Microbial fertilizer and application thereof
Microbial fertilizer composed of Bacillus cereus RBC-1 and Klebsiella acidogenetic bacteria LZST 2022-02 solves the problems of high cost and seed germination inhibition in traditional saline-alkali land improvement methods, and achieves saline-alkali soil improvement and plant growth promotion, with environmentally friendly economic benefits.
Patent Information
- Application Number
- CN202511334011.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-18
- Publication Date
- 2025-12-12
AI Technical Summary
In existing technologies, traditional methods for improving saline-alkali soil are costly, prone to repetition, and cause secondary pollution. Microbial remediation methods have inconsistent effects, and seed germination is inhibited under saline-alkali stress. There is a lack of effective microbial fertilizer solutions.
Microbial fertilizer composed of Bacillus cereus RBC-1 and Klebsiella acidogeneticus LZST 2022-02 is prepared as an aqueous or powder formulation by mixing with liquid or solid forms and combining with ingredients such as glycerol, trehalose, gelatin and diatomaceous earth. It is used to improve saline-alkali soil and promote seed germination.
It significantly reduces the toxic effects of salt and alkali on seed germination, promotes plant germination under saline-alkali conditions, improves saline-alkali soil, reduces soil alkali content, enhances plant growth capacity in saline-alkali environments, and is low-cost and environmentally friendly.
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Figure CN121109215A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of microbial technology, specifically relating to a microbial fertilizer and its application. Background Technology
[0002] Soil salinization is a global ecological problem. It not only damages soil structure and reduces soil fertility, but also causes soil compaction and increased osmotic pressure, ultimately leading to reduced crop yields and even food security issues. Therefore, the improvement and rational development and utilization of saline-alkali land are key factors in promoting increased grain production.
[0003] Currently, traditional methods for improving saline-alkali soil, such as physical salt leaching and chemical amendments, suffer from problems such as high cost, repetitive processes, and secondary pollution. Microbial remediation, on the other hand, increases the organic matter content in the soil through microbial communities, improves the root microenvironment of plants, and thus improves crop yield and quality. Microbial remediation is simple to operate, easy to promote, environmentally friendly, and highly sustainable, and has a broader application prospect.
[0004] While there are numerous reports on microbial fertilizers in existing technologies, the application effects vary significantly depending on the strain composition. Since seed germination is the starting point and crucial stage of plant growth and development, it is particularly sensitive to environmental influences, and salt-alkali stress inhibits seed germination. Therefore, the research and development of microbial fertilizers that can not only improve saline-alkali land and promote crop growth, but also promote seed germination under saline-alkali conditions is of great significance. Summary of the Invention
[0005] To address the shortcomings of existing technologies, this invention provides a microbial fertilizer and its application.
[0006] The present invention adopts the following technical solution: A microbial fertilizer comprising Bacillus cereus RBC-1, with accession number CCTCC NO:M 2021024, and Klebsiella acidogenic bacteria LZST 2022-02, with accession number CGMCC NO.24286.
[0007] Preferably, the formulations of the above-mentioned microbial fertilizers include liquid microbial fertilizers and solid microbial fertilizers.
[0008] The preparation method of the above-mentioned microbial fertilizer includes the following steps: Microbial fertilizer was prepared by mixing Bacillus cereus RBC-1 bacterial solution with Klebsiella acidogenic bacteria LZST 2022-02 bacterial solution.
[0009] Preferably, the effective viable count of Bacillus cereus RBC-1 bacterial suspension is 1.0 × 10⁻⁶. 10 cfu / mL or higher; The effective viable count of Klebsiella acidogenic bacteria LZST 2022-02 was 1.0 × 10⁻⁶. 9 cfu / mL or higher.
[0010] Further preferred, the effective viable count of Bacillus cereus RBC-1 bacterial suspension is (1.0~2.3)×10⁻⁶. 10 cfu / mL; The effective viable count of Klebsiella acidogenic bacteria LZST 2022-02 bacterial suspension was (1.0~2.5)×10⁻⁶. 9 cfu / mL.
[0011] Preferably, a microbial fertilizer is prepared by mixing Bacillus cereus RBC-1 bacterial solution and Klebsiella acidogenic bacteria LZST 2022-02 bacterial solution at a volume ratio of 1:(0.8~1). Alternatively, Bacillus cereus RBC-1 bacterial solution and Klebsiella acidogenic bacteria LZST 2022-02 bacterial solution are mixed at a volume ratio of 1:(0.8~1), and then 6~8% glycerol, 2~3% trehalose and 0.5~1% gelatin are added by mass fraction to obtain microbial fertilizer. Alternatively, a bacterial solution of Bacillus cereus RBC-1 and a bacterial solution of Klebsiella pneumoniae LZST 2022-02 can be mixed at a volume ratio of 1:(0.8~1) to obtain a bacterial solution; 45~50 parts by weight of bacterial solution, 43~50 parts by weight of diatomaceous earth, 2~3 parts by weight of sodium dodecylbenzenesulfonate, 3~4 parts by weight of dispersant NNO, 1~2 parts by weight of vitamin C and 1~2 parts by weight of calcium carbonate can be mixed and spray-dried to obtain microbial fertilizer.
[0012] A preferred method for preparing Bacillus cereus RBC-1 bacterial suspension includes the following steps: (1) Inoculate Bacillus cereus RBC-1 into LB liquid medium and culture at 25-30℃ and 180-200 rpm until OD. 600 A value of 0.8~0.9 is used as the seed solution; (2) Inoculate the seed liquid into the fermentation medium at a volume ratio of 5% to 7%, and culture at 25 to 30°C and 180 to 200 rpm for 24 to 27 hours to obtain Bacillus cereus RBC-1 bacterial solution.
[0013] A preferred method for preparing Klebsiella acidogenic bacteria culture LZST 2022-02 includes the following steps: (1) Inoculate Klebsiella acid-producing bacteria LZST 2022-02 into LB liquid medium and culture at 25-30℃ and 180-200 rpm until OD. 600 A value of 0.8~0.9 is used as the seed solution; (2) Inoculate the seed liquid into the fermentation medium at a volume ratio of 5% to 7%, and culture at 25 to 30°C and 180 to 200 rpm for 24 to 27 hours to obtain the Klebsiella acid-producing bacterial solution LZST 2022-02.
[0014] More preferably, each liter of LB liquid culture medium contains: 10g peptone, 5g yeast extract, 10g sodium chloride, and water as the solvent; The fermentation medium consists of the following components per liter: 50g soybean meal powder, 40g corn starch, 30g glucose, 1g magnesium sulfate, 1.5g potassium dihydrogen phosphate, 0.2g manganese sulfate, and water as the solvent, with a pH of 7-7.5.
[0015] The above-mentioned microbial fertilizers are used in any of the following applications, including: (1) Improvement of saline-alkali soil; (2) Promotes seed germination (3) Promote plant growth.
[0016] Preferably, the application of microbial inoculants in promoting seed germination in saline-alkali soils; Application of microbial fertilizers in promoting plant growth in saline-alkali soils.
[0017] Preferably, when applying microbial fertilizer, the dilution ratio is 500 times or more.
[0018] Further preferred, when applying microbial fertilizer, the dilution ratio is 500 to 1000 times.
[0019] Preferably, the plants include wheat, clover, tobacco, and chili peppers.
[0020] The beneficial effects of the present invention include at least the following: 1. The microbial fertilizer provided by this invention can significantly reduce the toxic effects of salt and alkali on seed germination and promote the germination of plant seeds under saline-alkali conditions. This effect is of great significance for the cultivation of crops in saline-alkali environments. Furthermore, the effect of the 1000-fold diluted solution of the microbial fertilizer provided by this invention is even better, and the application cost of this microbial fertilizer is low.
[0021] 2. The microbial fertilizer provided by this invention has good colonization in saline-alkali soil and good salt and alkali tolerance. It can effectively reduce the alkali content in the soil and can be used to improve saline-alkali soil. It has a high stress resistance and growth promotion effect on plants and can promote plant growth in saline-alkali soil. It has a good application prospect in agricultural industry. Attached Figure Description
[0022] Figure 1 The growth curves of Bacillus cereus RBC-1 seed culture at different salt concentrations are shown.
[0023] Figure 2 The growth curves of Klebsiella acidogenic strain LZST 2022-02 seed culture under different salt concentrations are shown.
[0024] Figure 3 Growth curves of Bacillus cereus RBC-1 and Klebsiella acidogenic bacteria LZST 2022-02 composite seed culture at different salt concentrations.
[0025] Figure 4 The effect of the compound microbial fertilizer of Bacillus cereus and Klebsiella acidogeneticus provided in this invention on wheat seed germination under salt stress is shown in the figure.
[0026] Figure 5 The effect of the compound microbial fertilizer of Bacillus cereus and Klebsiella acidogeneticus provided in this invention on the germination of clover seeds under salt stress is shown in the figure.
[0027] Figure 6 The figure shows the effect of the compound microbial fertilizer of Bacillus cereus and Klebsiella acidogeneticus provided by the present invention on the growth of Nicotiana benthamiana under salt stress.
[0028] Figure 7 The effect of the compound microbial fertilizer of Bacillus cereus and Klebsiella acidogeneticus provided by the present invention on the growth of chili peppers under salt stress is shown in the figure.
[0029] Figure 8 The effect of the compound microbial fertilizer of Bacillus cereus and Klebsiella acidogeneticus provided by the present invention on the growth of chili peppers under salt stress is shown in the figure. Detailed Implementation
[0030] The present invention will be further described in detail below with reference to specific embodiments. The given embodiments are only for illustrating the present invention, but the scope of protection of the present invention is not limited thereto. Unless otherwise specified, the experimental methods in the following embodiments are conventional methods. Unless otherwise specified, the materials and reagents used in the following embodiments are commercially available.
[0031] Bacillus cereus RBC-1 (accession number CCTCC NO: M 2021024) and Klebsiella oxytoca LZST 2022-02 (accession number CGMCC No. 24286) have been disclosed in patent literature and are considered prior art.
[0032] Example 1 Colonization of microbial agents in saline-alkali soil Six salt-tolerant candidate bacteria strains preserved in the laboratory were inoculated into LB liquid medium and cultured at 30°C with shaking for 24 h. After centrifugation at 4000 rpm for 10 min, the bacterial cells were collected, washed three times with sterile physiological saline, and resuspended to a concentration of 1×10⁻⁶. 8 CFU / mL; Take 500g of self-made saline-alkali soil (pH 9.0, salt content 0.48%) and dispense it into sterile petri dishes, 100g per dish. Spray 10mL of bacterial suspension evenly onto the soil surface, cover with plastic wrap, poke holes for ventilation, and place in a 30℃ incubator. Weigh and replenish water daily to maintain soil moisture content at 20%. Subsequently, take soil samples regularly at sampling time points: 0h, day 1, day 3, and day 7. Weigh 1g of soil sample, add 9mL of sterile physiological saline, vortex for 10min, let stand for 2min, and take the supernatant for serial dilution. Spread the supernatant onto screening LB medium (containing 50μg / mL nystatin) and incubate at 30℃ for 48h. Count the number of typical colonies. Colony survival rate (%) = bacterial count on day 7 ÷ initial bacterial count × 100%. The test results are shown in Table 1.
[0033] Table 1
[0034] According to experimental data, Bacillus cereus RBC-1 and Klebsiella acidogenic bacteria LZST 2022-02 showed better colonization in saline-alkali soil.
[0035] Example 2 Alkali reduction ability Six salt-tolerant candidate bacteria preserved in the laboratory were inoculated onto LB solid medium (pH=10) and cultured at 30°C. The growth status of the strains was observed every 24 hours. Strains capable of growing on alkaline plates were selected, and single colonies were picked with an inoculation loop and inoculated into LB liquid medium (pH=10) and cultured at 30°C. The pH of the fermentation broth and the growth status of the strains were measured every 12 hours. The alkalinity-reducing ability was calculated using the following formula: η .
[0036] In the formula: η is the alkalinity-reducing ability of the strain, pH before is the original pH value of the unfermented culture medium, and pH after is the pH value of the culture medium after fermentation.
[0037] The experimental results are shown in Table 2.
[0038] Table 2
[0039] Based on experimental data, the three strains of Bacillus cereus RBC-1, Klebsiella acidogenic bacteria LZST 2022-02, and Bacillus megaterium showed good alkalinity reduction ability. The inventors found that Bacillus cereus RBC-1 and Klebsiella acidogenic bacteria LZST2022-02 had better alkalinity reduction ability and colonization in saline-alkali soil.
[0040] Example 3 The preparation of Bacillus cereus RBC-1 seed culture, Klebsiella acidogenic bacteria LZST 2022-02 seed culture, and compound microbial seed culture includes the following steps: The composition of the culture medium is shown in Tables 3, 4 and 5.
[0041] Table 3
[0042] Table 4
[0043] Table 5
[0044] (1) Activation of bacterial strains: Bacillus cereus RBC-1 was activated on LB solid medium and incubated upside down in a 25℃ incubator for 24 hours to obtain single colonies for later use; Klebsiella acidogenic LZST 2022-02 was activated on LB solid medium and incubated upside down in a 28℃ incubator for 24 hours to obtain single colonies for later use; (2) Preparation of seed culture: A single colony of activated Bacillus cereus RBC-1 was inoculated into LB liquid medium and cultured at 25℃ and 200rpm for 12h to obtain seed culture. The OD of the seed culture was... 600 The value is 0.8-0.9.
[0045] Single colonies of activated Klebsiella acidogenic bacteria (LZST 2022-02) were inoculated into LB broth and cultured at 28°C and 200 rpm for 12 hours to obtain a seed culture. The OD of the seed culture was... 600 The value is 0.8-0.9.
[0046] (3) Fermentation preparation: The Bacillus cereus RBC-1 seed culture was inoculated into the fermentation medium for fermentation. The initial pH of the fermentation medium was 7-7.5, the liquid volume was 30L / 50L, the seed culture inoculation was 5% by volume, the fermentation temperature was 30℃, the fermentation speed was 180r / min, and the fermentation time was 24h, thus obtaining the Bacillus cereus RBC-1 fermentation broth, with an effective viable count of 2.3×10⁻⁶. 10 cfu / mL.
[0047] The seed culture of Klebsiella acidogenic LZST 2022-02 was inoculated into fermentation medium for fermentation. The initial pH of the fermentation medium was 7-7.5, the liquid volume was 30L / 50L, the inoculation amount of the seed culture was 5% by volume, the fermentation temperature was 30℃, the fermentation speed was 180 r / min, and the fermentation time was 24 h, yielding the Klebsiella acidogenic LZST 2022-02 fermentation broth with an effective viable count of 2.5 × 10⁻⁶ cells / day. 9 cfu / mL.
[0048] The compound seed liquid is a mixture of Bacillus cereus RBC-1 fermentation broth and Klebsiella acidogenic bacteria LZST 2022-02 fermentation broth at a volume ratio of 1:1.
[0049] Salt tolerance test on the growth of Bacillus cereus RBC-1 seed culture, Klebsiella acidogenic LZST 2022-02 seed culture, Bacillus cereus RBC-1 seed culture, and Klebsiella acidogenic LZST 2022-02 composite seed culture under different salinity conditions: LB liquid medium was used as the basal medium, with NaCl added at mass fractions of 5%, 8%, 10%, and 13%, respectively. After activating the seed culture in LB liquid medium, the Bacillus cereus RBC-1 seed culture, Klebsiella acidogenic LZST 2022-02 seed culture, Bacillus cereus RBC-1 seed culture, and Klebsiella acidogenic LZST 2022-02 1:1 composite seed culture were inoculated at a volume fraction of 2% into pre-prepared LB medium with different salt concentration gradients, with three replicates for each salt concentration gradient. The inoculated culture medium was placed in a shaker at 28℃ and 180 rpm. Every two hours, the absorbance of the culture medium was measured at 600 nm, and a growth curve was plotted. Figure 1 , Figure 2 and Figure 3 The inventors discovered that mixing Bacillus cereus RBC-1 with Klebsiella pneumoniae LZST 2022-02 resulted in better salt tolerance.
[0050] Example 4 Preparation of microbial fertilizers: Microbial fertilizers are available in liquid and powder forms. Aqueous microbial fertilizer I: The fermentation broth of Bacillus cereus RBC-1 prepared in Example 3 was mixed with the fermentation broth of Klebsiella pneumoniae LZST 2022-02 at a volume ratio of 1:1, and then used as aqueous microbial fertilizer I.
[0051] Aqueous microbial fertilizer II: The fermentation broth of Bacillus cereus RBC-1 prepared in Example 3 was mixed with the fermentation broth of Klebsiella pneumoniae LZST 2022-02 at a volume ratio of 1:1. Then, 8% glycerol, 2% trehalose and 0.5% gelatin were added by mass fraction to prepare aqueous microbial fertilizer II.
[0052] The purpose of adding glycerol, trehalose, and gelatin to the liquid microbial fertilizer II is to protect the activity of the microbial agent during storage and extend the shelf life of the microbial fertilizer.
[0053] Powdered microbial fertilizer: The fermentation broth of Bacillus cereus RBC-1 prepared in Example 3 was mixed with the fermentation broth of Klebsiella acidogenic bacteria LZST 2022-02 at a volume ratio of 1:1 to obtain the bacterial solution.
[0054] By weight, 45 parts of bacterial solution, 50 parts of diatomaceous earth, 3 parts of sodium dodecylbenzenesulfonate, 3 parts of dispersant NNO, 1 part of vitamin C and 1 part of calcium carbonate are mixed and spray-dried to produce a wettable powder, which is the powdered microbial fertilizer.
[0055] Example 5 Effects of aqueous microbial fertilizer I on wheat seed germination under salt stress (1) Wheat seed pretreatment: During the germination period, the petri dish germination method is adopted. First, wheat seeds of uniform size and full grains are disinfected by soaking in 35% hydrogen peroxide for 5 minutes, and then washed with distilled water until the pungent odor disappears.
[0056] (2) Seed dressing: The experiment was designed with 3 treatments. The CK group (blank) took the rinsed seeds and placed them in a petri dish with 2 layers of filter paper. 100 seeds were used in each treatment. The CK group (blank) took 1.5 mL of distilled water for seed dressing. Treatment 1 took 1.5 mL of 500 times diluted liquid microbial fertilizer for seed dressing. Treatment 2 took 1.5 mL of 1000 times diluted liquid microbial fertilizer for seed dressing. After 60 minutes, the wheat seeds were taken out and dried for later use.
[0057] (3) Line the seed germination box with filter paper and add 9 mL of the prepared saline-alkali solution (see Table 6) to completely wet the filter paper. Arrange the treated wheat seeds neatly on the filter paper, using it as a germination bed. Simulate the germination environment by avoiding light. Record the number of germinated seeds in each group. All treatments were designed with 3 biological replicates. Germination test results: Germination rate = [number of germinated seeds on day 7 / total number of seeds in the petri dish] × 100%, Germination potential = [number of germinated seeds on day 3 / total number of seeds in the petri dish] × 100%.
[0058] (4) Experimental results are shown in Table 7: The application of aqueous microbial fertilizer I significantly improved the germination rate and germination potential of wheat, and significantly reduced the toxic effects of salt and alkali on seed germination. The effect of the 1000-fold dilution was even better, and it had a significant promoting effect on the early germination of wheat. Figure 4 .
[0059] Table 6. Preparation methods of saline-alkali solutions
[0060] Table 7
[0061] Example 6 Effects of aqueous microbial fertilizer I on clover seed germination under salt stress (1) Pretreatment of clover seeds: During the germination period, the petri dish germination method is adopted. First, place clover seeds of uniform size and full grains in a petri dish with two layers of filter paper, soak them in distilled water for 60 minutes, and then take out the clover seeds and dry them for later use.
[0062] (2) Seed dressing: The experiment was designed with 3 treatments. The CK group (blank) took the rinsed seeds and placed them in a petri dish with 2 layers of filter paper. Each treatment had 400 seeds. The CK group (blank) took 1.5 mL of distilled water for seed dressing. Treatment 1 took 1.5 mL of 500 times diluted liquid microbial fertilizer for seed dressing. Treatment 2 took 1.5 mL of 1000 times diluted liquid microbial fertilizer for seed dressing. After 60 minutes, the clover seeds were taken out and dried for later use.
[0063] (3) Line the seed germination box with filter paper and add 9 ml of the prepared saline-alkali solution (see Table 6) to completely wet the filter paper. Arrange the treated clover seeds neatly on the filter paper, using the filter paper as the germination bed, and simulate the germination environment by avoiding light. Record the number of germinated seeds in each group. All treatments were designed with 3 biological replicates. Germination rate was determined as follows: Germination rate = [Number of germinated seeds on day 7 / Total number of seeds in the petri dish] × 100%.
[0064] Table 8
[0065] (4) The experimental results are shown in Table 8: The application of aqueous microbial fertilizer I significantly improved the germination rate of clover and effectively reduced the toxic effects of salt and alkali on seed germination. The effect of the 1000-fold dilution was even better, and it significantly promoted the early germination of clover. Figure 5 .
[0066] Example 7 Effects of aqueous microbial fertilizer I on tobacco growth promotion under salt stress Pot experiment: The test plant was *Nicotiana benthamiana*, and the experimental soil was sandy saline-alkali soil from a vegetable garden with a pH of 7.65 and a salt content of 4.13 g / kg. *Nicotiana benthamiana* plants that had grown for 20 days and exhibited uniform growth were selected for treatment. Three treatments were set up: one control group (no fertilizer) was watered with 100 ml; one group was treated with 1 g urea per pot and watered with 100 ml; and one group was treated with 100 mL of a 1000-fold diluted liquid microbial fertilizer. Each treatment had four replicates. Plant height was measured on days 10 and 18 after treatment to observe the effect on tobacco growth. The experimental results showed that the 1000-fold diluted liquid microbial fertilizer had a significant growth-promoting effect on *Nicotiana benthamiana* and improved its tolerance to saline-alkali stress (see Table 9). Figure 6 .
[0067] Table 9
[0068] Example 8 Effects of aqueous microbial fertilizer II on pepper growth promotion under salt stress Pot experiment: The test plant was the screw pepper, and the experimental soil was sandy saline-alkali soil from a vegetable garden with a pH of 7.50 and a salt content of 4.01 g / kg. Peppers with uniform growth after 20 days were selected for treatment. Five treatments were set up: one control group (no fertilizer) watered with 100 ml; one group treated with 1 g urea per pot and watered with 100 ml; one group treated with 100 mL of a 500-fold diluted liquid microbial fertilizer; one group treated with 100 mL of a 1000-fold diluted liquid microbial fertilizer; and one group treated with 100 mL of a 2000-fold diluted liquid microbial fertilizer. Each treatment had 10 replicates. Plant height was measured 11 days after treatment to observe the effect on pepper growth. The experimental results showed that the 500-fold and 1000-fold diluted liquid microbial fertilizers significantly promoted pepper growth and improved tolerance to saline-alkali stress (see Table 10). Figure 7 , Figure 8 .
[0069] Table 10
[0070] The microbial fertilizer provided by this invention can significantly reduce the toxic effects of salt and alkali on seed germination and promote seed germination under saline-alkali conditions. This effect is of great significance for the cultivation of crops in saline-alkali environments. The microbial fertilizer provided by this invention exhibits good establishment in saline-alkali soils and also demonstrates excellent salt and alkali tolerance, effectively reducing the alkali content in the soil and thus serving as a soil amendment for saline-alkali soils. It also shows high stress resistance and growth-promoting effects on plants, promoting their growth in saline-alkali soils. The 1000-fold dilution of the microbial fertilizer provided by this invention further enhances its effectiveness. This microbial fertilizer has low application costs and shows great promise for application in the agricultural industry.
Claims
1. A microbial fertilizer, characterized in that, It contains Bacillus cereus RBC-1, with accession number CCTCC NO:M2021024 and Klebsiella acidogenetic LZST 2022-02, with accession number CGMCC NO.24286.
2. The microbial fertilizer as described in claim 1, characterized in that, The formulations of the microbial fertilizer include liquid microbial fertilizer and solid microbial fertilizer.
3. The method for preparing the microbial fertilizer according to claim 1, characterized in that, Includes the following steps: Microbial fertilizer was prepared by mixing Bacillus cereus RBC-1 bacterial solution with Klebsiella acidogenic bacteria LZST 2022-02 bacterial solution.
4. The preparation method according to claim 3, characterized in that, The effective viable count of Bacillus cereus RBC-1 bacterial suspension was 1.0 × 10⁻⁶. 10 cfu / mL or higher; The effective viable count of Klebsiella acidogenic bacteria LZST 2022-02 was 1.0 × 10⁻⁶. 9 cfu / mL or higher; Preferably, the effective viable count of Bacillus cereus RBC-1 bacterial suspension is (1.0~2.3)×10⁻¹⁰. 10 cfu / mL; The effective viable count of Klebsiella acidogenic bacteria LZST 2022-02 bacterial suspension was (1.0~2.5)×10⁻⁶. 9 cfu / mL.
5. The preparation method according to claim 3, characterized in that, Microbial fertilizer was prepared by mixing Bacillus cereus RBC-1 bacterial solution and Klebsiella acidogenic bacteria LZST 2022-02 bacterial solution at a volume ratio of 1:(0.8~1). Alternatively, Bacillus cereus RBC-1 bacterial solution and Klebsiella acidogenic bacteria LZST 2022-02 bacterial solution are mixed at a volume ratio of 1:(0.8~1), and then 6~8% glycerol, 2~3% trehalose and 0.5~1% gelatin are added by mass fraction to obtain microbial fertilizer. Alternatively, a bacterial solution of Bacillus cereus RBC-1 and a bacterial solution of Klebsiella pneumoniae LZST 2022-02 can be mixed at a volume ratio of 1:(0.8~1) to obtain a bacterial solution; 45~50 parts by weight of bacterial solution, 43~50 parts by weight of diatomaceous earth, 2~3 parts by weight of sodium dodecylbenzenesulfonate, 3~4 parts by weight of dispersant NNO, 1~2 parts by weight of vitamin C and 1~2 parts by weight of calcium carbonate can be mixed and spray-dried to obtain microbial fertilizer.
6. The preparation method according to claim 3, characterized in that, The method for preparing Bacillus cereus RBC-1 bacterial suspension includes the following steps: (1) Inoculate Bacillus cereus RBC-1 into LB liquid medium and culture at 25-30℃ and 180-200 rpm until OD. 600 A value of 0.8~0.9 is used as the seed solution; (2) Inoculate the seed culture into the fermentation medium at a volume ratio of 5% to 7%, and culture at 25 to 30°C and 180 to 200 rpm for 24 to 27 hours to obtain Bacillus cereus RBC-1 bacterial culture; A preferred method for preparing Klebsiella acidogenic bacteria culture LZST 2022-02 includes the following steps: (1) Inoculate Klebsiella acid-producing bacteria LZST 2022-02 into LB liquid medium and culture at 25-30℃ and 180-200 rpm until OD. 600 A value of 0.8~0.9 is used as the seed solution; (2) Inoculate the seed culture into the fermentation medium at a volume ratio of 5% to 7%, and culture at 25 to 30°C and 180 to 200 rpm for 24 to 27 hours to obtain Klebsiella acid-producing bacterial culture LZST 2022-02; Preferably, each liter of LB liquid culture medium contains: 10g peptone, 5g yeast extract, 10g sodium chloride, and water as the solvent; The fermentation medium consists of the following components per liter: 50g soybean meal powder, 40g corn starch, 30g glucose, 1g magnesium sulfate, 1.5g potassium dihydrogen phosphate, 0.2g manganese sulfate, and water as the solvent, with a pH of 7-7.
5.
7. The use of the microbial fertilizer according to any one of claims 1-2 or the microbial fertilizer prepared by the method according to any one of claims 3-6 in any of the following, including: (1) Improvement of saline-alkali soil; (2) Promotes seed germination (3) Promote plant growth.
8. The application as described in claim 7, characterized in that, Application of microbial inoculants in promoting seed germination in saline-alkali soils; Application of microbial fertilizers in promoting plant growth in saline-alkali soils.
9. The application as described in claim 7, characterized in that, When applying microbial fertilizer, the dilution ratio should be 500 times or more; Preferably, when applying microbial fertilizer, the dilution ratio is 500 to 1000 times.
10. The application as described in claim 7, characterized in that, The plants mentioned include wheat, clover, tobacco, and chili peppers.