Method for improving soil structure using Brevibacillus laterus
Microbial organic fertilizer prepared by Bacillus brevis, combined with iron-based soybean peptone, solved the shortcomings of soil structure improvement and pest reduction in the existing technology, achieved the effect of soil structure improvement and pest control, and at the same time reduced pesticide residues and atmosphere Risk of ozone layer destruction.
Patent Information
- Application Number
- CN202411354657.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-27
- Publication Date
- 2024-12-27
- Estimated Expiration
- 2044-09-27
AI Technical Summary
The prior art has shortcomings in the use of microbial organic fertilizer to improve soil structure and reduce pests and diseases, and may lead to soil microbial ecological imbalance and atmospheric ozone layer damage.
Bacillus lateral breach was used to improve soil structure. Bacillus lateral breach was inoculated into culture medium to form spores and adsorb them into the matrix to prepare microbial organic fertilizer, and apply it to the soil, combined with the use of iron-based soybean peptone To improve nutritional value and bacterial absorption efficiency.
It significantly improves crop yield, improves soil structure, reduces pests and diseases, reduces pesticide residues, improves fertilizer utilization, and increases the fresh weight of tomatoes per plant and the effect of preventing and treating diseases through iron-based soybean peptone.
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field of microbial pesticides, in particular to a method for improving soil structure by utilizing Brevibacillus laterus. Background Art
[0002] Bacillus brevis usually contains one or more paraspores and can produce a variety of metabolites. Its spore preparations are widely used. In the agricultural field, they are usually used to produce nematicides and microbial fertilizers to prevent and control the occurrence and development of plant diseases and insect pests.
[0003] The Chinese patent with the authorization announcement number CN100579940C provides a method for preparing a biological organic fertilizer for preventing and controlling nematode damage and its use. The method for preparing a biological organic fertilizer for preventing and controlling nematode damage is carried out according to the following steps: ①, the stage of configuring biological organic materials; ②, the stage of high-temperature fermentation and composting of biological organic materials; ③, the stage of adding the pseudo-Paecilomyces lilacinus agent to expand the culture; ④, the secondary fermentation stage; ⑤, granulation; ⑥, quality inspection; ⑦, packaging and warehousing.
[0004] The Chinese patent with the authorization announcement number CN101456761B discloses a method for preparing microbial organic fertilizer for preventing and controlling crop wilt. The current method is likely to cause ecological imbalance of soil microorganisms and damage to the atmospheric ozone layer. The invention method is specifically as follows: first, a liquid fermentation culture method is used to prepare a biocontrol bacteria seed liquid, and then the hydrolyzate of domestic waste is used as a matrix, the biocontrol bacteria seed liquid is added to the matrix, and the matrix is fully mixed and fermented for 5 to 7 days; pig manure is mixed with filler, decomposed pig manure compost, and bamboo vinegar stock solution, and a pig manure compost product is obtained by static forced ventilation. The fermented matrix and the pig manure compost product are fully mixed in a volume ratio of 1:3 to 5 to prepare a biological organic fertilizer containing biocontrol bacteria.
[0005] The Chinese patent with the authorization announcement number CN102173952B discloses an organic fertilizer for preventing continuous cropping of tomatoes, and its raw materials include the following components and weight ratios: 20-30% of earthworm castings, 25-35% of crushed corn stalks, 25-35% of rice bran, and 10-15% of vermiculite. The preparation method is to weigh earthworm castings, crushed corn stalks, rice bran, and vermiculite respectively according to the weight ratio; fully mix the crushed corn stalks, rice bran, and vermiculite, and wet them with water to make the moisture content of the compost material 50-60%; then pile the mixed materials into a narrow upper and wide lower trapezoidal pile in a greenhouse or plastic greenhouse, and carry out natural aerobic fermentation or fermentation with fermentation bacteria; and fully mix the fermented corn stalks, rice bran, and vermiculite mixture with earthworm castings.
[0006] The organic fertilizers prepared by the above patents and prior art are still insufficient in improving soil structure, alleviating pests and diseases, and increasing crop yields. Summary of the invention
[0007] In order to solve the above problems, the present invention provides a method for improving soil structure using Brevibacillus laterus, and the operating steps are as follows:
[0008] S1: inoculate Brevibacillus laterus X10 into 80-100 mL of culture medium to obtain strains;
[0009] S2: Inoculate 1-2% of the inoculum into 200-300 mL of liquid nutrient broth medium and perform spore staining every 2 hours to check spore formation.
[0010] S3: When the number of bacteria reaches 2×10 8 cfu / mL, when the spore formation rate reaches 80%, it is adsorbed into the matrix at a ratio of 30-35% to obtain microbial organic fertilizer;
[0011] S4: Apply microbial organic fertilizer in the hole, mix it with the soil, place the tomato seedlings in the hole, cover with soil, and water.
[0012] Furthermore, the culture temperature of S1 is 30-35°C and the culture time is 24-48h.
[0013] Furthermore, the preparation method of the medium of S1 is:
[0014] Add 5-10 parts of peptone, 2-5 parts of yeast extract, 5-10 parts of NaCl, and 0.6-1.3 parts of agar powder to 800-1000 parts of water, pH 7, and sterilize at 100-120℃ for 20-30 minutes.
[0015] Furthermore, the culture temperature of S2 is 30-35°C and the culture time is 48-72h.
[0016] Furthermore, the preparation method of the liquid nutrient broth medium of S2 is:
[0017] Add 1-4 parts of beef, 3-6 parts of iron-based soy peptone, 2-5 parts of glucose, 2-5 parts of NaCl, and 10-15 parts of agar powder to 800-1000 parts of water, pH is 7, and sterilize at 100-120℃ for 20-30 minutes.
[0018] Furthermore, the preparation method of the iron-based soy peptone is:
[0019] Before the reaction begins, 24-48 parts of soy peptone (CAS: 91079-46-8) are added to a stirring kettle containing 500-1000 parts of water, and then 0.01-0.1 parts of ferric acrylate, 0.04-0.5 parts of rose oxide, and 1-3 parts of sodium bicarbonate are added. After stirring for reaction, water is removed by distillation to obtain iron-based soy peptone.
[0020] Furthermore, the stirring rate is 300-500 rpm.
[0021] Furthermore, the stirring reaction time is 100-150 min.
[0022] Furthermore, the matrix of S3 includes: 40-50 parts of chicken manure, 10-20 parts of bran, 5-10 parts of soybean cake powder, 5-10 parts of peat, 5-10 parts of vermiculite, 5-10 parts of (NH4)2SO4, 1-5 parts of KH2PO4, and 0.1-1 parts of K2SO4.
[0023] Furthermore, in S4, 10-20 portions of microbial organic fertilizer are applied to each tomato seedling.
[0024] Reaction mechanism: NH2 in soy peptone undergoes amino-propylene addition reaction with ferric acrylate, and further undergoes the same reaction with the propylene group of rose ether, finally forming iron-based soy peptone; in this process, the iron atom acts as a coordination center and forms a coordination bond with the nitrogen atom of the amino acid residue in soy peptone, increasing the stability and functionality of the protein; the formation of this structure lays the foundation for the subsequent biological activity.
[0025] From a chemical point of view, iron complexes and tetrahydropyran functional groups play multiple roles in the culture medium. Iron is a trace element necessary for the growth of many bacteria, and its presence in soy peptone improves the nutritional value of soy peptone and promotes the efficiency of absorption by Bacillus laterus. At the same time, the tetrahydropyran functional group, as a stable ring structure, can provide additional hydrogen bonding sites, enhance the interaction with the surface molecules of the bacteria, and thus promote the absorption and utilization of nutrients.
[0026] Compared with the prior art, the method of improving soil structure by using Brevibacillus laterus of the present invention has the following significant effects:
[0027] 1. The microbial organic fertilizer prepared by the present invention can promote the growth of plant roots and enhance the absorption capacity of the roots, thereby increasing crop yields, because the Brevibacillus laterosporus therein can colonize the root system of plants;
[0028] 2. The microbial organic fertilizer prepared by the present invention can improve soil structure, improve loose soil, solve the soil compaction phenomenon, thereby activating the soil and improving fertilizer utilization;
[0029] 3. The microbial organic fertilizer prepared by the present invention can inhibit the reproduction of pathogens inside and outside the plant, reduce pests and diseases, and reduce pesticide residues. It is a green and environmentally friendly organic fertilizer;
[0030] 4. The culture medium is added with iron-based soy peptone. The addition of iron complexes improves the quality of the nutritious gravy and can effectively increase the fresh weight of a single tomato plant. DETAILED DESCRIPTION
[0031] In order to further explain the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the following is a detailed description in combination with embodiments and comparative examples:
[0032] 1. After the tomatoes are harvested, weigh the fresh weight of each plant;
[0033] 2. 30 days after transplanting tomatoes, check the incidence of tomato bacterial wilt and calculate the control effect: control effect = (number of tomatoes without bacterial wilt / total number of tomatoes) × 100%.
[0034] Example 1: A method for improving soil structure using Brevibacillus laterus, the operating steps are as follows:
[0035] S1: Inoculate Brevibacillus laterus X10 into 80 mL of culture medium to obtain strains;
[0036] S2: Inoculate 1% of the culture medium into 200 mL of liquid nutrient broth and perform spore staining every 2 hours to check spore formation.
[0037] S3: When the number of bacteria reaches 2×10 8 cfu / mL, when the spore formation rate reaches 80%, 30% is adsorbed into the matrix to obtain microbial organic fertilizer;
[0038] S4: Apply microbial organic fertilizer in the hole, mix it with the soil, place the tomato seedlings in the hole, cover with soil, and water.
[0039] The culture temperature of S1 is 30°C and the culture time is 24h.
[0040] The preparation method of the S1 culture medium is:
[0041] Add 5 g of peptone, 2 g of yeast extract, 5 g of NaCl, and 0.6 g of agar powder to 800 g of water, pH 7, and sterilize at 100°C for 20 min.
[0042] The culture temperature of S2 is 30°C and the culture time is 48 hours.
[0043] The preparation method of the liquid nutrient broth medium of S2 is as follows:
[0044] Add 1g beef, 3g iron-based soy peptone, 2g glucose, 2g NaCl, and 10g agar powder to 800g water, pH 7, and sterilize at 100℃ for 20min.
[0045] The preparation method of the iron-based soy peptone is:
[0046] Before the reaction begins, 24 g of soy peptone (CAS: 91079-46-8) is added to a stirring kettle containing 500 g of water, and then 0.01 g of ferric acrylate, 0.04 g of rose ether, and 1 g of sodium bicarbonate are added. After stirring for reaction, water is distilled off to obtain iron-based soy peptone.
[0047] The stirring rate is 300 rpm.
[0048] The reaction time is 100 min.
[0049] The matrix of S3 includes: 40g chicken manure, 10g bran, 5g soybean cake powder, 5g peat, 5g vermiculite, 5g (NH4)2SO4, 1gKH2PO4, and 0.1gK2SO4.
[0050] In the S4, 10 g of microbial organic fertilizer was applied to each tomato seedling.
[0051] Example 2: A method for improving soil structure using Brevibacillus laterus, wherein the operating steps are as follows:
[0052] S1: Inoculate Brevibacillus laterus X10 into 85 mL of culture medium to obtain strains;
[0053] S2: Inoculate 1.5% of the culture medium into 240 mL of liquid nutrient broth and perform spore staining every 2 hours to check spore formation.
[0054] S3: When the number of bacteria reaches 2×10 8 cfu / mL, when the spore formation rate reaches 80%, it is adsorbed into the matrix at a ratio of 32% to obtain microbial organic fertilizer;
[0055] S4: Apply microbial organic fertilizer in the hole, mix it with the soil, place the tomato seedlings in the hole, cover with soil, and water.
[0056] The culture temperature of S1 is 32° C. and the culture time is 32 h.
[0057] The preparation method of the S1 culture medium is:
[0058] Add 6 g of peptone, 3 g of yeast extract, 6 g of NaCl, and 0.8 g of agar powder to 850 g of water, pH 7, and sterilize at 105°C for 25 min.
[0059] The culture temperature of S2 is 32° C. and the culture time is 58 h.
[0060] The preparation method of the liquid nutrient broth medium of S2 is as follows:
[0061] Add 2g beef, 4g iron-based soy peptone, 3g glucose, 3g NaCl, and 12g agar powder to 850g water, pH 7, and sterilize at 105°C for 25min.
[0062] The preparation method of the iron-based soy peptone is:
[0063] Before the reaction begins, 32 g of soy peptone (CAS: 91079-46-8) is added to a stirring kettle containing 600 g of water, and then 0.05 g of ferric acrylate, 0.2 g of rose ether, and 2 g of sodium bicarbonate are added. After stirring for reaction, water is distilled off to obtain iron-based soy peptone.
[0064] The stirring rate is 350 rpm.
[0065] The reaction time is 110 min.
[0066] The matrix of S3 includes: 43g chicken manure, 13g bran, 6g soybean cake powder, 6g peat, 6g vermiculite, 6g (NH4)2SO4, 2gKH2PO4, and 0.5gK2SO4.
[0067] In the S4, 13 g of microbial organic fertilizer was applied to each tomato seedling.
[0068] Example 3: A method for improving soil structure using Brevibacillus laterus, the operating steps are as follows:
[0069] S1: Inoculate Brevibacillus laterus X10 into 95 mL of culture medium to obtain strains;
[0070] S2: Inoculate 1.5% of the culture medium into 280 mL of liquid nutrient broth and perform spore staining every 2 hours to check spore formation.
[0071] S3: When the number of bacteria reaches 2×10 8 cfu / mL, when the spore formation rate reaches 80%, 34% is adsorbed into the matrix to obtain microbial organic fertilizer;
[0072] S4: Apply microbial organic fertilizer in the hole, mix it with the soil, place the tomato seedlings in the hole, cover with soil, and water.
[0073] The culture temperature of S1 is 34° C. and the culture time is 42 h.
[0074] The preparation method of the S1 culture medium is:
[0075] Add 9 g of peptone, 4 g of yeast extract, 9 g of NaCl, and 1 g of agar powder to 950 g of water, pH 7, and sterilize at 115°C for 25 min.
[0076] The culture temperature of S2 is 34°C and the culture time is 68h.
[0077] The preparation method of the liquid nutrient broth medium of S2 is as follows:
[0078] Add 3g beef, 5g iron-based soy peptone, 4g glucose, 4g NaCl, and 14g agar powder to 950g water, pH 7, and sterilize at 115°C for 25min.
[0079] The preparation method of the iron-based soy peptone is:
[0080] Before the reaction begins, 42 g of soy peptone (CAS: 91079-46-8) is added to a stirring kettle containing 900 g of water, and then 0.08 g of ferric acrylate, 0.4 g of rose ether, and 2 g of sodium bicarbonate are added. After stirring for reaction, water is distilled off to obtain iron-based soy peptone.
[0081] The stirring rate is 450 rpm.
[0082] The reaction time is 140 min.
[0083] The matrix of S3 includes: 48g chicken manure, 18g bran, 9g soybean cake powder, 9g peat, 9g vermiculite, 9g (NH4)2SO4, 4gKH2PO4, and 0.8gK2SO4.
[0084] In the S4, 18 g of microbial organic fertilizer was applied to each tomato seedling.
[0085] Example 4: A method for improving soil structure using Brevibacillus laterus, the operating steps are as follows:
[0086] S1: inoculate Brevibacillus laterus X10 into 100 mL of culture medium to obtain strains;
[0087] S2: Inoculate 2% of the culture medium into 300 mL of liquid nutrient broth and perform spore staining every 2 hours to check spore formation.
[0088] S3: When the number of bacteria reaches 2×10 8 cfu / mL, when the spore formation rate reaches 80%, 35% is adsorbed into the matrix to obtain microbial organic fertilizer;
[0089] S4: Apply microbial organic fertilizer in the hole, mix it with the soil, place the tomato seedlings in the hole, cover with soil, and water.
[0090] The culture temperature of S1 is 35° C. and the culture time is 48 h.
[0091] The preparation method of the S1 culture medium is:
[0092] Add 10 g of peptone, 5 g of yeast extract, 10 g of NaCl, and 1.3 g of agar powder to 1000 g of water, pH 7, and sterilize at 120°C for 30 min.
[0093] The culture temperature of S2 is 35°C and the culture time is 72h.
[0094] The preparation method of the liquid nutrient broth medium of S2 is as follows:
[0095] Add 4g beef, 6g iron-based soy peptone, 5g glucose, 5g NaCl, and 15g agar powder to 1000g water, pH 7, and sterilize at 120℃ for 30min.
[0096] The preparation method of the iron-based soy peptone is:
[0097] Before the reaction begins, 48 g of soy peptone (CAS: 91079-46-8) is added to a stirring kettle containing 1000 g of water, and then 0.1 g of ferric acrylate, 0.5 g of rose ether, and 3 g of sodium bicarbonate are added. After stirring for reaction, water is distilled off to obtain iron-based soy peptone.
[0098] The stirring rate is 500 rpm.
[0099] The reaction time is 150 min.
[0100] The matrix of S3 includes: 50g chicken manure, 20g bran, 10g soybean cake powder, 10g peat, 10g vermiculite, 10g (NH4)2SO4, 5gKH2PO4, and 1gK2SO4.
[0101] In the S4, 20 g of microbial organic fertilizer was applied to each tomato seedling.
[0102] Comparative Example 1: No iron-based soy peptone was added, and the rest was the same as in Example 1.
[0103] Comparative Example 2: No iron acrylate was added, and the rest was the same as in Example 1.
[0104] Comparative Example 3: Rose oxide was not added, and the other steps were the same as in Example 1.
[0105] Table 1: Test results of various embodiments and comparative examples
[0106]
[0107] Through the data analysis of the above examples and comparative examples, the iron-based soybean peptone prepared by the present invention can effectively increase the fresh weight of a single plant; the microbial organic fertilizer prepared by the present invention can significantly improve the prevention and treatment effect of tomato bacterial wilt.
[0108] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technical personnel in this field can make some changes or modify the technical contents disclosed above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.
Claims
1. A method for improving soil structure using Brevibacillus laterus, characterized in that: The operation steps are: S1: inoculate Brevibacillus laterus X10 into 80-100 mL of culture medium to obtain strains; S2: Inoculate 1-2% of the inoculum into 200-300 mL of liquid nutrient broth medium and perform spore staining every 2 hours to check spore formation. S3: When the number of bacteria reaches 2x10 8 cfu / mL, when the spore formation rate reaches 80%, it is adsorbed into the matrix at a ratio of 30-35% to obtain microbial organic fertilizer; S4: Apply the microbial organic fertilizer to the hole, mix it with the soil, place the tomato seedlings in the hole, cover with soil, and water; The preparation method of the liquid nutrient broth medium of S2 is as follows: Add 1-4 parts of beef, 3-6 parts of iron-based soy peptone, 2-5 parts of glucose, 2-5 parts of NaCl, and 10-15 parts of agar powder to 800-1000 parts of water, pH 7, and sterilize at 100-120℃ for 20-30min; The preparation method of the iron-based soy peptone is: Before the reaction starts, 24-48 parts of soybean peptone are added to a stirring kettle containing 500-1000 parts of water, and then 0.01-0.1 parts of ferric acrylate, 0.04-0.5 parts of rose ether and 1-3 parts of sodium bicarbonate are added. After stirring for reaction, water is distilled off to obtain iron-based soybean peptone.
2. The method for improving soil structure using Brevibacillus laterus according to claim 1, characterized in that: The culture temperature of S1 is 30-35°C and the culture time is 24-48h.
3. The method for improving soil structure using Brevibacillus laterus according to claim 1, characterized in that: The preparation method of the S1 culture medium is: Add 5-10 parts of peptone, 2-5 parts of yeast extract, 5-10 parts of NaCl, and 0.6-1.3 parts of agar powder to 800-1000 parts of water, pH 7, and sterilize at 100-120℃ for 20-30 minutes.
4. The method for improving soil structure using Brevibacillus laterus according to claim 1, characterized in that: The culture temperature of S2 is 30-35°C and the culture time is 48-72h.
5. The method for improving soil structure using Brevibacillus laterus according to claim 1, characterized in that: The stirring rate is 300-500 rpm.
6. The method for improving soil structure using Brevibacillus laterus according to claim 1, characterized in that: The stirring reaction time is 100-150 minutes.
7. The method for improving soil structure using Brevibacillus laterus according to claim 1, characterized in that: The substrate of S3 includes: 40-50 parts of chicken manure, 10-20 parts of bran, 5-10 parts of soybean cake powder, 5-10 parts of peat, 5-10 parts of vermiculite, 5-10 parts of (NH4)2SO4, 1-5 parts of KH2PO4, and 0.1-1 parts of K2SO4.
8. The method for improving soil structure using Brevibacillus laterus according to claim 1, characterized in that: In the S4, 10-20 portions of microbial organic fertilizer are applied to each tomato seedling.