Bacillus velezensis E09 strain and application thereof

Abstract

The invention discloses a bacillus velezensis E09 strain and application thereof, the strain is preserved in Guangdong Microbial Culture Collection Center (GDMCC) on June 26, 2025, and the preservation number is GDMCC No: 66598. The invention further discloses a method for preparing the bacillus velezensis E09 strain. Researches show that the E09 strain has the effect of inhibiting banana bacterial fruit rot pathogen BG43 (Dickeya fangzhongdai BG43) newly found in China and has a broad-spectrum inhibition effect on eight plant pathogenic fungi, the inhibition rate is 40%-67%, and the effect of multiple prevention through one strain is achieved. The E09 strain has multiple functions of growth promotion, salt stress resistance, phosphate solubilization, potassium solubilization and broad-spectrum bacteriostasis, and a biological control strain which is stable in bacteriostatic effect, efficient and environment-friendly is provided for green prevention and control of agricultural and forestry plant diseases.

Description

Technical Field

[0001] This invention relates to the field of agricultural microbiology, specifically to a strain of Bacillus belyssus E09 and its applications. Background Technology

[0002] banana( Muse Banana (spp.) is a plant belonging to the genus Musa in the family Musaceae. It is not only the world's fourth largest fruit producer but also a key food crop after rice, wheat, and corn. my country is the world's second largest banana producer and the world's largest banana consumer; the security of its banana industry is directly related to the livelihoods of tens of millions of banana farmers and the development of the national economy. However, banana production has long been threatened by both fungal diseases (such as banana wilt and banana anthracnose) and bacterial diseases (such as banana fruit rot). Among these, bacterial fruit rot of banana is a newly discovered high-risk banana fruit disease (First Report of Bacterial Fruit Rot of Banana Caused by...). Dickey fangzhongdai In China. Ms. Xinting Liu, Mr. Chong Ma, Dr. Yiming Zhu, and Prof. Erxun Zhou, Plant Disease, https: / / doi.org / 10.1094 / PDIS-04-25-0745-PDN), whose pathogenic bacteria ( Dickeya fangzhongdai BG43 can infect unripe bananas through wounds or natural openings, causing browning and rotting of the banana fruit. In severe cases, the incidence rate can reach 20%, and it will cause serious problems for the post-harvest storage and preservation of bananas, resulting in significant economic losses.

[0003] Currently, chemical pesticides remain the mainstream technology for the prevention and control of bacterial fruit rot in bananas. Although commonly used copper preparations (such as copper hydroxide) and antibiotics (such as kasugamycin) have certain antibacterial effects, they have significant limitations: (1) Pathogens are prone to developing drug resistance, and the field control efficacy is declining year by year; (2) Copper ions are prone to accumulate in the soil, destroying the microbial community and causing heavy metal pollution; (3) The abuse of antibiotics may lead to the spread of drug-resistant genes, resulting in the growth of drug-resistant bacteria, which seriously threatens public health and safety; (4) Chemical agents are difficult to penetrate the waxy layer of the fruit and have limited effect on the already infected pathogens.

[0004] Biocontrol bacteria are microorganisms derived from the ecological environment. As fungicides, they can reduce the use of chemical pesticides, protect the ecological balance, are environmentally friendly, are less likely to develop resistance, and promote the sustainable development of agriculture. As an environmentally friendly alternative technology, biocontrol agents have shown potential in the control of diseases in other crops. Existing research focuses on the antagonistic effects of biocontrol bacteria such as Bacillus (e.g., Bacillus subtilis) and Pseudomonas on plant fungal diseases, but their control efficacy against the newly discovered bacterial fruit rot pathogen BG43, a Gram-negative bacterium, is generally insufficient. In addition, there are three major bottlenecks in the application of existing biocontrol agents in bananas: (1) the target strain screening system is imperfect, and there is a lack of specific antagonistic bacteria against fruit rot pathogens; (2) the colonization ability of biocontrol bacteria is insufficient, making it difficult to form a stable biofilm on the fruit surface; (3) the functional mechanism research is fragmented, and the synergistic effects of multiple growth promotion and disease resistance have not been systematically analyzed. Summary of the Invention

[0005] The purpose of this invention is to overcome the above-mentioned defects and deficiencies in the prior art and to provide a strain of Bacillus belyssus E09 ( Bacillus velezensis E09) strain.

[0006] A second objective of the present invention is to provide the fermentation broth of the Bacillus berberis E09 strain.

[0007] A third objective of this invention is to provide a microbial preparation containing the Bacillus belyssus strain E09 or its fermentation broth.

[0008] The fourth object of the present invention is to provide the application of Bacillus belyssus strain E09 or its fermentation broth or microbial preparation.

[0009] The above-mentioned objective of the present invention is achieved through the following technical solution: This invention provides a strain of Bacillus belyssus E09 ( Bacillus velezensis The strain E09 was deposited on June 26, 2025, at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), with accession number GDMCC No: 66598. The address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province, 510070, China.

[0010] This invention successfully isolated a new strain, E09, through targeted screening of rhizosphere soil microorganisms from healthy plants. The strain exhibits single colonies that are round, milky-white, with a smooth, moist surface and wrinkled protrusions around the edges. Gram-positive rod-shaped cells are confirmed by 16S rRNA alignment and phylogenetic analysis to be *Bacillus belye* (…). Bacillus velezensis ), named Bacillus belyss E09 ( Bacillus velezensisStrain E09. This strain was deposited on June 26, 2025, at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), with accession number GDMCC No: 66598. It is classified and named as follows: Bacillus from Velez E09, the address of which is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province, Postcode: 510070.

[0011] The present invention also provides the fermentation broth of the above-mentioned Bacillus vesiculosus strain E09.

[0012] The present invention also provides a microbial preparation containing the above-mentioned Bacillus berberis E09 strain or fermentation broth.

[0013] This invention shows that the Bacillus berleis strain E09 has the ability to secrete indoleacetic acid (IAA), which can promote banana biomass growth. It can also secrete siderophores (siderophiles) to compete with pathogens for iron ions in the environment; it has nitrogen-fixing and inorganic phosphorus-solubilizing effects; it has stable biofilm formation, aggregation and motility, and strong colonization ability, thus forming a stable biofilm on the surface of banana fruits; it can secrete amylase, nitrate reductase, catalase, protease, and cellulase. Meanwhile, the E09 strain has natural tolerance to salt stress ≤9%, making it applicable in salt-stressed environments (such as saline-alkali land). Furthermore, the E09 strain has broad-spectrum antibacterial activity against the banana bacterial fruit rot pathogen BG43 (… Dickeya fangzhongdai BG43) and 8 important plant pathogenic fungi [(Anthracnose fungus of banana) Colletotrichum musae ), fruit anthracnose ( Colletotrichum fructicola ), Fusarium oxysporum Cuban specialized race 4 ( Fusarium oxysporum f. sp. from Cuba Race 4), Curvularia crescentis ( Curvularia lunate Fusarium ( ) Fusarium falcate ), Flathead anthrax bacteria ( Colletotrichum truncatum ), Siamese anthrax bacteria ( Colletotrichum siamense ) and Higgins anthrax ( Colletotrichum higginsianum [This] has an inhibitory effect, with an inhibition rate of 40%–67%, achieving the goal of "one bacterium, multiple defenses," breaking through the limitation of traditional biocontrol bacteria's "one bacterium, one target." Simultaneously, the fermentation broth of strain E09 can effectively delay the aging of banana bacterial fruit rot pathogen BG43 in detached banana fruit. Dickey fangzhongdai The Bacillus berberis strain E09 of this invention exhibits significant effects in controlling banana bacterial fruit rot, including preventing peel blackening caused by BG43 and inhibiting the rotting process. This means the strain possesses multiple functions, including promoting growth, resisting environmental stress, and providing broad-spectrum disease prevention.

[0014] Therefore, the present invention provides the above-mentioned Bacillus berberis E09 strain, fermentation broth, or microbial preparation for the prevention and control of bacterial fruit rot pathogen BG43 in bananas ( Dickeya fangzhongdai BG43) or preparations for controlling bacterial fruit rot of bananas (BG43) Dickeya fangzhongda Applications of BG43 products.

[0015] This invention also provides the above-mentioned Bacillus vesiculosus E09 strain, fermentation broth, or microbial preparation in the prevention and control of bacterial fruit rot pathogen BG43 in bananas ( Dickeya fangzhongdai BG43) and banana fruit anthracnose fungus or preparations for controlling banana bacterial fruit rot fungus BG43 ( Dickeya fangzhongda The application of BG43 and the banana anthracnose fungus in products; the banana anthracnose fungus is *Anthracnose moniliforme* (BG43). Colletotrichum musae ) and fruit anthracnose ( Colletotrichum fructicola ).

[0016] This invention also provides the application of the above-mentioned Bacillus vesiculosus E09 strain, fermentation broth, or microbial preparation in the prevention and control of plant diseases or in the preparation of products for the prevention and control of plant diseases; the pathogen of the plant disease is Bacillus thuringiensis BG43 (banana bacterial fruit rot). Dickeya fangzhongdai BG43), Banana anthrax bacteria ( Colletotrichum musae ), fruit anthracnose ( Colletotrichum fructicola And any one or more of the following six plant pathogenic fungi: Fusarium oxysporum Cuban specialized race 4 ( Fusarium oxysporum f. sp. from Cuba Race 4), Curvularia crescentis ( Curvularia lunate Fusarium ( ) Fusarium falcate ), Flathead anthrax bacteria ( Colletotrichum truncated ), Siamese anthrax bacteria ( Colletotrichum siamense ) and Higgins anthrax ( Colletotrichum Higginsian ).

[0017] This invention also provides the application of the above-mentioned Bacillus vesiculosus E09 strain, fermentation broth, or microbial preparation in promoting plant growth and controlling plant diseases, or in the preparation of products that promote plant growth and control plant diseases; the pathogen of the plant disease is Bacillus thuringiensis BG43 (banana bacterial fruit rot). Dickeya fangzhongdai BG43), Banana anthrax bacteria ( Colletotrichum musae ), fruit anthracnose ( Colletotrichum fructicola And any one or more of the following six plant pathogenic fungi: Fusarium oxysporum Cuban specialized race 4 ( Fusarium oxysporum f.sp. from Cuba Race 4), Curvularia crescentis ( Curvularia lunate Fusarium ( ) Fusarium sickle-shaped ), Flathead anthrax bacteria ( Colletotrichum truncatum ), Siamese anthrax bacteria ( Colletotrichum Siamese ) and Higgins anthrax ( Colletotrichum higginsianum ).

[0018] This invention also provides the application of the above-mentioned Bacillus vesiculosus E09 strain, fermentation broth, or microbial preparation in promoting plant growth and controlling plant diseases under salt stress, or in the preparation of products that promote plant growth and control plant diseases under salt stress; wherein the salt stress environment is ≤9% salt stress; and the pathogen of the plant disease is Bacillus thuringiensis BG43 (banana bacterial fruit rot). Dickeya fangzhongdai BG43), Banana anthrax bacteria ( Colletotrichum musae ), fruit anthracnose ( Colletotrichum fructicola And any one or more of the following six plant pathogenic fungi: Fusarium oxysporum Cuban specialized race 4 ( Fusarium oxysporum f. sp. from Cuba Race 4), Curvularia crescentis ( Curvularia lunate Fusarium ( ) Fusarium falcate ), Flathead anthrax bacteria ( Colletotrichum truncated ), Siamese anthrax bacteria ( Colletotrichum siamense ) and Higgins anthrax ( Colletotrichum Higginsian ).

[0019] This invention also provides a method for promoting plant growth and preventing plant diseases, comprising applying the above-mentioned Bacillus berberis E09 strain, fermentation broth, or microbial preparation; wherein the pathogen of the plant disease is Bacillus thuringiensis BG43 (a type of banana bacterial fruit rot pathogen). Dickeya fangzhongdai BG43), Banana anthrax bacteria ( Colletotrichum musae ), fruit anthracnose ( Colletotrichum fructicola And any one or more of the following six plant pathogenic fungi: Fusarium oxysporum Cuban specialized race 4 ( Fusarium oxysporum f. sp. from Cuba Race 4), Curvularia crescentis ( Curvularia lunate Fusarium ( ) Fusarium falcate ), Flathead anthrax bacteria ( Colletotrichum truncated ), Siamese anthrax bacteria ( Colletotrichum siamense ) and Higgins anthrax ( Colletotrichum Higginsian ).

[0020] Furthermore, the plant is a banana, corn, soybean, turmeric, chili pepper, or a cruciferous plant.

[0021] Compared with the prior art, the present invention has the following beneficial effects: This invention discloses a strain of Bacillus vesiculus E09 and its applications. The strain was deposited at the Guangdong Provincial Microbial Culture Collection Center (GDMCC) on June 26, 2025, with accession number GDMCC No: 66598. Research shows that the Bacillus vesiculus E09 strain promotes banana growth and is tolerant to salt stress. It exhibits significant inhibitory effects on BG43, a bacterial fruit rot pathogen of banana, and two types of banana anthracnose pathogens (Anthracnose moniliforme and Anthracnose citrinum). Furthermore, it shows broad-spectrum inhibitory effects on six plant pathogenic fungi, including Fusarium oxysporum race 4 (Cubanus cocovenenans), Curvularia crescentis, Fusarium oxysporum, Anthracnose stenoptera, Anthracnose siamense, and Anthracnose hyamniotes, with inhibition rates ranging from 40% to 67%. This achieves the goal of "one strain, multiple defenses," overcoming the limitations of traditional biocontrol bacteria that target only one pathogen per target. The E09 strain possesses multiple functions, including promoting growth, resisting environmental stress, and broad-spectrum disease prevention, providing a stable, efficient, and environmentally friendly biological control strain for the green prevention and control of diseases in agricultural and forestry plants. Attached Figure Description

[0022] Figure 1 The colony morphology of Bacillus belyssus strain E09 on LB agar plates.

[0023] Figure 2 Gram staining results for Bacillus vesiculosus strain E09.

[0024] Figure 3 The results of physiological and biochemical plate assays for Bacillus belyssus strain E09 are shown.

[0025] Figure 4 A phylogenetic tree for Bacillus belyss E09 strain constructed based on the 16S rRNA gene.

[0026] Figure 5 The growth of Bacillus belyssus strain E09 on salt stress plates.

[0027] Figure 6 Bacillus vesiculosus strain E09 and BG43, a bacterial fruit rot pathogen of bananas ( Dickey fangzhongdai The inhibitory effect of BG43 in plate confrontation and banana inoculation experiments.

[0028] Figure 7 The inhibitory effect of Bacillus vesiculosus strain E09 on plate confrontation with eight important plant pathogenic fungi was investigated.

[0029] Figure 8 This is a data analysis of the inhibitory effects of Bacillus belyssus strain E09 on plate confrontation with eight important plant pathogenic fungi. Detailed Implementation

[0030] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0031] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0032] Example 1 Isolation and Identification of Strains 1. Source and isolation culture of the strain The E09 strain was isolated and purified from the rhizosphere soil of healthy plants using a gradient dilution method. The E09 strain culture was streaked onto LB solid medium (5 g yeast extract, 10 g tryptone, 10 g NaCl, 1000 mL water, 15 g agar, sterilized at 121℃ for 20 min) using an inoculation loop and cultured at 37℃ for 24 h. The colony color and morphological characteristics were observed and recorded, and cell morphology was observed by microscopy and Gram staining.

[0033] 2. Morphological and physiological / biochemical identification Colony characteristics: After incubation at 37°C for 24 h on LB agar plates, strain E09 forms round, milky-white colonies (2–3 mm in diameter), with a smooth, moist surface and radially wrinkled edges (as shown in the image). Figure 1 ).

[0034] Microscopic observation: Gram staining revealed Gram-positive bacilli, with a cell size of (0.5–0.7) μm × (1.1–1.9) μm (e.g. Figure 2 ).

[0035] Physiological and biochemical characteristics: as shown in Table 1 and Figure 3 As shown, strain E09 was positive for MR test, negative for VP test, positive for starch hydrolysis test, positive for nitrate reduction test, and negative for indole test. Strain E09 can promote banana biomass growth by secreting indoleacetic acid (IAA), secrete siderophores (ferophiles) to compete with pathogens for iron ions in the environment; it has nitrogen-fixing and inorganic phosphorus-degrading functions; it has stable biofilms, exhibits aggregation and motility, and strong colonization ability; it can secrete amylase, nitrate reductase, catalase, protease, and cellulase.

[0036] Table 1 Physiological and biochemical characteristics of strain E09

[0037] 3. Molecular biological identification Genomic DNA was extracted from strain E09 using a boiling water bath method with bacterial suspension. PCR amplification was performed using universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTACGACTT-3') for bacterial 16S rRNA. The PCR reaction mixture consisted of 1 μL of DNA template and 0.5 μL each of forward and reverse primers. Yes 10 μL of polymerase was added, and ddH2O was added to a final volume of 20 μL. The PCR amplification program was as follows: 95℃ pre-denaturation for 5 min, 95℃ denaturation for 30 s, 55℃ annealing for 30 s, 72℃ extension for 1 min 30 s, for 35 cycles, followed by a final extension at 72℃ for 10 min. The PCR products were analyzed by 1% agarose gel electrophoresis. The PCR amplification products of 16S rRNA were sent to Shanghai Sangon Biotech Co., Ltd. for sequencing. The sequencing results were submitted to the NCBI database for BLAST homology analysis, and then sequence alignment and phylogenetic tree construction were performed using MEGA 7.0 software.

[0038] Results: BLAST comparison analysis in the database showed that strain E09 was similar to Bacillus belye (… Bacillus velezensis The homology between the two rRNAs reached 100%. A phylogenetic tree constructed using 16S rRNA was used. Figure 4 Analysis showed that strain E09 was related to Bacillus belye ( Bacillus velezensis Based on the fact that it belongs to the same branch, and combined with morphological and biomolecular identification, strain E09 was classified as Bacillus bereaves (B. bereaves). Bacillus velezensis ), named Bacillus belyss E09 ( Bacillus velezensis The strain (E09) was deposited on June 26, 2025, at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), with accession number GDMCC No: 66598. The address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province, 510070, China.

[0039] Example 2 Salt stress test of Bacillus belyss E09 strain The E09 strain originated from the rhizosphere microenvironment of healthy plants. The NaCl content in normal LB solid medium (5 g yeast extract, 10 g tryptone, 10 g NaCl, 1000 mL water, 15 g agar, sterilized at 121℃ for 20 min) was 1%. The NaCl content of the LB solid medium was varied to 5% (50 g / L), 6% (60 g / L), 7% (70 g / L), 8% (80 g / L), 9% (90 g / L), and 10% (100 g / L) to prepare salt stress LB plates with different salt concentrations. Single colonies of the E09 strain were incubated overnight at 37℃ and 200 rpm in LB liquid medium (5 g yeast extract, 10 g tryptone, 10 g NaCl, 1000 mL water, sterilized at 121℃ for 20 min). Add 5 μL of the solution to the center of each salt stress treatment LB agar plate and incubate at 37°C for 24 h. If colonies grow, strain E09 is tolerant of this salt concentration; if no colonies grow, strain E09 is not tolerant of this salt concentration.

[0040] The results are as follows Figure 5 As shown, strain E09 grew colonies on a 9% salt stress plate but not on a 10% salt stress plate. Therefore, strain E09 has natural tolerance to salt stress ≤9%.

[0041] Example 3 Preparation of fermentation broth of Bacillus belyssus strain E09 The E09 strain stored at -80℃ was activated, and the activated colonies were picked and cultured on LB solid medium at 37℃ for 24 hours.

[0042] Pick a single colony and place it on LB liquid medium. Incubate at 37°C and 180 r / min for 24 h to obtain the seed culture.

[0043] The seed culture containing Bacillus vesiculus strain E09 was added to LB liquid medium at a volume ratio of 1:1000, and the mixture was incubated overnight at 37°C and 180 r / min to obtain the fermentation broth of Bacillus vesiculus strain E09.

[0044] Example 4: Determination of the antibacterial effect of Bacillus belyssus strain E09 1. The inhibitory effect of strain E09 on bacterial fruit rot pathogens of bananas. The Bacillus berberis E09 strain prepared in Example 3 was used in the perforated inhibition zone method. The fermentation broth contained the indicator bacterium BG43 (a type of banana bacterial fruit rot pathogen). Dickeya fangzhongdaiWells were punched in LB solid medium plates (5 g yeast extract, 10 g tryptone, 10 g NaCl, 1000 mL water, 15 g agar, sterilized at 121℃ for 20 min) containing BG43 yeast extract. 20 μL of fermentation broth was added dropwise to each well of the solid medium, and an equal volume of liquid LB was added to the control group. Three replicates were performed. After incubation at 37℃ for 12 h, the inhibition status and the size of the inhibition zone were observed, and the diameter of the inhibition zone was measured and recorded.

[0045] The results are as follows Figure 6 As shown, Bacillus vesiculosus strain E09 is effective against BG43, a bacterial pathogen causing fruit rot in bananas. Dickey fangzhongdai BG43) Plate antibacterial zone ≥15mm.

[0046] 2. Antifungal effect of strain E09 against eight important plant pathogenic fungi, including banana. The two-point confrontation method was employed, using the fermentation broth of Bacillus leuciscus E09 strain prepared in Example 3. 5 μL of the E09 strain culture was then reacted with eight important plant pathogenic fungi [Anthracis chinensis (Bacillus anthracis)]. Colletotrichum musae ), fruit anthracnose ( Colletotrichum fructicola ), Fusarium oxysporum Cuban specialized race 4 ( Fusarium oxysporum f. sp. from Cuba Race 4), Curvularia crescentis ( Curvularia lunate Fusarium ( ) Fusarium falcate ), Flathead anthrax bacteria ( Colletotrichum truncatum ), Siamese anthrax bacteria ( Colletotrichum siamense ) and Higgins anthrax ( Colletotrichum higginsianum The mycelial cakes were plated in pairs on PDA solid medium (200g potato, 20g glucose, 15g agar, 1000mL water, sterilized at 121℃ for 20 min), with an equal volume of liquid LB added to the control group. Three replicates were set up. After incubation at 28℃ until the fungal colonies in the control group reached the LB drop position, the fungal colony radius of the treatment group and the control group was measured and recorded.

[0047] The formula for calculating the antibacterial rate is as follows: Inhibition rate = [(Control colony radius - Treatment colony radius) / Control colony radius] × 100%.

[0048] The results are shown in Table 2 and... Figure 7~8 As shown, the Bacillus vesiculosus E09 strain is effective against the key pathogenic fungus of banana anthracnose (Bacillus planus). Colletotrichum musae with fruit anthracnose Colletotrichum fructicola ) and Fusarium oxysporum Cuban specialized type 4 physiological race ( Fusarium oxysporum f. sp. from CubaRace 4), Curvularia crescentis ( Curvularia lunate Fusarium ( ) Fusarium falcate ), Flathead anthrax bacteria ( Colletotrichum truncated ), Siamese anthrax bacteria ( Colletotrichum siamense ) and Higgins anthrax ( Colletotrichum Higginsian All of them have an inhibitory effect, with an inhibition rate of 40% to 67%.

[0049] Table 2. Inhibition rate of strain E09 against eight common important plant pathogenic fungi of banana.

[0050] Example 5: Efficacy determination of Bacillus vesiculosus strain E09 against bacterial fruit rot in bananas. Pierce the peel of green bananas with a 5mm diameter hollow tube, creating circular holes. Make four holes per banana. Collect single colonies of strain E09 and the banana bacterial fruit rot pathogen BG43. Dickeya fangzhongdai BG43 was added separately to LB liquid medium and incubated overnight at 37°C and 200 rpm. The OD of each bacterial culture was adjusted. 600 The control group received 40 μL of E09 + 40 μL of BG43 per banana well (base 1.0), while the control group received 40 μL of LB + 40 μL of BG43 per banana well. The bananas were placed in plastic trays lined with damp gauze and incubated for 3 days in the dark at 75% humidity and 28°C.

[0051] The results are as follows Figure 6 As shown, the fermentation broth of strain E09 can effectively delay the blackening of the peel and inhibit the rotting process in detached banana fruits, and is effective against the bacterial fruit rot pathogen of bananas (…). Dickeya fangzhongdai The control effect of BG43 on bacterial fruit rot of bananas is significant.

[0052] In summary, this invention isolated a *Bacillus belye* strain E09 that promotes growth and increases salt stress tolerance in bananas. The E09 strain can effectively prevent bacterial fruit rot in bananas caused by *Bacillus ventricosa* BG43. Simultaneously, it inhibits eight important plant pathogenic fungi, including *Anthracnose fungus*, and can be used as a biocontrol agent to control various plant diseases. The E09 strain of this invention possesses multiple functions, including growth promotion, resistance to environmental stress, and broad-spectrum disease control, providing a stable, efficient, and environmentally friendly biological control strain for the green prevention and control of agricultural and forestry plant diseases.

[0053] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. 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.

Claims

1. A strain of Bacillus belyssus E09 ( Bacillus velezensis strain E09, characterized in that, The strain was deposited at the Guangdong Provincial Microbial Culture Collection Center (GDMCC) on June 26, 2025, with accession number GDMCC No: 66598.

2. The fermentation broth of Bacillus vesiculosus strain E09 as described in claim 1.

3. A microbial preparation, characterized in that, Contains the Bacillus berberis E09 strain as described in claim 1 or the fermentation broth as described in claim 2.

4. The *Bacillus vesiculosus* strain E09 of claim 1, the fermentation broth of claim 2, or the microbial preparation of claim 3 in controlling bacterial fruit rot pathogen BG43 in bananas (… Dickeya fangzhongdai BG43) or preparations for controlling bacterial fruit rot of bananas (BG43) Dickeya fangzhongda Applications of BG43 products.

5. The *Bacillus vesiculosus* strain E09 of claim 1, the fermentation broth of claim 2, or the microbial preparation of claim 3 in controlling bacterial fruit rot pathogen BG43 in bananas (… Dickeya fangzhongdai BG43) and banana fruit anthracnose fungus or preparations for controlling banana bacterial fruit rot fungus BG43 ( Dickeya fangzhongda The application of BG43 and the banana anthracnose fungus in products; the banana anthracnose fungus is *Anthracnose moniliforme* (BG43). Colletotrichum musae ) and fruit anthracnose ( Colletotrichum fructicola ).

6. The use of the Bacillus vesiculosus E09 strain of claim 1, the fermentation broth of claim 2, or the microbial preparation of claim 3 in the prevention and control of plant diseases or in the preparation of products for the prevention and control of plant diseases; wherein the pathogen of the plant disease is Bacillus thuringiensis BG43 (banana bacterial fruit rot pathogen). Dickeya fangzhongdai BG43), Banana anthrax bacteria ( Colletotrichum musae ), fruit anthracnose ( Colletotrichum fructicola And any one or more of the following six plant pathogenic fungi: Fusarium oxysporum Cuban specialized race 4 ( Fusarium oxysporum f.sp. cubense Race 4), Curvularia crescentis ( Curvularia lunata Fusarium ( ) Fusarium falciforme ), Flathead anthrax bacteria ( Colletotrichum truncatum ), Siamese anthrax bacteria ( Colletotrichum siamense ) and Higgins anthrax ( Colletotrichum higginsianum ).

7. The use of the Bacillus vesiculosus E09 strain of claim 1, the fermentation broth of claim 2, or the microbial preparation of claim 3 in promoting plant growth and controlling plant diseases, or in the preparation of products that promote plant growth and control plant diseases; wherein the pathogen of the plant disease is Bacillus thuringiensis BG43 (banana bacterial fruit rot pathogen). Dickeya fangzhongdai BG43), Banana anthrax bacteria ( Colletotrichum musae ), fruit anthracnose ( Colletotrichum fructicola And any one or more of the following six plant pathogenic fungi: Fusarium oxysporum Cuban specialized race 4 ( Fusarium oxysporum f. sp. cubense Race 4), Curvularia crescentis ( Curvularia lunata Fusarium ( ) Fusarium falciforme ), Flathead anthrax bacteria ( Colletotrichum truncatum ), Siamese anthrax bacteria ( Colletotrichum siamense ) and Higgins anthrax ( Colletotrichum higginsianum ).

8. The application of the Bacillus vesiculosus E09 strain of claim 1, the fermentation broth of claim 2, or the microbial preparation of claim 3 in promoting plant growth and controlling plant diseases under salt stress, or in the preparation of products that promote plant growth and control plant diseases under salt stress; wherein the salt stress environment is ≤9% salt stress; and the pathogen of the plant disease is Bacillus thuringiensis BG43 (banana bacterial fruit rot pathogen). Dickeya fangzhongdai BG43), Banana anthrax bacteria ( Colletotrichum musae ), fruit anthracnose ( Colletotrichum fructicola And any one or more of the following six plant pathogenic fungi: Fusarium oxysporum Cuban specialized race 4 ( Fusarium oxysporum f.sp. cubense Race 4), Curvularia crescentis ( Curvularia lunata Fusarium ( ) Fusarium falciforme ), Flathead anthrax bacteria ( Colletotrichum truncatum ), Siamese anthrax bacteria ( Colletotrichum siamense ) and Higgins anthrax ( Colletotrichum higginsianum ).

9. A method for promoting plant growth and preventing plant diseases, characterized in that, The *Bacillus berberis* strain E09 of claim 1, the fermentation broth of claim 2, or the microbial preparation of claim 3 are applied; the pathogen of the plant disease is *Bacillus ventriclis* BG43, the causal agent of banana bacterial fruit rot. Dickeya fangzhongdai BG43), Banana anthrax bacteria ( Colletotrichum musae ), fruit anthracnose ( Colletotrichum fructicola And any one or more of the following six plant pathogenic fungi: Fusarium oxysporum Cuban specialized race 4 ( Fusarium oxysporum f.sp. cubense Race 4), Curvularia crescentis ( Curvularia lunata Fusarium ( ) Fusarium falciforme ), Flathead anthrax bacteria ( Colletotrichum truncatum ), Siamese anthrax bacteria ( Colletotrichum siamense ) and Higgins anthrax ( Colletotrichum higginsianum ).

10. The application according to any one of claims 6 to 8 or the method according to claim 9, characterized in that, The plants mentioned are bananas, corn, soybeans, turmeric, peppers, or cruciferous plants.

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