Bacillus TRMB57781 as well as preparation and application thereof

Abstract

The invention discloses a bacillus TRMB57781. The preservation number of the bacillus TRMB57781 is CGMCC (China General Microbiological Culture Collection Center) No.36226; the paenibacillus TRMB57781 is used for antagonizing plant pathogenic bacteria or is used for preparing a cyclic lipopeptide antibiotic Fusaricidin B. The paenibacillus TRMB57781 has the advantages that the paenibacillus TRMB57781 can be The invention further discloses a preparation of the paenibacillus TRMB57781, and the preparation of the paenibacillus TRMB57781 is prepared from the paenibacillus TRMB57781. The bacillus TRMB57781 preparation can be used for preventing and treating pear fire blight of rosaceae plants, not only can achieve the prevention and treatment effect equivalent to that of chemical prevention and treatment, but also can avoid ecological and safety problems caused by the chemical prevention and treatment.

Description

Technical Field

[0001] This invention relates to the field of novel strains of Bacillus subtilis and their applications. Specifically, it relates to a strain of Bacillus subtilis TRMB57781, its preparations, and its applications. Background Technology

[0002] Pear fire blight is a highly destructive bacterial disease caused by *Erwinia amylovora*. It poses a serious threat to pears and many other Rosaceae plants globally, causing significant economic losses. Since its initial discovery in North America in the 19th century, pear fire blight has spread rapidly and is now widely distributed in many countries and regions across Europe, Asia, Africa, South America, and Oceania. Its pathogenic mechanism primarily involves infecting the leaves, flowers, fruits, buds, branches, main branches, trunk, and roots of the pear. In naturally infected pear plants in the field, the flowers are the first to show symptoms, which appear after flowering in April. The pathogen spreads from the initially infected organs to perennial branches and can further spread to the trunk and roots, eventually leading to the death of the pear plant.

[0003] China is an important origin of pear plants and one of the three major cultivated fruits. As a major producer, China makes a significant contribution to the agricultural economy. Among them, the Korla fragrant pear, originating from southern Xinjiang, is of excellent quality and is a geographical indication agricultural product of Xinjiang. Income from Korla fragrant pears alone accounts for more than 30% of the per capita net income of farmers and herdsmen. However, in recent years, pear blight has also appeared in the Korla fragrant pear growing areas of southern Xinjiang. Outbreaks of pear blight not only lead to a sharp decline in the yield of the current season's fruit but also have a long-term negative impact on the overall vigor and lifespan of the fruit trees, posing a threat to the industry. Control measures are needed to ensure the stable development of the fragrant pear industry.

[0004] Traditionally, the control of pear fire blight has relied primarily on chemical fungicides, such as streptomycin and oxytetracycline. Currently, agricultural streptomycin has shown significant effectiveness in inhibiting the pathogen, with efficacy ranging from approximately 60% to 97%, similar to that of kasugamycin. While chemical control can effectively suppress the growth and spread of the pathogen in the short term, the long-term and excessive use of chemical agents has led to a series of serious problems. First, the pathogen's resistance to chemical fungicides is constantly increasing, causing a gradual decline in control effectiveness, requiring continuously increased dosages or the use of newer agents, further increasing control costs and difficulty. Second, chemical residues pollute the soil, water bodies, and other ecological environments, disrupting the ecological balance and affecting the survival and reproduction of non-target organisms. Furthermore, high doses of chemical residues may also harm human health and pose food safety risks. Therefore, finding safe, efficient, and sustainable biological control methods to replace or partially replace chemical control has become a research hotspot and urgent need in the field of pear fire blight control.

[0005] Biological control aims to eliminate diseases naturally by using organisms known as biocontrol agents (BCAs), which are harmless to human health and the environment (as they are natural components of the soil microbiome). These characteristics lead to a reduction in the use of agrochemicals and a balance in crop production within the ecosystem. Bacillus bacteria, due to their unique biological characteristics, show great potential in the field of plant disease biological control. Bacillus can produce a variety of antibacterial substances, such as lipopeptides, proteins, fats, and polysaccharides. These substances exert their antibacterial effect through mechanisms such as inhibiting pathogen growth, interfering with pathogen metabolism, and disrupting pathogen cell walls or cell membranes. Simultaneously, Bacillus has excellent colonization capabilities, rapidly colonizing plant rhizosphere and plant surfaces to form dominant microbial communities that compete with pathogens for nutrients and living space, thereby inhibiting pathogen infection. Furthermore, Bacillus can produce various plant growth regulators during metabolism, such as auxins, cytokinins, and gibberellins, promoting plant growth and development and enhancing plant resistance. Bacillus has advantages such as ease of cultivation, low fermentation cost, and environmental friendliness, making it a research focus and popular biocontrol resource in the field of biological control. Summary of the Invention

[0006] Therefore, the technical problem to be solved by the present invention is to provide a Bacillus subtilis TRMB57781 and its preparation and application, so as to solve the problems of unsatisfactory biological control methods and environmentally unfriendly chemical control methods for pear fire blight.

[0007] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0008] A type of spore-forming bacterium, TRMB57781, belonging to the Paenibacilluspeoriae family, was deposited on October 9, 2025, at the China General Microbiological Culture Collection Center (CGMCC), with the accession number CGMCC No. 36226. The address of the depository is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0009] An application of Bacillus subtilis TRMB57781, specifically its use in antagonizing plant pathogens.

[0010] The above-mentioned Bacillus subtilis TRMB57781 was used to treat plant pathogens, specifically Erwinia amyloliquefaciens.

[0011] An application of Bacillus subtilis TRMB57781, specifically its use in the preparation of the cyclic lipopeptide antibiotic Fusaricidin B.

[0012] The application of the aforementioned Bacillus subtilis TRMB57781, and the method for preparing Fusaricidin B using Bacillus subtilis TRMB57781, are as follows:

[0013] Step (1): Inoculate Bacillus subtilis TRMB57781 into the fermentation medium to obtain the fermentation broth; the formula of the fermentation medium is: glucose 24.27 g / L, cottonseed meal 29.05 g / L, beef extract 3 g / L, NH4NO3 4 g / L, L-leucine 2.3 g / L, KH2PO4 3 g / L, NaCl 3 g / L, FeSO4 0.02 g / L; the fermentation culture conditions are 37 ℃, 180 r / min shaking culture for 48 h, and sterilization is performed after fermentation to obtain sterilized fermentation broth;

[0014] Step (2): Take the sterile fermentation filtrate, adjust the pH to 2.0 with 12 mol / L HCl, let it stand at 4 ℃ for 12 h, and then centrifuge at 4 ℃ and 12000 r / min for 20 min. Collect the precipitate and dissolve it with sterile water to obtain crude lipopeptide extract.

[0015] Step (3): The crude lipopeptide extract was further separated and purified by chromatographic separation to obtain Fusaricidin B.

[0016] A preparation of Bacillus subtilis TRMB57781 was prepared using Bacillus subtilis TRMB57781.

[0017] The above-mentioned Bacillus subtilis TRMB57781 preparation is a fermentation broth obtained by inoculating Bacillus subtilis TRMB57781 into a fermentation medium. The fermentation medium formula is as follows: glucose 24.27 g / L, cottonseed meal 29.05 g / L, beef extract 3 g / L, NH4NO3 4 g / L, L-leucine 2.3 g / L, KH2PO4 3 g / L, NaCl 3 g / L, FeSO4 0.02 g / L. The fermentation conditions are 37 ℃, shaking culture at 180 r / min for 6 days, and storage at 4 ℃ after fermentation.

[0018] The aforementioned Bacillus subtilis TRMB57781 preparation also includes substances that inhibit Erwinia amylase activity and / or substances that can maintain or activate Bacillus subtilis TRMB57781.

[0019] Application of a Bacillus subtilis preparation TRMB57781: The above-mentioned Bacillus subtilis preparation TRMB57781 was used for the prevention and control of fire blight in pear plants of the Rosaceae family.

[0020] For the application of the above-mentioned Bacillus subtilis TRMB57781 preparation, the Rosaceae plants are apple, hawthorn, peach, plum or pear; the Bacillus subtilis TRMB57781 preparation is diluted 50-100 times and then sprayed onto the Rosaceae plants.

[0021] The technical solution of the present invention achieves the following beneficial technical effects:

[0022] The fermentation broth of Bacillus subtilis TRMB57781 in this invention can inhibit pear fire blight. When it is made into a preparation for the prevention and control of pear fire blight, it can achieve the effect of chemical control while avoiding the ecological and safety problems caused by chemical control. This provides new biocontrol strain resources and theoretical basis for the biological control of pear fire blight, promotes the application and development of biological control technology in the pear industry, reduces the use of chemical pesticides, and realizes the green and sustainable development of the pear industry. Attached Figure Description

[0023] Figure 1A Colony morphology of TRMB57781 strain on LB agar medium in this embodiment of the invention;

[0024] Figure 1B Morphological images of TRMB57781 cells and spores under a scanning electron microscope at 1000× resolution in this embodiment of the invention;

[0025] Figure 1C In this embodiment of the invention, a phylogenetic tree of TRMB57781 and 24 closely related Bacillus type strains and outgroups is presented based on the 16S rRNA gene sequence (the values ​​at the nodes represent confidence levels, and the scale bar represents a 0.01 nucleotide substitution rate).

[0026] Figure 2 The circular genome map of Bacillus subtilis TRMB57781 in this embodiment of the invention; from the outside to the inside, it consists of antibiotic resistance gene CARD (purple), tmRNA (blue), rRNA (brown), tRNA (light purple), clustered regularly spaced short palindromic repeat CRISPR (light green), coding gene CDS (pink), GC content (black), GC Skew⁻ (dark purple), and GC Skew⁺ (dark green);

[0027] Figure 3 In this embodiment of the invention, genome similarity analysis is performed; the left triangular matrix is ​​a genome similarity heatmap (color scales represent similarity, red indicates high similarity, and blue indicates low similarity), and the right side is an adjacency phylogenetic tree constructed based on genome differences;

[0028] Figure 4The dilution curves of the pangenome and core genome in this embodiment of the invention; the blue curve represents the pangenome, which shows an upward trend (gradually slowing down) as the number of genomes increases, reflecting the expansion of the species' total gene pool; the red curve represents the core genome, which shows a downward trend (eventually stabilizing) as the number of genomes increases, reflecting the contraction and stabilization of the species' conserved gene clusters.

[0029] Figure 5 Upset plot of gene cluster intersection analysis of Paenibacillus strains in this embodiment of the invention; the right vertical axis "Set Size" represents the total number of gene clusters of each strain (range 0-4000); the left vertical axis "Intersection Size" represents the number of intersections of different gene families (Intersection Size), reflecting the distribution pattern of shared gene families; the black dots in the middle point matrix represent the corresponding strains participating in the formation of the gene family intersection, and the connecting lines represent the sharing pattern of gene families among multiple strains;

[0030] Figure 6 In this embodiment of the invention, the antiSMASH software, through the MIBiG comparison module, presents the comparison results of the secondary metabolic biosynthesis gene cluster of strain TRMB57781 (top) with the MIBiG reference cluster BGC0001152.5 (bottom, Fusaricidin B biosynthesis gene cluster in Paenibacillus polymyxa);

[0031] Figure 7A Comparison of antibacterial activity of TRMB57781 cultured in different culture media in this invention embodiment (actual photos of inhibition zones).

[0032] Figure 7B Comparison of antibacterial activity of TRMB57781 cultured in N3 medium with different carbon sources in the embodiments of the present invention (actual photos of inhibition zones).

[0033] Figure 7C Comparison of antibacterial activities of TRMB57781 cultured in N3 medium with different nitrogen sources in the embodiments of the present invention (actual photos of inhibition zones).

[0034] Figure 8A Comparison of antibacterial activity (inhibition zone diameter) of TRMB57781 cultured in different culture media in this embodiment of the invention.

[0035] Figure 8B Comparison of antibacterial activity (inhibition zone diameter) of TRMB57781 cultured in N3 medium with different carbon sources in this invention embodiment.

[0036] Figure 8CComparison of antibacterial activity (inhibition zone diameter) of TRMB57781 cultured in N3 medium with different nitrogen sources in this invention embodiment.

[0037] Figure 9 The infection phenotypes on day 3 of the in vitro inflorescence experiment in this embodiment of the invention are shown in the top row as the pretreatment groups: a1 (sterile water, blank control), b1 (70% agricultural streptomycin sulfate, positive control), c1 (TRMB57781 fermentation stock), d1 (fermentation broth diluted 1:50), e1 (fermentation broth diluted 1:100), and f1 (fermentation broth diluted 1:500). The bottom row shows the disease phenotypes of the corresponding pretreatment groups after inoculation with the pathogen (Erwinia amylovora).

[0038] Figure 10 In this embodiment of the invention, the number of rotten inflorescences in different treatment groups is statistically analyzed using biaxial bar charts and line graphs. The left vertical axis represents the total number of inflorescences, and the right vertical axis represents the rot rate (%). The purple bar chart is calculated using the formula: number of rotten inflorescences / total number of inflorescences * 100%. a: sterile water blank control; b: 70% agricultural streptomycin sulfate soluble powder positive control; c: TRMB57781 fermentation stock solution; d: fermentation broth diluted 50 times; e: fermentation broth diluted 100 times; f: fermentation broth diluted 500 times.

[0039] Figure 11 In this embodiment of the invention, the activity of TRMB57781 fermentation broth on detached leaves was determined; the treatment group was the same as... Figure 10 The treatment conditions corresponding to af were used to record the changes in leaf phenotype at the initial morphology, 1 day, 3 days, and 5 days.

[0040] Figure 12 This invention presents a combined bar chart (unit: mm) showing the statistical combination of the number of rotten inflorescences in different treatment groups. The treatment groups are the same. Figure 10 The treatment conditions corresponding to af, and the quantitative results of leaf lesion length under different treatments (mean ± error).

[0041] Figure 13 The infection phenotype of Korla fragrant pear seedlings on day 21 of the pear fire blight prevention experiment in this embodiment of the invention; the treatment group was the same as... Figure 10 The processing conditions corresponding to 'af' in the middle;

[0042] Figure 14 Statistics on lesion length (unit: mm) of each treatment group on day 21 of the pear blight prevention experiment in this embodiment of the invention; treatment groups were the same Figure 10 The treatment conditions corresponding to af, and the quantitative results of the length of branch lesions under different treatments (mean ± error).

[0043] Figure 15The infection phenotype of Korla fragrant pear seedlings on day 21 of the pear blight treatment experiment in this embodiment of the invention; the treatment group was the same as... Figure 10 The processing conditions corresponding to 'af' in the middle;

[0044] Figure 16 Statistics on lesion length (unit: mm) of each treatment group on day 21 of the pear blight treatment experiment in this embodiment of the invention; treatment groups were the same Figure 10 The corresponding treatment conditions in the middle af, and the quantitative results of lesion length under different treatments (mean ± error).

[0045] Figure 17 In this embodiment of the invention, the antibacterial activity of the crude extract of TRMB57781 fermentation product against pear fire blight pathogen was determined by the Oxford cup method (1: low polarity or non-polar crude extract; 2: protein crude extract; 3: medium polarity crude extract; 4: lipopeptide crude extract).

[0046] Figure 18 In this embodiment of the invention, the crude lipopeptide extract was analyzed using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (TOF MS) electrospray ionization positive ion mode (ES+).

[0047] Preservation instructions for Bacillus subtilis TRMB57781:

[0048] Deposit date: October 9, 2025

[0049] Accession number: CGMCC No. 36226;

[0050] Preservation institution: China General Microbiological Culture Collection Center, abbreviated as CGMCC;

[0051] Address of the depository: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Detailed Implementation

[0052] The Bacillus TRMB57781 used in this embodiment was isolated from wastewater from the Alar Chemical Plant. Its taxonomic position was determined by multiphase classification identification technology, and its biocontrol potential for controlling pear fire blight was systematically studied.

[0053] 1. Activation of pathogens and test strains

[0054] Erwinia amylovora and strain TRMB57781 were both deposited in the strain preservation room of the Key Laboratory of Biological Resources Conservation and Utilization of Xinjiang Production and Construction Corps in the Tarim Basin (preserved in frozen glycerol tubes at -80 ℃). Strain TRMB57781 was deposited with the China General Microbiological Culture Collection Center (CGMCC) on October 9, 2025, with accession number CGMCC No. 36226. For activation, the frozen strains were aseptically inoculated into 90 mm Petri dishes containing LysogenyBroth (LB) solid medium. Both Erwinia amylovora and strain TRMB57781 were cultured at 37 ℃ for 24–48 h. After single colonies formed, a single colony was picked and inoculated into 50 mL LB liquid medium (250 mL Erlenmeyer flask) and cultured at 37 ℃ with shaking at 120 rpm for 24–48 h to obtain a logarithmic growth phase bacterial suspension.

[0055] 2. Polymorphic taxonomic identification and whole-genome sequencing analysis

[0056] Strain classification and identification were performed using a multiphasic taxonomic approach. Morphological observation and recording of colony characteristics were conducted, and surface ultrastructure was analyzed using scanning electron microscopy. Physiological and biochemical tests included a catalase test, with the production of bubbles in a freshly prepared 3wt% H₂O₂ solution as the positive criterion; an oxidase test using API test strips; carbon and nitrogen source metabolic profiles analyzed using the BIOLOG system; and molecular biological identification based on CTAB method for extracting genomic DNA, amplifying the 16S-rRNA fragment using primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGCTACCTTGTTACGACTT-3'). The amplified products were bidirectionally sequenced by Shanghai Sangon Biotech Co., Ltd., and compared with high-similarity sequences in the EzBioCloud database, in conjunction with 24 reference strains and the E. coli type strain E. coli K-12 MG1655. T As an outgroup to calibrate evolutionary relationships, a phylogenetic tree was constructed using the maximum likelihood method with MEGA X software, and the Bootstrap method was repeated 1000 times to confirm the reliability of the nodes.

[0057] Whole-genome analysis was performed using next-generation sequencing (NGS). Single colonies were picked and inoculated into TSB liquid medium (17 g / L tryptone, 5 g / L soybean papain hydrolysate, 5 g / L sodium chloride, 2.5 g / L glucose monohydrate, 2.5 g / L dipotassium hydrogen phosphate, pH 7.2-7.5) from Beijing Aoboxing Biotechnology Co., Ltd. After incubation at 37 ℃ and 120 r / min for 24 h, the bacterial cells were collected by centrifugation at 8000 r / min. The whole genome of TRM57781 was sequenced, assembled, and annotated by Nanjing Paisennuo Gene Technology Co., Ltd. using the Illumina Hiseq 2000 sequencing platform. Ten strains with the highest sequence similarity to TRMB57781 16S rRNA were selected by NCBI-BLAST alignment. Their whole-genome reference sequences were downloaded for pan-genome differential analysis. Simultaneously, antiSMASH 8.00 was used to predict the biosynthetic gene clusters of TRMB57781, and IPGA comparative genomics analysis was performed. Genome assembly was performed using SPAdes, and the genome was annotated using Bakta v1.8.2 (database version v5.0, light mode) to identify functional elements such as CDS, tRNA, and rRNA. Antibiotic resistance-related characteristics were predicted using the CARD RGI tool integrated into the Proksee platform, and genomic resistance potential was analyzed.

[0058] 3. Evaluation of the antibacterial activity of TRMB 57781 against Erwinia amylovora, optimization of fermentation medium, and preparation of optimal fermentation broth.

[0059] The antibacterial activity of TRMB57781 against Erwinia amylovora was evaluated using the plate confrontation method. Single colonies of Erwinia amylovora were picked and inoculated into LB broth and cultured at 37 ℃ and 180 r / min until OD57781 was reached. 600 =0.5, take 100 μL of bacterial suspension and spread it evenly on the surface of LB agar plate using a sterile spreader. Use a sterile punch (6 mm in diameter) to punch holes in the pathogen agar plate and inject 200 μL of TRMB57781 strain fermentation broth into the holes. After incubation at 37 ℃ for 24 hours, measure the diameter of the inhibition zone to preliminarily verify the antibacterial activity.

[0060] To screen high-yield culture media, experiments were conducted based on antagonistic activity. The experimental groups included TSB, N1 (6 g / L yeast extract, 10 g / L fructose, 3 g / L soybean flour, 0.20 g / L MnSO4, pH 7.2–7.5, sterilized at 121 ℃ for 30 min), N2 (13.74 g / L corn flour, 15.00 g / L fish meal, 15.73 g / L yeast extract, 5.00 g / L corn steep liquor, 0.59 g / L MgSO4, 0.30 g / L KH2PO4, pH 7.0–7.6, sterilized at 121 ℃ for 30 min), and N3 (24.27 g / L glucose, 29.05 g / L soybean flour, 3 g / L beef extract, 4 g / L NH4NO3, 2.3 g / L L-leucine, 3 g / L KH2PO4, NaCl 3 g / L). (Sterilized at 121 ℃ for 30 min with 0.02 g / L FeSO4 and 0.02 g / L FeSO4). LB medium was used as a control group. During the experiment, 10 mL culture media were dispensed into sterile glass sample bottles, inoculated with TRMB57781 strain, and cultured with shaking at 37 ℃ and 180 r / min for 72 h. Subsequently, the pathogen suspension was spread onto plates and perforated, with 200 μL of fermentation broth injected into each well. The plates were then incubated at 37 ℃ for 24 h. The mean diameter of the inhibition zone was determined through three replicate experiments to identify the optimal basal medium.

[0061] In the carbon source optimization stage, glucose in the basal medium was replaced with equal masses of cellulose, xylose, and fructose; in the nitrogen source optimization stage, soybean meal was replaced with equal amounts of cottonseed meal, corn steep liquor powder, and peanut meal. After incubation at 37 ℃ and 180 r / min for 60 h, the average diameter of the inhibition zone was measured through three sets of replicate experiments to screen the optimal carbon and nitrogen source combination, and finally construct the fermentation medium system.

[0062] Seed culture was prepared using LB liquid medium, and cultured with shaking at 37 ℃ and 180 r / min for 24 h to prepare 300 mL of seed culture. The optimal fermentation broth was prepared using N3S medium (glucose 24.27 g / L, cottonseed meal 29.05 g / L, beef extract 3 g / L, NH4NO3 4 g / L, L-leucine 2.3 g / L, KH2PO4 3 g / L, NaCl 3 g / L, FeSO4 0.02 g / L, sterilized at 121 ℃ for 30 min). Large-scale fermentation was carried out in a 100 L fermenter containing 30 L of N3S medium: after sterilization, 2% seed culture was flame-inoculated, and the culture parameters were set at 37 ℃ and 180 r / min for 6 days. After fermentation, the culture was stored at 4 ℃ for later use.

[0063] 4. Evaluation of the biocontrol effect of TRMB57781 fermentation broth on different parts of detached pear material.

[0064] (1) Experiment on the activity of TRMB57781 fermentation broth on detached inflorescences

[0065] The experimental materials were obtained from the orchard of Tarim University, specifically from branches of fragrant pear trees with flower buds. Hydroponics was performed in tissue culture bottles using a sterile aqueous solution containing 3 wt% sucrose. After all the flowers on the branches with flower buds had bloomed, the fermentation solution was sprayed onto the inflorescences to ensure overall hydration. The treated inflorescences were then placed in an artificial climate cultivation chamber at 25 ℃ and 75% relative humidity for 24 h, followed by spraying with pathogenic bacteria. The control group consisted of: water instead of the fermentation solution as a negative control; and mixtures of 72% agricultural streptomycin sulfate soluble powder (sprayed at a ratio of 1:5000 according to the instructions), fermentation solution, and sterile water at ratios of 1:50, 1:100, and 1:500 as positive controls. Each experimental group contained 20 flowers, and the experiment was repeated three times. The results were observed after three days.

[0066] (2) Experiment on the activity of TRMB57781 fermentation broth on detached leaves

[0067] Healthy, disease-free leaves of roughly the same size were harvested from the fragrant pear orchard at Tarim University. In a clean bench, the surface of the harvested leaves was punctured three times with a sterile bamboo skewer. The leaves were then inoculated with the pathogen via a fungal inoculation method and placed in a 30°C constant temperature incubator. Once symptoms appeared, the fermentation solution was applied with a cotton swab. The disease progression was dynamically observed at regular intervals, and the condition was recorded at 1, 3, and 5 days post-inoculation, with the size of the affected area tallied. Control treatments were included: a negative control consisting of water instead of the fermentation solution; and positive controls including 72% agricultural streptomycin sulfate soluble powder (sprayed according to instructions) and mixtures of fermentation solution and sterile water at ratios of 1:50, 1:100, and 1:500.

[0068] (3) Experiment on the prevention and treatment of pear fire blight in potted plants by TRMB57781 fermentation broth

[0069] A. Pre-experiment treatment of test seedlings

[0070] Fifty Korla fragrant pear seedlings were cultivated in a greenhouse in March and April 2024 at the Southern Xinjiang Modern Agriculture Research Institute of Tarim University in Alar, Xinjiang. The Korla fragrant pear seedlings used in the experiment were healthy, disease-free, one-year-old seedlings purchased from the local market in 2024. They were planted in pots (20 cm high, 16 cm inner diameter), watered thoroughly every 7 days, and fertilized with compound fertilizer once a month. Before the experiment, the seedlings were pruned to ensure they were of uniform height, and new shoots were allowed to grow to 10-20 cm before use. Five days before inoculation with the pathogen, excessively long or short branches were pruned, leaving only current-year branches with relatively uniform growth.

[0071] B. Experiment on the prevention and treatment of pear blight in potted plants treated with TRMB57781 fermentation broth

[0072] In the prevention experiment, the tested Korla fragrant pear seedlings were thoroughly and evenly sprayed with the prepared fermented bacterial solution, and after 48 hours, the inoculation concentration was 1×10⁻⁶. 9 The pathogen was inoculated at cfu / mL and then wrapped with plastic wrap to retain moisture. In the treatment experiment, the test pear seedlings were first inoculated with the pathogen, wrapped with plastic wrap to retain moisture for 48 hours, and then sprayed with fermented bacterial solution on the branches. Both types of experiments included control treatments: the negative control was treated with water instead of fermented bacterial solution; the positive controls included 72% agricultural streptomycin sulfate soluble powder (sprayed according to the instructions) and mixtures of fermented bacterial solution and sterile water at ratios of 1:50, 1:100, and 1:500. Each experiment was repeated three times, with 10 seedlings in each group. The observation indicators included the total number of branches, the number of diseased branches (total number of diseased branches in the treatment experiment), and the severity of the disease (the proportion of lesions to the total length of the branch). The treatment experiment also included the treatment effect during the incubation period. The preventive effect was observed for 1 day, 7 days, 14 days, and 21 days after application; the latent period of the therapeutic effect was measured for 1 day, 4 days, 7 days, 14 days, and 21 days after spraying the fermentation liquid.

[0073] 5. Isolation and purification of active metabolites of TRMB57781

[0074] Strains TRMB57781 were inoculated into LB liquid medium and cultured under shaking conditions for 48 h. After the OD value of the bacterial culture reached 0.8-1.0, crude extracts of the antagonistic substances were obtained using three different extraction methods, and their biological activities were determined.

[0075] (1) Hydrochloric acid precipitation method: 150 mL of sterile fermentation filtrate was adjusted to pH 2.0 with 12 mol / L HCl, and after standing at 4℃ for 12 h, it was centrifuged at 4℃ and 12000 r / min for 20 min. The precipitate was collected and dissolved in sterile water to obtain crude lipopeptide extract.

[0076] (2) Ammonium sulfate saturated precipitation method: slowly add ammonium sulfate to the sterile filtrate until saturation, and selective precipitation is achieved by destroying the hydration membrane through high salt concentration. After standing at 4 ℃ for 12 h, centrifuge at 4 ℃ and 12000 r / min for 20 min, collect the precipitate and dissolve to obtain crude protein extract.

[0077] (3) Organic solvent extraction method: Add methanol-ethyl acetate mixed solvent to the filtrate at a ratio of 1:3, extract by ultrasonication for 4 h, take the upper layer, concentrate by rotary evaporation, dissolve the extract, and separate the polar and non-polar compounds.

[0078] Activity was assessed using the plate perforation method. LB plates containing the pathogen Erwinia amylovora were perforated, and 200 μL of crude extract treated with a sterile filter membrane was added. After incubation at 37 °C for 4 days, the diameter of the inhibition zone was measured.

[0079] Lipopeptide bioactive substances were extracted and purified using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (TOF MS). An ACQUITY UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) was used. The mobile phase was 0.1% formic acid (A) - methanol (B) = 10:90; the flow rate was 0.6 mL / min. -1 Column temperature: 30 ℃, injection volume: 0.1-1.0 μL; mass spectrometry using electrospray ionization (ESI), scanning range m / z 100~1500 in positive ion detection mode; capillary voltage: 3.0 kV for positive ions / 2.5 kV for negative ions, nozzle voltage: 40 V, drying gas flow rate: 800 L / h, ion source temperature: 120 ℃, collision gas: argon, collision energy: 20 ~ At 50 eV, the collected experimental data were corrected in real time using the Lock Mass pathway combined with isotope fitting (i-FIT) analysis.

[0080] 6. Data Processing and Analysis

[0081] After initial data processing in Excel, statistical analysis was performed using SPSS 24.0 software. The threshold for determining significance was set as (p < 0.05), and the significance test was conducted using Duncan's multiple range test. Graphs and charts were generated using GraphPad Prism 8.0 and Origin 2024 software.

[0082] 7. Results and Discussion

[0083] (1) Polymorphic taxonomic identification of strain TRMB57781

[0084] The colony characteristics were recorded by morphological observation. Single colonies formed on LB plates (30 ℃, 48 h incubation) were milky white with a slight gray tinge, regularly round in shape, with neat edges, a waxy and smooth, slightly raised surface, a diameter ranging from 1.0 to 1.5 mm, and a viscous texture. When picked up, a stringy phenomenon could be observed. Figure 1A Combined with scanning electron microscopy analysis of the surface ultrastructure, under 1000× magnification, the bacteria are rod-shaped, uniform in size (2.0–3.5 μm long, 0.6–0.8 μm wide), densely packed, and have fine texture on the surface. Figure 1B ).

[0085] Combined with 24 reference strains and the Escherichia coli model strain E. coli K-12 MG1655 T As an outgroup for calibrating evolutionary relationships, a phylogenetic tree was constructed using the maximum likelihood method in MEGA X software, and the node reliability was confirmed by repeated validation 1000 times using Bootstrap. The results showed that TRMB57781 is related to the type strain of *Paenibacillus*, *P. peoriae* DSM 8537. T High support clustering (bootstrap value 95) showed that the two species shared 99.3% 16S rRNA sequence similarity (meeting the intragenus interspecific classification threshold of ≥98.7%), leading to a preliminary classification of them into the genus *Paenibacillus*. Figure 1C ).

[0086] Comparison of physiological and biochemical characteristics (Table 1) shows that strain TRMB57781 produces acid from D-mannose (negative, while closely related species are all positive), L-fucose and myo-inositol (both positive, while most closely related species are negative), and has significant metabolic differences from closely related species of the genus Paenibacillus (P. peoriae, P. ottowii, etc.), further supporting its taxonomic uniqueness from the perspective of physiological function.

[0087] Table 1. Comparison of physiological and biochemical characteristics between strain TRMB57781 and phylogenetic-related strains.

[0088]

[0089] Classification: "+" represents positive; "-" represents negative; "Ne" represents not detected; data source: BacDive (https: / / doi.org / 10.1093 / nar / gkab961).

[0090] (2) Genome analysis of TRMB57781 and antibacterial potential of strain TRMB57782

[0091] The assembled genome, based on whole-genome next-generation sequencing results, contains 606 contigs with a total length of 5,900,343 bp. Bakta annotated a total of 5,396 genomic features, including 5,209 CDS (coding sequences), 116 tRNAs, 8 rRNAs, and 50 ncRNA regions.

[0092] Proksee's RGI, based on the CARD database, predicted 4,788 antibiotic resistance-related traits (covering resistance genes, mutation sites, etc.). See details... Figure 2 .

[0093] Comparative genomics results were obtained from whole-genome data analysis using IPGA software, and ANI cluster analysis was performed ( Figure 3 The strain TRMB57781 clustered with Paenibacillus polymyxa ATCC 842ᵀ and Paenibacillus skribbensis AM49ᵀ, indicating a close phylogenetic relationship. It showed a dark blue color (low similarity) with strains such as Paenibacillus rhizovicinus 14171Rᵀ, suggesting a more distant phylogenetic relationship. The pan-genome and core genome dilution curves ( Figure 4 The trend reveals the diversity and conservation of the species' genome, quantifies the "conservative-variable" genome segment, and suggests that the species maintains basic survival and adapts to complex environments through genetic diversity by adopting a "conservative core + variable pangenome" model, while still having untapped genetic potential.

[0094] In gene cluster intersection analysis, such as Figure 5 As shown, the Upset diagram system reveals the gene cluster sharing patterns of 11 Paenibacillus strains, accurately identifying three types of genetic units: core gene clusters shared by all strains (corresponding to species-conserved functional modules), unique gene clusters unique to each strain (functionally associated niche specialization), and strain groups with close gene sharing. These results, at the gene cluster level, confirm that Paenibacillus possesses a "conserved core-diverse pan-genetic" genomic structure, providing direct evidence for elucidating the genetic basis of its niche differentiation and functional innovation.

[0095] Secondary metabolite synthesis gene clusters were predicted using antiSMASH and compared with the MIBiG database: the Fusaricidin B synthesis gene cluster in strain TRMB57781 showed 97% similarity to the homology cluster of Paenibacillus polymyxa, and the module structure was highly conserved, suggesting the potential ability of this strain to synthesize Fusaricidin B in its metabolic pathway (see...). Figure 6 ).

[0096] (2) Evaluation of antibacterial activity of TRMB57781 and optimization of optimal fermentation medium

[0097] A. Evaluation of the antibacterial activity of TRMB57781 in different culture media

[0098] Using *Erwinia amylovora* as the target strain, the antibacterial activity of TRMB57781 was evaluated using the plate confrontation method. TRMB57781 showed good antibacterial activity. The antibacterial activities of different bacterial culture media (TSB, LB, N1, N2, and N3) were statistically analyzed. The results showed that after culturing at 37 ℃ for 24 h, the inhibition zone diameter was measured. The results are as follows: Figure 7A and Figure 8A The average diameters of the inhibition zones on TSB, LB, N1, N2, and N3 culture media were measured to be 10.74 mm, 11.06 mm, 12.90 mm, 11.32 mm, and 15.64 mm, respectively. Therefore, N3 was selected as the basal culture medium.

[0099] B. Screening of fermentation media with optimal C and N sources for TRMB57781

[0100] Using *Erwinia amylovora* as the target strain, the antibacterial activity of TRMB57781 was evaluated using the plate confrontation method, and the optimal culture medium and carbon and nitrogen sources were screened. Under constant temperature incubation at 37℃ for 24 h, the antibacterial effect of this strain in five bacterial culture media (TSB, LB, N1, N2, N3) was compared, and the average diameter of the inhibition zone was measured. Figure 7A The diameters of the media were 10.74 mm, 11.06 mm, 12.90 mm, 11.32 mm, and 15.64 mm, respectively. The N3 medium group had the largest inhibition zone, so N3 was selected as the basal medium for subsequent optimization.

[0101] Based on the basal medium N3, further carbon source screening was conducted: cellulose, xylose, and fructose were selected as candidate carbon sources, and compared with glucose in the original medium. After incubation at 37 ℃ for 24 h, the average diameter of the inhibition zone in each group was compared ( Figure 7B and Figure 8B The inhibition zones were 9.86 mm (cellulose), 9.66 mm (xylose), 13.26 mm (fructose), and 13.56 mm (glucose), respectively. The glucose group had the largest inhibition zone, indicating that it was most conducive to the synthesis of antibacterial metabolites by the strain, and was therefore identified as the optimal carbon source.

[0102] Nitrogen sources were screened based on the optimal carbon source: cottonseed meal, corn steep liquor powder, and peanut meal were selected as candidate nitrogen sources. Results are shown below. Figure 7C and Figure 8C As shown in the figure, compared with the soybean meal in the original culture medium, under the same culture conditions, the average diameters of the inhibition zones were 12.20 mm (cottonseed meal), 8.64 mm (corn steep liquor powder), 9.24 mm (peanut meal), and 10.12 mm (soybean meal). The inhibition zone in the cottonseed meal group was significantly larger than that in the other groups, indicating that it provided the best nitrogen source for bacterial growth and the expression of antibacterial metabolites; therefore, it was determined to be the optimal nitrogen source.

[0103] (3) Evaluation of the biocontrol effect of TRMB57781 fermentation broth on different parts of pear detachment material

[0104] The results of the activity assay of TRMB57781 fermentation broth on detached inflorescences showed that ( Figure 9 and Figure 10 Typical flower rot symptoms were observed in the nectaries, anthers, and calyxes of both the control group and the undiluted bacterial solution treatment group, indicating that the undiluted bacterial solution did not reduce the infection effect. However, the number of diseased inflorescences in the fermentation broth treatment group (especially the 1:50 and 1:100 dilution groups) was significantly lower than that in the control group. The 1:50 dilution group had a control efficacy index of 84.62%, which was better than that of agricultural streptomycin (69.48%), with a difference of 15.14%.

[0105] like Figure 11 and Figure 12 As shown, the activity assay on detached leaves indicated that the control effect of the bacterial solution may decrease over time due to volatilization. Overall, agricultural streptomycin showed better control effect, but the 1:100 dilution of the bacterial solution achieved control efficacy of 72.84 and 75.49 on days 1 and 3, respectively, which is close to streptomycin's 81.58 and 80.23, suggesting its biocontrol value. This dilution is the optimal choice to balance antibacterial effect and plant safety, while streptomycin, although highly effective in the short term, still needs to be monitored for its side effects.

[0106] In the pot experiment, preventive treatment ( Figure 13 and Figure 14The results showed that potted plants sprayed with different concentrations of fermentation broth after infection with Erwinia amylovora showed a significant reduction in lesion length compared to the control group, with effects comparable to agricultural streptomycin, indicating its good biological preventative effect; treatment ( Figure 15 and Figure 16 In this study, spraying fermentation liquid after inoculation with pathogens significantly improved the length of lesions, with effects comparable to agricultural streptomycin, demonstrating its excellent biological therapeutic effect.

[0107] (4) Preliminary study on extraction of secondary metabolites and antibacterial active substances of TRMB57781

[0108] To isolate Fusaricidin B from Paenibacillus peoriae TRMB57781, a complete chain of evidence, from substance separation to functional association, was constructed following the process of "targeted extraction - precise identification - gene corroboration." Based on the lipopeptide characteristics of Fusaricidin B, the crude extract from the fermentation broth was treated with HCl precipitation (ammonium sulfate saturated precipitation and organic solvent extraction yielded relatively small amounts of Fusaricidin B, resulting in insignificant activity; furthermore, organic reagent extraction of Fusaricidin B easily damages the target compound's structure, mainly because prolonged soaking in methanol solvent during large-scale fermentation can lead to inactivation; while hydrochloric acid precipitation, utilizing point precipitation at pH 2, has high extraction efficiency in small amounts of fermentation broth, showing significant activity). This enriched the target component and removed water-soluble impurities, laying the foundation for subsequent detection. High-resolution time-of-flight mass spectrometry (HR-TOF MS, ES⁺ mode) analysis showed… Figure 18 Of the 130 candidate molecular formulas, only one matched the theoretical characteristics of Fusaricidin B: the measured mass-to-charge ratio of 897.5766 deviated from the theoretical mass of 897.5773 by -0.7 mDa (-0.8 ppm), the deduced degree of unsaturation was 9.5, consistent with its polycyclic structure, thus chemically confirming the target compound. Genome-wide analysis and antiSMASH validation showed that the Fusaricidin B synthesis gene, labeled as a Polyketide+NRP type lipopeptide, had 100% sequence similarity to the reference sequence, confirming that the strain possesses the genetic basis for synthesizing this compound. The chemical detection and genetic evidence corroborated each other, clearly demonstrating that Paenibacillus peoriae TRMB57781 can produce Fusaricidin B, and that it is directly associated with anti-amylopectin activity, forming a closed loop of evidence and providing support for research on the antibacterial mechanism.

[0109] In summary, this embodiment isolated a bacterium, TRMB57781, with good antibacterial activity against Erwinia amylovora from wastewater of the Alar Chemical Plant. It was deposited in the strain preservation room of the Key Laboratory of Biological Resources Conservation and Utilization of the Xinjiang Production and Construction Corps in the Tarim Basin. Morphologically, it was regularly round with neat edges and a viscous texture. Under scanning electron microscopy, the bacteria were rod-shaped, uniform in size, densely packed, and had a fine texture on the surface. TRMB57781 is similar to the type strain of the genus Paenibacillus, Paenibacillus peoriae DSM 8537. T High-support clustering, with 99.3% 16S rRNA sequence similarity between the two strains, preliminarily classifies them as belonging to the genus *Paenibacillus*. Comparison of physiological and biochemical characteristics revealed that strain TRMB57781 exhibits significant metabolic differences in acid production from D-mannose (negative, closely related species are all positive), L-fucose, and myo-inositol (both positive, closely related species are mostly negative), compared to closely related species of the genus *Paenibacillus* (*P. peoriae*, *P. ottowii*, etc.), further supporting its taxonomic uniqueness from a physiological function perspective.

[0110] Whole-genome sequencing and assembly revealed that strain TRMB57781 has a genome size of 5.90 Mb (containing 606 contigs), with 5396 gene signatures obtained through Bakta annotation (including 5209 CDS, 116 tRNAs, 8 rRNAs, and 50 ncRNAs). Proksee-RGI, based on the CARD database, predicted 4788 antibiotic resistance-related signatures, suggesting its environmental adaptation potential. In comparative genomics analysis, ANI clustering showed that this strain is closely related to Paenibacillus polymyxa ATCC 842ᵀ and Paenibacillus kribbensis AM49ᵀ, but genetically distant from Paenibacillus rhizovicinus 14171Rᵀ. The pangenome-core genome dilution curve revealed a "conserved core + variable pangenome" architecture, which ensures both basic metabolic conservation and expands ecological adaptation space through pangenome diversity, and also indicates untapped genetic potential. Upset analysis of gene cluster intersections elucidated gene-sharing patterns in 11 Paenibacillus strains, precisely identifying core gene clusters (species-conserved functions), unique gene clusters (niche specialization), and closely sharing gene groups. This gene cluster-level validation of the aforementioned structure provides direct evidence for elucidating the genetic basis of niche differentiation and functional innovation. Furthermore, antiSMASH combined with secondary metabolism prediction from the MIBiG database showed that the Fusaricidin B synthetic gene cluster in TRMB57781 exhibited 97% sequence similarity to the Paenibacillus polymyxa homology cluster, with conserved module structure, suggesting its potential ability to synthesize this metabolite.

[0111] In the plate confrontation method for evaluating the antibacterial activity of TRMB57781 and the optimization of the fermentation medium, the inhibition zone diameter formed in the N3 medium group was larger than that in other experimental groups, and therefore it was selected as the basal medium. Further carbon and nitrogen source screening experiments showed that glucose was the optimal carbon source, as it achieved the maximum average inhibition zone diameter. Among the different nitrogen source experimental groups, the cottonseed meal group had the highest inhibition zone diameter, thus it was identified as the optimal nitrogen source, and the best fermentation medium, N3S, was confirmed.

[0112] Evaluation of the biocontrol efficacy of different parts of detached pear materials showed that in the detached inflorescence experiment, the TRMB57781 fermentation broth caused typical flower rot symptoms in the nectaries, anthers, and calyxes of both the control group and the undiluted broth treatment group, indicating that the undiluted broth did not reduce the infection effect. The number of diseased inflorescences in the fermentation broth treatment groups (especially the 1:50 and 1:100 dilution groups) was significantly lower than that in the control group. The 1:50 dilution group achieved a control efficacy index of 84.62%, which was 15.14% higher than that of agricultural streptomycin (69.48%). The detached leaf experiment showed that the control effect of the broth may decrease over time due to volatilization. Overall, agricultural streptomycin showed better control efficacy, but the 1:100 dilution broth showed poor control on days 1 and 3. The control efficacy of the two drugs reached 72.84 and 75.49, respectively, which is close to that of streptomycin (81.58 and 80.23), indicating their control value. This dilution is the optimal choice to balance antibacterial effect and plant safety. Although streptomycin is highly effective in the short term, its side effects still need to be monitored. In the pot experiment, after infection with Erwinia amylovora, the length of lesions in the preventive treatment group was significantly reduced compared with the blank control group, with an effect comparable to agricultural streptomycin, demonstrating a good biological prevention effect. After inoculation with the pathogen, the length of lesions in the treatment group was significantly improved by spraying fermentation liquid, with an effect comparable to agricultural streptomycin, showing an excellent biological treatment effect.

[0113] The isolation of Fusaricidin B from *Paenibacillus peoriae* TRMB57781 followed a "targeted extraction-precise identification-genetic corroboration" process: lipopeptide components were enriched by HCl precipitation, and HR-TOF MS (ES⁺) confirmed one matching Fusaricidin B (m / z = 897.5766, deviation from theoretical value -0.7 mDa, unsaturation degree 9.5) among 130 candidate molecular formulas. Whole-genome and antiSMASH validation showed that its synthetic gene was 100% homologous to the reference sequence. Chemical and genetic evidence confirms that this strain can produce Fusaricidin B and is directly associated with its antibacterial activity against *Erwinia amylase*, providing support for research on its antibacterial mechanism.

[0114] This embodiment identifies the strain as *Paenibacillus peoriae*, belonging to the genus *Paenibacillus*, through polyphasic taxonomy combined with morphological characteristics, physiological and biochemical properties, and 16S rRNA gene sequence analysis. *Paenibacillus peoriae* TRMB57781 possesses excellent biocontrol potential, providing a new strain resource for the green control of pear fire blight and offering preliminary exploration of its biocontrol and antibacterial mechanisms.

Claims

1. A bacilloid TRMB57781, characterized by, It was preserved in China General Microbiological Culture Collection Center on October 09, 2025, and the preservation number is CGMCC No. 36226.

2. Use of a Paenibacillus sp. TRMB57781, characterized in that The use of Paenibacillus sp. TRMB57781 in antagonizing plant pathogenic bacteria according to claim 1.

3. Use of Paenibacillus campinasensis TRMB57781 according to claim 2, characterized in that, The plant pathogenic bacteria is Erwinia amylovora.

4. Use of a Paenibacillus sp. TRMB57781, characterized in that The use of Paenibacillus sp. TRMB57781 in preparing Fusaricidin B according to claim 1.

5. Use of Paenibacillus campinasensis TRMB57781 according to claim 4, characterized in that, The method for preparing Fusaricidin B by using Paenibacillus sp. TRMB57781 is as follows: Step (1), inoculate Paenibacillus sp. TRMB57781 into fermentation medium to obtain fermentation broth; the formula of the fermentation medium is: glucose 24.27 g / L, cottonseed powder 29.05 g / L, beef extract 3 g / L, NH4NO3 4 g / L, L-leucine 2.3 g / L, KH2PO4 3 g / L, NaCl 3 g / L, FeSO4 0.02 g / L; the fermentation culture condition is 37℃, 180 r / min oscillation culture for 48 h, sterilize after fermentation, and obtain sterilized fermentation broth; Step (2), take the sterilized fermentation filtrate, adjust to pH 2.0 with 12 mol / L HCl, stand at 4℃ for 12 h, centrifuge at 4℃, 12000 r / min for 20 min, collect the precipitate, and dissolve with sterile water to obtain lipopeptide crude extract; Step (3), further separate and purify the lipopeptide crude extract by chromatography to obtain Fusaricidin B.

6. A preparation of Paenibacillus sp. TRMB57781 characterized in that, Paenibacillus sp. TRMB57781 is prepared according to claim 1.

7. The Paenibacillus-like TRMB57781 preparation according to claim 6, characterized in that, The fermentation broth obtained by inoculating Paenibacillus sp. TRMB57781 into fermentation medium; the formula of the fermentation medium is: glucose 24.27 g / L, cottonseed powder 29.05 g / L, beef extract 3 g / L, NH4NO3 4 g / L, L-leucine 2.3 g / L, KH2PO4 3 g / L, NaCl 3 g / L, FeSO4 0.02 g / L; the fermentation culture condition is 37℃, 180 r / min oscillation culture for 6 d, and store at 4℃ after fermentation.

8. The Paenibacillus-like TRMB57781 preparation according to claim 6, characterized in that, It also includes substances with inhibitory activity on Erwinia amylovora and / or substances capable of maintaining or activating Paenibacillus sp. TRMB57781.

9. Use of a preparation of Paenibacillus sp. TRMB57781 characterized in that, The Paenibacillus sp. TRMB57781 preparation according to any one of claims 6-8 is used for the prevention and treatment of Rosaceae plant fire blight.

10. Use of the Paenibacillus-like strain TRMB57781 preparation according to claim 9, characterized in that, The Rosaceae plant is apple, hawthorn, peach, plum or pear; the Paenibacillus sp. TRMB57781 preparation is diluted 50-100 times and sprayed on the Rosaceae plant by spraying.

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