A composition for inhibiting wheat pathogens, its preparation method and application

Abstract

This invention relates to a composition for inhibiting wheat pathogens, its preparation method, and its application. Specifically, it relates to a composition for inhibiting wheat pathogens, the components of which include *Bacillus belye* (…). Bacillus velezensis The invention comprises a microbial agent, a Zr-metal-organic framework material, and a protective agent; its components, by mass parts, are: 10-50 parts of Bacillus belyssioides inoculant, 10-80 parts of Zr-metal-organic framework, and 10-30 parts of protective agent. This invention can simultaneously solve two major problems in wheat production: rampant disease and saline-alkali soil stress, and has promising application prospects and significant economic and ecological value.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and in particular to a composition for inhibiting wheat pathogens, its preparation method, and its application. Background Technology

[0002] As a major global food crop, the safe production of wheat is crucial to national food strategies. However, modern intensive wheat cultivation faces two major challenges. First, wheat is susceptible to various pathogens throughout its growth cycle, such as Fusarium graminearum, which causes Fusarium head blight. Fusarium graminearum Fusarium pseudobulb, which causes stem base rot, Fusarium pseudograminearum Rhizoctonia solani, which causes sheath blight ( Rhizoctonia solan i) and Rhizoctonia granatum ( Rhizoctonia cerealis ), and Botrytis cinerea, which causes leaf spot disease ( Botrytis cinerea Currently, the control of these diseases still heavily relies on chemical pesticides. Long-term, excessive use of chemical pesticides not only easily leads to pesticide resistance in pathogens, causing a decline in control effectiveness, but also results in a series of problems such as pesticide residues, environmental pollution, disruption of soil microecological balance, and harm to human and animal health. Therefore, developing efficient, safe, and environmentally friendly biological control methods to replace or reduce the use of chemical pesticides has become an urgent need for sustainable agricultural development. Secondly, globally, the area of ​​saline-alkali land is expanding, becoming a significant factor limiting agricultural production. High salinity and alkalinity environments cause osmotic stress and ion toxicity to wheat plants, inhibiting seed germination, hindering root development, leading to stunted growth, yellowing leaves, and ultimately severe yield reduction. Traditional methods for improving saline-alkali land, such as water conservancy projects and soil replacement, have limitations such as high cost, long cycle, and difficulty in maintaining lasting effects. Although some microbial agents have been tried to alleviate plant salinity and alkalinity stress, most strains have problems such as single function, poor colonization ability in saline-alkali stress, and unstable activity, resulting in less than ideal effects in practical applications.

[0003] In summary, current technologies lack a comprehensive solution that can effectively address both wheat pathogen infestation and high salinity stress simultaneously. Current biocontrol agents often have narrow antimicrobial spectra and insufficient stability and efficacy under adverse conditions; while some growth-promoting agents lack significant control effects against multiple important diseases. Therefore, there is an urgent need in this field for a microbial preparation that combines broad-spectrum and highly effective antimicrobial activity with potent salinity-tolerant growth-promoting properties to ensure healthy wheat growth and yield under both normal and adverse conditions. Summary of the Invention

[0004] Based on the above problems, this invention has made an innovative discovery of the control effect of Bacillus belye in wheat pathogens. Moreover, this strain has high salt and alkali tolerance and relatively stable tolerance. It can not only improve the salt and alkali tolerance of wheat but also promote wheat growth. This solves the problems of unstable effect and intolerance to high salt and alkali in the actual application of biocontrol bacteria for the control of wheat pathogens.

[0005] This invention provides a composition for inhibiting wheat pathogens, the components of which include Bacillus belye (…). Bacillus velezensis Inoculants, Zr-metal-organic framework materials, and protective agents.

[0006] Furthermore, its components, by mass parts, are: 10-50 parts of Bacillus belysin inoculum, 10-80 parts of Zr-organic metal framework, and 10-30 parts of protectant.

[0007] Furthermore, the Bacillus berberis inoculant is a mixture of four Bacillus berberis strains, namely Bacillus berberis LTS-AF1 (… Bacillus velezensis LTS-AF1), Bacillus belysinus LIS-AF2 ( Bacillus velezensis LIS-AF2), Bacillus belysinus LTS-AF3 ( Bacillus velezensis LTS-AF3), Bacillus belysinus LTS-AF4 ( Bacillus velezensis LTS-AF4); the effective viable count of the Bacillus vesiculosus agent is ≥10. 10 CFU / mL.

[0008] The Bacillus berberis LTS-AF1 ( Bacillus velezensis LTS-AF1), its accession number is CCTCC NO: M 2025378;

[0009] The Bacillus berberis LTS-AF2 ( Bacillus velezensis LIS-AF2), its accession number is CCTCC NO: M 2025379;

[0010] The Bacillus berberis LTS-AF3 ( Bacillus velezensis LTS-AF3), its accession number is CCTCC NO: M 2025380;

[0011] The Bacillus berberis LTS-AF4 ( Bacillus velezensis LTS-AF4), its accession number is CCTCC NO: M 2025381;

[0012] All of the above-mentioned Bacillus belyes are deposited at the China Center for Type Culture Collection, located at Wuhan University, Wuhan, China, on March 5, 2025.

[0013] Furthermore, the *Bacillus vesiculosus* inoculum is composed of *Bacillus vesiculosus* LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 bacterial solutions, with a volume ratio of (2~3):(2~5):(1~5):(1~4). In each bacterial solution, the effective viable count of *Bacillus vesiculosus* strain is ≥10. 9 CFU / mL.

[0014] Furthermore, the Zr-metal-organic framework material includes at least one of UiO-66 dry powder, UiO-66-OH, and UiO-66-COOH.

[0015] Furthermore, the protective agent includes trehalose and / or maltodextrin.

[0016] Furthermore, the dosage form of the composition is a solid powder or a liquid formulation.

[0017] Furthermore, the pathogens include Fusarium graminearum, Fusarium, Fusarium pseudograminearum, Rhizoctonia solani, Rhizoctonia graminearum, and Botrytis cinerea.

[0018] The present invention also provides a method for preparing the above-described composition for inhibiting wheat pathogens, comprising the following steps:

[0019] 1) Preparation of Bacillus vesiculus inoculum: Bacillus vesiculus LTS-AF1, LTS-AF2, LTS-AF3 and LTS-AF4 were fermented and cultured separately, and their bacterial solutions were mixed in volume ratio to obtain Bacillus vesiculus inoculum.

[0020] 2) Permeation adsorption: The bacterial agent prepared in step 1) is mixed with a suspension of Zr-metal-organic framework material and treated under vacuum conditions to obtain a mixture;

[0021] 3) In-situ embedding: Add the protectant solution to the mixture prepared in step 2), stir and mix evenly to obtain a composition that inhibits wheat pathogens.

[0022] Preferably, in step 1), the fermentation and cultivation of Bacillus belyssus LTS-AF1, LTS-AF2, LTS-AF3 and LTS-AF4 further includes the addition of inducers, wherein the inducers include sodium stearate and / or nano-ferric oxide; the amount of sodium stearate added as the inducer is 0.1%-0.5% of the total weight of the Bacillus belyssus inoculum, and the amount of nano-ferric oxide added as the inducer is 0.05%-0.1% of the total weight of the Bacillus belyssus inoculum.

[0023] Preferably, in step 2), the vacuum condition is 25-35 kPa, and the processing time is 5-10 min.

[0024] Preferably, in step 3), the stirring time is 5-10 minutes and the embedding temperature is ≤30℃.

[0025] The present invention further provides the application of the above-described composition for inhibiting wheat pathogens in the preparation of formulations for the prevention and control of wheat diseases.

[0026] The present invention further provides the application of the above-described composition for inhibiting wheat pathogens in enhancing wheat's tolerance to abiotic stress, wherein the abiotic stress includes at least one of high temperature, low temperature, acid-base stress, or high salt stress.

[0027] The present invention further provides the application of the above-described composition for inhibiting wheat pathogens in promoting wheat growth, wherein the application is carried out in a saline-alkali environment and is carried out throughout the entire growth cycle of wheat by means of foliar spraying, soil application or inter-row irrigation.

[0028] Furthermore, wheat germination rate can be improved by applying the fertilizer to the soil or soaking seeds during the sowing period; and / or wheat disease resistance can be improved by applying the fertilizer to the soil and spraying the seeds during the flowering period.

[0029] Beneficial effects

[0030] This invention successfully provides a multifunctional microbial preparation that integrates broad-spectrum antibacterial properties, stress resistance and growth promotion, environmental friendliness, and high stability. It can simultaneously solve two major problems in wheat production: rampant disease and saline-alkali soil stress. It has good application prospects and huge economic and ecological value.

[0031] The four specific Bacillus belyssus strains (LTS-AF1 to LTS-AF4) and their combinations used in this invention exhibit strong inhibitory activity against a variety of key pathogens of wheat, including Fusarium graminearum, Fusarium pseudograminearum, Rhizoctonia solani, Rhizoctonia graminearum, and Botrytis cinerea. This demonstrates an excellent synergistic effect among the strains, providing a comprehensive and efficient biological control solution.

[0032] The composition of this invention, as a microbial preparation, can directly inhibit pathogens, providing an effective alternative or supplement to chemical pesticides. Its large-scale application will significantly reduce the amount of chemical pesticides used in the field, thereby reducing pesticide residues, environmental pollution, and delaying the development of pathogen resistance, which aligns with the development direction of green and sustainable agriculture.

[0033] The composition of this invention exhibits strong salt and alkali tolerance and effectively enhances the survival and growth capacity of wheat under salt and alkali stress. Under severe salt and alkali stress, soaking seeds using this invention increases germination rate; during the mid-growth stage, it significantly improves plant height, fresh weight, and survival rate, with a significant increase in survival rate even in severely saline-alkali soils. This indicates that this invention effectively alleviates the osmotic stress and ion toxicity of salt and alkali on wheat, ensuring normal crop growth under adverse conditions. Even under normal, stress-free conditions, this invention promotes wheat growth. This growth-promoting effect is even more pronounced under salt and alkali stress. By improving root development and enhancing nutrient absorption, this invention lays a solid foundation for increasing wheat biomass and final yield.

[0034] The composition of this invention can activate wheat's own defense system. This invention not only directly inhibits pathogens but also systematically enhances wheat's disease resistance, forming a biological protective barrier from the inside out. Through a unique Zr-metal-organic framework (UiO-66 series) carrier and in-situ embedding technology, the live bacteria in this invention are efficiently immobilized and protected. Furthermore, it exhibits excellent environmental adaptability and long-lasting effects, greatly ensuring the product's shelf life and field application effectiveness. Attached Figure Description

[0035] Figure 1 OD concentration of Bacillus vesiculosus strains at different fermentation times 600 Change diagram;

[0036] Figure 2 Viable bacteria count diagram of the solid powder of this invention;

[0037] Figure 3 Viable bacteria count diagram of the liquid formulation of this invention;

[0038] Figure 4 A graph showing the change in viable bacterial count in the formulation of this invention under different salinity and alkalinity conditions. Detailed Implementation

[0039] To make the objectives, technical solutions, and advantages of this application clearer, a more detailed description is provided below. However, it should be understood that the description herein is merely for explaining this application and is not intended to limit its scope.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of this application. All reagents and instruments used herein are commercially available, and the characterization methods involved can be found in relevant descriptions in the prior art, and will not be repeated here.

[0041] To further understand this application, the following detailed description is provided in conjunction with the preferred embodiments.

[0042] Example 1

[0043] This embodiment provides a composition for inhibiting wheat pathogens, the components of which include Bacillus belye (…). Bacillus velezensis Inoculants, Zr-metal-organic framework materials, and protective agents.

[0044] As a further preferred embodiment, the components, by mass parts, are: 10-50 parts of Bacillus belysin inoculum, 10-80 parts of Zr-organic metal framework, and 10-30 parts of protective agent.

[0045] Preferably, the amount of Bacillus belye inoculum is 10-50 parts, for example, 10, 15, 20, 25, 30, 35, 40, 45, 50 parts or any value between any two values;

[0046] The Zr-metal-organic framework material is 10-80 parts, for example, it can be 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80 parts or any value between any two values;

[0047] The protective agent is 10-30 parts, for example, it can be 10, 11, 12, 13, 14, 13, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 parts or any value between any two numbers;

[0048] This invention is a synergistic "three-in-one" system consisting of an active ingredient (microbial agent), a functional carrier (Zr-metal-organic framework), and a stabilizer (protectant). The microbial agent is responsible for the core functions of antibacterial and growth-promoting; the Zr-metal-organic framework, through adsorption and encapsulation, plays a role in fixing, protecting, and slowing down the release of microorganisms, greatly improving the survival time and efficiency of the microbial agent in the field; the protectant protects the activity of microorganisms during processing and storage, preventing damage such as dehydration.

[0049] As a further preferred embodiment, the Bacillus berberis inoculant is a mixture of four Bacillus berberis strains, namely Bacillus berberis LTS-AF1 (… Bacillus velezensis LTS-AF1), Bacillus belysinus LIS-AF2 ( Bacillus velezensis LIS-AF2), Bacillus belysinus LTS-AF3 ( Bacillus velezensis LTS-AF3), Bacillus belysinus LTS-AF4 ( Bacillus velezensis LTS-AF4); the effective viable count of the Bacillus vesiculosus agent is ≥10. 10 CFU / mL.

[0050] The Bacillus berberis LTS-AF1 ( Bacillus velezensis LTS-AF1), its accession number is CCTCC NO: M 2025378;

[0051] The Bacillus berberis LTS-AF2 ( Bacillus velezensis LIS-AF2), its accession number is CCTCC NO: M 2025379;

[0052] The Bacillus berberis LTS-AF3 ( Bacillus velezensis LTS-AF3), its accession number is CCTCC NO: M 2025380;

[0053] The Bacillus berberis LTS-AF4 ( Bacillus velezensis LTS-AF4), with accession number CCTCC NO: M 2025381.

[0054] All of the above-mentioned Bacillus belyes are deposited at the China Center for Type Culture Collection, located at Wuhan University, Wuhan, China, on March 5, 2025.

[0055] Preferably, the Bacillus berberis inoculant is composed of a mixture of Bacillus berberis LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 bacterial solutions, wherein the volume ratio of the LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 bacterial solutions is (2~3):(2~5):(1~5):(1~4), and the effective viable count of Bacillus berberis strain in each bacterial solution is ≥10. 9 CFU / mL.

[0056] As a further preferred embodiment, the Zr-metal-organic framework material includes at least one of UiO-66 dry powder, UiO-66-OH, and UiO-66-COOH.

[0057] Uio-66, as a Zr-metal-organic framework, can provide a highly efficient and stable multifunctional carrier. In practical applications, it has both slow-release and microbial protection functions, which can enhance biocontrol agents and improve soil water and fertilizer utilization. The strength of the Zr-O bond gives UiO-66 excellent chemical and thermal stability. It maintains its structural integrity even in acidic, alkaline, or high-temperature soil environments and will not release harmful metal ions, which meets the requirements of sustainable agricultural development.

[0058] The hydroxyl (-OH) and carboxyl (-COOH) groups on the surface of Uio-66 are polar, hydrogen-bonding functional groups that can form multi-point hydrogen bonds and electrostatic adsorption with the amino, hydroxyl, and teichoic acid groups on the Bacillus cell wall, significantly improving the adsorption rate of cells on solid carriers. The carboxyl groups partially dissociate into -COO- at pH 6-8, generating strong electrostatic attraction with the positive potential (amino groups) on the cell surface, further enhancing the immobilization effect. The hydroxyl groups increase surface hydrophilicity, ensuring good dispersion of the carrier in liquid culture media and preventing particle aggregation that would reduce the cell contact area.

[0059] Therefore, the selected UiO-66 series material carrier not only provides physical protection, but its Zr-O bonds also offer excellent chemical stability, making it suitable for agricultural environments. Furthermore, the -OH and -COOH functional groups can tightly bind to the bacterial cell wall through hydrogen bonds and electrostatic interactions, increasing the loading rate. Its abundant porous structure and surface functional groups (-OH, -COOH) can efficiently load and immobilize the bacteria, enabling slow release of the bacterial agent and prolonging the action time. Simultaneously, the carrier itself is non-toxic and harmless, does not affect bacterial activity, and works synergistically with the bacterial strain, ultimately resulting in a significantly larger actual inhibition zone diameter than that of free bacterial solutions and traditional carriers such as sodium alginate.

[0060] As a further preferred embodiment, the protective agent includes trehalose and / or maltodextrin.

[0061] Trehalose is a recognized excellent bioprotectant that can stabilize proteins and membrane structures during dehydration; maltodextrin acts as a filler and film-forming agent to help form solid powders.

[0062] As a further preferred embodiment, the dosage form of the composition is a solid powder or a liquid formulation.

[0063] As a further preferred embodiment, the pathogens include Fusarium graminearum, Fusarium, Fusarium pseudograminearum, Rhizoctonia solani, and Botrytis cinerea.

[0064] Example 2

[0065] This embodiment provides a method for preparing the composition for inhibiting wheat pathogens described in Example 1, comprising the following steps:

[0066] 1) Preparation of Bacillus vesiculus inoculum: Bacillus vesiculus LTS-AF1, LTS-AF2, LTS-AF3 and LTS-AF4 were fermented and cultured separately, and their bacterial solutions were mixed in volume ratio to obtain Bacillus vesiculus inoculum.

[0067] 2) Permeation adsorption: The bacterial agent prepared in step 1) is mixed with a suspension of Zr-metal-organic framework material and treated under vacuum conditions to obtain a mixture;

[0068] 3) In-situ embedding: Add the protectant solution to the mixture prepared in step 2), stir and mix evenly to obtain a composition that inhibits wheat pathogens.

[0069] As a further preferred embodiment, in step 1), the fermentation and culture of Bacillus belyssus LTS-AF1, LTS-AF2, LTS-AF3 and LTS-AF4 further includes the addition of inducers, the inducers including sodium stearate and / or nano ferric oxide; the amount of sodium stearate added as the inducer is 0.1%-0.5% of the total weight of the Bacillus belyssus inoculum, and the amount of nano ferric oxide added as the inducer is 0.05%-0.1% of the total weight of the Bacillus belyssus inoculum.

[0070] The sodium stearate inducer can reduce the surface tension of the liquid and improve the dispersion of oxygen and substrate, while the nano-ferric oxide provides electronic bridging and stabilizes the redox potential of the system. The two work synergistically to accelerate the aerobic metabolic pathway and improve fermentation efficiency.

[0071] As a further preferred embodiment, in step 2), the vacuum condition is 25-35 kPa, and the processing time is 5-10 min.

[0072] As a further preferred embodiment, in step 3), the stirring time is 5-10 min and the embedding temperature is ≤30℃.

[0073] Preferably, the method for preparing the solid powder of the composition of the present invention includes the following steps:

[0074] 1) Preparation of bacterial agent: *Bacillus belye* LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 were fermented and cultured separately, with inducers added during fermentation. The fermented *Bacillus belye* bacterial cultures LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 were then mixed in a volume ratio of (2~3):(2~5):(1~5):(1~4). After mixing, the mixture was centrifuged and concentrated, and the supernatant was removed to obtain the *Bacillus belye* bacterial agent, at which point the viable count was ≥10⁻⁶. 10CFU / g, take 10-50 parts of Bacillus vesiculosus inoculum by weight.

[0075] 2) Permeation and adsorption: Add 10-80 parts by mass of Zr-metal-organic framework material to deionized water and sonicate for 2-10 min (power 200W) to obtain a uniform nanoparticle suspension; slowly add the bacterial agent prepared in step 1) to the above suspension and stir magnetically for 5-10 min to allow the bacterial strain to fully contact UiO-66 through surface adsorption. Then place it in a vacuum chamber of 25-35 kPa and evacuate for 5-10 min to allow the bacterial agent to penetrate into the material pores. After restoring normal pressure, stir for another 20-40 min to obtain a mixture.

[0076] 3) In-situ encapsulation: Take 10-30 parts by weight of the protective agent and place it in deionized water. Dissolve it completely in a water bath, cool it, and then add it to the mixture prepared in step 2). Continue stirring for 5-10 minutes to obtain a suspension, keeping the temperature ≤30℃ during this process. Spray dry the suspension at a feed temperature ≤30℃. After drying, collect the dry powder and vacuum dry it to remove residual moisture. The yield is approximately 90%, thus obtaining the solid powder composition for inhibiting wheat pathogens.

[0077] Preferably, the method for preparing the liquid formulation of the composition of the present invention includes the following steps:

[0078] 1) Preparation of bacterial culture: Bacillus belye LTS-AF1, LTS-AF2, LTS-AF3 and LTS-AF4 were fermented and cultured separately, with inducers added during the fermentation process; the fermented Bacillus belye bacterial cultures LTS-AF1, LTS-AF2, LTS-AF3 and LTS-AF4 were mixed in a volume ratio of (2~3):(2~5):(1~5):(1~4), and then concentrated by centrifugation until the viable count was ≥10. 10 CFU / mL, take 10-50 parts of Bacillus vesiculosus inoculum by weight.

[0079] 2) Permeation and adsorption: Add 10-80 parts by mass of Zr-metal-organic framework material to deionized water and sonicate for 2-10 min (power 200W) to obtain a uniform nanoparticle suspension; slowly add the bacterial agent prepared in step 1) to the above suspension and stir magnetically for 5-10 min to allow the bacterial strain to fully contact UiO-66 through surface adsorption. Then place it in a vacuum chamber of 25-35 kPa and evacuate for 5-10 min to allow the bacterial agent to penetrate into the material pores. After restoring normal pressure, stir for another 20-40 min to obtain a mixture.

[0080] 3) In-situ embedding: Take 10-30 parts of the protectant by mass and place it in deionized water. Dissolve it completely in a water bath. After cooling, add it to the mixture prepared in step 2) and continue stirring for 5-10 min. During stirring, add 0.1 mL of Tween-80 and adjust the pH to 6-8 with NaHCO3. Make up the volume to 100 mL with deionized water and continue stirring at low speed for 2-5 min to obtain the liquid formulation of the composition that inhibits wheat pathogens. Store the liquid formulation in a light-proof bottle.

[0081] Example 3

[0082] This embodiment provides the application of the composition for inhibiting wheat pathogens described in Example 1 in the preparation of an agent for the prevention and control of wheat diseases.

[0083] This embodiment also provides the application of the composition for inhibiting wheat pathogens described in Example 1 in enhancing wheat's tolerance to abiotic stress, wherein the abiotic stress includes at least one of high temperature, low temperature, acid-base stress, or high salt stress.

[0084] This embodiment also provides the application of the composition for inhibiting wheat pathogens described in Example 1 in promoting wheat growth. The application is carried out in a saline-alkali environment and is carried out throughout the entire growth cycle of wheat by means of foliar spraying, soil application or inter-row irrigation.

[0085] As a further preferred embodiment, the wheat germination rate is improved by applying the fertilizer to the soil or soaking the seeds during the sowing period; and / or by applying the fertilizer to the soil and spraying the seeds during the flowering period to improve the wheat's disease resistance.

[0086] Example 4

[0087] Experimental Example 1: Preparation of Microbial Agent

[0088] 1) Activation of strains: Take out the Bacillus belye: LTS-AF1, LTS-AF2, LTS-AF3 and LTS-AF4 stored in the -80℃ freezer, thaw them on ice, and then use a pipette to take about 5 μL of each strain and streak them evenly on LB solid medium in a clean bench. Place the plates in an incubator at 32℃ and invert them for incubation.

[0089] 2) Seed culture preparation (each of the four strains was prepared according to the experimental steps below, which will not be repeated here):

[0090] a) Primary seed culture: When single colonies grow on solid LB plates, pick up the single colonies with a pipette tip in a clean bench and inoculate them into test tubes containing beef extract peptone medium. Place the test tubes in a shaker at 30 ℃ and shake at 180 rpm for incubation.

[0091] b) Secondary seed culture: After the primary seed culture has been shaken and cultured in a shaker for about 12 h, take 10% of the primary seed culture in a clean bench and inoculate it into an Erlenmeyer flask containing 500 mL of liquid beef extract peptone medium. Place it in a shaker at 30℃ and shake at 180 rpm for about 8 h. Take it out and prepare to inoculate it into a 10 L fermenter.

[0092] 3) Microbial culture: After sterilization and cooling of a 10L fermenter containing fermentation medium, pump in all 500mL of secondary seed culture, and add sodium stearate and nano ferric oxide to a final concentration of 0.2% and 0.08% respectively. Then carry out fermentation in the fermenter. Sodium stearate can reduce the surface tension of the liquid and improve the dispersion of oxygen and substrate. Nano ferric oxide provides electron bridging and stabilizes the redox potential of the system. The two work together to accelerate the aerobic metabolic pathway and improve the fermentation efficiency.

[0093] Samples were taken every 2 hours to determine the OD of the bacterial agent. 600 like Figure 1 As shown, the OD of the fermentation broth after 42 hours of experimentation... 600 The value tends to stabilize, and the viable count is ≥10. 9 CFU / mL, fermentation complete, remove the inoculum.

[0094] Experimental Example 2: Disease Resistance of Bacillus belye

[0095] Take solid PDA medium and draw a cross in the center of the plate. Take the fermented Bacillus beryl bacterial suspensions LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 from Experiment 1, mixed with Bacillus beryl bacterial agent in a volume ratio of 2:3:3:4, and a Bacillus beryl mixed solution (Bacillus beryl mixed solution is Bacillus beryl LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 fermented without the addition of inducers, in a volume ratio of 2:3:3:4). 10 μL of each of the mixed solutions were used to inoculate 2-4 bacterial cultures at symmetrical locations 2-3 cm from the edge of the plate around the center point. A drop was also placed in the blank area of ​​the streak line. After drying in a clean bench, pathogenic bacterial discs (Fusarium graminearum, Fusarium graminearum, Fusarium pseudograminearum, Rhizoctonia solani, Rhizoctonia solani, Botrytis cinerea) were inoculated at the center point of the opposing plates. Plates inoculated only with pathogenic bacteria served as the control group. Each treatment was repeated three times. The plates were inverted and incubated at 30°C for 5-10 days, and the growth of the bacterial strains was observed.

[0096] The experimental results are shown in Table 1. Bacillus belyss LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 all showed good inhibitory effects against Fusarium graminearum, Fusarium, Fusarium pseudograminearum, Rhizoctonia solani, Rhizoctonia solani, and Botrytis cinerea. Under the condition that the number of viable bacteria in Bacillus belysss was comparable to that in the bacterial solution of a single species, the inhibitory effect on various pathogens was better, indicating that the compound bacterial solution produced a synergistic inhibitory effect on pathogenic microorganisms.

[0097] Table 1: Inhibition rate (%) of Bacillus belysinensis strain and its inoculum against pathogens.

[0098]

[0099] Experimental Example 3

[0100] A composition (solid powder) for inhibiting wheat pathogens comprises 10g of Bacillus belye inoculum, 80g of Zr-organic metal framework material, and 5g of trehalose and 5g of maltodextrin as protectants.

[0101] Taking the preparation of 100g as an example, please refer to the following steps:

[0102] 1) Preparation of inoculum: The fermented bacterial broths LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 from Experiment 1 were mixed in a volume ratio of 2:2:5:4. After mixing, the mixture was centrifuged and concentrated to remove the supernatant, yielding the Bacillus belyssus inoculum. At this point, the viable count was ≥10⁻⁶. 10 CFU / g, use 10g of Bacillus vesiculosus agent according to mass.

[0103] 2) Permeation and adsorption: Add 80g of UiO-66-OH to 30mL of deionized water and sonicate for 5min (power 200W) to obtain a uniform nanoparticle suspension; slowly add 10g of Bacillus belye inoculum to the above suspension and stir magnetically for 10min to allow the inoculum to fully contact UiO-66 through surface adsorption. Then place it in a 30kPa vacuum chamber and evacuate for 5min to allow the Bacillus belye inoculum to permeate into the material pores. After restoring to normal pressure, stir for another 30min to obtain a mixture.

[0104] 3) In-situ encapsulation: Dissolve 5g of trehalose and 5g of maltodextrin in 20mL of deionized water in a 40℃ water bath for 10min until completely dissolved. After cooling, add the dissolved trehalose and continue stirring for 5-10min to obtain a suspension, maintaining the temperature ≤30℃ during this process. Spray dry the suspension at a feed temperature ≤30℃. After drying, collect the dry powder and vacuum dry it to remove residual moisture. The yield is approximately 90%, yielding about 100g of solid powder.

[0105] Test Example 4

[0106] A composition (liquid formulation) for inhibiting wheat pathogens comprises 10 mL of Bacillus belye inoculum, 10 g of Zr-organic metal framework material, and 10 g of trehalose and 5 g of maltodextrin as protectants.

[0107] Taking the preparation of 100mL as an example, please refer to the following steps:

[0108] 1) Preparation of microbial agents: The fermented bacterial solutions LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 from Experiment 1 were mixed in a volume ratio of 3:3:2:1, and then concentrated by centrifugation until the viable cell count was ≥10. 10 CFU / mL was used to obtain Bacillus vesiculosus inoculum. 10 mL of Bacillus vesiculosus inoculum was taken.

[0109] 2) Permeation and adsorption: Take 10g of UiO-66-COOH and add it to 30mL of deionized water. Sonicate for 5min (power 200W) to obtain a uniform nanoparticle suspension. Slowly add 10mL of Bacillus belye inoculum to the above suspension and stir magnetically for 10min to allow the inoculum to fully contact UiO-66 through surface adsorption. Then place it in a 30kPa vacuum chamber and evacuate for 8min to allow the inoculum to penetrate into the material pores. After restoring normal pressure, stir for another 30min to obtain a mixture.

[0110] 3) In-situ embedding: Take 10g of trehalose and 5g of maltodextrin and place them in 40mL of deionized water. After dissolving completely in a 40℃ water bath for 10min, cool and add them to the compound solution and continue stirring for 5-10min. During stirring, add 0.1mL of Tween-80 and adjust the pH to 7.0 with NaHCO3. Make up the volume to 100mL with deionized water and continue stirring at low speed for 2min-5min to obtain 100mL of liquid formulation. Store the liquid formulation in a light-proof bottle.

[0111] Test Example 5: Composition Stability Test

[0112] (1) Temperature test

[0113] 100g of the solid powder from Example 3 and 100ml of the liquid preparation from Example 4 were placed in environments of 5℃, 20℃, 30℃, and 45℃ for 180 days, respectively. Samples were taken every 15 days, and plate counting was performed according to the standard GB 4789.2-2022 to calculate the viable count of the bacterial strains. The experimental results are as follows: Figure 2 , Figure 3 As shown, within the temperature range of 5-45℃, the viable bacterial count of both solid powder and liquid formulations remained above 10 for 180 days. 9 The CFU / mL (g) concentration indicates high bacterial activity and product stability.

[0114] (2) Salt and alkali test

[0115] LB medium with salt concentrations of 2%, 4%, 6%, and 8%, and LB medium with pH values ​​of 3.0, 6.0, and 9.0 were prepared respectively. The solid powder from Experiment 3 and the liquid formulation from Experiment 4 were inoculated into the medium at an initial concentration of 1 × 10⁻⁶. 7 CFU / mL, each group was repeated three times, and incubated in a shaker at 30℃ for 24 h. After the incubation, samples were taken, and plate counting was performed according to the standard GB 4789.2-2022 to calculate the viable count of the bacterial strain. The experimental results are as follows: Figure 4 As shown, the bacterial count increased to varying degrees under different salinity and alkalinity conditions. Even under extreme conditions such as 8% NaCl, pH 3.0, and pH 9.0, the bacterial strains still exhibited trace growth with viable counts >102. 7 The CFU / mL(g) indicates that both the solid powder and liquid formulations have high activity.

[0116] Comparative Example 1

[0117] A composition for inhibiting wheat pathogens (solid powder, total mass 100g) comprises 10g of Bacillus belye mixed solution (prepared by the same method as the mixed solution in Example 2), 80g of Zr-organic metal framework material, and 5g of trehalose and 5g of maltodextrin as protectants.

[0118] Taking the preparation of 100g as an example, please refer to the following steps:

[0119] 1) Preparation of the mixed solution: The *Bacillus belyssioides* bacterial cultures without added inducers, LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4, were mixed in a volume ratio of 2:2:5:4. After mixing, the mixture was centrifuged and concentrated to remove the supernatant, yielding the *Bacillus belyssioides* mixed solution with a viable count ≥10⁻⁶. 10 CFU / g, use 10g of Bacillus belye mixed solution by mass. Other components, dosages and preparation methods are the same as in Experiment 3.

[0120] Comparative Example 2

[0121] A composition for inhibiting wheat pathogens (solid powder, total mass 100g) comprising 100g of Zr-metal-organic framework material.

[0122] Comparative Example 3

[0123] An inhibitory preparation for wheat pathogens (solid powder, total mass 100g) comprises 100g of Bacillus berberis inoculum. Its preparation method is the same as step 1 in Example 3).

[0124] Comparative Example 4

[0125] A composition for inhibiting wheat pathogens (solid powder, total mass 100g) comprises 10g of Bacillus bellis inoculum, 80g of sodium alginate, and 5g of trehalose and 5g of maltodextrin as protectants. Other components, dosages, and preparation methods are the same as in Example 3.

[0126] Comparative Example 5

[0127] A composition for inhibiting wheat pathogens (solid powder, total mass 100g) comprises 50g of Bacillus bellis inoculum, 25g of trehalose as a protectant, and 25g of maltodextrin. Other components, dosages, and preparation methods are the same as in Example 3.

[0128] Comparative Example 6

[0129] A composition for inhibiting wheat pathogens (solid powder, total mass 100g) comprises 20g of Bacillus belye inoculum and 80g of Zr-metal-organic framework material. Other components, dosages, and preparation methods are the same as in Example 3.

[0130] Comparative Example 7

[0131] A composition for inhibiting wheat pathogens (solid powder, total mass 100g) comprises 10g of Bacillus belye inoculum (a mixture of fermented bacterial broths LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 from Example 1 in a volume ratio of 4:1:5:5), 80g of Zr-organic metal framework material, and 5g of trehalose and 5g of maltodextrin as protectants. Other components, dosages, and preparation methods are the same as in Example 3.

[0132] Comparative Example 8

[0133] A composition for inhibiting wheat pathogens (solid powder, total mass 100g) comprises 60g of Bacillus belye inoculum, 5g of Zr-organic metal framework material, and 15g of trehalose and 20g of maltodextrin as protectants. Other components, dosages, and preparation methods are the same as in Example 3.

[0134] Example 5 Antibacterial Test

[0135] Preparation of fungal mycelial cakes: Select fungal pathogens such as Fusarium graminearum, Fusarium graminearum, Fusarium pseudograminearum, Rhizoctonia solani, Rhizoctonia solani, and Botrytis cinerea (5mm×5mm) and inoculate them onto PDA plates. Incubate at 30℃ for 5-7 days until the mycelium has covered the plate. Use a sterile punch to make mycelial cakes with a diameter of 5mm from the edge of the activated fungal plate for later use.

[0136] Preparation of bacterial plates: Place one fungal disc in the center of PDA medium. Using sterile forceps, gently place four sterile Oxford cups around the disc. Inoculate the corresponding 50 μL formulation into the Oxford cups. The reagents for the experimental groups are designed as follows:

[0137] Control group 1: Sterile water;

[0138] Control group 2: Sterilized UiO-66 (Comparative Example 2);

[0139] Control group 3: Bacillus vesiculosus preparation with sodium alginate as microcarrier (i.e., using the composition of Comparative Example 4).

[0140] Control group 4: using the composition of Comparative Example 1;

[0141] Control group 5: The composition of Comparative Example 5 was used;

[0142] Control group 6: The composition of comparative example 6 was used;

[0143] Control group 7: The composition of Comparative Example 7 was used;

[0144] Control group 8: The composition of comparative example 8 was used;

[0145] Experimental Group 1: Bacillus belysin inoculum 1.0*10 7 CFU / mL (using the composition of Comparative Example 3);

[0146] Experimental Group 2: Solid powder prepared with sterile water (using the composition of Experimental Example 3), bacterial concentration 1.0*10⁻⁶ 7 CFU / mL;

[0147] Experimental Group 3: Liquid preparation made with sterile water (using the composition from Experimental Example 4), bacterial concentration 1.0*10⁻⁶ 7 CFU / mL;

[0148] The plates were incubated at 30℃ for 2-3 days, and the diameter of the inhibition zone was observed and measured. The experimental results are shown in Table 2. The diameter of the inhibition zone of both the solid and liquid formulations was larger than that of the free composite bacterial solution, and the antibacterial effect was significantly better than that of the free Bacillus beryl inoculum. Although the antibacterial effect of the Bacillus beryl preparation with sodium alginate as the microcarrier was better than that of the Bacillus beryl inoculum, its antibacterial activity was still lower than that of the composition of the present invention with UiO-66 as the carrier. The UiO-66 carrier had no inhibition zone, indicating that the UiO-66 carrier itself had no inhibitory effect on pathogens, and the antibacterial effect was entirely contributed by Bacillus beryl and its metabolites. When UiO-66 was combined with Bacillus beryl, the antibacterial effect was significantly improved. This is closely related to the increased loading rate of Bacillus beryl on the surface of the modified UiO-66. After loading, the number of viable Bacillus beryl on the surface and the bacterial activity were higher than those of the composite bacterial solution and the sodium alginate composite formulation, resulting in a significant improvement in its antibacterial activity.

[0149] The inhibitory effect of control group 4 was lower than that of experimental group 2, which indicates that even if the formulation ratio and components are the same as those in experimental example 3, the absence of an inducer will severely reduce the antibacterial effect of the composition. This suggests that the inducer is crucial for Bacillus belyssus to function.

[0150] Although control groups 5 and 6 showed a slight improvement in inhibitory effect compared to experimental group 1, they still lagged far behind experimental groups 2 and 3. Therefore, the optimal effect can only be achieved by using Zr-metal organic frames and protectants in combination. The absence of any component will prevent the highest antibacterial efficiency from being achieved. Meanwhile, the experimental data of control groups 7 and 8 show the importance of the compound ratio of various components.

[0151] Table 2: Formulation Inhibition Test (Diameter, mm)

[0152]

[0153] Example 6: Wheat germination under salt stress

[0154] Seeds were soaked in 5% sodium hypochlorite solution for 3 minutes, then rinsed three times with sterile distilled water and air-dried on sterile filter paper. The experimental group used a solution diluted to 10... 7In Experiment 4 (CFU / mL), the liquid formulation prepared by the present invention was used to soak seeds for 12 hours, while the control group was soaked in water for 12 hours. Each group was repeated three times, with 30 seeds per replicate. The seeds were evenly spread on 9cm×9cm filter paper petri dishes, and the same volume of nutrient solution, mildly saline-alkali nutrient solution, moderately saline-alkali nutrient solution, and severely saline-alkali nutrient solution were poured on them respectively. The petri dishes were placed in a constant temperature incubator at 20℃ with 12h / 24h light and cultured for 7 days. The germination of wheat was recorded, and the results are shown in Table 3. The germination rate of wheat increased to varying degrees under different salt concentrations after soaking in the liquid formulation of the present invention. The germination-promoting effect of the liquid formulation on wheat was positively correlated with the degree of salt and alkali stress: the more severe the salt and alkali stress, the more obvious the effect of the agent in improving the germination rate, especially in the severely saline-alkali environment. This indicates that the slow-release effect of the liquid formulation plays an important role in alleviating stress in high salt and alkali environments.

[0155] Table 3: Wheat seed germination rate (%) under different treatments

[0156]

[0157] Example 7: Wheat growth under salt and alkali stress

[0158] After mixing vermiculite and potting soil in a 1:1 ratio, weigh out the same amount of soil to ensure that each pot has the same weight. Add water to fully saturate the soil for two days, ensuring the same amount of water is used, and weigh out the water, pots, and soil to ensure they are of consistent weight. On the 10th day after emergence, select seedlings of uniform germination size and size and inoculate them into pots. Every 7 days, water with the same volume of (A) nutrient solution, (B) slightly saline-alkali solution, (C) moderately saline-alkali solution, and (D) heavily saline-alkali solution, respectively. The experiment was designed as follows:

[0159] (A) Nutrient solution group:

[0160] Control group A1: Watered with nutrient solution once every 7 days;

[0161] Control groups A2-A7: Using the compositions of control groups 3-8 in Example 5, the corresponding composition containing 10 was applied every 7 days. 7 A nutrient solution containing CFU / mL solid formulation was applied and sprayed onto the leaves at a concentration of 10 CFU / mL. 7 Preparations with corresponding CFU / mL concentrations;

[0162] Experimental group A1: Watered every 7 days with a solution containing 10 7 The nutrient solution of the solid formulation prepared in Experiment Example 3 (CFU / mL) was sprayed onto the leaves at a concentration of 10... 7 The liquid formulation prepared in CFU / mL test example 4;

[0163] (B) Mildly saline-alkali group:

[0164] Control group B1: Watered with a mild saline-alkali nutrient solution every 7 days;

[0165] Control groups B2-B7: Using the compositions of control groups 3-8 in Example 5, the corresponding composition containing 10 was applied every 7 days. 7 A mild saline-alkali nutrient solution containing CFU / mL solid formulation was sprayed onto the leaves. 7 Preparations with corresponding CFU / mL concentrations;

[0166] Experimental group B1: Watered every 7 days with a solution containing 10 7 In Experiment 3, a slightly saline-alkali nutrient solution containing a solid formulation was prepared and sprayed onto the leaves at a concentration of 10 CFU / mL. 7 The liquid formulation prepared in CFU / mL test example 4;

[0167] (C) Moderate salinity group:

[0168] Control group C1: Watered with a moderately saline-alkaline nutrient solution every 7 days;

[0169] Control groups C2-C7: Using the compositions of control groups 3-8 in Example 5, water was applied every 7 days with the corresponding composition containing 10 7 A moderately saline nutrient solution containing CFU / mL solid formulation was applied as a foliar spray. 7 Preparations with corresponding CFU / mL concentrations;

[0170] Experimental group C1: Watered every 7 days with a solution containing 10 7 In Experiment 3, a medium-saline nutrient solution containing CFU / mL of the solid formulation was prepared and sprayed onto the leaves at a concentration of 10 CFU / mL. 7 The liquid formulation prepared in CFU / mL test example 4;

[0171] (D) Severely saline-alkali group:

[0172] Control group D1: Watered with a highly saline-alkali nutrient solution every 7 days;

[0173] Control groups D2–D7: Using the compositions of control groups 3–8 from Example 5, the corresponding composition containing 10% was applied every 7 days. 7 A highly saline-alkali nutrient solution containing CFU / mL solid formulation was sprayed onto the leaves. 7 Preparations with corresponding CFU / mL concentrations;

[0174] Experimental group D1: Watered every 7 days with a solution containing 10 7 In Experiment 3, a solid formulation of highly saline-alkali nutrient solution was prepared and sprayed onto the leaves at a concentration of 10 CFU / mL. 7 The liquid formulation prepared in CFU / mL test example 4;

[0175] The experiment was divided into 8 groups, with 30 plants in each group (3 replicates * 10 plants). Plant height and survival rate were measured every 7 days before watering, and fresh weight was calculated. The experiment lasted for 28 days, and the average value of each experimental group was statistically analyzed.

[0176] Table 4: Wheat growth status in each experimental group of group (A)

[0177]

[0178] Table 5: Wheat growth status in each experimental group of group (B)

[0179]

[0180] Table 6: Wheat growth status in each experimental group of group (C)

[0181]

[0182] Table 7: Wheat growth status in each experimental group of group (D)

[0183]

[0184] The experimental results are shown in Table 4-7:

[0185] ① No salt or alkali stress (control group A1 vs. experimental group A1)

[0186] The plant height and fresh weight of experimental group A1 were significantly higher than those of control group A1, with plant height increasing by 19.6% and fresh weight increasing by 18.6%, and the survival rate was 100%; indicating that the preparation has a promoting effect on wheat growth under non-stress conditions.

[0187] ② Under mild salt and alkali stress (control group B1 vs. experimental group B1)

[0188] Mild salt-alkali stress caused the plant height and fresh weight of the control group B1 to decrease by 16.8% and 27.9% respectively compared with the control group A1, and the survival rate was reduced to 85.1%, indicating that mild salt-alkali stress had an inhibitory effect on wheat. The plant height and fresh weight of the experimental group B1 increased by 21.6% and 37.1% respectively compared with the control group B1, and the survival rate increased by 11.0%, which was close to the level of the no-stress group, indicating that the preparation can effectively alleviate mild salt-alkali stress.

[0189] ③ Under moderate salt-alkali stress (control group C1 vs. experimental group C1)

[0190] Moderate salt-alkali stress significantly inhibited wheat growth. In the control group C1, plant height and fresh weight decreased by 30.7% and 52.3% respectively compared to the control group A1, with a survival rate of only 63.4%, exhibiting stunted growth and slow development. In the experimental group C1, plant height and fresh weight increased by 22.0% and 58.5% respectively compared to the control group C1, with a survival rate increasing by 22.0%, and all indicators were superior to those of the control group B1 under mild salt-alkali stress.

[0191] This indicates that the formulation can still function under moderate saline-alkali stress, possibly by improving soil structure and enhancing wheat's resistance to saline-alkali ions (Na+) through the extracellular polysaccharides produced by Bacillus. + Cl - It has the ability to repel ions and reduce ion toxicity.

[0192] ④ Under severe salt and alkali stress (control group D1 vs. experimental group D1)

[0193] Under severe saline-alkali stress, the plant height and fresh weight of the control group D1 decreased by 51.2% and 73.3% respectively compared with the control group A1, and the survival rate was only 30%. Wheat growth was severely inhibited, and some plants showed yellowing and wilting of leaves. The plant height and fresh weight of the experimental group D1 increased by 36.9% and 60.9% respectively compared with the control group D1, and the survival rate increased by 28.6%, indicating that the preparation can still play a role under severe saline-alkali stress, but it is still lower than that of the moderate stress group. This shows that the preparation has a limited effect on alleviating severe saline-alkali stress, but it can significantly delay the damage caused by stress.

[0194] The results on plant height and fresh weight showed that as the degree of salt and alkali stress intensified (mild → severe), the growth-promoting effect of the formulation on wheat (increase in plant height and fresh weight) showed an upward trend (increase in plant height: 21.6% → 36.9%; increase in fresh weight: 37.1% → 60.9%), indicating that the "alleviating effect" of the formulation was relatively more significant under higher salt and alkali stress.

[0195] Survival rate results showed that the formulation could reduce wheat mortality caused by salt-alkali stress. In particular, the survival rate increase was greater under moderate and severe stress (22% and 28.6%) than under mild stress (11.0%), indicating that the formulation could enhance the stress resistance and survival ability of wheat.

[0196] The results of the control groups showed that although the control groups A2-A7, B2-B7, C2-C7, and D2-D7 were slightly higher in plant height, fresh weight, and survival rate compared with the control groups A1, B1, C1, and D1, respectively, their effects were worse than those of the experimental groups A1, B1, C1, and D1. This further illustrates that the combined formulation in this patent has a more significant and obvious advantage in wheat growth under salt and alkali stress.

[0197] Example 8: Disease resistance of wheat under salt-alkali stress

[0198] In Experiment 7, at the end of the 28-day experiment, the activity of polyphenol oxidase (PPO) was measured from wheat leaves and roots. Polyphenol oxidase is involved in the oxidation of phenolic substances in plants, enhances the degree of cell wall lignification, and can directly inhibit pathogen infection. It is an important disease resistance-related enzyme.

[0199] Table 8: Polyphenol oxidase activity (U / mg) in each experimental group

[0200]

[0201] The experimental results are shown in Table 8:

[0202] Under salt-alkali stress, the PPO activity in the control group increased with the degree of stress (control group A1 → control group D1), indicating that salt-alkali stress may indirectly induce the basic disease resistance of wheat, but the increase is limited.

[0203] The activity of PPO enzyme in roots was lower than that in leaves, but the increase in PPO in roots after treatment with the formulation (e.g., the increase in C1 in the experimental group was 37.7% compared to the control group) was higher than that in leaves (29.3%). This indicates that as the direct contact site with salt and alkali stress, the formulation may preferentially enhance the expression of disease resistance-related enzymes in the roots, thereby reducing the infection of root pathogens (such as root rot).

[0204] The composition of the present invention can significantly enhance the PPO enzyme activity of wheat under salt and alkali stress, and the increase in enzyme activity is more significant with the increase in stress intensity, indicating that the formulation enhances the adaptability and disease resistance of wheat to salt and alkali stress by strengthening the disease resistance metabolic pathway.

[0205] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A composition for inhibiting wheat pathogens, characterized in that, Its components include Bacillus belesiensis ( Bacillus velezensis The bacterial agent, Zr-organic metal framework material, and protective agent; its components, by mass parts, are: 10-50 parts of Bacillus berlesi bacterial agent, 10-80 parts of Zr-organic metal framework, and 10-30 parts of protective agent; the Bacillus berlesi bacterial agent is a mixture of four Bacillus berlesi strains, namely Bacillus berlesi LTS-AF1 (… Bacillus velezensis LTS-AF1), Bacillus belysinus LIS-AF2 ( Bacillus velezensis LIS-AF2), Bacillus belysinus LTS-AF3 ( Bacillus velezensis LTS-AF3), Bacillus belysinus LTS-AF4 ( Bacillus velezensis LTS-AF4); the effective viable count of the Bacillus vesiculosus agent is ≥10. 10 CFU / mL; The composition is used to simultaneously prevent and control wheat diseases and enhance wheat's tolerance to salt and alkali stress. The Zr-metal-organic framework material includes at least one of UiO-66 dry powder, UiO-66-OH, and UiO-66-COOH; The protective agent includes trehalose and / or maltodextrin; Preparation of Bacillus berleis inoculum: Bacillus berleis LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 were fermented and cultured separately, and their bacterial solutions were mixed in a specific volume ratio to obtain Bacillus berleis inoculum. The volume ratio of the Bacillus berleis LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 bacterial solutions was (2~3):(2~5):(1~5):(1~4). In each bacterial solution, the effective viable count of the Bacillus berleis strain was ≥10. 9 CFU / mL.

2. The composition according to claim 1, characterized in that, The Bacillus berberis LTS-AF1 ( Bacillus velezensis LTS-AF1), with accession number CCTCCNO: M 2025378; The Bacillus berberis LTS-AF2 ( Bacillus velezensis LIS-AF2), its accession number is CCTCCNO: M 2025379; The Bacillus berberis LTS-AF3 ( Bacillus velezensis LTS-AF3), its accession number is CCTCCNO: M 2025380; The Bacillus berberis LTS-AF4 ( Bacillus velezensis LTS-AF4), its accession number is CCTCCNO: M 2025381; All of the above-mentioned Bacillus belyes are deposited at the China Center for Type Culture Collection, located at Wuhan University, Wuhan, China, on March 5, 2025.

3. The composition according to claim 1, characterized in that, The composition is in the form of a solid powder or a liquid preparation; the pathogens include Fusarium graminearum, Fusarium, Fusarium pseudograminearum, Rhizoctonia solani, Rhizoctonia graminearum, and Botrytis cinerea.

4. A method for preparing a composition for inhibiting wheat pathogens according to any one of claims 1-3, characterized in that, Includes the following steps: 1) Preparation of Bacillus vesiculus inoculum: Bacillus vesiculus LTS-AF1, LTS-AF2, LTS-AF3 and LTS-AF4 were fermented and cultured separately, and their bacterial solutions were mixed in volume ratio to obtain Bacillus vesiculus inoculum. 2) Permeation adsorption: The bacterial agent prepared in step 1) is mixed with a suspension of Zr-metal-organic framework material and treated under vacuum conditions to obtain a mixture; 3) In-situ embedding: Add the protectant solution to the mixture prepared in step 2), stir and mix evenly to obtain a composition that inhibits wheat pathogens; In step 1), the fermentation and cultivation of Bacillus belyssus LTS-AF1, LTS-AF2, LTS-AF3, and LTS-AF4 further includes the addition of inducers, which include sodium stearate and / or nano-ferric oxide; the amount of sodium stearate added as an inducer is 0.1%-0.5% of the total weight of the Bacillus belyssus inoculum, and the amount of nano-ferric oxide added as an inducer is 0.05%-0.1% of the total weight of the Bacillus belyssus inoculum. In step 2), the vacuum condition is 25-35 kPa, and the processing time is 5-10 min. In step 3), the stirring time is 5-10 minutes, and the embedding temperature is ≤30℃.

5. The use of a wheat pathogen-inhibiting composition according to any one of claims 1-3 in the preparation of an agent for the prevention and control of wheat diseases.

6. The use of a composition for inhibiting wheat pathogens according to any one of claims 1-3 in enhancing wheat's tolerance to abiotic stress, characterized in that, The stresses include at least one of high temperature, low temperature, acid-base stress, or high salt stress.

7. The use of a composition for inhibiting wheat pathogens according to any one of claims 1-3 in promoting wheat growth, characterized in that, The application is carried out in saline-alkali environments and is carried out throughout the entire growth cycle of wheat through foliar spraying, soil application, or inter-row irrigation.

8. The application according to claim 7, characterized in that, Apply the pesticide during the wheat sowing period by soil application or seed soaking to improve wheat germination rate; and / or apply it during the wheat flowering period by soil application and foliar spraying to improve wheat disease resistance.

Images