A strain of Bacillus belyssus B05 and its application in the control of soybean root rot

Abstract

This invention discloses a Bacillus velezensis strain B05 and its application in controlling soybean root rot. The Bacillus velezensis strain B05 has the accession number CCTCC NO: M 2026071. Strain B05 significantly inhibits the growth of Fusarium and other pathogens. This invention is the first to screen specific genes associated with the superior antibacterial phenotype of strain B05 and clarify their sequence characteristics. Compared with other Bacillus velezensis strains, these specific genes increase the inhibition rate of strain B05 against Fusarium to over 90%, providing targeted genes for functional optimization of Bacillus velezensis and reducing the blindness of genetic engineering.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, specifically to a strain of Bacillus velezensis B05 and its related specific genes, and their application in the prevention and control of soybean root rot and the preparation of antibacterial agents. Background Technology

[0002] Soybean root rot is a prevalent disease in soybean-producing areas of my country, particularly in fields with continuous cropping, severely impacting soybean yield. *Bacillus belyssae*, an important biocontrol microorganism, is widely studied and applied due to its antibacterial and growth-promoting functions. However, existing *Bacillus belyssae* strains exhibit varying antibacterial spectra. Current research focuses on the inhibitory effects against one or two soybean root rot pathogens, lacking the exploration and research of strains with broad-spectrum antibacterial effects against multiple pathogens, thus failing to meet practical production needs. Furthermore, current research on genes related to the antibacterial function of *Bacillus belyssae* is insufficient, especially regarding specific genes between different strains and their association with antibacterial phenotypes. This hinders the precise optimization of strain performance through genetic engineering and makes it difficult to effectively protect and target core biocontrol genes. Summary of the Invention

[0003] In view of the fact that the antitoxin genes and transcriptional regulatory genes of Bacillus belyssus reported in the prior art are different from the specific gene sequences screened in this invention, and have not been confirmed to be associated with the high antibacterial phenotype, thus failing to solve the technical problem of blind optimization of the function of existing biocontrol bacteria, the purpose of this invention is to provide a Bacillus belyssus strain B05 and its application in the control of soybean root rot.

[0004] The objective of this invention can be achieved through the following technical solutions:

[0005] In the first aspect, this invention seeks protection for a strain of Bacillus velezensis, B05, which was deposited on January 12, 2026, at the China Center for Type Culture Collection (CCTCC), Wuhan University, China, with accession number CCTCC NO: M 2026071.

[0006] Strain B05 colonies are oval, milky white in color, with irregular edges and an opaque surface. The bacteria are rod-shaped, produce spores, and are Gram-positive.

[0007] The genome of strain B05 is a circular DNA molecule, 3,228,435 bp in length, with an average gene length of 908.38 bp. Coding genes account for 88.15% of the entire genome; the ratio of guanine (G) to cytosine (C) in the genome is 46%, and the predicted total number of genes is 3906, including 3810 coding sequences (CDS), 11 rRNA genes, 84 tRNA genes, and 1 non-coding RNA (ncRNA) gene. Its whole-genome characteristics can serve as a basis for identifying the source of specific genes.

[0008] This invention identifies specific genes associated with the superior antibacterial phenotype of Bacillus belyssus B05. These specific genes are defined as those unique to strain B05 and associated with its superior antibacterial phenotype, excluding redundant specific genes without phenotypic association. Specifically, these include the following core specific genes:

[0009] 1. Antitoxin genes ctg00005_02140 (yobK_1), ctg00014_03611 (yezG_1), ctg00014_03615 (yezG_3), ctg00014_03617 (yezG_5), and ctg00002_01034 (ywqK): These genes encode toxin-neutralizing proteins YobK, YezG, and YwqK, which are associated with the superior stress resistance phenotype of B05. They can neutralize the toxicity of endotoxins such as YobL, YeeF, and Ywqj produced by external stress, maintain cellular homeostasis, and regulate stress response, thereby indirectly supporting the antibacterial and stress-resistant phenotype of the strain. They are associated genes with high antibacterial activity of B05.

[0010] 2. Transcriptional regulatory genes ctg00008_02898 (immR), ctg00004_01893 (sphR_1), and ctg00004_01899 (comA_1): These genes encode proteins such as the HTH-type transcriptional regulator ImmR, the alkaline phosphatase synthesis transcriptional regulator SphR, and the transcriptional regulator ComA. They are associated with the superior antibacterial phenotype of B05 and are speculated to regulate the expression of pathogen inhibition-related genes by participating in specific metabolic pathways. Therefore, they are associated genes with the high antibacterial activity of B05.

[0011] 3. Toxin-regulating enzyme genes ctg00008_02899 (immA_1) and ctg00019_03884 (immA_2): These genes encode proteins such as the metallopeptidase ImmA and are associated with the superior antibacterial phenotype of B05. It is speculated that they can participate in the population regulation of bacterial growth inhibition and virulence attenuation by regulating the toxin-antitoxin system, and are associated genes of high antibacterial activity of B05.

[0012] 4. Specific genes related to the synthesis of secondary metabolites: Among the 8 BGCs, genes specific to B05 (such as bacillolysin, macrolide H and other genes related to synthesis) are associated with the excellent antibacterial phenotype of B05 and are speculated to be involved in the synthesis of antibacterial active secondary metabolites, directly contributing to the high antibacterial function.

[0013] The results showed that strain B05 possesses a specific gene selected from at least one of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, and SEQ ID NO:19. The protein sequences encoded by these specific genes are shown in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, and SEQ ID NO:20, respectively; the nucleotide sequences of each specific gene and the amino acid sequences of their encoded proteins are detailed in the sequence listing. These specific genes are unique to the aforementioned *Bacillus belyssiensis* strain B05, and strain B05 exhibits an inhibition rate of ≥90% against *Fusarium*.

[0014] Secondly, the present invention seeks protection for the use of the above-mentioned Bacillus belyssus strain B05 in at least one of the following (a1)-(a3):

[0015] (a1) Promoting soybean growth or preparing microbial preparations for promoting soybean growth;

[0016] (a2) Inhibit the pathogenic fungi causing root rot or prepare antibacterial agents for inhibiting the pathogenic fungi causing root rot;

[0017] (a3) Controlling soybean root rot or preparing microbial preparations for controlling soybean root rot.

[0018] Furthermore, the pathogen causing root rot is at least one of Fusarium oxysporum, Fusarium solanum, Fusarium graminearum, Rhizoctonia solani, and Phytophthora soybeanii.

[0019] Thirdly, the present invention claims protection for a microbial preparation, the active ingredient of which comprises at least one of the hyphae, spores and fermentation broth of the aforementioned Bacillus berberis strain B05.

[0020] Fourthly, the present invention claims protection for a method for promoting soybean growth by applying the aforementioned Bacillus berberis strain B05 or the aforementioned microbial preparation to the crop or the crop growth environment.

[0021] Fifthly, the present invention claims protection for a method for preventing soybean root rot, which involves applying the aforementioned Bacillus berberis strain B05 or the aforementioned microbial preparation to soybeans or the soybean growing environment.

[0022] In the technical solution of this invention, the application methods include seed treatment, root irrigation, soil mixing, or foliar spraying.

[0023] In a sixth aspect, the present invention claims protection for a specific gene associated with the superior antibacterial phenotype of Bacillus belyssus strain B05, the nucleotide sequence of which is selected from at least one of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17 and SEQ ID NO:19.

[0024] In a seventh aspect, the present invention seeks protection for biological materials related to the aforementioned specific gene, wherein the biological material is at least one of the following (b1)-(b4):

[0025] (b1) The protein encoded by the specific gene, wherein the amino acid sequence of the protein is selected from at least one of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18 and SEQ ID NO:20;

[0026] (b2) An expression cassette containing the specific gene;

[0027] (b3) A recombinant vector containing the specific gene or a recombinant vector containing the expression cassette (b2);

[0028] (b4) A recombinant microorganism containing the specific gene, a recombinant microorganism containing the expression cassette of (b2), or a recombinant microorganism containing the recombinant vector of (b3).

[0029] Eighthly, the present invention claims protection for the use of the aforementioned specific gene or the aforementioned biological material in at least one of the following (c1)-(c2):

[0030] (c1) Enhances the antibacterial activity of Bacillus belyssus;

[0031] (c2) Prepare antibacterial agents for the pathogens causing root rot.

[0032] In the technical solution of this invention, the antibacterial agent prepared by strain B05 is used to simultaneously inhibit *Fusarium oxysporum*, *Fusarium solanum*, *Fusarium graminearum*, *Rhizoctonia solani*, and *Phytophthora sacchariformis*, exhibiting excellent antibacterial phenotype. Specifically, the excellent antibacterial phenotype is as follows: the inhibition rate against *Rhizoctonia solani* reaches 86.56%. The inhibition rates against *Fusarium oxysporum*, *Fusarium solanum*, *Fusarium graminearum*, and *Phytophthora sacchariformis* are all ≥90%, and the pathogen colony diameter is ≤0.85 cm.

[0033] In a specific embodiment of the present invention, the vector is a prokaryotic expression vector or a eukaryotic expression vector, selected from any one of pET-28a, pUC19, and pBI121, but not limited thereto.

[0034] In specific embodiments of the present invention, the microorganism is Bacillus, Escherichia coli, or yeast, but is not limited thereto.

[0035] In a specific embodiment of the present invention, the fermentation method of the Bacillus berleis strain B05 is as follows: Bacillus berleis strain B05 is cultured in LB solid medium at 37°C for 24 hours, and the culture is repeated twice. A single colony of Bacillus berleis is picked and placed in an Erlenmeyer flask containing LB broth liquid medium, and cultured on a shaker at 37°C and 180 rpm for 24 hours.

[0036] The fermentation broth was centrifuged at 8000 rpm for 10 minutes and filtered repeatedly three times using a 0.22 μm disposable sterile filter membrane. The final fermentation broth was a supernatant free of bacteria.

[0037] The beneficial effects of this invention are:

[0038] The Bacillus berreatus strain B05 of this invention has a broad spectrum of inhibition against pathogens causing soybean root rot, and exhibits strong inhibitory effects against pathogens such as Fusarium oxysporum, Fusarium solanum, Fusarium graminearum, Rhizoctonia solani, and Phytophthora soybeanii.

[0039] The Bacillus berberis strain B05 of this invention has functions such as nitrogen fixation and iron production, which can promote the growth of soybean plants.

[0040] This invention is the first to screen specific genes associated with the superior antibacterial phenotype of strain B05 and clarify their sequence characteristics. Compared with other Bacillus belyss strains, these specific genes increase the inhibition rate of strain B05 against Fusarium oxysporum, Fusarium solanum, Fusarium graminearum, Rhizoctonia solani, and Phytophthora soymilk to over 90%, and the inhibition rate against Rhizoctonia solani reaches 86.56%. This provides targeted genes for the functional optimization of Bacillus belyss and reduces the blindness of genetic engineering modification.

[0041] The recombinant vectors and recombinant microorganisms constructed based on the specific genes exhibit superior antibacterial activity compared to the original strains, and also demonstrate higher stability, which can further expand the application scenarios of biocontrol microorganisms.

[0042] The discovery and protection of these specific genes provide new gene resources for the development of more targeted and stable antibacterial agents, which has significant agricultural application value and economic significance.

[0043] The *Bacillus belys* strain B05 of this invention can inhibit the growth of *Fusarium* and other pathogens (such as *Rhizoctonia solani* and *Phytophthora sacchariformis*). The stock solution of strain B05 showed an inhibition rate of 94.24% against *Fusarium solanum*, 90.10% against *Fusarium oxysporum*, 90.38% against *Fusarium graminearum*, 86.56% against *Rhizoctonia solani*, and 94.32% against *Phytophthora sacchariformis*. Through comparative analysis, it was found that the *Bacillus belys* strain described in this application currently exhibits the broadest antibacterial spectrum against major root rot pathogens and the best inhibitory effect against several root rot pathogens. The results compared with existing reports are as follows:

[0044] Comparative document 1: "CN 116103202 A Bacillus belye B6 with biocontrol activity, biocontrol agent and its application" shows that Bacillus belye has an inhibition rate of 69.9% against Fusarium solanum rot, 62.96% against Fusarium oxysporum, 58.99% against Fusarium graminearum, 80.59% against Rhizoctonia solani, 100% against Sclerotinia sclerotiorum, 71.56% against Coccidioides praecoxib, and 48.76% against Botrytis cinerea. The inhibition rate against key pathogens of root rot (such as Fusarium) is generally lower than that of strain B05 in this application. Its application is mainly for the control of other plant diseases, and the antibacterial spectrum against root rot pathogens has not been systematically evaluated.

[0045] Comparative document 2, "CN 115074287 A Bacillus belye JBNH 101 and its culture medium, fermentation product preparation method, formulation and application", only provides antibacterial experiments against Fusarium laminarum and Rhizoctonia solani, lacks specific data on antibacterial effect, and cannot evaluate its efficacy; it is only effective against a few pathogens, and its antibacterial spectrum is significantly narrower than that of this application.

[0046] Comparative document 3, "CN 115786218 A A strain of Bacillus belye and its application," shows that Bacillus belye exhibits an inhibition rate of 75.47% against Rhizoctonia solani, 69.11% against Phytophthora soybeanis, and 75.47% against Alternaria alternata. The overall inhibition rate against root rot-related pathogens is lower than the measured effect of strain B05 in this application; the number of pathogens evaluated is also limited, failing to comprehensively cover the main root rot pathogens.

[0047] Comparative document 4, "CN 113215031 A Bacillus belye 19573-3 and its application," mentions Bacillus belye inhibiting Fusarium oxysporum and Phytophthora spp., but provides no specific data. It only provides a qualitative description of its antibacterial activity, lacking quantitative data to support its effectiveness; the pathogen coverage is low, and its potential for controlling root rot is not systematically evaluated.

[0048] Meanwhile, the fermentation broth of strain B05 can promote soybean plant growth. Its application in soybean cultivation reduces the production of pathogens, lowers the incidence of soybean root rot, and promotes soybean plant growth, thus demonstrating great potential for application in soybean and other crop production. Attached Figure Description

[0049] Figure 1 This study aims to isolate, screen, and identify Bacillus belyssus B05. A shows the homologous gene comparison of six candidate strains; BD shows the phenotype and quantitative analysis of the antagonistic relationship between the six Bacillus belyssus strains and Fusarium solani; E shows the colony morphology of strain B05; and F shows the Gram staining results of strain B05.

[0050] Figure 2 The plate antagonistic effect of Bacillus bereaves strain B05 stock solution on five different pathogens (Fusarium solani, Fusarium oxysporum, Fusarium graminearum, Rhizoctonia solani, and Phytophthora sacchariformis).

[0051] Figure 3 The results show the growth promotion and extracellular enzyme activity detection results of Bacillus belyssus strain B05; where A is nitrogen-free medium; B is inorganic phosphate cell medium; C is CAS detection medium; D is extracellular casein detection medium; E is extracellular amylase detection medium; F is extracellular cellulase detection medium; and G is extracellular xylanase detection medium.

[0052] Figure 4 The study investigated the greenhouse control effect of Bacillus vesiculosus strain B05 on soybean root rot and its growth-promoting effect on soybean plants.

[0053] Information on the preservation of biological materials

[0054] Bacillus velezensis strain B05 is deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, China, with accession number CCTCC NO: M 2026071 and deposit date of January 12, 2026. Detailed Implementation

[0055] The present invention will be further described below with reference to the accompanying drawings and specific embodiments. This detailed description is not intended to limit the present invention, but rather to provide a more detailed description of some aspects, features and embodiments of the present invention.

[0056] Unless otherwise specified, all materials and reagents used in the following examples are commercially available. Where specific techniques or conditions are not specified, they should be performed in accordance with the techniques or conditions described in the literature or the product instructions.

[0057] Example 1: Isolation, screening and identification of Bacillus belye B05

[0058] Isolation and identification of antagonistic strains: Strains were isolated from plant rhizosphere soil samples from major soybean producing areas in my country; after serial dilution of the samples, they were spread on LB agar plates and incubated at 37℃ for 24-48 h. Single colonies were picked for purification culture to obtain multiple bacterial strains; plate confrontation experiments were used to verify the antibacterial effect of each strain on Fusarium solani.

[0059] The results showed that six strains (B05, CO1, CO2, D09, F08, and F10) exhibited antagonistic activity. Figure 1 (B in the text). Among them, strain B05 had a colony diameter of 1.78 cm and an inhibition rate of 75.23%, showing a significantly better antagonistic effect against Fusarium solani than all other strains. Figure 1 (C and D in the text).

[0060] 16S rDNA sequence alignment and phylogenetic analysis: To identify antagonistic bacterial strains, whole-genome DNA was extracted from microorganisms using the TIANamp bacterial DNA kit (Tiangen Biotech (Beijing) Co., Ltd.) as a PCR template. The 16S rRNA sequence of the strains was amplified using primers 27F / 1492R. The amplified products were sent to Shanghai Sangon Biotech Co., Ltd. for sequencing, and the sequences were compared with the National Center for Biotechnology Information (NCBI) database for homology analysis.

[0061] Based on 16S rDNA sequence analysis, all six antagonistic bacterial strains were identified as *Bacillus belye*. Homology comparison of the complete genomes of the six antagonistic bacteria revealed a total of 3,439 shared genes. In unique gene analysis, strain B05 possessed the most unique genes, reaching 141, demonstrating stronger gene specificity than other strains. Figure 1 (A) Based on the antibacterial activity and genetic specificity of the mixture of 6 antagonistic bacterial strains, strain B05 was selected for further research on the inhibition mechanism.

[0062] The isolated strain B05 was inoculated onto LB agar plates and incubated at 28°C for 1 day. Single colony morphology was then observed. (See also...) Figure 1In strain E, the colonies are oval, slightly raised, milky white in color, with irregular edges, an opaque surface, and some wrinkles. After Gram staining, strain B05 was observed under an oil immersion optical microscope; the results are shown in [reference needed]. Figure 1 In the F group, strain B05 is rod-shaped and purple, indicating that strain B05 is a Gram-positive bacterium.

[0063] Strain B05 was deposited at the China Center for Type Culture Collection on January 12, 2026, with accession number CCTCC NO: M 2026071.

[0064] Activation of Bacillus belyssus B05: Bacillus belyssus B05 was cultured in LB solid medium at 37°C for 24 h, and then passaged twice. Single colonies were picked using an inoculation loop and placed in an Erlenmeyer flask containing LB broth liquid medium. The culture was then incubated at 37°C and 180 rpm on a shaker for 24 h to obtain the working culture of B05.

[0065] Five pathogens causing soybean root rot were activated on PDA plates. After culturing for 5 days, the bacterial blocks were inoculated into the center of a 50% PDA + 50% LB mixed solid plate. 5 μL of activated Bacillus vesiculosus B05 solution was inoculated 2 cm to the left and right of the inoculated plate. Each treatment was repeated at least six times. After culturing at 28℃ for 3-5 days, the antagonistic effect was observed, the diameter of the inhibition zone was measured, and the inhibition rate was calculated.

[0066] The formula for calculating the inhibition rate is:

[0067]

[0068] The results are as follows Figure 2 As shown, *Bacillus belyss* B05 exhibits varying degrees of inhibitory effects against *Fusarium solani*, *Fusarium oxysporum*, *Fusarium graminearum*, *Rhizoctonia solani*, and *Phytophthora sacchariformis*, with antagonistic effects superior to the other five isolated *Bacillus belyss* strains. The inhibition rate of the original culture medium of B05 against the pathogens was investigated. The results showed that the original culture medium of B05 achieved an inhibition rate of 94.24% against *Fusarium solani*, 90.10% against *Fusarium graminearum*, 90.38% against *Fusarium graminearum*, 86.56% against *Rhizoctonia solani*, and 94.32% against *Phytophthora sacchariformis*.

[0069] Example 2: Growth promotion and extracellular enzyme activity detection of Bacillus belyssus B05

[0070] Nitrogen fixation capacity assay of strain B05: Strain B05 was activated using LB agar plates, and single colonies were picked and cultured in LB broth at 37°C for 1 day to prepare OD. 600=0.7-1.0 working bacterial culture. Take 5 μL and inoculate it onto nitrogen-free medium, and incubate at 28℃ for 3-5 days. If the bacteria can grow normally, it indicates that the strain has the ability to fix nitrogen.

[0071] Phosphate-solubilizing ability of strain B05 was tested: strain B05 was activated using LB agar plates, and single colonies were picked and cultured in LB broth medium at 37°C for 1 day to prepare OD. 600 =0.7-1.0 working bacterial culture. Take 5 μL and inoculate it in the center of the inorganic phosphorus cell culture medium. Incubate at 28°C for 5-7 days. The appearance of a transparent zone around the bacteria indicates that the bacteria have the ability to dissolve inorganic phosphorus; the size of the transparent zone can be used to determine the strain's ability to utilize inorganic phosphorus.

[0072] Siderophore production capacity assay: B05 strain was activated using LB agar plates. Single colonies were picked and cultured in LB broth at 37°C for 1 day to prepare a working bacterial suspension with an OD600 of 0.7-1.0. 5 μL of this suspension was inoculated into the center of siderophore detection medium (CAS) and incubated at 28°C for 5-7 days. The orange-yellow halo forming around the bacteria indicates their ability to produce siderophores; the size of the halo can be used to determine the strain's siderophore production capacity.

[0073] Extracellular enzyme activity assay: Strain B05 was activated using LB solid plates. Single colonies were picked and cultured in LB broth at 37°C for 1 day to prepare working bacterial suspensions with OD600 = 0.7-1.0. 5 μL of each suspension was inoculated onto cellulase, xylanase, amylase, and protease detection media, and incubated at 28°C for 3-5 days. The appearance of a clear zone around the bacteria indicated enzyme activity; the size of the clear zone could be used to determine the level of enzyme activity.

[0074] The results are as follows Figure 3 As shown, strain B05 can grow normally on nitrogen-free medium ( Figure 3 In the case of A), no phosphorus solubilization zone was formed on the inorganic phosphorus medium. Figure 3 (B in the text) induces an orange-yellow halo on the CAS detection medium. Figure 3 (C in the figure). Meanwhile, strain B05 produced large transparent regions (DG in the figure) on the detection media for extracellular proteases, starch hydrolases, cellulases, and xylanases, indicating that strain B05 has the ability to produce extracellular proteases, starch hydrolases, cellulases, and xylanases.

[0075] Example 3: Control and growth-promoting effects of Bacillus belyi B05 on soybean root rot.

[0076] Soybean seeds (Mengdou 1137) were washed with pure water and then sterilized in 75% (v / v) ethanol solution for 3 minutes, followed by sterilization in 1% sodium hypochlorite solution for 3 minutes. They were then rinsed repeatedly with sterile water before use. The seeds were planted in sterilized vermiculite and grown under greenhouse conditions. Subsequent experimental procedures were conducted after the seedling stage. Two treatment groups were set up, with 5 pots in each group and each treatment replicated 3 times.

[0077] Bacillus belye B05 was cultured in LB broth at 28°C with shaking at 180 rpm for 24 h, then centrifuged to collect the precipitate and diluted to 0 D with sterile water. 600 =0.7-1.0 working bacterial solution, ready for use.

[0078] Spores of Fusarium solani grown on PDA plates were eluted with sterile water and diluted to 10. 5 CFU / mL, ready for use.

[0079] After the soybean plants reach the seedling stage, the working bacterial solution of strain B05 is dispensed into containers and used for root irrigation at a rate of 10 mL per pot. After waiting for 1 day, Fusarium spores are added following the same procedure, with a control group not receiving spores.

[0080] Three to five weeks after treatment, observe the changes in soybean seedlings, count the number of plants affected by root rot, and at the same time, count the biomass of each soybean plant: plant height, root length, aboveground weight, and underground weight.

[0081] The results of the experiment on the control of soybean root rot and its effect on promoting growth by Bacillus vesiculosus B05 are shown in [the table below]. Figure 4 After applying the working bacterial solution of strain B05, the growth of soybean plants was observed. Soybean plants treated with the biocontrol strain B05 were taller, and the number of plants with root rot was significantly lower. Statistical data showed that, compared with the control group, soybean plants treated with Bacillus belyceta var. B05 had increased plant height, root length, aboveground weight, and underground weight by 17.47%, 14.06%, 55.81%, and 62.6%, respectively. Figure 4 (B and C in the text). Untreated plants showed blackening and rotting roots, yellowing leaves, and a 100% incidence of root rot. Figure 4 In the A section, the disease rate of plants treated with B05 was 37.5%, and the inhibition rate of strain B05 against soybean root rot caused by Fusarium was 62.5%. Strain B05 has an effective inhibitory effect on soybean root rot, can alleviate the disease, has a significant control effect on soybean root rot, and also has a significant growth-promoting effect.

[0082] The above-mentioned research examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above-mentioned research examples. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

[0083] Example 4: Screening of B05-specific genes and phenotypic association analysis

[0084] Whole genome extraction: The whole genome DNA of the six antagonistic strains in Example 1 was extracted using a bacterial genomic DNA extraction kit (Tiangen Biotech Co., Ltd., Beijing). The specific steps were described in the kit instructions.

[0085] Homology alignment analysis: The whole genome sequences of the six strains were aligned using BLAST software, with a similarity threshold of 95%, to screen for shared and specific genes. The results showed that the six strains shared a total of 3439 genes, while strain B05 had 141 unique genes.

[0086] Specific gene function annotation: The 141 specific genes of B05 were compared with the NR, COG, and KEGG databases for function annotation. Based on the annotation information, specific genes related to antibacterial activity were screened (i.e., the core specific genes protected in this invention, such as YobK, YezG, YwqK, ImmR, ImmA, etc.); specifically including the following core specific genes:

[0087] 1. Antitoxin genes ctg00005_02140 (yobK_1) (as shown in SEQ ID NO:1), ctg00014_03611 (yezG_1) (as shown in SEQ ID NO:3), ctg00014_03615 (yezG_3) (as shown in SEQ ID NO:5), ctg00014_03617 (yezG_5) (as shown in SEQ ID NO:7), and ctg00002_01034 (ywqK) (as shown in SEQ ID NO:9) encode toxin-neutralizing proteins YobK, YezG, and YwqK; the amino acid sequences of the proteins encoded by the above antitoxin genes are shown in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, and SEQ ID NO:10, respectively.

[0088] 2. Transcriptional regulatory genes ctg00008_02898 (immR) (as shown in SEQ ID NO:11), ctg00004_01893 (sphR_1) (as shown in SEQ ID NO:13), and ctg00004_01899 (comA_1) (as shown in SEQ ID NO:15) encode HTH-type transcriptional regulator ImmR, alkaline phosphatase synthesis transcriptional regulator SphR, and transcriptional regulator ComA, etc. The amino acid sequences of the proteins encoded by the above transcriptional regulatory genes are shown in SEQ ID NO:12, SEQ ID NO:14, and SEQ ID NO:16, respectively.

[0089] 3. The toxin-regulating enzyme genes ctg00008_02899 (immA_1) (as shown in SEQ ID NO:17) and ctg00019_03884 (immA_2) (as shown in SEQ ID NO:19) can encode proteins such as the metallopeptidase ImmA. The amino acid sequences of the proteins encoded by the above-mentioned toxin-regulating enzyme genes are shown in SEQ ID NO:18 and SEQ ID NO:20, respectively.

[0090] Validation of the association between specific genes and antibacterial phenotype: The expression level of the target specific gene in strain B05 during co-culture with Fusarium solani was detected by qRT-PCR.

[0091] Results: The expression levels of target-specific genes (SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17 and SEQ ID NO:19) in strain B05 were significantly upregulated after 24 h of co-culture; demonstrating that the target-specific genes are associated with the superior antibacterial phenotype of B05.

[0092] Example 5: Application in the preparation of microbial inoculants

[0093] 1. Preparation of microbial inoculum: The Bacillus belye strain B05 from Example 1 was inoculated into LB liquid medium and cultured at 37°C and 180 rpm for 12 h to obtain a bacterial suspension; an appropriate amount of adjuvants (such as Tween 80 and glycerol) was added to the bacterial suspension and stirred evenly to prepare the inoculum; wherein, the concentration of the strain was 10. 8 -10 9 CFU / mL.

[0094] 2. Validation of antibacterial effect: Soybean seedlings were selected and divided into experimental and control groups. The experimental group was sprayed with the prepared inoculant, while the control group was sprayed with an equal amount of LB liquid culture medium and adjuvants. After 24 hours, a mixed inoculant solution of *Fusarium solani*, *Fusarium oxysporum*, *Fusarium graminearum*, *Rhizoctonia solani*, and *Phytophthora sojae* (10 μL) was sprayed. 6 The fungicide (CFU / mL) was placed in an incubator at 25℃ and 80% humidity for 7 days. Plant disease incidence was observed, and the disease index and control effect were calculated. Results showed that the disease index of the experimental group was significantly lower than that of the control group, with a control effect exceeding 65%, proving that this antibacterial agent can effectively control soybean root rot.

[0095] Example 6: Functional Verification of Specific Genes

[0096] To further clarify the functional association between the specific genes screened in this invention and the excellent antibacterial phenotype of Bacillus bereaves B05, homologous recombination technology was used to knock out some representative specific genes in a targeted manner to construct gene deletion mutants. The inhibitory effects of the mutants and the original strains on Fusarium were compared by plate confrontation experiments.

[0097] Specifically, several representative genes from the antitoxin gene, transcriptional regulatory genes, and genes related to secondary metabolite synthesis (such as yobK_1, immR, immA_1, and some secondary metabolic synthesis genes) were selected, and knockout vectors were constructed for each. Gene-deleted mutant strains were obtained through double-crossover homologous recombination. After PCR and sequencing verification confirmed the successful deletion of the target gene, the antibacterial activity of each mutant strain against Fusarium was determined according to the aforementioned plate confrontation experiment method.

[0098] The above results confirm that the specific genes screened in this invention (such as the antitoxin genes, transcriptional regulatory genes, and toxin regulatory enzyme genes shown in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17 and SEQ ID NO:19) are key functional genes for Bacillus belesiensis B05 to maintain a high antibacterial phenotype, and their integrity plays an important role in the excellent antibacterial activity of the strain.

[0099] sequence list

[0100] ctg00005_02140 (yobK_1):

[0101] SEQ ID NO:1

[0102] ATGACTTATCATGAAATTGAAGAATTTATTAAAGAAAACGCTGAAGATGATGATTTTACAGGCGGAATAAGTGAAGAAAAAATCAAAGAGATACAAAATCGTTTAAATGTAAAACTTCCCGAGAGTTATAAGTGGTTTTTAAATAAATATGGCTCCGGTGGAATTTACGGGGTAAATATACTAGGCCATGGGAAAGTTTCTGCAAGGGTTGTTACAGTTACTGAGGAATATAGAACTTATTATAATTTAAGTGACGGTATTATCGTTATAGAAGATGTAGATGAATTCTCTTATTGCCTTGATACAAACAAAATGGAAAACGACGAATGTCCAGTAATAGTTTGGGCAAATGAAGCCGGTTATGGTGACAAGGTTGCAGATAACTTTCTGCAATTTTTCTTGGAAGAGCTTGAAGAAGCAAAAGAAAATTGGGAAGAGGACGAAGATTGGGACGATTAA

[0103] SEQ ID NO:2

[0104] MTYHEIEEFIKENAEDDDFTGGISEEKIKEIQNRLNVKLPESYKWFLNKYGSGGIYGVNILGHGKVSARVVTVTEEYRTYYNLSDGIIVIEDVDEFSYCLDTNKMENDECPVIVWANEAGYGDKVADNFLQFFLEELEEAKENWEEDEDWDD

[0105] ctg00014_03611(yezG_1)

[0106] SEQ ID NO:3

[0107] ATGGAGACAGAAAAAATGGGCCAATTATATCAAAAAATAGCAGAACTAATTAATGAAATGATTCCAGCAGAATGGAATAAAGTTGCTCTATACGCAGAGATTTTTAGAATGACTCTAGAGAAGTATACTTTTTCTTTAATACAAAAAATAGTGAAGCGTTCATTTATTCACATGACATTCCGGAACATTATCAAGTAAGTGAACAAATTTATGATGACTTGTTAATCGAACTACAAGATTTATTTGATGAACTAAGAGATGAATGGAAAGCGAATAACTCAGAGTGTGGACTAACCTTACACTCAAGCTAGAGCATACCGGAAAGTTTTCAATTGACTATGATTATGATGATGTTATTGCTTCAGAACTAAACGGTACTCAACGTCAAGTAGTTTGGGAATATAAACATTTAGGTTTGGTTCCTACTGGTAAGAAAAACAGGGAGTTCTTGGAACGCTATCTCAATTCGGAAGAAGAATAA

[0108] SEQ ID NO:4

[0109] METEKMGQLYQKIAELINEMIPAEWNKVALYAEILDDSREVYFFFNTKNSEAFIYSHDIPEHYQVSEQIYDDLLIELQDLFDELRDEWKANNSDVWTNLTLKLEHTGKFSIDYDYDDVIASELNGTQRQVVWEYKHLGLVPTGKKNREFLERYLNSEEE

[0110] ctg00014_03615(yezG_3)

[0111] SEQ ID NO:5

[0112] ATGGAAACTACTAAAATGGAACAACTATATCAAAAGATTGCTGAAAAAATAAATGATATGATTCCCTCCGAATGGATGAGGTTACTCTATACGCCGAAATTTTAGATGACTCCAGAGAAGTTTATTTTTTCTTTAACACACATGAAGACGAAGAATATATTTATTCACACGATATTCCAGAACATTTTCAAGTAAGTAATCGAATTTACAAAAATATGCTACTAGACCTTCAAGATTTATTTGATGAATTAAGGGATGAATTTAAAAAGCAATCCTCAGATATTTGGACTAACCTCACACTAAATTTAGATAGAAACGGTGAATTCTCAATTGATTATGATTATGAAGATGTACTCGCTTCAAAATATACTGATACTGAGCGACAAGTCATTTGGGAATATAAGCACTTGGGAATTACGCCAATAAGCGAGAGTAGCAGAAAGTTCCTTGAACAATATCTAAATGAACAATGA

[0113] SEQ ID NO:6

[0114] METTKMEQLYQKIAEKINDMIPSEWMRVTLYAEILDDSREVYFFFNTHEDEEYIYSHDIPEHFQVSNRIYKNMLLDLQDLFDELRDEFKKQSSDIWTNLTLNLDRNGEFSIDYDYEDVLASKYTDTERQVIWEYKHLGITPISESSRKFLEQYLNEQ

[0115] ctg00014_03617(yezG_5)

[0116] SEQ ID NO:7

[0117] GTGGAAACTGAAAAGATGGGAGTACTATATCAAAAAATCGCTGAGCAAATAAATGAAATTATCCCCTCTGAGTGGGGGAAGGTTGCTCTTTATGCCGAAATTTTAGATGATTCTTCAGAAGTTTACTTCTTCTTTACTACTCCACAAAATCAGGAATATATATATTCACATGATATTCCAGAACACTTTAACGTTGATGAAGATATTTATGACGAACTTCTATATGAATTGCATGATTACTTTGAAGAGTTACGGGATGAATTTAAAACAAACAACGAGGACTTATGGACAAACCTCACTTTACATCTCGAACGAAGCGGTAAATTCTCAATTGACTACGATTACACCGATGTCATCGCCTCAGACTTAAACGGAACACAACGACAAGTGATATGGGAATATCAAAATTTAGGGATATTGCCTGAAGATGAGAACGATAAAGAATTTGTAATAAGCTACTTCGGTCTATAA

[0118] SEQ ID NO:8

[0119] METEKMGVLYQKIAEQINEIIPSEWGKVALYAEILDDSSEVYFFFTTPQNQEYIYSHDIPEHFNVDEDIYDELLYELHDYFEELRDEFKTNNEDLWTNLTLHLERSGKFSIDYDYTDVIASDLNGTQRQVIWEYQNLGILPEDENDKEFVISYFGL

[0120] ctg00002_01034(ywqK)

[0121] SEQ ID NO:9

[0122] ATGGAAAATGAATATGATATGAAGAATATAAAAGTAATAGGAGTGGACTTTGAGGATTTATGGTTCTCTTCGGTCAGTGATGAGATACTAGATAATCCTGAAGACGAAAATGGCCAGCCGTTTACAGGGCTGGCTTACGAATTATATCCAAATGGACAAATCATCTATTTTACTAAATATAAGAATGGGCTGGCTCATGGTTTAACAAGTGAATTTTATGAAAACGGGAATAAAAAAAGTGAAAAAGAATATAGATATGGGCAATTGCATGGTAAAAGTATAATTTGGTTTGAAAATGGAAGAAAAAAAAAGTGAGCAACAGTATGAGCATTCCATACTAATTTGCGAAAAGAGTTGGGATGAAGAAGGCAATCTATTAAATCAGTATGAGCTAGATACAAGTAGTCCTCACTTTGAAATACTAGAATCGAGAAGAGACTCACATTAATTTAGGTCGAGAGTGA

[0123] SEQ ID NO:10

[0124] MENEYDMKNIKVIGVDFEDLWFSSVSDEILDNPEDENGQPFTGLAYELYPNGQIIYFTKYKNGLAHGLTSEFYENGNKKSEKEYRYGQLHGKSIIWFENGRKKSEQQYEHSILICEKSWDEEGNLLNQYELDTSSPHFEILESRRETHINLGRE

[0125] ctg00008_02898(immR)

[0126] SEQ ID NO:11

[0127] ATGACAATTAGGTCAAAGACTAAAAATATGTAGAGAAAACTTAGGTTGGAATCAGCAACAAGCTGCTAATAAAATCGGAATATCCAAAAACACATTATCTAATTATGAAAGAGATTACAGAGACCCCGATACTTCTACTTTGAAAAAGATCAGTGACGTTTACAATGTAACAACAGATTATTTATTAGGAAAAGAACAGGTGAACCAAAACAAAGAATCTAATGACAATAATCTGCTTAACAAAACAATTAACGAAGCCATCCAAGAATTAAAGGATGAAGATACACTCCTTTTTATGAATAACGGTGAATTCGATGAAGAAACTGCTTTATTAATTAAGAAAGCATTAAAAAACGGAATTAAATTCATAGACGAGTTAAAAAAGAAAGAGTGA

[0128] SEQ ID NO:12

[0129] MTLGQRLKICRENLGWNQQQAANKIGISKNTLSNYERDYRDPDTSTLKKISDVYNVTTDYLLGKEQVNQNKESNDNNLLNKTINEAIQELKDEDTLLFMNNGEFDEETALLIKKALKNGIKFIDELKKKE

[0130] ctg00004_01893(sphR_1)

[0131] SEQ ID NO:13

[0132] ATGAATAACATTTTATACATTGAAGATGACCAAGAAATTGGCCAATTTGTAAAAGGTGATTTAGAAGATCGGGGCTATATGATTATATGGCTCACTTCTTCCTATAATTACGAAACATATATAGAAAAGGCTGATCTGATCGTGCTTGATATAATGATGCCCGGTTTA GACGGATTTACAATCGGTCAAAGGATGAAAAACACATCCGCAGATCCCGCTCTTGCTTTTAACAGCCAGAACGGGTCTGGAAGATAAGCTGAAAGGCTTAGGATTTGCGGATGATTACGTGACAAAGCCCTTTCATCCCGATGAACTGGCTGCTCGAATCGAAGTA TTATTAAGAAGATTTGACAAGACTGCCATTCATGAATTGAAGCTAAAACACTTAAGTGTTCACTTGAAAGAAAAAAGAATTTTAAATCATCAAAGTCATGAGGAAATATTTTTAACTGATAAGCAGTTAAGGATTTTTTTGCTTTTGCTTAGACATCCCAATCAAATATTAACAAAGGAACAAATTTATGAAAATATTTGGGAACAGCCTTATATCGAAGGGGATAAGACGTTGATGGTACATATCCGGCATTTACGGCAAAAAATCGAGATCAATCCAAATGAGCCGGAAATTCTTGAAACAATCAGAGGGATTGGATACAGGATCAAACAATGA

[0133] SEQ ID NO:14

[0134] MNNILYIEDDQEIGQFVKGDLEDRGYMIIWLTSSYNYETYIEKADLIVLDIMMPGLDGFTIGQRMKKTHPQIPLLLLTARTGLEDKLKGLGFADDYVTKPFHPDELAARIE VLLRRFDKTAIHELKLKHLSVHLKEKRILNHQSHEEIFLTDKQLRIFLLLLRHPNQILTKEQIYENIWEQPYIEGDKTLMVHIRHLRQKIEINPNEPEILETIRGIGYRIKQ

[0135] ctg00004_01899(comA_1)

[0136] SEQ ID NO:15

[0137] ATGATAAAGATTCTATTAATTGATGATCATATTGGTGTTGCTCAAGGGACAAAAGCCATTCTTGAAAAAAGCAATAAAATGGGAGTAACAATATTGTCTTGCTGTAAAGAAGTACTTAATCACCTTAAGCATTACGAGTATGACTTACTTTTACTTGATTTATATATGCCGGAGTTAAATGGAATGGAATGGAATTGTCAAAAATGCTTCTTAGAGAGAGTCCCGATCAAAAAATTATAATTTATACTGGTTTTGATATCTCTGCGCATTTCAATCTTTTGGTTGAAGTTGGTGTTTCCGGTTCATAAGTAAGTCTTCAACAGAAGAGCATATGATA AAAGTAATTGAAAGTGTCATTGAGGGAGATACCATTATTCCTACTCATCTCTTTTAAACAATTAAGAAGAACAGAAGCAAATACGTTCAACATTGATAAGCTTGAAGATAGAATAAGAGATATTACTTTAAATGAAAGAGAACAAGATATCTTAGCGGGCGTAGCGGAGGGAATGACAAACCGGGAATTATCAGCAAAGCTGTTGATCAGTCAAAGAGCGGTTGAATACATACTTACTGGTGTGTACAACAAGCTCGGTGTAAAATCCAGAACAGAGGCACTGATCAAAGCAAATCGATATTCATTAATTTCTATGAAAACTATATATGAGTAA

[0138] SEQ ID NO:16

[0139] MIKILLIDDHIGVAQGTKAILEKSNKMGVTILSCCKEVLNHLKHYEYDLLLLDLLIMPELNGMELSKMLLRESPDQKIIIYTGFDISAHFNLLVEVGVSGFISKSSTEEHM IKVIESVIEGDTIIPTHLFKLRRTEANTFNIDKLEDRIRDITLNEREQDILAGVAEGMTNRELSAKLLISQRAVEYLTGVYNKLGVKSRTEALIKANRYSLISMKTIE

[0140] ctg00008_02899(immA_1)

[0141] SEQ ID NO:17

[0142] GTGATTATCATTTATACAAGTAGAAATAAAACATAAATCGTTAGCTGTAATAAAAGAATATGGCACAAATAACGTTTATCAAATATGTGAATCTTTGGGTATTTACATCTTAAAAAATAATCTAGGAC AAGCAAATGGATTTCTTCAATTTGACGAACCCACAAATCAATATCTAATTCATATCAATTTTAAAATACGAACACTTTGTAATTGCCCACGAGTTAGGGCATTATTACCTACACAAATCTCAAC ACTTTTAAGATTCTGAACTGTTCAAGTGCCCTGAAAGATAAACTTGAACATCAAGCAAATCTTTTTGCCTCAGAACTGTTACTCACAGACAGAATTCATGATGCCATCCCCATCATTAGAATTCA CTCAAGAGCAAGTGGCGTCTTATTTTGAAGTGCCTTTATCTGCGGTTGTCGATTATAAATTCGCGCAGCCCGGTAACACCCTGAACAGTGTATTCAATTTCTGTAATAACCAGCTTAAAGCGTTTGGGTAA

[0143] SEQ ID NO:18

[0144] MIIIYTSRRIKHKSLAVIKEYGTNNVYQICESLGIYILKNNLGQANGFLQFDEPTNQYLIHINENLKYEHFVIAHELGHYYLHKNLNTFKILNCSSALKDKLEHQANLFASELLLTDRMIHDAIPIIREFTQEQVASYFEVPLSAVVDYKFAQPGNTLNSVFNFCNNQLKAFG

[0145] ctg00019_03884(immA_2)

[0146] SEQ ID NO:19

[0147] TTGATAAAAGCAGCTGTGCAAAGACTAATTAAAAAGTATAAAACCAGTAATCCTTATGAGCTTGCATCATACATAAATATAAATGTTATTCCATGGAACTTGCATCATGAAATAATGGGTTTTTATAAGTATGATAAGCGAAATAAATATATCGTTATCAATTCCAACCTAAACCAGGCAGAAAGAACTTTTGTGTGCTCCCATGAATTAGGGCATGCACAGTTACACCCACGGGCAAATACACCATTTATGAAAGAGCGTACTCTTTTCTCAGTTGATAAATATGAGGTTGAGGCAAATACCTTTGCGGTTGAGCTCCTTCTTCCCGATTGGGTAGTAAGCCAATATAAAAATACTGAATTCACCCTTGATGATATAGCTGTCATGAATGGGGTTCCTGCAGAGTTAGCCCACCTAAAAGACCTATCAGAGCTAAAAAATTTTTAG

[0148] SEQ ID NO:20

[0149] MIKAAVQRLIKKYKTSNPYELASYININVIPWNLHHEIMGFYKYDKRNKYIVINSNLNQAERTFVCSHELGHAQLHPRANTPFMKERTLFSVDKYEVEANTFAVELLLPDWVVSQYKNTEFTLDDIAVMNGVPAELAHLKDLSELKNF。

Claims

1. A strain of Bacillus velezensis, B05, characterized in that, The preservation number of the Bacillus belyssus strain B05 is CCTCC NO: M 2026071.

2. The Bacillus belyssus strain B05 according to claim 1, characterized in that, This strain B05 has a specific gene selected from at least one of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17 and SEQ ID NO:

19.

3. The Bacillus belyssus strain B05 according to claim 1 or 2 is used in at least one of the following (a1)-(a3): (a1) Promoting soybean growth or preparing microbial preparations for promoting soybean growth; (a2) Inhibit the pathogenic fungus causing soybean root rot or prepare antibacterial agents for inhibiting the pathogenic fungus causing root rot; (a3) Controlling soybean root rot or preparing microbial preparations for controlling soybean root rot.

4. The application according to claim 3, characterized in that, The pathogen causing root rot is at least one of Fusarium oxysporum, Fusarium solanum, Fusarium graminearum, Rhizoctonia solani, and Phytophthora soybeanii.

5. A microbial preparation, characterized in that, The active ingredient of the microbial preparation comprises at least one of the hyphae, spores and fermentation broth of the Bacillus berberis strain B05 as described in claim 1 or 2.

6. A method for promoting soybean growth, characterized in that, The Bacillus berberis strain B05 of claim 1 or 2 or the microbial preparation of claim 5 is applied to soybeans or the soybean growing environment.

7. A method for preventing and controlling soybean root rot, characterized in that, The Bacillus berreatus strain B05 of claim 1 or 2 or the microbial preparation of claim 4 or 5 is applied to soybeans or the soybean growing environment.

8. A specific gene associated with the superior antibacterial phenotype of Bacillus belyssus strain B05, characterized in that, The nucleotide sequence of the specific gene is selected from at least one of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17 and SEQ ID NO:19; the specific gene is unique to the Bacillus belyssus strain B05 of claim 1, and the B05 strain has an inhibition rate of ≥90% against Fusarium.

9. A biomaterial related to the specific gene of claim 8, characterized in that, The biomaterial is at least one of the following (b1)-(b4): (b1) The protein encoded by the specific gene, wherein the amino acid sequence of the protein is selected from at least one of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18 and SEQ ID NO:20; (b2) An expression cassette containing the specific gene; (b3) A recombinant vector containing the specific gene or a recombinant vector containing the expression cassette (b2); (b4) A recombinant microorganism containing the specific gene, a recombinant microorganism containing the expression cassette of (b2), or a recombinant microorganism containing the recombinant vector of (b3).

10. The use of the specific gene of claim 8 or the biomaterial of claim 9 in at least one of the following (c1)-(c2): (c1) Enhances the antibacterial activity of Bacillus belyssus; (c2) Prepare antibacterial agents for the pathogens causing root rot.

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