na2HPO4: 5 g / L, MgSO4: 1.5 g / L) for 24 h to obtain a Rhodotorula mucilaginosa activation solution.
[0048] 4. The Rhodotorula mucilaginosa activation solution and the Candida tropicalis activation solution were inoculated into the detoxified hydrolysis liquor at a volume ratio of 1:3 and a total inoculation amount of 4%, and cultured at 160 rpm and 30°C for 24 h to obtain a fermentation product. The fermentation product was centrifuged at 5000 rpm for 5 min, and the precipitate was collected, washed with deionized water, and spray-dried to obtain a fermentation preparation.
[0049] 5. The fermentation preparation, Bacillus licheniformis, and Bacillus mycoides were mixed at a mass ratio of 4:1:2 to obtain a composite microbial agent.
[0050] 6. The composite microbial agent was mixed with a base fertilizer (urea 21 kg, superphosphate 24 kg, potassium sulfate 25 kg, nitro humic acid 2 kg, ammonium nitrate 5 kg, ferrous sulfate 6 kg) at a proportion of 2% to obtain a bio-fertilizer.
[0051] Comparative Example 4
[0052] 1. 100 g of crushed corncob was mixed with 300 mL of acid solution (1% v / v sulfuric acid and 2% v / v phosphoric acid), and heated at 130°C for 1 h to obtain a hydrolysis liquor. The hydrolysis liquor was filtered, and after the first filtration, the residue was washed with 180 mL of water, and the second filtration was performed. The two filtrates were combined to obtain the corncob hydrolysis liquor.
[0053] 2. Candida tropicalis was cultured in a seed medium (3% w / v malt extract powder) for 24 h to obtain a Candida tropicalis activation solution.
[0054] Rhodotorula mucilaginosa was cultured in a seed medium (glucose: 15 g / L, (NH4)2SO4: 2 g / L, yeast powder: 1 g / L, KH2PO4: 7 g / L, Na2HPO4: 5 g / L, MgSO4: 1.5 g / L) for 24 h to obtain a Rhodotorula mucilaginosa activation solution.
[0055] 3. The Rhodotorula mucilaginosa activation solution and Candida tropicalis activation solution were inoculated into the corncob hydrolysate at a total inoculation amount of 4% and a volume ratio of 1:3, and cultured at 160 rpm and 30°C for 24 h to obtain a fermentation product. The fermentation product was centrifuged at 5000 rpm for 5 min, and the precipitate was collected, washed with deionized water, and spray-dried to obtain a fermentation preparation.
[0056] 4. The fermentation preparation, Bacillus cereus, and Bacillus mycoides were mixed at a mass ratio of 4:1:2 to obtain a composite microbial agent.
[0057] 5. The composite microbial agent was mixed with a base fertilizer (urea 21 kg, superphosphate 24 kg, potassium sulfate 25 kg, nitro humic acid 2 kg, ammonium nitrate 5 kg, ferrous sulfate 6 kg) at a proportion of 2% to obtain a bio-fertilizer.
[0058] Comparative Example 5
[0059] 1. 100 g of crushed corncob was mixed with 300 mL of acid solution (1% v / v sulfuric acid and 2% v / v phosphoric acid), and heated at 130°C for 1 h to obtain a hydrolysis solution. The hydrolysis solution was suction-filtered, and after the first suction filtration, the filter residue was washed with 180 mL of water, and the second suction filtration was performed. The two filtrates were combined to obtain a corncob hydrolysate.
[0060] 2. Candida tropicalis was cultured in a seed medium (3% w / v malt extract powder) for 24 h to obtain a Candida tropicalis activation solution.
[0061] Rhodotorula mucilaginosa was cultured in a seed medium (glucose: 15 g / L, (NH4)2SO4: 2 g / L, yeast powder: 1 g / L, KH2PO4: 7 g / L, Na2HPO4: 5 g / L, MgSO4: 1.5 g / L) for 24 h to obtain a Rhodotorula mucilaginosa activation solution.
[0062] 3. The Rhodotorula mucilaginosa activation solution and Candida tropicalis activation solution were inoculated into the corncob hydrolysate at a total inoculation amount of 4% and a volume ratio of 1:3, and cultured at 160 rpm and 30°C for 24 h to obtain a fermentation product. The fermentation product was centrifuged at 5000 rpm for 5 min, and the precipitate was collected, washed with deionized water, and spray-dried to obtain a fermentation preparation.
[0063] 4. The fermentation preparation, Bacillus licheniformis, and Bacillus amyloliquefaciens were mixed at a mass ratio of 4:1:2 to obtain a composite microbial agent.
[0064] 5. The complex microbial agent is mixed with the base fertilizer (urea 21 kg, superphosphate 24 kg, potassium sulfate 25 kg, nitro humic acid 2 kg, ammonium nitrate 5 kg, ferrous sulfate 6 kg) at a ratio of 2% to obtain a biological fertilizer.
[0065] Performance test:
[0066] 1. The growth promoting effect of the complex microbial agent prepared in Example 1 and Comparative Examples 1-3 on cucumbers was studied, and the specific experiment was as follows:
[0067] The complex microbial agent was diluted with sterile normal saline to 1 x 10 5 CFU / mL to prepare a complex microbial agent liquid. A sterile filter paper was placed at the bottom of a sterile culture dish, and 5 cucumber seeds were evenly placed in each culture dish. The following liquids were added to the culture dishes:
[0068] Treatment group: 2 mL of diluted complex microbial agent liquid with a concentration of 1 x 10 5 CFU / mL was added.
[0069] Control group: 2 mL of sterile water was added.
[0070] Three repeated culture dishes were set up for each treatment group and control group.
[0071] All culture dishes were placed in a 30°C constant temperature incubator, kept in a dark environment, and cultured for 5 days. After 5 days, the culture dishes were transferred to a light incubator, set to a temperature of 25°C, a light intensity of 3000 lx, and a light cycle of 16 h light / 8 h dark, and continued to be cultured for 10 days.
[0072] Seed germination: record the germination rate (germination rate = number of germinated seeds / total number of seeds x 100%) and observe the germination time (time from start of culture to seed emergence).
[0073] Seedling growth indicators: measure seedling height (distance from root to top of seedling), root length (main root length), root fresh weight, aboveground fresh weight, and other indicators.
[0074] Table 1 Biological fertilizer promoting growth test results on cucumbers
[0075] Treatment group Average taproot length (cm) Average shoot length (cm) Average fresh weight (g) Example 1 8.94 7.2 0.34 Comparative Example 1 7.47 6.01 0.21 Comparative Example 2 7.21 6.15 0.24 Comparative Example 3 6.89 5.31 0.20
[0076] The results are shown in Table 1. Compared with Comparative Examples 1-3, the biological fertilizer of Example 1 has better growth promoting effect on cucumbers, indicating that the synergistic effect of Candida tropicalis and Rhodotorula mucilaginosa can be used to ferment suitable substances for crop growth. Although activated carbon detoxification can remove some inhibitory substances that are not conducive to the growth of yeast, the use of Candida tropicalis and Rhodotorula mucilaginosa for co-fermentation can improve the tolerance of the bacteria to inhibitory substances, and the amount of bacteria obtained by fermentation of the hydrolysate after activated carbon detoxification is close. Moreover, the two types of yeast can be fermented in non-detoxified hydrolysate to produce substances that promote crop growth, which is beneficial to increasing crop yield.
[0077] 2. Tomato was planted in a test field in Xinxiang City, Henan Province, and the soil was prone to tomato collar and root rot. The biological fertilizers prepared in Example 1 and Comparative Examples 1-5 were applied to the tomato seedlings after 30 days of growth, with 100 tomato seedlings in each experimental group, and the fertilizer application rate was 100 kg / acre. The control group did not apply fertilizer, and the watering frequency was once every 5 days.
[0078] The incidence rate and disease prevention rate of tomato collar and root rot were calculated according to the following formula:
[0079] Incidence rate = number of diseased plants in each group / total number of plants in each group x 100%.
[0080] Disease prevention rate = (control group incidence rate - experimental group incidence rate) / control group incidence rate x 100%.
[0081] Table 2 Incidence rate and disease prevention rate of tomato collar and root rot
[0082] Incidence of disease (%) Disease control (%) Example 1 5 91 Comparative Example 1 20 79 Comparative Example 2 18 82 Comparative Example 3 25 70 Comparative Example 4 16 82 Comparative Example 5 23 77 Control group 52 /
[0083] The results are shown in the above table. As can be seen from Example 1 and Comparative Examples 1-3, the use of Candida tropicalis and Rhodotorula mucilaginosa in combination with Bacillus licheniformis and Bacillus mycoides can effectively prevent and control tomato collar and root rot. As can be seen from Example 1, Comparative Example 4 and Comparative Example 5, the use of the fermentation preparation of the present application in combination with Bacillus licheniformis and Bacillus mycoides has better effect on preventing and controlling tomato collar and root rot.
[0084] 3. Rice Twoyou 8106 was planted in a rice planting field in Linyi City, Shandong Province, and the soil was cadmium-contaminated soil with a cadmium content of 0.37-0.40 mg / kg. The biological fertilizers prepared in Example 1 and Comparative Examples 1-5 were applied to the soil, and the fertilizer application rate was 50 kg / acre. Ten kg of urea per mu was applied as topdressing at the jointing stage. After the rice matured, the grains were harvested and the yield was calculated.
[0085] Table 3 Rice yield
[0086]
[0087]
[0088] Table 4 Selenium and cadmium content in rice
[0089] Selenium content (mg / kg) Cadmium content (mg / kg) Example 1 0.45 0.07 Comparative Example 1 0.32 0.25 Comparative Example 2 0.38 0.38 Comparative Example 3 0.28 0.22 Comparative Example 4 0.35 0.77 Comparative Example 5 0.27 0.18
[0090] The results are shown in Tables 3 and 4, and it can be seen that the fermentation preparation of the application, combined with Bacillus licheniformis and Bacillus gellan, can effectively increase the yield of rice, increase the selenium content, and inhibit the enrichment of cadmium.
[0091] Although embodiments of the present application have been shown and described, it is to be understood that various modifications, substitutions, replacements and changes can be made to these embodiments without departing from the principles and spirit of the present application, and the scope of the present application is defined by the appended claims and their equivalents.
Claims
1. A method of preparing a fermentation preparation for use in a biofertilizer, characterized by, The method comprises the following steps: S1, corn cob is crushed and then hydrolyzed to obtain a hydrolysis liquor; S2, the hydrolysis liquor is inoculated with Rhodotorula mucilaginosa activation liquid and Candida tropicalis activation liquid to perform fermentation, and a fermentation liquor is obtained; S3, the fermentation liquor is centrifuged, and the precipitate is collected and spray-dried to obtain a fermentation preparation.
2. The method of claim 1, wherein the fermentation preparation for the biofertilizer is prepared by the steps of, The hydrolysis treatment comprises: mixing corn cob powder with an acid solution in a mass-volume ratio of 1:2-4 to obtain a mixture; and performing acid hydrolysis treatment on the mixture at a temperature of 120-140℃ for 0.5-2h to obtain the hydrolysis liquor; the acid solution is 0.5-2% sulfuric acid and 2-4% phosphoric acid.
3. The method of claim 1, wherein the fermentation preparation for the biofertilizer is prepared by the steps of, The inoculation amount of the Rhodotorula mucilaginosa activation liquid and the Candida tropicalis activation liquid is in a volume ratio of 1:2-4.
4. A complex microbial agent, characterized in that, The method comprises: The fermentation preparation, Bacillus licheniformis, and Bacillus mycoides.
5. The complex microbial agent according to claim 4, characterized in that, The mass ratio of the fermentation preparation, Bacillus licheniformis, and Bacillus mycoides is 3-5:1:2-3.
6. A biofertilizer characterized in that, The method comprises: The composite microbial agent and the base fertilizer.
7. The biofertilizer according to claim 6, characterized in that, The base fertilizer comprises 12-15 parts of urea, 20-24 parts of phosphate fertilizer, 20-26 parts of potassium fertilizer, 1-5 parts of nitro humic acid, 1-5 parts of ammonium nitrate, and 1-6 parts of ferrous sulfate.
8. The biofertilizer according to claim 6, characterized in that, The addition amount of the composite microbial agent is 1-3% of the base fertilizer.