Bacillus velezensis with double effects of disease resistance and growth promotion and application thereof

Abstract

The application belongs to the field of agricultural microorganism technology, and particularly relates to a bacillus velezensis with double effects of disease resistance and growth promotion and application thereof. The strain number is BVZX-1. The bacillus velezensis BVZX-1 strain has been preserved in the China General Microbiological Culture Collection Center, the preservation date is November 7, 2025, the preservation address is No. 1, Xili, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC No. 36546. The bacillus velezensis BVZX-1 has effects of disease resistance, growth promotion, seed germination promotion and seed aging delay.

Description

Technical Field

[0001] This invention belongs to the field of agricultural microbial technology, specifically relating to a Bacillus belye with dual functions of disease resistance and growth promotion and its application. Background Technology

[0002] Aspergillus niger ( Aspergillus niger ), neatly arranged small sclerotia ( Sclerotium rolfsii Sacc.), Fusarium oxysporum ( Fusarium oxysporum Harmful fungi such as Aspergillus niger, white mold, and Fusarium oxysporum are the main causes of peanut diseases. These fungi are found in many plants, such as soybeans, cotton, tomatoes, and onions. Typically, Aspergillus niger, white mold, and Fusarium oxysporum can reduce peanut yield by more than 10%, and in severe outbreaks, yield reductions can reach as high as 80%, even leading to total crop failure. Given the limited arable land resources and the lack of high-yield, high-quality, resistant varieties, conventional control measures such as crop rotation, deep plowing, and planting disease-resistant varieties are insufficient to effectively control peanut white mold. Therefore, disease control still relies primarily on chemical fungicides. However, long-term use of chemical pesticides not only pollutes the environment, causes pesticide residues, and promotes drug resistance in pathogens, but also negatively impacts soil biodiversity. Biological control, with its significant advantages of high efficiency, eco-friendliness, and sustainable development, has become an important means of controlling harmful fungi.

[0003] Bacillus ( Bacillus Due to its rapid reproduction, high metabolic rate, strong colonization ability, spore production, and high efficiency, *Bacillus belye* is widely used in the field of plant disease control. *Bacillus belye* has multiple effects in the biological control of plants; its fermentation products, such as ituronine, panthenol, and surfactants, have antibacterial effects, inhibiting and antagonizing pathogenic bacteria. Therefore, *Bacillus belye* is effective against corn leaf blight. ] It also has good inhibitory effects on eight pathogenic bacteria of ginseng, including Alternaria alternata, Botrytis cinerea, Plasmodium ginsenoside, Plasmodium ginsenoside, Phytophthora infestans, and Phytophthora infestans, as well as on apple and pear fire blight, cucumber powdery mildew, and downy mildew.

[0004] Studies by Sun Meng et al. have found that polyketide antibiotics produced by Bacillus mainly include difenocil, macrolides, and bacitracin, which can interfere with protein synthesis by binding to the ribosomal subunits of pathogens. Some polyketide antibiotics can interact with the cell membrane of pathogens, disrupting membrane integrity. Studies by Wu Fengkang et al. have found that the fermentation broth and crude enzyme solution of Bacillus belye F41-14 have good control effects on cotton Verticillium wilt. Bacillus belye has been extensively studied in the control of plant blight, demonstrating effective and broad-spectrum control functions. Summary of the Invention

[0005] This invention isolated a highly effective antagonistic endophytic bacterium, *Bacillus belyssiensis* BVZX-1, from peanut seed material (Huayu 25). Morphological characteristics and multigene phylogenetic analysis were then performed. The antibacterial properties of BVZX-1 bacterial suspension and sterile filtrate were tested using a confrontation culture method. Pot experiments were conducted to determine the growth-promoting and disease-resistant abilities of BVZX-1. BVZX-1 exhibited significant antagonistic activity against three plant pathogens, including *Fusarium oxysporum*. This strain possesses division-promoting functions and can significantly improve peanut germination rate. This invention confirms that *Bacillus belyssiensis* BVZX-1 possesses both disease-resistant and growth-promoting effects, providing a theoretical basis and technical support for the sustainable development of peanut disease control.

[0006] The first objective of this invention is to provide a Bacillus belesiensis. (Bacillus velezensis) The strain number is BVZX-1. The Bacillus belyes BVZX-1 strain has been deposited at the China General Microbiological Culture Collection Center (CGMCC) on November 7, 2025. The deposit address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, and the accession number is CGMCC No. 36546.

[0007] Furthermore, the aforementioned Bacillus belye (Bacillus velezensis) Its 16S nucleotide sequence is shown in SEQ ID NO.1, and its rpob sequence is shown in SEQ ID NO.2.

[0008] The present invention also includes a microbial preparation comprising the above-mentioned Bacillus belye. (Bacillus velezensis) The strain, fermentation broth and / or sterile filtrate, and the microbial agents made from the strain.

[0009] The present invention also includes a bacterial suspension, wherein the effective viable count of Bacillus belyceae BVZX-1 in the bacterial suspension is 1 × 10⁻⁶. 6 ~1×10 8 The bacterial suspension is prepared by activating Bacillus belyssus BVZX-1, picking a single colony and inoculating it into the culture medium, and culturing for 16-32 h.

[0010] Furthermore, the temperature for culturing the bacterial suspension is controlled at 28–32°C, and the rotation speed is controlled at 150–200 rpm.

[0011] The present invention also includes the application of Bacillus belye BVZX-1 or its microbial preparation in inhibiting the growth of Aspergillus niger, Fusarium oxysporum, and Sclerotium sclerotiorum.

[0012] The present invention also includes the application of Bacillus vesiculosus BVZX-1 or its microbial preparations in the biological control of plant diseases caused by Aspergillus niger, Fusarium oxysporum, and Sclerotium sclerotiorum.

[0013] Furthermore, the plant diseases mentioned are peanut crown rot, peanut fruit rot, and peanut white mold.

[0014] The present invention also includes the application of Bacillus vesiculosus BVZX-1 or its microbial preparation in promoting the growth of naturally aging peanuts.

[0015] The present invention also includes the application of Bacillus vesiculosus BVZX-1 or its microbial preparation in promoting peanut seed germination and delaying seed aging.

[0016] The present invention also includes the application of the fermentation broth of Bacillus vesiculosus BVZX-1 in improving the germination potential, germination rate and germination index of peanut seeds.

[0017] The present invention also includes the application of Bacillus vesicularis BVZX-1 in the prevention and control of peanut fruit rot caused by mixed infection of Fusarium oxysporum and Pythium spp.

[0018] The beneficial effects of this invention are as follows: 1) The seed endophytic bacterium BVZX-1 can solve two key bottleneck problems in peanut production: low seed vigor and soil-borne disease pressure. Its dual function makes it an important tool for integrated crop management. 2) BVZX-1 treatment significantly improved the germination index and germination rate of low-vitality seeds. This vitality recovery indicates that applying BVZX-1 can help seedlings quickly overcome the "sensitive window" during which weak seedlings are susceptible to damping-off disease, reducing the risk of seedling loss in the field; 3) Live Bacillus belye BVZX-1 cells consistently outperformed cell-free filtrate in inhibiting pathogens (especially Aspergillus niger and Sclerotium truncatum), highlighting a colonization-dependent mechanism in addition to antibacterial activity; 4) Peanut fruit rot is usually caused by a combination of fungi (such as Fusarium) and oomycetes (such as Pythium), which is a major challenge in peanut production. BVZX-1 showed high control effect against this combination of fungi in pot experiments. This result is of great significance. This finding supports the strategy of using "native" seed endophytic fungi for integrated disease management. 5) BVZX-1 significantly promotes plant growth and pod yield, consistent with the characteristics of plant growth-promoting rhizosphere bacteria (PGPR). Notably, its protective effect extends from the rhizosphere to the developing pods underground, indicating that BVZX-1 can successfully colonize or exert functional activity in the pod sphere.

[0019] In summary, Bacillus belyeis BVZX-1 is a dual-function endophytic bacterium whose mechanism of action relies on colonization (driven by live bacteria) rather than simple metabolite secretion. Agronomically, it plays two key roles: (1) as a seed quality enhancer, significantly restoring the viability of low-vitality seeds and improving seedling establishment potential; (2) as a highly effective biological fungicide, reducing the severity of pod rot by 85.8% in soil environments. These findings confirm that BVZX-1 is a highly promising biological initiator and soil / seed inoculant, providing a sustainable alternative to chemical pesticides for peanut production. Attached Figure Description

[0020] Figure 1 This is a colony morphology diagram of Bacillus belyssus BVZX-1. Figure 2 Phylogenetic tree of strain BVZX-1 based on 16S rDNA sequence; Figure 3. Phylogenetic tree of strain BVZX-1 based on the rpoB gene; Figure 4 Antibacterial spectrum of Bacillus belyssus BVZX-1; Figure 5 Graphs showing the antibacterial effects of different treatments of strain BVZX-1 against Aspergillus niger; Figure 6 The antibacterial effect of different treatments on *Rhizoctonia solani* strain BVZX-1; Figure 7 The antibacterial effect of different treatments of strain BVZX-1 on Fusarium oxysporum; Figure 8 The control effect of BVZX-1 on peanut fruit rot caused by Fusarium oxysporum and Pythium spp. in a soil inoculation pot experiment is shown in the figure. Figure 9 Two graphs showing the rate of rotten fruit under different treatments. Detailed Implementation

[0021] Embodiments of this embodiment will now be described in more detail with reference to the accompanying drawings. While some embodiments of this embodiment are shown in the drawings, it should be understood that this embodiment can be implemented in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided to provide a more thorough and complete understanding of this embodiment. It should be understood that the accompanying drawings and embodiments are for illustrative purposes only and are not intended to limit the scope of protection of this embodiment.

[0022] The Bacillus belysinus provided in this embodiment of the invention ( Bacillus velezensisThe strain BVZX-1, described as Bacillus belye, has been deposited at the China General Microbiological Culture Collection Center (CGMCC) on November 7, 2025. The deposit address is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, and the accession number is CGMCC No. 36546.

[0023] Test strains Biocontrol strain: Bacillus belye ( Bacillus svelezensis Code name BVZX-1.

[0024] Target pathogenic fungi: Sclerotium sclerotiorum ( Sclerotium rolfsii Sacc Fusarium oxysporum ( Fusarium oxysporum ), Aspergillus niger ( Aspergillus niger ).

[0025] The Bacillus belyssus BVZX-1 used in the strain test was isolated and preserved from peanut variety HY25. Peanut Fusarium strain, peanut Aspergillus niger strain, and Sclerotium sclerotiorum were all isolated from the regional test materials and preserved at the Shandong Peanut Research Institute.

[0026] plant materials The plant materials used were Huayu 25, Liaohua 3, Nongda R271, and HY6628, which were bred by the Shandong Peanut Research Institute.

[0027] Test culture medium The LB and PDA culture media used in the experiment were provided by Beijing Solarbio Science & Technology Co., Ltd., and the formulations were based on the methods described above. Example 1

[0028] 1. Isolation and Identification of Biocontrol Bacteria Peanut seed treatment: Twenty peanut seeds were selected and disinfected in an ultra-clean workbench by soaking in 10% sodium hypochlorite for 3 minutes, then in alcohol for 1 minute, and finally rinsed three times with sterile water. To ensure better release of the peanut seed bacteria, the peanuts were divided. Surgical blades, tweezers, scissors, and other tools were disinfected with alcohol after each use, and repeatedly flaked with an alcohol lamp. The peanut seeds were placed on the sterilized workbench, separated along the kernel length, crushed, and placed in a 50ml Erlenmeyer flask. The mixture was shaken at 150 rpm for 3 hours, then centrifuged in 10ml centrifuge tubes. The supernatant was collected, and 150μL of the supernatant solution was spread onto LB solid medium and incubated at 28℃ for 1-2 days. Once clear bacterial zones appeared, colonies were picked, purified, and stored in glycerol at -80℃. Subsequent confrontation experiments were conducted to verify the results, and an endophytic biocontrol strain was selected and named BVZX-1.

[0029] Identification of strain BVZX-1 BVZX-1 was picked and streaked onto LB agar, then incubated in the dark at 28°C for 24 hours. The colony morphology of BVZX-1 on LB agar was then observed. Figure 1 As shown. Using the total genomic DNA extracted from the bacterial genomic DNA extraction kit from Beijing Solarbio Science & Technology Co., Ltd. as a template, PCR amplification was performed using the universal 16S rDNA primer 27F (5'-AGAGTTTGATCMTGGCTCAG-3' / 1492R (5'-TACGGYTACCTTGTTACGACTT-3') and the rpoB gene primer 2292F / 3354R (5-AGGTCAACTAGTTCAGTATG-GAC-3'5'-AAGAACCGTAACCGGCAACTT-3) [P3]. The target fragment was amplified by PCR. The PCR reaction system (30 μL) consisted of: 1 μL DNA, 1 μL each of forward and reverse primers (10 μmol / L), 15 μL 2×TaqPlus PCR Mix (TIANGEN), and 12 μL ddH2O. PCR reaction cycles: 95℃ for 5 min; 95℃ for 30 s, 55.8℃ for 30 s, 72℃ for 1.5 min, 35 cycles; 72℃ for 5 min. After passing 2% agarose gel electrophoresis, the reaction products were sent to Qingdao NIO Science & Technology Biotechnology Co., Ltd. for sequencing. The sequencing results were analyzed and compared with the 16S rDNA and rpob sequences of standard strains in the NCBI database, and a multi-gene phylogenetic tree was constructed using the 16S rDNA and rpob sequences.

[0030] The 16S primer gene DNA and rpoB primer gene fragment of BVZX-1 were obtained by PCR amplification. Sequencing and splicing yielded effective gene sequences of 1421 bp (as shown in SEQ ID NO.1) and 550 bp (as shown in SEQ ID NO.2), respectively. After alignment with NCBI, 8-13 sequence files of type strains with similar lengths to the sample sequence were selected and downloaded. Multiple sequence alignment was performed using MEGA software, and a phylogenetic tree (Bootstrap ≥ 1000) was constructed using the neighbor-joining (NJ) method. BLAST analysis showed that BVZX-1 and... Bacillus velezensis PQ813691.1 clustered on the same branch. In summary, based on the morphological characteristics and polygenic phylogenetic analysis of strain BVZX-1, it was determined that BVZX-1 is *Bacillus belyes* (e.g., *Bacillus beryl*). Figure 2 (As shown in Figure 3).

[0031] Example 2: Inhibition rate of strain BVZX-1 against three pathogens The antibacterial effect of strain BVZX-1 was determined using three pathogens—Aspergillus niger, Sclerotinia sclerotiorum, and Fusarium oxysporum—as target bacteria. The confrontation experiment results showed that strain BVZX-1 had good inhibitory effects on all three pathogens, indicating that strain BVZX-1 has good inhibitory activity against common peanut pathogens (see Figure 4). Figure 4 In Table 1, A represents the antibacterial effect of Aspergillus niger, B represents the antibacterial effect of Sclerotium sclerotiorum, and C represents the antibacterial effect of Fusarium oxysporum. The inhibition rates are shown in Table 1.

[0032] Table 1. Inhibition rate of strain BVZX-1 against three pathogens

[0033] Example 3: Inhibitory effects of bacterial strains, fermentation broth, and sterile filtrate on pathogens. 1. Preparation of fermentation broth Preparation of fermentation broth, sterile filtrate, and bacterial cells: 5 mL of BVZX-1 seed culture was inoculated into a 250 mL Erlenmeyer flask containing 100 mL of culture medium and cultured at 30 °C and 180 r / min for 24 h with shaking to obtain fermentation broth; the fermentation broth was centrifuged at 4 °C and 4000 r / min for 1 h, and the precipitate obtained by centrifugation was the bacterial cells; the supernatant was filtered through a 0.22 µm pore size filter membrane, and centrifuged again at 4 °C overnight to obtain sterile filtrate.

[0034] Aspergillus niger, Sclerotium sclerotiorum, and Fusarium oxysporum, each with a diameter of 4 mm, were inoculated into the center of each PDA plate containing different fermentation broths, sterile filtrate, and bacterial cells. PDA plates containing sterile liquid culture medium were used as blank controls. The plates were incubated in the dark at 23°C. Each treatment was repeated 3 times. When the pathogenic fungal colonies reached 2 / 3 of the control plate, the colony diameter was measured, and the inhibition rate was calculated.

[0035] Inhibitory effects of two strains, fermentation broth, and sterile filtrate on pathogens. 2.1 Inhibitory effect of strain BVZX-1 on Aspergillus niger Depend on Figure 5 As shown in Table 2, the sterile filtrate had a poor inhibitory effect on Aspergillus niger, while the fermentation broth had a moderate inhibitory effect. The bacterial cells, however, showed the best inhibitory effect. Figure 5 In the diagram, A represents CK, B represents bacterial cells, C represents fermentation broth, and D represents sterile filtrate.

[0036] Table 2. Inhibition rate and colony diameter of strain BVZX-1 against Aspergillus niger.

[0037] 2.2 Inhibitory effect of strain BVZX-1 on *Sclerotium sclerotiorum* Depend on Figure 6As shown in Table 3, the sterile filtrate had a poor inhibitory effect on *Sclerotium moniliforme*, while the fermentation broth had a moderate inhibitory effect. The bacterial cells, however, showed the best inhibitory effect.

[0038] Figure 6 A: Neat sclerotium CK, B: bacterial cells, C: fermentation broth, D: sterile filtrate.

[0039] Table 3. Inhibition rate and colony diameter of strain BVZX-1 against *Sclerotium truncatum*.

[0040] 2.3 Inhibitory effect of strain BVZX-1 on Fusarium oxysporum Depend on Figure 7 As shown in Table 4, the sterile filtrate had a poor inhibitory effect on Fusarium oxysporum, while the fermentation broth showed a better inhibitory effect. The bacterial cells exhibited the best inhibitory effect, reaching 83%. Figure 7 In the diagram, A represents CK, B represents bacterial cells, C represents sterile filtrate fermentation broth, and D represents fermentation broth.

[0041] Table 4. Inhibition rate and colony diameter of strain BVZX-1 against Fusarium oxysporum.

[0042] Example 4: Pot test on the control effect of fruit rot disease 1. Experimental Materials and Methods This embodiment verifies the control effect and growth-promoting effect of Bacillus vesicularis BVZX-1 fermentation broth on peanut fruit rot through a pot experiment. The specific experimental process is as follows: Fermentation broth preparation: Pick a single colony of Bacillus belye BVZX-1 and inoculate it into an Erlenmeyer flask containing 200 mL of LB liquid medium. Incubate in a constant temperature shaking incubator at 28℃ and 200 r / min for 36 h to obtain BVZX-1 fermentation broth for later use.

[0043] Experimental Design: The experiment included a BVZX-1 treatment group and a water control group (CK group), with three replicates per group, each with three pots arranged randomly. The peanut variety used was Huayu 6628. Healthy peanut seeds were disinfected with NaClO solution and then sown in pots with a height of 20cm, an inner diameter of 15cm, and a bottom diameter of 12cm, with three seeds sown in each pot. The seedlings were then conventionally cultivated until they were robust.

[0044] Treatment method: All experimental pots were inoculated with a mixed pathogenic fungus of Fusarium oxysporum and Pythium spp. in advance to construct a peanut fruit rot disease model. Subsequently, each pot in the BVZX-1 treatment group was watered with 30 mL of the prepared fermentation broth, while the CK group was watered with an equal amount of water as a blank control.

[0045] Cultivation and Investigation: The experiment was conducted outdoors at natural temperatures, with a light cycle of 16 hours of light / 8 hours of darkness. Weeds were regularly removed from the pots to avoid interference from other biological stresses on the experimental results. After the peanuts matured, seed testing was conducted, and the incidence of fruit rot (including incidence rate, disease index, and rot rate) and plant growth parameters (main stem height, lateral branch length, number of branches per plant, and number of fruits per plant) were systematically investigated. The control effects were calculated and differential analyses were performed.

[0046] 2. Analysis of disease resistance effect The results of the pot experiment showed that ( Figure 8 , Figure 9 ), Figure 8 In this context, 'a' represents the BVZX-1 treatment group. Figure 8 Group b represents the control group (CK). The Bacillus vesicle BVZX-1 fermentation broth showed significant control over peanut pod rot caused by mixed infection of Fusarium oxysporum and Pythium spp. Compared to the CK group, the severity of peanut pod rot in the BVZX-1 treatment group was significantly reduced, with all disease indicators showing significant decreases (P<0.01): the incidence of pod rot decreased from 56.54% to 15.79%, achieving a control effect of 71.8%; the disease index decreased from 52.48 to 7.44; and the rate of rotten pods decreased from 56.0% to 15.8%, a reduction of 71.8% (Tables 5 and 6).

[0047] Phenotypic observation showed that the peanut rot was severe in the CK group, while the number of rotten peanuts in the BVZX-1 treatment group was extremely small. The difference between the two groups was highly significant, further confirming that the BVZX-1 fermentation liquid can effectively inhibit the occurrence and spread of peanut rot, and has outstanding disease control effect.

[0048] Table 5. Analysis of Variance for Different Treatments

[0049] Note: P < 0.05 3. Analysis of the effects of promoting fertility Depend on Figure 8 It can be seen that the BVZX-1 fermentation broth has a significant growth-promoting effect on peanut plants of Huayu 6628. Compared with the CK group, the peanut plants in the BVZX-1 treatment group grew vigorously, and all growth parameters were significantly improved (Table 6): the main stem height increased from 16.00 cm to 29.07 cm, the lateral branch length increased from 20.07 cm to 34.93 cm, the number of branches per plant increased from 10.50 to 16.53, the number of fruits per plant increased from 51.33 to 95.00, the number of healthy fruits increased significantly, and the overall growth of the plants was significantly better than that of the control group.

[0050] Table 6. Effects of BVZX-1 treatment on growth and pod characteristics of potted peanuts inoculated with fruit rot pathogen.

[0051] 4. Conclusion This pot experiment demonstrates that Bacillus vesicles BVZX-1 fermentation broth has dual effects: on the one hand, it can significantly inhibit the occurrence of peanut fruit rot, effectively reduce the rate of rotten fruit, the incidence rate and the disease index, with a control effect of 71.8%, and excellent control performance; on the other hand, it can significantly promote the growth and development of peanut plants, increase key growth indicators such as plant height, number of branches and number of fruits, and help peanuts achieve healthy and high yield.

[0052] Example 5: Germination of peanut seeds after treatment with strain BVZX-1 Effects on peanut seed germination After surface disinfecting peanut seeds with 75% ethanol for 30 seconds, rinse them thoroughly with sterile water and blot dry with filter paper. Randomly select 360 seeds each of HY25, Liaohua 3, and Nongda 08123 varieties, and place 60 seeds into 50mL beakers. Add 30mL of BVZX-1 fermentation broth to each beaker (OD 600 = 1×10⁻⁶). 8 CFU / mL. The control group was treated with sterile water and soaked for 3 hours at room temperature (approximately 25°C). To prevent mold contamination and interference with test results, all experimental tools, including distilled water, filter paper, petri dishes, and tweezers, were sterilized using an autoclave. Two layers of cut filter paper were placed in a 90mm petri dish and moistened with sterile distilled water. The treated seeds were then evenly arranged in the petri dish and labeled. Each experiment was repeated three times, and the seeds were incubated at a constant temperature of 25°C. The number of germinating seeds was observed and recorded every 24 hours. The germination period began on day 1 of placement in the seedbed. Based on previous experiments, peanut germination is considered a 7-day cycle. Germination was defined as the emergence of a normal radicle in the germinated seed; a radicle longer than half the length of the seed was considered a normal germinated seed.

[0053] Germination potential GE = (Number of germinated seeds within 3 days / Total number of seeds tested) × 100%; Germination rate GP = (Number of germinated seeds within 7 days / Total number of seeds tested) × 100%; Germination index GI = ∑(Gt / Dt), where Gt refers to the number of germinated plants on day t; Dt refers to the corresponding number of germination days.

[0054] As shown in Table 7, the number of seeds germinated after treatment with the fermentation broth of strain BVZX-1 was higher than that of seeds treated with sterile water. Compared with the control, the fermentation broth of strain BVZX-1 promoted the germination of peanut seeds to varying degrees and improved the various germination indicators of different seeds.

[0055] Table 7 Germination indicators of three peanut varieties after soaking for 3 hours.

[0056] Note: Identical lowercase letters indicate no significant difference (P>0.05), while different lowercase letters indicate a significant difference (P<0.05). As shown in Table 7, the number of seeds germinated after treatment with the fermentation broth of strain BVZX-1 was higher than that of seeds treated with sterile water, showing an overall upward trend: the treatment groups of all varieties were significantly higher than the control group in terms of germination potential, germination rate and germination index, indicating that soaking for 3 hours has a general promoting effect on the germination performance of these three peanut varieties.

[0057] Table 7 lists the average values ​​and increases (Δ) of germination parameters for each variety. The BVZX-1 treatment significantly improved germination parameters for all varieties, with germination potential increasing by 15.56–17.78 percentage points, germination rate by 7.78–14.44 percentage points, and germination index by 10.68–15.34 units compared to the control group. Huayu 25 exhibited the highest absolute values ​​of germination potential, germination rate, and germination index in both the treatment and control groups. However, the magnitude of the response varied among different varieties: Nongda R271 showed the largest increases in germination rate (14.44 percentage points) and germination index (15.34 units), while Liaohua 3 and Huayu 25 showed the largest increases in germination potential (both 17.78 percentage points). This difference in response among varieties is consistent with the significant interaction between germination index treatment and variety observed in the analysis of variance (Table 8).

[0058] Example 6 Restoring the vitality of naturally aged peanut seeds Before performing the analysis of variance, we validated the assumptions of the statistical model. The Shapiro-Wilk test showed that the residuals of germination potential, germination rate and germination index conformed to a normal distribution (P>0.05), and the Levene test confirmed that the variances of the experimental groups were homogeneous (P>0.05). Therefore, all data were analyzed using the original data.

[0059] Two-way ANOVA showed that the BVZX-1 treatment had a highly significant positive effect on all measured germination parameters (Table 8). The treatment factor explained the largest proportion of variance in the model, and the partial eta-squared (η²p) values ​​for all indicators exceeded 0.80, indicating a strong biological effect (Table 8). Variety factors also had significant main effects on germination potential (P=0.022), germination rate (P=0.001), and germination index (P<0.001), but the degree of influence was less than that of the treatment factor (Table 8). The treatment × variety interaction had a significant effect on the germination index (F...). 2,12=4.84, P=0.029), while having no significant effect on germination potential and germination rate (P>0.05) (Table 8), indicating that the promoting effect of BVZX-1 on germination potential and germination rate is consistent among different varieties, while the effect on germination rate varies with genotype.

[0060] Table 8. Results of two-way ANOVA on the effects of BVZX-1 treatment and variety on peanut seed germination parameters.

Claims

1. A type of Bacillus belesii ( Bacillus velezensis ), characterized by: The strain number is BVZX-1. The Bacillus belyes BVZX-1 strain has been deposited at the China General Microbiological Culture Collection Center (CGMCC) on November 7, 2025. The deposit address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, and the accession number is CGMCC No. 36546.

2. The Bacillus belesiensis as described in claim 1 ( Bacillus velezensis ), characterized by: Its 16S nucleotide sequence is shown in SEQ ID NO.1, and its rpob sequence is shown in SEQ ID NO.

2.

3. A microbial preparation, characterized in that: The microbial preparation includes the Bacillus berberis as described in claim 1. Bacillus velezensis ( ) strains, fermentation broth and / or sterile filtrate, and microbial agents made from such strains.

4. The application of Bacillus vesiculosus BVZX-1 or its microbial preparation as described in claim 1 in inhibiting the growth of Fusarium oxysporum, Sclerotium sclerotiorum, and Aspergillus niger.

5. The application of Bacillus belyssus BVZX-1 or its microbial preparation as described in claim 1 in the biological control of plant diseases caused by Sclerotium sclerotiorum, Aspergillus niger, and Fusarium oxysporum.

6. The application as described in claim 5, characterized in that: The plant diseases mentioned are peanut white mold, peanut crown rot, and peanut fruit rot.

7. The application of Bacillus vesicularis BVZX-1 as described in claim 1 in the prevention and control of peanut fruit rot caused by mixed infection of Fusarium oxysporum and Pythium spp.

8. The application of Bacillus vesiculosus BVZX-1 or its microbial preparation as described in claim 1 in promoting the growth of naturally aging peanuts.

9. The application of the fermentation broth of Bacillus vesiculosus BVZX-1 as described in claim 1 in improving the germination potential, germination rate, and germination index of peanut seeds.

10. The application of Bacillus belye BVZX-1 or its microbial preparation as described in claim 1 in promoting peanut seed germination and delaying seed aging.

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