Bacillus laterosporus HP-2 and its application

Abstract

The present application relates to the field of microorganisms, and specifically discloses a Bacillus spporidia HP‑2 and an application thereof; a Bacillus spporidia HP‑2, which is deposited in the China Center for Type Culture Collection with a deposit number of CCTCCNO.M20221789; a biological fertilizer comprising the following raw materials: soybean meal, potassium phosphate, diammonium phosphate, EDTA chelated magnesium, EDTA chelated calcium, EDTA chelated iron, zinc sulfate, humic acid, biological fiber, breathable filler, and Bacillus spporidia HP‑2 bacterial liquid; the Bacillus spporidia HP‑2 bacterial liquid is prepared from Bacillus spporidia HP‑2; Bacillus spporidia HP‑2 has the advantages of high temperature resistance and alkali resistance, and can increase crop yield after application.

Description

Technical Field

[0001] The present application relates to the field of microorganisms, and more specifically, to Bacillus laterosporus HP-2 and applications thereof. Background Art

[0002] Bacillus laterosporus is a Gram-positive bacterium belonging to the genus Bacillus. It can form spores to resist adverse environments. It has strong vitality and adaptability and can survive and reproduce under a variety of environmental conditions. Bacillus laterosporus has wide application value in agriculture, environmental protection, biological control and other fields.

[0003] Bacillus laterosporus can decompose organic matter in the soil, release nutrients needed by plants, and promote plant growth. It can also inhibit and kill plant pathogens, reduce the probability of crop diseases, and increase crop yield and quality. It can also degrade oils, proteins, etc., reduce environmental pollution, and further ensure the quality of crops in the soil.

[0004] However, the activity of Bacillus laterosporus is affected by temperature and pH value. In alkaline soil, the activity of Bacillus laterosporus decreases, and in high temperature environment in summer, the activity is also easily reduced, which can easily affect the effect of Bacillus laterosporus in the soil and may also affect the quality and yield of crops.

[0005] Therefore, how to obtain a high temperature and alkali-resistant Bacillus laterosporus so that it can be applied in the soil to promote the growth of crops and increase crop yields is a problem to be solved. Summary of the invention

[0006] In order to obtain a high temperature and alkali resistant Bacillus laterosporus and apply it in the soil to promote the growth of crops and increase crop yields, the present application provides a Bacillus laterosporus HP-2 and its application.

[0007] In a first aspect, the present application provides a Bacillus laterosporus HP-2, which adopts the following technical solution:

[0008] The invention discloses a Bacillus spporidia HP-2. The Bacillus spporidia HP-2 is deposited in China Center for Type Culture Collection with the deposit number of CCTCCNO.M20221789.

[0009] By adopting the above technical scheme, Bacillus laterosporus HP-2 has the advantages of good high temperature resistance and alkali resistance, and the activity of Bacillus laterosporus HP-2 is not easily affected by external environmental factors during storage and use. When Bacillus laterosporus HP-2 is used as a biofertilizer and added to the soil, it can promote the growth of crops and improve the quality and yield of crops.

[0010] In a second aspect, the present application provides a biofertilizer, which adopts the following technical solution:

[0011] A biological fertilizer comprises the following raw materials in parts by weight:

[0012] 30-50 parts of soybean meal, 25-35 parts of potassium phosphate, 25-35 parts of ammonium dihydrogen phosphate, 0.0001-0.0003 parts of EDTA chelated magnesium, 0.0001-0.0005 parts of EDTA chelated calcium, 0.0001-0.0005 parts of EDTA chelated iron, 0.0001-0.0008 parts of zinc sulfate, 0.1-0.25 parts of humic acid, 1-5 parts of biological fiber, 2-8 parts of breathable filler, 50-70 parts of Bacillus laterosporus HP-2 bacterial liquid; the Bacillus laterosporus HP-2 bacterial liquid is made of Bacillus laterosporus HP-2.

[0013] By adopting the above technical scheme, materials such as soybean meal, bio-fiber and breathable filler can easily absorb the bacterial liquid of Bacillus laterosporus HP-2, so that Bacillus laterosporus HP-2 is evenly dispersed, and Bacillus laterosporus HP-2 can be slowly released in the soil. Combined with the high activity and high temperature and alkali resistance of Bacillus laterosporus HP-2, it can decompose organic matter for a long time and supplement the nutrients needed for the growth of crops, thereby promoting the growth and reproduction of crops in the soil.

[0014] The soybean meal, biological fiber and breathable filler are matched together. The better roughness and connected pores of the soybean meal are combined with the internal pore structure of the biological fiber and breathable filler to make the biofertilizer have better ventilation effect, ensuring that Bacillus laterosporus can come into contact with oxygen when applied in the soil, making Bacillus laterosporus HP-2 have higher activity while ensuring the growth and reproduction of Bacillus laterosporus HP-2.

[0015] Soybean meal and bio-fiber are combined. Soybean meal contains a high content of protein, which can provide the carbon and nitrogen sources required for the growth of Bacillus laterosporus HP-2. The bio-fiber can also be decomposed and utilized by Bacillus laterosporus HP-2 in the later stage, thereby further ensuring the activity of Bacillus laterosporus HP-2, as well as the growth rate and reproduction rate of Bacillus laterosporus HP-2.

[0016] Soybean meal, bio-fiber and breathable filler are combined. Soybean meal is the product after oil extraction and has a hydrophobic effect. Due to the large amount of soybean meal added, its hydrophobic effect can prevent excessive attachment of water in the soil. In addition, the pore structure of bio-fiber and breathable filler, combined with the pore effect of soybean meal, can ensure the circulation of rainwater, so that the soil has better air permeability, ensuring the activity of Bacillus laterosporus HP-2, thereby ensuring the reproduction of Bacillus laterosporus HP-2 at a higher content in the soil and promoting the growth of crops.

[0017] Preferably, the preparation method of the Bacillus laterosporus HP-2 bacterial solution is as follows:

[0018] Inoculate Bacillus laterosporus HP-2 into NA solid culture medium, and culture for 8-12 hours at 30-35° C. and pH 6.8-7.2 to obtain activated bacteria;

[0019] The activated bacteria are transferred to the expansion culture medium according to the inoculation amount of 4-8%, cultured at 30-35°C and 80-100rpm for 6-8 days, and the supernatant is collected after centrifugation to obtain the bacterial liquid.

[0020] By adopting the above technical scheme, after the Bacillus laterosporus HP-2 is activated by the NA solid culture medium, it is expanded in the expansion culture medium, so that the bacterial liquid contains a high content of Bacillus laterosporus HP-2, and the Bacillus laterosporus HP-2 has a high activity, and has the advantages of high temperature resistance and alkali resistance. Even in the hot summer, the activity of Bacillus laterosporus HP-2 is not easily affected, so that the Bacillus laterosporus HP-2 can still decompose organic matter in the hot summer to ensure the growth of crops; even if used in alkaline soil, the activity of Bacillus laterosporus HP-2 is not affected, and the organic matter can still be decomposed to produce carbon source and nitrogen source substances, further ensuring the growth of crops.

[0021] Preferably, the NA solid culture medium comprises the following raw materials: 4-6 g / L sodium chloride, 8-12 g / L beef extract, 8-12 g / L peptone, 15-25 g / L agar, and the balance water, and the volume is fixed to 1 L.

[0022] Preferably, the expansion culture medium comprises the following raw materials: 5-10 g / L glucose, 10-15 g / L tryptone, 2-8 g / L yeast extract, 4-8 g / L sodium chloride, 2-4 g / L potassium chloride, and the balance water, and the volume is fixed to 1 L.

[0023] Preferably, the content of Bacillus laterosporus HP-2 in the bacterial solution is 2.52×10 10 ~4.76×10 10 cfu / mL.

[0024] By adopting the above technical scheme, the raw material ratio of the NA solid culture medium and the expansion culture medium is limited, so that the Bacillus laterosporus HP-2 can be quickly activated and expanded, and the bacterial liquid contains a higher content of Bacillus laterosporus HP-2.

[0025] Preferably, the soybean meal is made of soybean meal, molasses and casein in a mass ratio of 1:0.1-0.2:0.1-0.3.

[0026] By adopting the above technical scheme, the viscosity of molasses is utilized to facilitate casein to adhere to the surface of soybean meal, the water insolubility of casein is utilized to prevent it from absorbing excessive moisture, and casein contains amino and carboxyl groups and has hydrophilicity, thereby ensuring the circulation of moisture on the surface of soybean meal. At the same time, the network structure formed by cross-linking casein and soybean meal is utilized to further improve the porosity of soybean meal, which is not only convenient for loading Bacillus laterosporus HP-2 and promoting the growth and reproduction of Bacillus laterosporus HP-2, but also has good air permeability, thereby ensuring the circulation of oxygen in the soil, allowing aerobic Bacillus laterosporus HP-2 to grow and reproduce rapidly, thereby promoting crop growth and increasing crop yield.

[0027] Preferably, the biofiber is composed of bamboo fiber and pectin-coated lignin in a mass ratio of 1:0.5-1.

[0028] By adopting the above technical scheme, chitosan, bamboo fiber, pectin, lignin and other substances can all supply the carbon source and nitrogen source substances required for the growth of Bacillus laterosporus HP-2, thereby ensuring the activity of Bacillus laterosporus HP-2, promoting the growth and reproduction of Bacillus laterosporus HP-2, increasing the content of Bacillus laterosporus HP-2 in the soil, and thus increasing the yield of crops.

[0029] Bamboo fiber and pectin coated lignin are combined. In alkaline soil, pectin is easily decomposed by alkali solution, so that lignin is gradually exposed. Lignin can also be dissolved in alkali solution. Through the use of alkali solution, the alkali content in the soil is reduced. When pectin and lignin are not decomposed, the adsorption effect of pectin and lignin is combined with the pores of lignin, and the adsorption effect of lignin and bamboo fiber on alkali solution to further reduce the alkali content in the soil. At the same time, with the good alkali resistance of Bacillus laterosporus HP-2, it is ensured that Bacillus laterosporus HP-2 can be highly active in alkaline soil, ensuring the growth of crops, thereby ensuring the yield of crops.

[0030] Preferably, the pectin coated lignin is prepared by loading lignin with a citric acid solution and then coating it with a pectin film liquid; the mass ratio of lignin, citric acid solution, and pectin film liquid is 1:0.4-0.8:0.5-1.

[0031] By adopting the above technical scheme, the adsorption effect of lignin is utilized to adsorb the citric acid solution, and then pectin is used for coating; when it rains, the alkali in the soil meets water and becomes alkaline solution, and the pectin gradually dissolves in the alkaline solution and is exposed to the citric acid solution, and the neutralization reaction of citric acid and alkali is utilized to reduce the alkali content in the soil, and the adsorption effect of lignin on the alkaline solution is combined to further reduce the free alkali content in the soil, and at the same time, the lignin is dissolved in the alkaline solution to further consume the alkali in the soil; the alkali content in the soil is reduced, and the good alkali resistance of Bacillus laterosporus HP-2 and the neutralization effect of acid produced by metabolism on alkali are combined to increase the yield of crops in the soil.

[0032] Preferably, the breathable filler consists of biochar and zeolite in a mass ratio of 1:0.5-1.

[0033] By adopting the above technical solution, biochar and zeolite are combined, and the pores of biochar and zeolite are utilized to improve soil permeability, ensure the activity of Bacillus laterosporus HP-2, and thus increase the yield of crops.

[0034] In summary, this application has the following beneficial effects:

[0035] 1. Bacillus laterosporus HP-2 has the advantages of good resistance to high temperature and alkali, and its activity is not easily affected by external environmental factors during storage and use. When Bacillus laterosporus HP-2 is used as a biofertilizer and added to the soil, it can promote the growth of crops and improve the quality and yield of crops.

[0036] 2. Use biofertilizer in alkaline soil in Northeast China. Utilize the air permeability of biofiber and breathable filler in biofertilizer to ensure the growth and reproduction of Bacillus laterosporus HP-2 while neutralizing the alkaline soil, thereby further promoting the growth of crops in the soil and increasing crop yields. DETAILED DESCRIPTION

[0037] The present application is further described in detail below in conjunction with embodiments.

[0038] Example

[0039] Example 1: Isolation and screening of Bacillus laterosporus HP-2:

[0040] (1) Test soil: Soil was collected from an orchard in Laoshan, Qingdao. The sampling site was a place where fruit trees had been planted for more than ten years. The collected soil was mixed, crushed, and weeds, stones, tree roots and other debris were removed. The soil was mixed evenly and placed in an ice bucket at 4°C for future use. The basic physical and chemical properties of the test soil are shown in Table 1.

[0041] Table 1 Physical and chemical properties of the tested soils

[0042]

[0043] (2) Culture medium:

[0044] NA solid culture medium: weigh 5 g of sodium chloride, 10 g of beef extract, 10 g of peptone, 20 g of agar, and 900 mL of water, heat to 60°C, mix and stir until completely dissolved, adjust the pH to 7.2, and add water to make the volume to 1 L; after packaging, sealing, and wrapping, sterilize at 121°C for 25 min, and let stand to cool to obtain NA solid culture medium;

[0045] (3) Isolation and screening of Bacillus laterosporus HP-2 strain: Weigh 10 g of the test soil sample, add 90 mL of sterile water, and shake at 30 °C and 112 r / min for 2 h to form a suspension. The suspension is diluted by gradient dilution to make 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 Take 10 μL of the dilutions of different gradients and spread them evenly on NA solid medium. Do three parallel replicates for each gradient and culture at 30℃ for 3 days. Select the strains with fast growth rate or transparent circles around them to obtain the primary screening strains.

[0046] (4) Purification: The primary screened strains were inoculated into NA solid medium for separation and purification, and the strains were stored at -40°C;

[0047] The purified strain was named Bacillus spporidia HP-2 and was deposited in the China Center for Type Culture Collection on November 14, 2022 with the accession number CCTCC No. M20221789 and the classification name Bacillus spporidia HP-2.

[0048] Identification of Bacillus laterosporus HP-2:

[0049] (1) Bacterial and colony morphology: The screened strains were subjected to Gram staining, capsule staining, and spore staining, and the bacterial morphology was observed under a microscope.

[0050] Bacillus laterosporus HP-2 is rod-shaped with rounded ends, arranged singly or in short chains, with a size of (0.5-0.8) μm × (1.0-2.5) μm; the colonies are round, with a moist and smooth surface, neat edges, opaque, slightly raised, soft texture, and easy to pick up;

[0051] (2) Antagonistic characteristics:

[0052] ① Gelatin liquefaction: Take gelatin culture medium and inoculate Bacillus laterosporus HP-2 by puncture inoculation method, and use no inoculation as blank control. After culturing at 35℃ for 48h, gently put the gelatin culture medium into a 4℃ refrigerator for 30min, and observe the changes of gelatin. The liquefaction indicates that Bacillus laterosporus HP-2 can produce extracellular enzymes of gelatinase; the experimental results show that the gelatin inoculated with Bacillus laterosporus HP-2 liquefies, indicating that Bacillus laterosporus HP-2 produces extracellular enzymes of gelatinase;

[0053] ② Starch hydrolysis experiment: Bacillus laterosporus HP-2 was inoculated on starch plate culture medium and cultured at 30℃ for 48h. Iodine solution was dripped directly on the culture surface to see if it turned blue to determine whether amylase was produced; if the experimental result showed blue, it indicated that Bacillus laterosporus HP-2 produced starch hydrolyzing enzyme;

[0054] ③ Sugar fermentation experiment: Weigh 0.5g of glucose, 0.5g of peptone, 0.5g of K2HPO4, and 100ml of water to prepare peptone culture medium, add 1% bromothymol blue and mix well, divide the culture solution into test tubes with a height of about 4cm, invert the Dulbecco's tubules in the tubes to fill them with culture solution, sterilize at 115℃ for 20min, inoculate the Bacillus laterosporus HP-2 strain that has been cultured for 10h into the culture medium with a puncture needle, and observe after culturing at room temperature for 1d, 3d and 5d. The color turns yellow and bubbles are generated, which is a positive reaction, indicating that the sugar can be fermented and utilized.

[0055] Application preparation example

[0056] Application Example of Preparation of Bacillus laterosporus HP-2 Bacterial Liquid The following raw materials are all commonly available in the market.

[0057] Application Preparation Example 1: The preparation method of Bacillus laterosporus HP-2 bacterial solution is as follows:

[0058] NA solid culture medium: weigh 5 g of sodium chloride, 10 g of beef extract, 10 g of peptone, 20 g of agar, and 900 mL of water, heat to 60°C, mix and stir until completely dissolved, adjust the pH to 7.2, and add water to make the volume to 1 L; after packaging, sealing, and wrapping, sterilize at 121°C for 25 min, and let stand to cool to obtain NA solid culture medium;

[0059] Expansion culture medium: weigh 8g glucose, 12g tryptone, 5g yeast extract, 6g sodium chloride, 4g potassium chloride, and 900mL water, mix and stir until all dissolved, adjust the pH to 7.2, and add water to make up to 1L; after packaging, sealing, and wrapping,

[0060] Sterilize at 121°C for 25 min, let stand and cool, and obtain expansion culture medium;

[0061] The Bacillus laterosporus HP-2 of Example 1 was inoculated into a NA solid culture medium at an inoculum size of 6%, and cultured at 32° C. and a pH of 7.0 for 10 h to obtain activated bacteria;

[0062] The activated bacteria were transferred to the expansion culture medium at an inoculum rate of 6%, and cultured at 32°C and 100 rpm for 7 days. The supernatant was collected after centrifugation to obtain the bacterial solution. The average content of Bacillus laterosporus HP-2 in the bacterial solution was 4.76×10 10 cfu / mL.

[0063] Application Preparation Example 2: The difference between this application preparation example and application preparation example 1 is:

[0064] NA solid medium: weigh 4 g of sodium chloride, 8 g of beef extract, 12 g of peptone, 15 g of agar, and 900 mL of water, heat to 60°C, mix and stir until completely dissolved, adjust the pH to 7.2, and add water to make the volume to 1 L; after packaging, sealing, and wrapping, sterilize at 121°C for 25 min, and let stand to cool to obtain NA solid medium;

[0065] Expansion culture medium: weigh 5g glucose, 10g tryptone, 2g yeast extract, 4g sodium chloride, 3g potassium chloride, and 900mL water, mix and stir until all dissolved, adjust the pH to 7.2, and add water to make up to 1L; after packaging, sealing, and wrapping,

[0066] Sterilize at 121°C for 25 min, let stand and cool, and obtain expansion culture medium;

[0067] The Bacillus laterosporus HP-2 of Example 1 was inoculated into a NA solid culture medium at an inoculum size of 6%, and cultured at 35° C. and a pH of 6.8 for 8 h to obtain activated bacteria;

[0068] The activated bacteria were transferred to the expansion culture medium at an inoculation rate of 4%, cultured at 35° C. and 80 rpm for 6 days, and the supernatant was collected after centrifugation to obtain the bacterial liquid.

[0069] Application Preparation Example 3: The difference between this application preparation example and application preparation example 1 is that:

[0070] NA solid culture medium: weigh 6 g of sodium chloride, 12 g of beef extract, 12 g of peptone, 25 g of agar, and 900 mL of water, heat to 60°C, mix and stir until completely dissolved, adjust the pH to 7.2, and add water to make the volume to 1 L; after packaging, sealing, and wrapping, sterilize at 121°C for 25 min, and let stand to cool to obtain NA solid culture medium;

[0071] Expansion culture medium: weigh 10g glucose, 15g tryptone, 8g yeast extract, 8g sodium chloride, 4g potassium chloride, and 900mL water, mix and stir until all dissolved, adjust the pH to 7.2, and add water to make up to 1L; after packaging, sealing, and wrapping,

[0072] Sterilize at 121°C for 25 min, let stand and cool, and obtain expansion culture medium;

[0073] The Bacillus laterosporus HP-2 of Example 1 was inoculated into a NA solid culture medium at an inoculum size of 6%, and cultured at 30° C. and a pH of 7.2 for 12 h to obtain activated bacteria;

[0074] The activated bacteria were transferred to the expansion culture medium at an inoculation rate of 8%, cultured at 30° C. and 100 rpm for 8 days, and the supernatant was collected after centrifugation to obtain the bacterial solution.

[0075] Application and preparation examples of soybean meal

[0076] The molasses in the following raw materials was purchased from Shandong Mingyue Chemical Co., Ltd.; other raw materials were all commercially available.

[0077] Application Preparation Example 4: Soybean meal is prepared by the following method:

[0078] 0.15 kg of molasses was evenly sprayed on the surface of 1 kg of soybean meal, and the average particle size of the soybean meal was 2 mm; then 0.2 kg of casein was added, and the casein was sieved through an 80-mesh sieve; the casein addition rate was 120 g / min, and during the addition process, the soybean meal was continuously stirred at a speed of 120 r / min. After being evenly mixed, it was dried and dispersed until the soybean meal did not stick to each other and agglomerated, thereby obtaining the finished soybean meal.

[0079] Application Preparation Example 5: This application preparation example differs from application preparation example 4 in that:

[0080] 0.1 kg of molasses was evenly sprayed on the surface of 1 kg of soybean meal, and the average particle size of the soybean meal was 2 mm; then 0.1 kg of casein was added, and the casein was sieved through an 80-mesh sieve; the casein addition rate was 120 g / min, and during the addition process, the soybean meal was continuously stirred at a speed of 120 r / min. After being evenly mixed, it was dried and dispersed until the soybean meal did not stick to each other and agglomerated, thereby obtaining the finished soybean meal.

[0081] Application Preparation Example 6: This application preparation example is different from application preparation example 4 in that:

[0082] 0.2 kg of molasses was evenly sprayed on the surface of 1 kg of soybean meal, and the average particle size of the soybean meal was 2 mm; then 0.3 kg of casein was added, and the casein was sieved through an 80-mesh sieve; the casein addition rate was 120 g / min, and during the addition process, the soybean meal was continuously stirred at a speed of 120 r / min. After being evenly mixed, it was dried and dispersed until the soybean meal did not stick to each other and agglomerated, thereby obtaining the finished soybean meal.

[0083] Application and preparation example of pectin coated lignin

[0084] The following raw materials are all commercially available.

[0085] Application Preparation Example 7: Pectin coated lignin is prepared by the following method:

[0086] 0.6 kg of citric acid solution was added to 1 kg of lignin, the adding speed of the citric acid solution was 200 g / min, and during the adding process, the mixture was continuously stirred at a speed of 80 r / min. The average diameter of the lignin was 0.5 mm, and the citric acid solution was a citric acid aqueous solution with a mass fraction of 1%. After the lignin completely absorbed the citric acid solution, the loaded lignin was obtained;

[0087] 1 kg of high-fat pectin was mixed with 3 kg of white sugar and 30 kg of water and stirred until the high-fat pectin was completely dissolved to obtain a pectin film liquid; 0.8 kg of pectin film liquid was evenly sprayed on the surface of the carrier lignin, and dried and dispersed until the lignin did not stick to each other and agglomerated to obtain a finished product of pectin coated lignin.

[0088] Application Preparation Example 8: This application preparation example differs from application preparation example 7 in that:

[0089] 0.4 kg of citric acid solution was added to 1 kg of lignin, the adding speed of the citric acid solution was 200 g / min, and during the adding process, the mixture was continuously stirred at a speed of 80 r / min. The average diameter of the lignin was 0.5 mm, and the citric acid solution was a citric acid aqueous solution with a mass fraction of 1%. After the lignin completely absorbed the citric acid solution, the loaded lignin was obtained;

[0090] 1 kg of high-fat pectin was mixed with 3 kg of white sugar and 30 kg of water and stirred until the high-fat pectin was completely dissolved to obtain a pectin film liquid; 0.5 kg of pectin film liquid was evenly sprayed on the surface of the carrier lignin, and the finished pectin-coated lignin was obtained after drying and dispersion until the lignin did not stick to each other and agglomerate.

[0091] Application Preparation Example 9: This application preparation example is different from application preparation example 7 in that:

[0092] 0.8 kg of citric acid solution was added to 1 kg of lignin, the adding speed of the citric acid solution was 200 g / min, and during the adding process, the mixture was continuously stirred at a speed of 80 r / min. The average diameter of the lignin was 0.5 mm, and the citric acid solution was a citric acid aqueous solution with a mass fraction of 1%. After the lignin completely absorbed the citric acid solution, the loaded lignin was obtained;

[0093] 1 kg of high-fat pectin was mixed with 3 kg of white sugar and 30 kg of water and stirred until the high-fat pectin was completely dissolved to obtain a pectin film liquid; 1 kg of pectin film liquid was evenly sprayed on the surface of the carrier lignin, and dried and dispersed until the lignin did not stick to each other and agglomerated to obtain a finished product of pectin coated lignin.

[0094] Application Examples

[0095] Application example 1: A biological fertilizer:

[0096] 40kg soybean meal, 30kg potassium phosphate, 30kg ammonium dihydrogen phosphate, 0.0002kg EDTA chelated magnesium, 0.0003kg EDTA chelated calcium, 0.0003kg EDTA chelated iron, 0.0005kg zinc sulfate, 0.2kg humic acid, 3kg bio-fiber, 5kg breathable filler, 60kg Bacillus laterosporus HP-2 bacterial liquid; the soybean meal is selected from the soybean meal prepared in Preparation Example 4; the Bacillus laterosporus HP-2 bacterial liquid is selected from the Bacillus laterosporus HP-2 bacterial liquid prepared in Preparation Example 1; the bio-fiber is composed of bamboo fiber in a mass ratio of 1:1 and pectin-coated lignin prepared in Preparation Example 7, and the average length of the bamboo fiber is 2mm; the breathable filler is composed of biochar and zeolite in a mass ratio of 1:1, and the average particle size of the biochar and zeolite is 0.3mm.

[0097] Application Example 2: The difference between this application example and application example 1 is:

[0098] 30kg soybean meal, 25kg potassium phosphate, 25kg ammonium dihydrogen phosphate, 0.0001kg EDTA chelated magnesium, 0.0001kg EDTA chelated calcium, 0.0001kg EDTA chelated iron, 0.0001kg zinc sulfate, 0.1kg humic acid, 1kg bio-fiber, 2kg breathable filler, 50kg Bacillus laterosporus HP-2 bacterial liquid; the soybean meal is selected from the soybean meal prepared in Preparation Example 5; the Bacillus laterosporus HP-2 bacterial liquid is selected from the Bacillus laterosporus HP-2 bacterial liquid prepared in Preparation Example 2; the bio-fiber is composed of bamboo fiber in a mass ratio of 1:0.5 and pectin-coated lignin prepared in Preparation Example 8; the breathable filler is composed of biochar and zeolite in a mass ratio of 1:0.5.

[0099] Application Example 3: The difference between this application example and application example 1 is:

[0100] 50kg soybean meal, 35kg potassium phosphate, 35kg ammonium dihydrogen phosphate, 0.0003kg EDTA chelated magnesium, 0.0005kg EDTA chelated calcium, 0.0005kg EDTA chelated iron, 0.0008kg zinc sulfate, 0.25kg humic acid, 5kg bio-fiber, 8kg breathable filler, 70kg Bacillus laterosporus HP-2 bacterial liquid; the soybean meal is selected from the soybean meal prepared in Preparation Example 6; the Bacillus laterosporus HP-2 bacterial liquid is selected from the Bacillus laterosporus HP-2 bacterial liquid prepared in Preparation Example 3; the bio-fiber is composed of bamboo fiber in a mass ratio of 1:1 and pectin-coated lignin prepared in Preparation Example 9; the breathable filler is composed of biochar and zeolite in a mass ratio of 1:1.

[0101] Application Example 4: The difference between this application example and application example 1 is:

[0102] Soybean meal is soybean meal without molasses and casein on the surface.

[0103] Application Example 5: The difference between this application example and application example 1 is:

[0104] The pectin-coated lignin is replaced by an equal mass of lignin in the biofiber.

[0105] Application Example 6: The difference between this application example and application example 1 is:

[0106] Preparation process of pectin coated lignin in biofiber without loading of citric acid solution.

[0107] Comparative Application Examples

[0108] Comparative Application Example 1: The difference between this comparative application example and application example 1 is:

[0109] No breathable fillers or biofibers are added to the biofertilizer raw materials.

[0110] Comparative Application Example 2: The difference between this comparative application example and application example 1 is:

[0111] No Bacillus laterosporus HP-2 bacterial solution was added to the bio-fertilizer raw materials.

[0112] Performance testing

[0113] 1. High temperature resistance test

[0114] The Bacillus laterosporus HP-2 bacterial solution prepared in Example 1 was placed at 80, 90, and 100° C. for 30 min, respectively. After treatment, the number of effective live bacteria in the bacterial solution was detected and calculated, and the data was recorded.

[0115] 2. Alkali resistance test

[0116] Sodium hydroxide solution was added to the Bacillus laterosporus HP-2 bacterial solution of Application Preparation Example 1 at a mass ratio of 1:0.2. The pH values ​​of the sodium hydroxide solutions were 8, 9, 10, and 11, respectively. After standing for 30 minutes, the number of effective live bacteria in the bacterial solution was detected and calculated, and the data was recorded.

[0117] Table 1 Bacillus laterosporus HP-2 detection table

[0118]

[0119] Combined with the data in Table 1, it can be seen that after high temperature treatment, the number of effective live bacteria in the bacterial solution tends to the initial effective live bacteria number, and Bacillus laterosporus HP-2 has good high temperature resistance; and under alkaline pH conditions, Bacillus laterosporus HP-2 in the bacterial solution still has a high number of live bacteria, indicating that Bacillus laterosporus HP-2 has good alkali resistance.

[0120] 3. Yield detection

[0121] The experimental fields are: Changchun City, Jilin Province, with soil pH of 7.8;

[0122] The 7 mu of land correspond to the biofertilizers prepared in Application Examples 1-6 and Comparative Application Examples 1-2.

[0123] Select corn of the same batch, same variety, and normal sowing, and apply different bio-fertilizers as base fertilizers to 7 acres of land. The application amount of bio-fertilizer is 50 kg per acre. During the growth process of corn, 15 kg of topdressing is added, and the amount and type of topdressing are the same. Record the yield of autumn harvest corn.

[0124] In the above experiments, except for the different biofertilizers, the other test conditions were the same, which was based on the single variable principle.

[0125] Table 2 Crop yield detection table

[0126] project Yield (kg / mu) Application Example 1 750 Application Example 2 710 Application Example 3 780 Application Example 4 640 Application Example 5 670 Application Example 6 690 Comparative application example 1 600 Comparative Application Example 2 540

[0127] It can be seen from Application Examples 1-3 and Table 2 that the biofertilizer prepared in the present application enables crops to still have a relatively high yield under alkaline soil conditions.

[0128] Combining Application Example 1 and Application Examples 4-6 and Table 2, it can be seen that the soybean meal in Application Example 4 is soybean meal with no molasses and casein on the surface. Compared with Application Example 1, the corn yield of Application Example 4 is lower than that of Application Example 1; this indicates that molasses and casein are combined with good air permeability to ensure the circulation of oxygen in the soil, so that aerobic Bacillus laterosporus HP-2 can grow and reproduce rapidly, thereby promoting crop growth and providing crop yields.

[0129] In Application Example 5, the pectin-coated lignin was replaced with an equal mass of lignin in the biofiber. In the preparation process of pectin-coated lignin in Application Example 6, no citric acid solution was loaded. Compared with Application Example 1, the corn yields of Application Examples 5 and 6 were lower than that of Application Example 1. This indicates that pectin is soluble in alkali solution, citric acid neutralizes the alkali, and the alkali content in the soil is controlled. Combined with the better alkali resistance of Bacillus laterosporus HP-2, the soil contains a higher content of nutrients, thereby promoting the growth of crops and increasing crop yields.

[0130] Combining Application Example 1 and Comparative Application Examples 1-2 and Table 2, it can be seen that no breathable filler and bio-fiber are added to the bio-fertilizer raw material of Comparative Application Example 1. Compared with Application Example 1, the corn yield of Comparative Application Example 1 is lower than that of Application Example 1; this indicates that good air permeability can promote the growth and reproduction of Bacillus laterosporus HP-2, thereby increasing crop yields, and that breathable fillers and bio-fibers can also be gradually decomposed and utilized by bacteria in the soil, further promoting the growth and reproduction of Bacillus laterosporus HP-2, thereby further increasing crop yields.

[0131] In comparative application example 2, no Bacillus laterosporus HP-2 bacterial solution was added to the bio-fertilizer raw materials. Compared with application example 1, the corn yield of comparative application example 2 was lower than that of application example 1, which indicated that the addition of Bacillus laterosporus HP-2 bacterial solution could decompose organic matter in the soil, increase the nutrients required for crop growth, and thus increase crop yield.

[0132] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. A Bacillus laterosporus ( Bacillus spporidia ) HP-2, characterized in that Bacillus laterosporus HP-2 is deposited in China Center for Type Culture Collection with the deposit number: CCTCCNO.M20221789.

2. A biological fertilizer, characterized in that: Contains the following raw materials in parts by weight: 30-50 parts of soybean meal, 25-35 parts of potassium phosphate, 25-35 parts of ammonium dihydrogen phosphate, 0.0001-0.0003 parts of EDTA chelated magnesium, 0.0001-0.0005 parts of EDTA chelated calcium, 0.0001-0.0005 parts of EDTA chelated iron, 0.0001-0.0008 parts of zinc sulfate, 0.1-0.25 parts of humic acid, 1-5 parts of biological fiber, 2-8 parts of breathable filler, and 50-70 parts of Bacillus laterosporus HP-2 bacterial liquid; the Bacillus laterosporus HP-2 bacterial liquid is prepared by the Bacillus laterosporus HP-2 described in claim 1.

3. A biological fertilizer according to claim 2, characterized in that: The content of Bacillus laterosporus HP-2 in the bacterial solution was 2.52×10 10 ~4.76×10 10 cfu / mL.

4. A biological fertilizer according to claim 2, characterized in that: The soybean meal is prepared from soybean meal, molasses and casein in a mass ratio of 1:0.1-0.2:0.1-0.

3.

5. A biological fertilizer according to claim 2, characterized in that: The biological fiber is composed of bamboo fiber and pectin-coated lignin in a mass ratio of 1:0.5-1.

6. A biological fertilizer according to claim 5, characterized in that: The pectin coated lignin is prepared by loading citric acid solution on lignin and then coating it with pectin film liquid; the mass ratio of lignin, citric acid solution and pectin film liquid is 1:0.4-0.8:0.5-1.

7. A biological fertilizer according to claim 2, characterized in that: The breathable filler is composed of biochar and zeolite in a mass ratio of 1:0.5-1.

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