Bacillus mucilaginosus HP-JZ2 and biological bacterial agent

Abstract

The application relates to the field of microorganisms, and particularly discloses a bacillus mucilaginosus HP-JZ2 and a biological bacterial agent, the bacillus mucilaginosus HP-JZ2 is preserved in the China Center for Type Culture Collection on November 14, 2022, and the preservation number is CCTCC NO.M20221792; the biological bacterial agent comprises the bacillus mucilaginosus HP-JZ2 bacterial agent; the bacillus mucilaginosus HP-JZ2 has the advantages that under the conditions of high temperature, high humidity, high acid and alkali and multiple microorganisms, the bacillus mucilaginosus HP-JZ2 still has good growth activity and reproduction activity; and the biological bacterial agent can be widely applied in the fields of agriculture, industry and the like.

Description

Technical Field

[0001] This application relates to the field of microbiology, and more specifically, to a Bacillus mucilaginosus HP-JZ2 and a biological agent. Background Technology

[0002] Bacillus mucilaginosus, also known as silicate bacteria, is characterized by its ability to decompose potassium and silicon in minerals such as feldspar and mica, as well as phosphorus in apatite. It also secretes plant growth stimulants and various enzymes, thereby enhancing crops' resistance to certain diseases.

[0003] Although Bacillus mucilaginosus has good stress resistance, its growth and reproduction activities can still be affected by high temperature, high humidity, and high acidity / alkalinity. Furthermore, Bacillus mucilaginosus is highly dependent on water, and too much or too little water can also affect its growth and reproduction activities. At the same time, under the complex microbial community conditions in the soil, Bacillus mucilaginosus is easily affected by other bacteria in the soil, and its growth and reproduction activities can be affected by competition or inhibition.

[0004] Therefore, how to provide a Bacillus subtilis strain with good resistance that still maintains good growth and reproductive activity under conditions of high temperature, high humidity, high acidity and alkali, and multiple microorganisms is a problem that needs to be solved. Summary of the Invention

[0005] In order to provide a Bacillus subtilis with better resistance, which can still maintain good growth and reproduction activity under high temperature, high humidity, high acid and alkali and multi-microorganism conditions, this application provides a Bacillus subtilis HP-JZ2 and a biological agent.

[0006] In a first aspect, this application provides a Bacillus mucilaginosus HP-JZ2, which adopts the following technical solution: A type of Bacillus mucilaginosus krassilnikov HP-JZ2 was deposited at the China Center for Type Culture Collection on November 14, 2022, with accession number CCTCCNO.M20221792.

[0007] By adopting the above technical solution, Bacillus mucilaginosus HP-JZ2 has good stress resistance and can still maintain good growth and reproduction activity under high temperature, high humidity, high acid and alkali and multi-microorganism conditions.

[0008] Secondly, this application provides a biological agent, which adopts the following technical solution: A biological agent comprising Bacillus mucilaginosus HP-JZ2 agent, wherein the Bacillus mucilaginosus HP-JZ2 agent is prepared from Bacillus mucilaginosus HP-JZ2.

[0009] By adopting the above technical solution, the bio-agent contains highly active Bacillus mucilaginosus HP-JZ2, enabling it to be used in soil remediation to decompose and transform pollutants in the soil, restoring soil health. The bio-agent can also be used as a bio-fertilizer, utilizing its nitrogen-fixing, phosphorus-solubilizing, and potassium-solubilizing abilities to improve soil fertility and promote crop growth. Furthermore, Bacillus mucilaginosus HP-JZ2 has a strong symbiotic ability in the soil, is not easily affected by other microorganisms, and can enhance plant absorption and utilization of nutrients, thereby improving crop yield and quality. Simultaneously, Bacillus mucilaginosus HP-JZ2 in the bio-agent can produce more beneficial substances such as extracellular polysaccharides, which can be applied in the food fermentation field to develop functional foods. Bacillus mucilaginosus HP-JZ2 can also produce highly bioactive substances such as antimicrobial peptides, which can be further applied in the fields of biopharmaceutical carriers or targeted drug delivery, broadening the application scenarios of the bio-agent while efficiently leveraging the role of Bacillus mucilaginosus HP-JZ2.

[0010] Preferably, the Bacillus mucilaginosus HP-JZ2 inoculum is prepared by the following method: Bacillus mucilaginosus HP-JZ2 was inoculated into slant culture medium at an inoculation amount of 5-8%, and activated at 30-32℃ for 2-3 days to obtain activated strains; The activated strain was inoculated into seed culture medium at an inoculation rate of 6-10%, and cultured in shake flasks at 30-32℃ for 8-14 hours to obtain the amplified strain. The amplified strain was inoculated into the fermentation medium at an inoculation rate of 5-8%, and fermented at 36-38℃ for 46-50 hours. The supernatant was then removed by centrifugation to obtain the bacterial agent.

[0011] By adopting the above technical solution, *Bacillus mucilaginosus* HP-JZ2 is first activated on an agar slant medium to promote its initial growth and reproduction and restore its own activity, which is beneficial to the expansion and growth of colonies and increases the activity of *Bacillus mucilaginosus* HP-JZ2. Then, it is inoculated into a seed medium for amplification to increase the cell content of *Bacillus mucilaginosus* HP-JZ2 and further promote its growth and reproduction. Finally, it is inoculated into a fermentation medium for fermentation treatment, which further stimulates the activity of *Bacillus mucilaginosus* HP-JZ2 and promotes the production of metabolites such as enzymes, organic acids and other bioactive substances. This makes the application of biological agents in agriculture, industry and medicine highly valuable and efficient in exerting the effects of biological agents.

[0012] Preferably, the slant culture medium comprises the following raw materials: potassium aluminosilicate 0.8-1 g / L, sodium aluminosilicate 0.2-0.3 g / L, sucrose 5-8 g / L, trehalose 1-2 g / L, disodium hydrogen phosphate 1-2 g / L, ferric chloride 0.004-0.006 g / L, calcium chloride 0.001-0.002 g / L, magnesium sulfate heptahydrate 0.4-0.5 g / L, agar 18-21.5 g / L, and water to a final volume of 1 L.

[0013] By adopting the above technical solution, sucrose and trehalose are combined. Sucrose has the advantages of rapid decomposition and high utilization efficiency, providing carbon source substances in the early stage of Bacillus mucilage HP-JZ2 growth, while trehalose can continuously provide carbon source energy required for the growth of Bacillus mucilage HP-JZ2, supporting the growth and metabolism of Bacillus mucilage HP-JZ2 in the long term. In addition, trehalose has the function of stabilizing the cell membrane and protein structure of Bacillus mucilage HP-JZ2, further protecting Bacillus mucilage HP-JZ2 from adverse external environmental effects and improving stress resistance. At the same time, trehalose can also act as a type molecule to induce Bacillus mucilage HP-JZ2 to produce metabolites, further enhancing the application value of Bacillus mucilage HP-JZ2.

[0014] Potassium aluminosilicate, sodium aluminosilicate, disodium hydrogen phosphate, ferric chloride, calcium chloride, magnesium sulfate heptahydrate, and other substances provide the necessary nutrients and trace elements for the growth of Bacillus mucilaginosus HP-JZ2, promoting the growth and reproduction of Bacillus mucilaginosus HP-JZ2 and increasing its activity and content.

[0015] Preferably, the seed culture medium comprises the following raw materials: glucose 2-4 g / L, molasses 5-8 g / L, trehalose 1-2 g / L, chitin 0.05-0.1 g / L, dipotassium hydrogen phosphate 0.4-0.6 g / L, magnesium sulfate heptahydrate 0.4-0.6 g / L, sodium chloride 0.1-0.35 g / L, calcium carbonate 0.5-1 g / L, and water to a final volume of 1 L.

[0016] By employing the above technical solution, glucose, molasses, and trehalose are combined. Glucose can be rapidly utilized by Bacillus mucilaginosus HP-JZ2, providing a carbon source to promote the growth and reproduction of Bacillus mucilaginosus HP-JZ2. Molasses and trehalose can continuously provide nutrients such as carbon dioxide to Bacillus mucilaginosus HP-JZ2, ensuring its growth and reproduction. Chitosan provides support for Bacillus mucilaginosus HP-JZ2, ensuring that it is in uniform contact with nutrients, thereby further promoting its growth and reproduction. Chitosan can also provide a nutrient source for bacterial growth, supporting the growth and metabolism of Bacillus mucilaginosus HP-JZ2.

[0017] Dipotassium hydrogen phosphate, magnesium sulfate heptahydrate, sodium chloride, and calcium sulfate provide nutrients and trace elements for the growth and reproduction of Bacillus mucilaginosus HP-JZ2, ensuring the activity of Bacillus mucilaginosus HP-JZ2 and increasing its content.

[0018] Preferably, the fermentation medium comprises the following raw materials: glucose 5-9 g / L, sucrose 2-3 g / L, isomalt oligosaccharide 0.2-0.5 g / L, yeast extract 0.1-0.3 g / L, ammonium sulfate 0.4-0.7 g / L, dipotassium hydrogen phosphate 1-1.5 g / L, calcium carbonate 0.4-0.7 g / L, ferric chloride 0.4-0.7 g / L, sodium chloride 0.1-0.3 g / L, magnesium sulfate heptahydrate 0.1-0.2 g / L, and water to a final volume of 1 L.

[0019] By employing the above-mentioned technical solution, glucose, sucrose, and isomaltooligosaccharides are combined. Glucose and sucrose can rapidly provide carbon and energy sources for Bacillus mucilaginosus HP-JZ2, while isomaltooligosaccharides can also be utilized during the fermentation process of Bacillus mucilaginosus HP-JZ2. By decomposing isomaltooligosaccharides, Bacillus mucilaginosus HP-JZ2 can obtain energy and carbon skeletons for synthesizing its own cellular substances and metabolites. Furthermore, isomaltooligosaccharides have low polymer content, which, after being absorbed by Bacillus mucilaginosus HP-JZ2, is beneficial to the growth and reproduction of Bacillus mucilaginosus HP-JZ2, improving fermentation efficiency and thus increasing the content of beneficial active substances.

[0020] The combination of yeast extract, ammonium sulfate, dipotassium hydrogen phosphate, calcium carbonate, ferric chloride, sodium chloride, and magnesium sulfate heptahydrate provides nutrients for the fermentation process and further promotes the fermentation of Bacillus mucilaginosus HP-JZ2 to produce beneficial active substances.

[0021] Preferably, the shaking speed during the shake flask culture is 180-200 r / min.

[0022] Preferably, the shaking speed during the fermentation process is 180-200 r / min.

[0023] By adopting the above technical solution, the dissolved oxygen content in the culture medium can be increased during shake flask culture, thereby meeting the oxygen requirements of Bacillus mucilaginosus HP-JZ2. By limiting the rotation speed, it is ensured that Bacillus mucilaginosus HP-JZ2 is in uniform contact with the culture medium, while ensuring the absorption of nutrients and the discharge of metabolic products, thus ensuring the activity and content of Bacillus mucilaginosus HP-JZ2.

[0024] In summary, this application has the following beneficial effects: 1. Bacillus mucilaginosus HP-JZ2 has good stress resistance and can still maintain good growth and reproduction activity under conditions of high temperature, high humidity, high acidity and alkali and multiple microorganisms.

[0025] 2. The bio-agent contains highly active Bacillus mucilaginosus HP-JZ2, enabling it to be used in soil remediation to decompose and transform pollutants in the soil, restoring soil health. It can also be used as a bio-fertilizer, utilizing its nitrogen-fixing, phosphorus-solubilizing, and potassium-solubilizing abilities to improve soil fertility and promote crop growth. Furthermore, Bacillus mucilaginosus HP-JZ2 exhibits strong symbiotic capabilities in the soil, is less susceptible to interference from other microorganisms, and enhances plant nutrient absorption and utilization, thereby increasing crop yield and quality. Simultaneously, Bacillus mucilaginosus HP-JZ2 in the bio-agent can produce numerous beneficial substances such as extracellular polysaccharides, which can be applied in the food fermentation field to develop functional foods. Bacillus mucilaginosus HP-JZ2 can also produce highly bioactive substances such as antimicrobial peptides, which can be further applied in the fields of biopharmaceutical carriers or targeted drug delivery, broadening the application scenarios of the bio-agent while efficiently leveraging the effects of Bacillus mucilaginosus HP-JZ2.

[0026] 3. The combination of trehalose and isomaltooligosaccharide provides Bacillus mucilaginosus HP-JZ2 with a continuous source of carbon energy for its growth. Furthermore, isomaltooligosaccharide has a low polymer content, which is beneficial to the growth and reproduction of Bacillus mucilaginosus HP-JZ2 after absorption, thereby improving fermentation efficiency and increasing the content of beneficial active substances. Detailed Implementation

[0027] The present application will be further described in detail below with reference to the embodiments. Example

[0028] Example 1: Isolation and purification of Bacillus mucilaginosus HP-JZ2: (1) Test soil: Soil was collected from Laoshan, Qingdao. The sampling site was a fruit and vegetable land that was more than 10 years old. The collected soil was mixed, crushed, and weeds, stones, tree roots and other debris were removed. The mixture was then placed in an ice bucket and stored at 4℃ for later use. The basic physicochemical properties of the test soil are shown in Table 1. Table 1 Physicochemical properties of the tested soils

[0029] (2) Isolation medium: 15g beef extract, 10g peptone, 15g sodium chloride, 18g agar, add 900mL water, stir under 60℃ water bath conditions until all agar and other substances are dissolved, then add water to make up to 1L, adjust pH to 7.2, then sterilize at 121℃ for 20min, cool, and obtain isolation medium; Liquid culture medium: 10g sucrose, 1g ammonium chloride, 1g disodium hydrogen phosphate, 0.5g magnesium sulfate heptahydrate, 0.005g ferric chloride, 0.3g calcium carbonate, add 900mL water and mix well. Then add water to make up to 1L, pH 7.2, mix well, and then sterilize at 121℃ for 25min. Cool to obtain liquid culture medium. (3) Isolation and screening of Bacillus mucilaginosus HP-JZ2: Weigh 10g of the soil sample, add 90mL of sterile physiological saline, and shake for 1h at 30℃ and 120r / min to form a suspension. The suspension is then serially diluted to prepare 10 -1 10 -2 10 -3 10 -4 10 -5 10 -6 Diluted at multiples of 10 times, 10 μL of each dilution was evenly spread on the isolation medium, with three replicates for each dilution. The culture was carried out at 30°C for 2 days, and single colonies with good growth were selected to obtain the screening cells. (4) Purification of Bacillus mucilaginosus HP-JZ2: The selected bacterial cells were streaked onto the isolation medium using the streak method and cultured at 37°C for 24 hours. The above steps were repeated until the purified strain was obtained.

[0030] The purified strain was named Bacillus mucilaginosus HP-JZ2 and deposited at the China Center for Type Culture Collection on November 14, 2022, with accession number CCTCCNO.M20221792. It was classified as Bacillus mucilaginosus krassilnikov HP-JZ2.

[0031] Identification of Bacillus mucilaginosus HP-JZ2: (1) Cell and colony morphology: After staining Bacillus mucilaginosus HP-JZ2, the cell morphology was observed under a microscope; The cells of Bacillus mucilaginosus HP-JZ2 are thick and long rods with rounded ends; the colonies on the culture medium are half-glass beads, smooth, moist and shiny, with neat and smooth edges, without forks or hairy protrusions.

[0032] Application and preparation examples of Bacillus mucilaginosus HP-JZ2 inoculant Application Preparation Example 1: The Bacillus mucilaginosus HP-JZ2 inoculum was prepared using the following method: Weigh out 1g of potassium aluminosilicate, 0.25g of sodium aluminosilicate, 6g of sucrose, 2g of trehalose, 2g of disodium hydrogen phosphate, 0.005g of ferric chloride, 0.002g of calcium chloride, 0.45g of magnesium sulfate heptahydrate, and 20g of agar. Mix the agar with 900mL of water and stir in a 60℃ water bath until the agar is completely dissolved. Then add potassium aluminosilicate, sodium aluminosilicate, sucrose, trehalose, disodium hydrogen phosphate, ferric chloride, calcium chloride, and magnesium sulfate heptahydrate and mix well. Add water to make up to 1L, pH 7.2, tilt to solidify, and then sterilize at 121℃ for 20min. Cool to obtain the slant culture medium. Weigh out 3g of glucose, 6g of molasses, 2g of trehalose, 0.08g of chitin, 0.5g of dipotassium hydrogen phosphate, 0.5g of magnesium sulfate heptahydrate, 0.25g of sodium chloride, 0.8g of calcium carbonate, and 900mL of water. Mix well, then add water to make up to 1L. The pH is 7.2. Sterilize at 121℃ for 20min, cool, and the seed culture medium is obtained. Weigh out 7g of glucose, 3g of sucrose, 0.4g of isomaltooligosaccharide, 0.2g of yeast extract, 0.5g of ammonium sulfate, 1.2g of dipotassium hydrogen phosphate, 0.5g of calcium carbonate, 0.5g of ferric chloride, 0.2g of sodium chloride, and 0.2g of magnesium sulfate heptahydrate. Mix them with 900mL of water and stir well. Then add water to make up to 1L. The pH is 7.2. Sterilize at 121℃ for 20min and cool to obtain the fermentation medium. Bacillus mucilaginosus HP-JZ2 was inoculated into slant culture medium at an inoculation amount of 7%, and activated at 30℃ for 2.5 days to obtain the activated strain; The activated strain was inoculated into seed culture medium at an inoculation amount of 8%, and cultured in a shake flask at 30℃ for 12 h at a shaker speed of 200 r / min to obtain the amplified strain. The amplified strain was inoculated into the fermentation medium at an inoculation amount of 7%, and fermented at 37°C for 48 hours with a shaking speed of 200 r / min. After centrifugation, the supernatant was removed to obtain the bacterial agent.

[0033] Application Preparation Example 2: The difference between this application preparation example and application preparation example 1 is that: Weigh out 0.8g potassium aluminosilicate, 0.2g sodium aluminosilicate, 5g sucrose, 1g trehalose, 1g disodium hydrogen phosphate, 0.004g ferric chloride, 0.001g calcium chloride, 0.4g magnesium sulfate heptahydrate, and 18g agar. Mix the agar with 900mL of water and stir in a 60℃ water bath until the agar is completely dissolved. Then add potassium aluminosilicate, sodium aluminosilicate, sucrose, trehalose, disodium hydrogen phosphate, ferric chloride, calcium chloride, and magnesium sulfate heptahydrate and mix well. Add water to make up to 1L, pH 7.2, tilt to solidify, and then sterilize at 121℃ for 20min. Cool to obtain the slant culture medium. Weigh out 2g of glucose, 5g of molasses, 1g of trehalose, 0.05g of chitin, 0.4g of dipotassium hydrogen phosphate, 0.4g of magnesium sulfate heptahydrate, 0.1g of sodium chloride, 0.5g of calcium carbonate, and 900mL of water. Mix well, then add water to make up to 1L. The pH is 7.2. Sterilize at 121℃ for 20min, cool, and the seed culture medium is obtained. Weigh out 5g of glucose, 2g of sucrose, 0.2g of isomaltooligosaccharide, 0.1g of yeast extract, 0.4g of ammonium sulfate, 1g of dipotassium hydrogen phosphate, 0.4g of calcium carbonate, 0.4g of ferric chloride, 0.1g of sodium chloride, and 0.1g of magnesium sulfate heptahydrate. Mix them with 900mL of water and stir well. Then add water to make up to 1L. The pH is 7.2. Sterilize at 121℃ for 20min and cool to obtain the fermentation medium. Bacillus mucilaginosus HP-JZ2 was inoculated into slant culture medium at an inoculation amount of 5%, and activated at 32℃ for 2 days to obtain the activated strain; The activated strain was inoculated into seed culture medium at a rate of 6%, and cultured in a shake flask at 32°C for 8 hours at a shaker speed of 180 r / min to obtain the amplified strain. The amplified strain was inoculated into the fermentation medium at an inoculation amount of 5%, and fermented at 38°C for 46 hours with a shaking speed of 180 r / min. After centrifugation, the supernatant was removed to obtain the bacterial agent.

[0034] Application Preparation Example 3: The difference between this application preparation example and application preparation example 1 is that: Weigh out 1g of potassium aluminosilicate, 0.3g of sodium aluminosilicate, 8g of sucrose, 2g of trehalose, 2g of disodium hydrogen phosphate, 0.006g of ferric chloride, 0.002g of calcium chloride, 0.5g of magnesium sulfate heptahydrate, and 21.5g of agar. Mix the agar with 900mL of water and stir in a 60℃ water bath until the agar is completely dissolved. Then add potassium aluminosilicate, sodium aluminosilicate, sucrose, trehalose, disodium hydrogen phosphate, ferric chloride, calcium chloride, and magnesium sulfate heptahydrate and mix well. Add water to make up to 1L, pH 7.2, tilt to solidify, and then sterilize at 121℃ for 20min. Cool to obtain the slant culture medium. Weigh out 4g of glucose, 8g of molasses, 2g of trehalose, 0.1g of chitin, 0.6g of dipotassium hydrogen phosphate, 0.6g of magnesium sulfate heptahydrate, 0.35g of sodium chloride, 1g of calcium carbonate, and 900mL of water. Mix well, then add water to make up to 1L. The pH is 7.2. Sterilize at 121℃ for 20min, cool, and the seed culture medium is obtained. Weigh out 9g of glucose, 3g of sucrose, 0.5g of isomaltooligosaccharide, 0.3g of yeast extract, 0.7g of ammonium sulfate, 1.5g of dipotassium hydrogen phosphate, 0.7g of calcium carbonate, 0.7g of ferric chloride, 0.3g of sodium chloride, and 0.2g of magnesium sulfate heptahydrate. Mix them with 900mL of water and stir well. Then add water to make up to 1L. The pH is 7.2. Sterilize at 121℃ for 20min and cool to obtain the fermentation medium. Bacillus mucilaginosus HP-JZ2 was inoculated into slant culture medium at an inoculation amount of 8%, and activated at 30℃ for 3 days to obtain the activated strain; The activated strain was inoculated into seed culture medium at a rate of 10%, and cultured in a shake flask at 30°C for 14 hours at a shaker speed of 200 r / min to obtain the amplified strain. The amplified strain was inoculated into the fermentation medium at an inoculation amount of 8%, and fermented at 36°C for 50 days with a shaking speed of 200 r / min. After centrifugation, the supernatant was removed to obtain the bacterial agent.

[0035] Application examples Application Example 1: A biological agent, in which Bacillus mucilaginosus HP-JZ2 agent prepared in Preparation Example 1 is added.

[0036] Application Example 2: A biological agent, in which Bacillus mucilaginosus HP-JZ2 agent prepared in Preparation Example 2 is added.

[0037] Application Example 3: A biological agent, in which Bacillus mucilaginosus HP-JZ2 bacterial agent prepared in Preparation Example 3 is added.

[0038] Comparative application examples Comparative Application Example 1: The difference between this comparative application example and Application Example 1 is that: Trehalose was not added to the slant culture medium and seed culture medium during the preparation of Bacillus mucilaginosus HP-JZ2 inoculum.

[0039] Comparative Application Example 2: The difference between this comparative application example and Application Example 1 is that: Isomalt oligosaccharide was not added to the fermentation medium during the preparation of Bacillus mucilaginosus HP-JZ2 inoculant.

[0040] Performance testing 1. Antagonism test The viable counts of Bacillus mucilaginosus HP-JZ2 in application examples 1-3 and comparative application examples 1-2 were measured and the data were recorded. The Bacillus mucilaginosus HP-JZ2 bacterial agent was treated at 45℃ for 12 min, and the number of viable bacteria was detected again and the data was recorded. The Bacillus mucilaginosus HP-JZ2 bacterial agent was treated for 1 day at a relative humidity of 95% and an ambient temperature of 35℃. The number of viable bacteria was then tested again and the data were recorded. Add Bacillus mucilaginosus HP-JZ2 inoculum to a sulfuric acid solution with a pH of 5.5 at a mass ratio of 1:0.5, treat for 1 day, and then test the viable bacteria count again and record the data. The HP-JZ2 inoculant was added to the soil, where actinomycetes and fungi were present. After 30 days, the number of viable Bacillus HP-JZ2 inoculants was tested again, and the data were recorded.

[0041] Table 2 Performance Test Table

[0042] As can be seen from Application Examples 1-3 and Table 2, the Bacillus mucilaginosus HP-JZ2 bacterial agent obtained in this application has a high number of viable bacteria and good resistance to high temperature, high humidity and high acidity. At the same time, it still has high growth and reproduction activity under various microbial competition conditions.

[0043] Combining Application Example 1 and Comparative Application Examples 1-2 with Table 2, it can be seen that trehalose in slant culture medium and seed culture medium, and isomalt oligosaccharide in fermentation culture medium, can promote the growth and reproduction of Bacillus mucilaginosus HP-JZ2, increase the viable count of Bacillus mucilaginosus HP-JZ2 inoculum, and also ensure the resistance of Bacillus mucilaginosus HP-JZ2, so that it can still have good activity and growth and reproduction under harsh conditions.

[0044] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A type of Bacillus mucilaginosus HP-JZ2, characterized in that, Bacillus mucilaginosus krassilnikov HP-JZ2 was deposited at the China Center for Type Culture Collection on November 14, 2022, with accession number CCTCCNO.M20221792.

2. A biological agent, characterized in that: The product contains Bacillus mucilaginosus HP-JZ2 bacterial agent, which is prepared using Bacillus mucilaginosus HP-JZ2 as described in claim 1.

3. The biological agent according to claim 2, characterized in that, The Bacillus mucilaginosus HP-JZ2 inoculum was prepared using the following method: Bacillus mucilaginosus HP-JZ2 was inoculated into slant culture medium at an inoculation amount of 5-8%, and activated at 30-32℃ for 2-3 days to obtain activated strains; The activated strain was inoculated into seed culture medium at an inoculation rate of 6-10%, and cultured in shake flasks at 30-32℃ for 8-14 hours to obtain the amplified strain. The amplified strain was inoculated into the fermentation medium at an inoculation rate of 5-8%, and fermented at 36-38℃ for 46-50 hours. The supernatant was then removed by centrifugation to obtain the bacterial agent.

4. The biological agent according to claim 3, characterized in that, The slant culture medium comprises the following raw materials: potassium aluminosilicate 0.8-1 g / L, sodium aluminosilicate 0.2-0.3 g / L, sucrose 5-8 g / L, trehalose 1-2 g / L, disodium hydrogen phosphate 1-2 g / L, ferric chloride 0.004-0.006 g / L, calcium chloride 0.001-0.002 g / L, magnesium sulfate heptahydrate 0.4-0.5 g / L, agar 18-21.5 g / L, and water to a final volume of 1 L.

5. A biological agent according to claim 3, characterized in that, The seed culture medium comprises the following ingredients: glucose 2-4 g / L, molasses 5-8 g / L, trehalose 1-2 g / L, chitin 0.05-0.1 g / L, dipotassium hydrogen phosphate 0.4-0.6 g / L, magnesium sulfate heptahydrate 0.4-0.6 g / L, sodium chloride 0.1-0.35 g / L, calcium carbonate 0.5-1 g / L, and water to a final volume of 1 L.

6. The biological agent according to claim 3, characterized in that, The fermentation medium comprises the following raw materials: glucose 5-9 g / L, sucrose 2-3 g / L, isomalt oligosaccharide 0.2-0.5 g / L, yeast extract 0.1-0.3 g / L, ammonium sulfate 0.4-0.7 g / L, dipotassium hydrogen phosphate 1-1.5 g / L, calcium carbonate 0.4-0.7 g / L, ferric chloride 0.4-0.7 g / L, sodium chloride 0.1-0.3 g / L, magnesium sulfate heptahydrate 0.1-0.2 g / L, and water to a final volume of 1 L.

7. A biological agent according to claim 3, characterized in that, The shaking speed during the shake flask culture process is 180-200 r / min.

8. A biological agent according to claim 3, characterized in that, The shaking speed during the fermentation process is 180-200 r / min.

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