Corn leaf endophyte and application thereof in inhibition of corn spider mites

Abstract

The invention discloses corn leaf endophyte and application thereof in inhibition of corn spider mites. The invention provides bacillus velezensis, the bacillus velezensis is bacillus velezensis 2025-FCW119-1M2, and the preservation number of the bacillus velezensis is CGMCC (China General Microbiological Culture Collection Center) No.33928. The bacillus velezensis provided by the invention is a bacillus velezensis strain. The invention also provides an application of the strain in any one of the following: A1) preventing plant diseases and insect pests; a2) treating plant diseases and insect pests; and A3) inhibiting phytopathogen. The endophyte with the activity of antagonizing the corn pathogenic bacteria is successfully separated from the corn leaves and is identified as the bacillus, a new microbial resource is provided for biological prevention and control of corn diseases, and the endophyte is expected to be developed into a biological prevention and control preparation, so that the use of chemical pesticides can be reduced, and the environmental pollution can be reduced.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and relates to endophytes in corn leaves and applications thereof in inhibiting corn spider mites. Background Art

[0002] Corn, one of the world's most important food crops, faces threats from various pests and diseases during its growth. Spider mites are a particularly common and serious pest, severely impacting corn production and yield. They are characterized by high frequency, widespread impact, and poor control effectiveness. However, traditional chemical control methods face numerous drawbacks, including environmental pollution, impacts on human and animal health, and the development of pesticide resistance in pests and pathogens. Therefore, finding safe, effective, and environmentally friendly control methods to address these issues has become a pressing issue in corn production and is of great significance to the sustainable development of agriculture.

[0003] In recent years, corn endophytes have attracted considerable attention as a novel biocontrol method. Their environmentally friendly nature demonstrates their significant advantages over other technologies. This suggests that endophytes may be a potential biocontrol agent for controlling the occurrence and spread of spider mites in corn. However, the mechanism of action of corn endophytes for controlling spider mites and their practical application still face numerous challenges. Currently, research on corn endophytes is still inadequate, especially the discovery and utilization of endophytes with highly effective antagonistic activity. Therefore, developing efficient methods for the isolation, identification, and application of corn endophytes is of great practical significance. Summary of the Invention

[0004] The technical problem solved by the present invention is to provide a new endophyte with high-efficiency antagonistic activity and capable of preventing and controlling plant diseases and insect pests.

[0005] In order to solve the above technical problems, the first aspect of the present invention provides Bacillus velezensis 2025-FCW119-1M2, which has a deposit number of CGMCC No.33928.

[0006] In a second aspect, the present invention provides a culture of the Bacillus Velez subtilis according to the first aspect.

[0007] The above-mentioned culture is obtained by culturing the Bacillus Velez subtilis in a microbial culture medium.

[0008] The culture includes living cells of the Bacillus Velez subtilis or its metabolites.

[0009] The microbial culture medium may be a solid culture medium or a liquid culture medium.

[0010] The term "culture" refers to any liquid or solid product (i.e., all materials within a culture vessel, i.e., fermentation products) containing a microbial population after artificial inoculation and cultivation. This refers to a product obtained by growing and / or amplifying microorganisms. It can be a biologically pure culture of the microorganism or contain a certain amount of culture medium, metabolites, and / or other components produced during the culture process. The term "culture" also includes subcultures obtained by subculturing microorganisms, which can be cultures of a single generation or a mixture of several generations.

[0011] The culture of Bacillus velezensis described in the first aspect above may include secondary metabolites, specifically secondary metabolites with antifungal and / or anti-mite efficacy, further specifically Dihydrocapsaicin (CAS 19408-84-5), Crocin (CAS 42553-65-1), Immunomycin (CAS 104987-12-4), Avermectin A1baglycone, Novobiocin (CAS 303-81-1), Avermectin A2b monosaccharide, AvermectinA1a monosaccharide (CAS 71828-13-2), Solavetivone (CAS 61167-63-3), Tyrosol (CAS 501-94-0), Ononin (CAS 486-62-4), fumarillin (CAS 23110-15-8), Ricinoleic acid acidmethylester (CAS 141-24-2), Nerol (CAS 106-25-2) and Cinnamic acid (CAS 140-10-3).

[0012] In a third aspect, the present invention provides a composition comprising the Bacillus Velez subtilis according to the first aspect or the culture according to the second aspect.

[0013] In a fourth aspect, the present invention provides a product comprising the Bacillus Velez subtilis described in the first aspect, the culture described in the second aspect, or the composition described in the third aspect.

[0014] The above-mentioned products may further include other agriculturally or horticulturally acceptable diluents, fillers, solvents, spontaneity promoters, carriers, emulsifiers, preservatives, dispersants, antifreeze agents, thickeners, adjuvants, or any combination thereof.

[0015] The products mentioned above are biological agents or biocontrol agents.

[0016] Furthermore, the biological agent is a microbial agent.

[0017] The products mentioned above have at least one of the following functions:

[0018] A1) Prevent plant diseases and insect pests;

[0019] A2) Treatment of plant diseases and insect pests;

[0020] A3) Inhibit plant pathogens.

[0021] In a fifth aspect, the present invention provides use of the Bacillus Velez subtilis described in the first aspect, the culture described in the second aspect, or the composition described in the third aspect in at least one of the following or in preparing a product having at least one of the following functions:

[0022] A1) Prevent plant diseases and insect pests;

[0023] A2) Treatment of plant diseases and insect pests;

[0024] A3) Inhibit plant pathogens.

[0025] In a sixth aspect, the present invention provides a method for preventing plant diseases and insect pests, comprising the following steps: applying the Bacillus Velez subtilis described in the first aspect, the culture described in the second aspect, the composition described in the third aspect, or the product described in the fourth aspect to plants, plant organs, or plant tissues to cultivate plants and prevent plant diseases and insect pests.

[0026] In the seventh aspect, the present invention provides a method for treating plant diseases and insect pests, comprising the following steps: applying the Bacillus Velez subtilis described in the first aspect, the culture described in the second aspect, the composition described in the third aspect, or the product described in the fourth aspect to plants, plant organs, or plant tissues to cultivate plants and achieve the treatment of plant diseases and insect pests.

[0027] In an eighth aspect, the present invention provides a method for inhibiting plant pathogens, comprising the steps of: applying the Bacillus Velez subtilis described in the first aspect, the culture described in the second aspect, the composition described in the third aspect, or the product described in the fourth aspect to plants, plant organs, or plant tissues to cultivate plants, thereby inhibiting plant pathogens;

[0028] Alternatively, a method for inhibiting plant pathogens comprises the following steps: subjecting the Bacillus Velez subtilis described in the first aspect, the culture described in the second aspect, the composition described in the third aspect, or the product described in the fourth aspect to plant pathogens to inhibit the plant pathogens.

[0029] In the above, the plant pests are spider mites.

[0030] The spider mite may be Tetranychus urticae.

[0031] In the above, the plant pathogens are Setosphaeria turcica (corn leaf spot fungus) (asexual form is Exserohilum turcicum), Bipolarismaydis (corn leaf spot) (asexual form is Bipolarismaydis), Pythium inflatum, Fusarium graminearum, and Fusarium verticillioides.

[0032] In the above, the action may include spraying.

[0033] In the above, the plant organ may include any one of roots, stems, and leaves.

[0034] In the above, the plant tissue may include any one of meristem, guard tissue, ground tissue, and conducting tissue.

[0035] Furthermore, the plant is selected from the Poaceae family.

[0036] Furthermore, the plant is selected from corn.

[0037] Experimental results of the present invention show that:

[0038] (1) Providing a new biocontrol resource: This study successfully isolated endophytes with antagonistic activity against corn pathogens from corn leaves and identified them as Bacillus sp. This provides a new microbial resource for the biocontrol of corn diseases. These endophytes are expected to be developed into biocontrol agents, thereby reducing the use of chemical pesticides and lowering environmental pollution.

[0039] (2) Accurate identification of endophytes: The molecular identification method is used to accurately determine the taxonomic status of endophytes, which helps to systematically study and utilize endophytes, and also provides a basis for the subsequent preservation and application of strains.

[0040] (3) Biological control: Conduct field endophyte liquid biological control experiments on spider mites, providing new microbial resources and technical support for the biological control of corn pests and diseases.

[0041] Preservation Instructions

[0042] Strain name: 2025-FCW119-1M2

[0043] Classification name: Bacillus velezensis

[0044] Depository: General Microbiology Center of China Culture Collection Administration

[0045] Abbreviation of depository institution: CGMCC

[0046] Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing

[0047] Deposit date: March 21, 2025

[0048] CGMCC registration number: CGMCC No.33928 BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 To determine the in vitro antagonistic activity of maize endophyte FCW119-1M2 against selected plant pathogens.

[0050] Figure 2 Identification of strain FCW119-1M2 and analysis of its antagonism to five corn fungal pathogens; A: Phylogenetic tree of strain FCW119-1M2 based on 16SrRNA sequence; B: ANI heat map of strain FCW119-1M2; C: Analysis of the antifungal rate of strain FCW119-1M2 against five corn pathogenic fungi.

[0051] Figure 3 Figure 5 shows the effect of spraying FCW119-1M2 on spider mites; A shows the damage of spider mites to 009 and B73 inbred lines after spraying FCW119-1M2; B shows the damage of spider mites on the tenth leaf of corn plants of 009 and B73 inbred lines, where a: 009 inbred line after spraying FCW119-1M2, b: 009 inbred line control, c: B73 inbred line after spraying FCW119-1M2, d: B73 inbred line control.

[0052] Figure 4 Metabolome analysis of strains FCW119-1M2 and BC225T1; A is principal component analysis; B is orthogonal partial least squares discriminant analysis (OPLS-DA) of strains FCW119-1M2 and BC225T1.

[0053] Figure 5 Volcano plot of differential metabolites between strains FCW119-1M2 and BC225T1.

[0054] Figure 6 Differential metabolites (avermectins) between strains FCW119-1M2 and BC225T1 are upregulated in black and downregulated in gray.

[0055] Figure 7 Schematic diagram of corn planting in field experiments. DETAILED DESCRIPTION

[0056] The present invention will be further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for further improvements by those skilled in the art and are not intended to limit the present invention in any way.

[0057] Unless otherwise specified, the experimental methods in the following examples are conventional methods and were performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials and reagents used in the following examples, unless otherwise specified, were all commercially available.

[0058] Unless otherwise specified, the quantitative tests in the following examples were performed three times, and the results were averaged.

[0059] Example 1: Isolation and identification of functional endophyte strains

[0060] 1. Isolation and culture of endophytes

[0061] (1) Material preparation: 330 mite-inoculated and 330 uninoculated maize leaves were obtained from the Maize Research Institute, Beijing Academy of Agricultural and Forestry Sciences. These leaves were collected from the Nanfan Base of the Maize Research Institute, Beijing Academy of Agricultural and Forestry Sciences, Yazhou District, Hainan Province (109°11′N, 18°39′E) in January 2022. LB, R2A, TSA, PDA, and MEA culture media, as well as sterile water, sterile mortars, centrifuge tubes, and other experimental equipment were prepared.

[0062] (2) Preparation of bacterial suspension: 0.1 g of corn leaves surface sterilized with 75% alcohol were placed in a sterile mortar, and an appropriate amount of sterile water was added to grind into a homogenate, and then transferred to a 5 mL sterile centrifuge tube to prepare 10 -1 -10 -4 Gradient bacterial suspension. The specific operation is: take 1mL from the original bacterial solution and add it to 9mL sterile water, mix well to obtain 10 -1 Gradient bacterial suspension, and so on, prepare 10 -2 , 10 -3 , 10 -4 Gradient bacterial suspension.

[0063] (3) Inoculation and culture: Take 0.1 mL of each gradient and inoculate it on LB, R2A, TSA (cultured at 37℃ for 1-2 days) and PDA, MEA (cultured at 28℃ for 3-5 days) plates respectively, with 3 replicates for each concentration. Place the inoculated plates upside down in the incubator for culture. After the colony grows stably, pick colonies with different morphologies for purification and culture. The purification method is as follows: inoculate the picked colonies on new corresponding culture medium plates, streak them using the four-zone streak method, pick single colonies after culture and streak them again for purification until pure cultured colonies are obtained. The purified colonies are numbered and stored at -80℃ with 25% glycerol for later use.

[0064] A total of 2,896 maize endophytes were screened from these 660 maize leaf samples. These included 1,018 bacterial strains in LB medium, 488 in R2A medium, 699 in TSA medium, 259 in PDA medium, 255 in MEA medium, and 177 fungal strains. These strains were carefully labeled and preserved.

[0065] 2. Confrontation plate test of 5 corn pathogens

[0066] (1) Endophyte culture: Three maize endophytes (FCW119-1M1, FCW119-1M2, and BC255T1) with significant antagonistic activity against five pathogenic fungi (Pythium graminearum, Pythium graminearum, and Pythium truncatum) were inoculated into sterile LB medium and cultured overnight at 37°C and 200 rpm / min to obtain an endophyte solution.

[0067] (2) Preparation of pathogen spore suspension: The mycelia of five corn pathogens (Setosphaeria turcica (asexual form: Exserohilum turcicum), Bipolaris maydis (asexual form: Bipolaris maydis), Pythium inflatum, Fusarium graminearum, and Fusarium verticillioides) purchased from Anhui Longping High-Tech Seed Co., Ltd. were inoculated into conical flasks containing sterile water and glass beads, and the flasks were shaken at 28°C and 200 rpm / min for 15 min to break the mycelia and release the spores. Spore suspensions of each pathogen were prepared. The spore suspensions were counted using a hemocytometer and the spore concentration was adjusted to an appropriate range.

[0068] (3) Plate preparation and experimental setup: 3 mL of each pathogen spore suspension (concentration of 1×10 7spores / mL) were added to sterile MEA medium (150 mL) at about 50°C, mixed evenly, and poured into sterile culture dishes for plate making. After the culture medium solidified, a sterile filter paper was placed on the surface of the culture medium. 5 μL of endophytic bacterial solution (concentration of 3×10 7 CFU / mL), and the control group was added with an equal amount of sterile LB medium, with 2 replicates for each group.

[0069] (4) Observation and calculation of results: Place the plate in a constant temperature incubator at 28°C for 48 hours and observe the formation of the inhibition zone around the filter paper. Use a vernier caliper to measure the radius of the inhibition zone of the control and experimental groups, and calculate the antifungal rate of the strain according to the formula.

[0070]

[0071] Where R represents the antifungal rate, r0 is the radius of the control group, and r1 is the radius of the experimental group.

[0072] The results are as follows Figure 1 、 Figure 2 C. As shown in Tables 1 and 2, strain FCW119-1M2 exhibited antifungal activity against all five corn pathogens, with inhibition rates exceeding 80%. FCW119-1M1 was less effective than strain FCW119-1M2. BC255T1 was only effective against Fusarium spp.

[0073] Table 1 is the measurement data of antifungal circle size

[0074]

[0075] Table 2 shows the calculation results of antifungal rate

[0076]

[0077] 3. Molecular identification of endophytes

[0078] (1) Gene amplification: PCR amplification was performed using bacterial DNA from FCW119-1M2 and BC255T1 as templates, using universal primers for the bacterial 16S rRNA gene (F: 5'-AGAGTTTGATCCTGGCTCAG-3' and R: 5'-GGTTACCTTGTTACGACTT-3'). The PCR reaction system included template DNA, primers, dNTPs, Taq DNA polymerase, and buffer. The PCR reaction conditions were: pre-denaturation at 95°C for 5 min, followed by 30 cycles of denaturation at 95°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 1 min, and a final extension at 72°C for 10 min.

[0079] (2) Sequencing and alignment: After the PCR amplification product is purified, it is sent to a sequencing company for sequencing. The measured 16S rRNA gene sequence is compared with a known bacterial gene sequence database (such as the NCBI GenBank database) and the taxonomic status of the strain is determined by analyzing the sequence similarity.

[0080] The results showed that the nucleotide sequence of the nucleic acid encoding the 16S rRNA of FCW119-1M2 was sequence 1.

[0081] Phylogenetic tree constructed by 16S rRNA of strain FCW119-1M2 and model strains ( Figure 2 A) showed that FCW119-1M2 was Bacillus, and ANI analysis showed that ( Figure 2 B) The genome similarity between FCW119-1M2 and Bacillus velezensis FZB42 is 97.35%, indicating that FCW119-1M2 is Bacillus velezensis.

[0082] BC255T1 is also a Bacillus sp.

[0083] FCW119-1M2, recorded as 2025-FCW119-1M2, was deposited in the General Microbiology Center of China Culture Collection Administration (CGMCC, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Postal Code: 100101) on March 21, 2025, with the accession number CGMCC No. 33928 and the classification name Bacillus velezensis.

[0084] Example 2: Spraying of endophyte solution and inoculation of pests and diseases

[0085] The maize inbred line B73 is described in the following document: The B73 maize genome: complexity, diversity, and dynamics; Patrick S. Schnable et al., 2009 Nov 20; 326(5956): 1112-5. doi: 10.1126 / science.1178534.

[0086] Maize inbred line 009 is described in the following literature: Pyramiding of favorable haplotypes of major QTLs for yield-related traits to improve maize (Zea mays L.) productivity; Xiaqing Wang et al., Agriculture Communications, Volume 3, Issue

[0087] 2,2025,100083,ISSN 2949-7981,

[0088] https: / / doi.org / 10.1016 / j.agrcom.2025.100083.

[0089] 1. Corn planting

[0090] Two maize inbred lines, B73 and 009, were selected and sown on November 24, 2023, at the Nanfan Base of the Maize Research Center of Beijing Academy of Agricultural and Forestry Sciences, Yazhou District, Sanya City, Hainan Province (109°11′N, 18°39′E). Figure 7 shown.

[0091] 2. Endophyte culture and dilution

[0092] The endophytic strain FCW119-1M2 with significant inhibitory effect was selected for field trials to evaluate its control effect on corn mites.

[0093] 50 μL of glycerol-preserved FCW119-1M2 bacterial solution from Example 1 (sterilized at 121°C for 15 min) was inoculated into 100 mL of LB medium and cultured at 37°C and 200 rpm / min for 24 h to prepare a seed solution. 8 mL of the seed solution was inoculated into 800 mL of sterilized LB medium and cultured for another 24 h under the same conditions to obtain a bacterial solution with a final concentration of 3.3 × 10 7 CFU / mL.

[0094] 3. Bacteria spraying and pest inoculation

[0095] Dilute the bacterial solution obtained in step 2 above 25-fold with sterile water. Spray the maize inbred lines every other day (10 L of diluted bacterial solution per plot) for three consecutive times. Then, inoculate healthy spider mites. Inoculation and phenotypic identification methods can be found in the Laboratory and Field Evaluation of Maize Resistance to the Two-spotted Spider Mite, Tetranychus urticae. A maize inbred line not sprayed with the endophytic strain FCW119-1M2 served as a control.

[0096] 4. Pest and disease survey

[0097] 20 days after inoculation, the degree of damage was assessed based on the distribution of spider mites on the corn leaves, and typical damage symptoms on the tenth leaf of each treatment group were recorded.

[0098] The results of spraying endophytes on mites are as follows: Figure 3 As shown in the figure, compared with the control group, the damage level of mites in maize inbred line 009 sprayed with the endophyte FCW119-1M2 was significantly reduced, while the damage level of mites in maize inbred line B73 sprayed with the endophyte FCW119-1M2 did not change significantly. Therefore, FCW119-1M2 has a strong inhibitory effect on mites, and its effect is particularly strong in maize 009.

[0099] The same method was used to detect the strain BC255T1 isolated in Example 1. The results showed that the strain had no antagonistic effect on corn mites.

[0100] 5. Metabolome analysis

[0101] The strain BC255T1, which is known to have no antagonistic effect on corn mites, was selected as the control strain for metabolomics and HPLC detection.

[0102] To further explore the mechanism of endophyte control of pests and diseases, two endophytes were analyzed using non-targeted metabolomics. The culture medium of FCW119-1M2 (FCW119-1M2 was cultured in LB medium at 37°C and 200 rpm / min for 24 h) and the culture medium of BC255T1 (BC255T1 was cultured in LB medium at 37°C and 200 rpm / min for 24 h) of Example 1 were frozen and transferred to EP tubes three times (300 μL, 300 μL, 400 μL) with 1000 μL of methanol-acetonitrile extract (methanol-acetonitrile volume ratio = 1:1) and vortexed for 30 seconds. The sample was placed on a steel column, milled at 45 Hz for 10 minutes, sonicated in an ice-water bath for 10 minutes, and allowed to stand at -20°C for 1 hour. The sample was centrifuged at 12,000 rpm for 15 minutes at 4°C. 500 μL of the supernatant was transferred to an EP tube, the extract dried in a vacuum concentrator, and reconstituted with 160 μL of extraction solution (acetonitrile-water volume ratio: 1:1). The sample was vortexed for 30 seconds, sonicated in an ice-water bath for 10 minutes, and centrifuged again at 12,000 rpm for 15 minutes at 4°C. 120 μL of the supernatant was transferred to a 2 mL injection vial. 10 μL of each sample was mixed to form a QC sample for analysis. Analyses were performed using a Waters UPLC Acquity I-Class PLUS ultra-high performance liquid chromatography coupled to a Waters UPLC Xevo G2-XS QTOF high-resolution mass spectrometer using a Waters Acquity UPLC HSS T3 column (1.8 μm, 2.1 x 100 mm). Mobile phase A in both positive ion mode (POS) and negative ion mode (NEG) consisted of 0.1% formic acid in water, and mobile phase B was 0.1% formic acid in acetonitrile. The injection volume was 2 μL, and elution was performed according to a specific gradient program at a flow rate of 400 μL / min. Raw data were acquired using MassLynx V4.2, and data processing, including peak extraction and alignment, was performed using Progenesis QI software. Identification was performed based on the Progenesis QI online METLIN database, public databases, and a self-built library. Theoretical fragment identification was also performed (fragment ion identification mass number deviation was less than 100 parts per million).After the raw peak area information was normalized by the total peak area, principal component analysis and Spearman correlation analysis were performed to determine the reproducibility of samples within the group and quality control samples. The classification and pathway information of the identified compounds were searched in the KEGG, HMDB, and LipidMaps databases, and the difference fold was calculated and compared. The significant p-value of each compound was calculated using the T test. OPLS-DA modeling was performed using the R language package ropls, and 200 permutation tests were performed to verify the reliability of the model. The model VIP value was calculated through multiple cross-validation. The difference fold, P value, and VIP value of the OPLS-DA model were combined to screen differential metabolites (screening criteria were FC ≥ 3, P value ≤ 0.01, and VIP ≥ 1). The hypergeometric distribution test was used to calculate the differential metabolites with significant KEGG pathway enrichment. The metabolomics data were analyzed using BMKCloud (www.biocloud.net).

[0103] These two endophytes were compared and their potential mechanisms of insect and disease resistance were explored through untargeted metabolomics. 2,498 metabolites were annotated, mainly including 213 carboxylic acids and their derivatives, 132 fatty acyl groups, 127 pregnenolone lipids, 88 glycerophospholipids, 88 steroids and steroid derivatives, and 82 organic oxygen compounds (Table 3). Principal component analysis (PCA) revealed clear separation between the different strains, indicating that the metabolites in different strains are significantly different, which is consistent with the antagonism experiments and field experiments ( Figure 4 A). Orthogonal partial least squares discriminant analysis (OPLS-DA) found that the values ​​of R2X, R2Y and Q2 were close to 1, with Q2>0.9, further verifying the reliability of the OPLS-DA model ( Figure 4 B) Differential metabolites can be screened based on variable projection importance analysis.

[0104] Table 3 Metabolite classification of FCW119-1M2 metabolome

[0105]

[0106]

[0107] By screening the differential metabolites, a total of 823 differentially expressed metabolites were identified between FCW119-1M2 and BC225T1. In addition, the number of upregulated (593) and downregulated (230) metabolites were visualized in the volcano plot ( Figure 5 ).

[0108] Table 4 shows the 14 differential metabolites of antifungal and anti-tetranychus mites screened from strain FCW119-1M2

[0109]

[0110] In order to reveal the mechanism by which these two endophytes can inhibit corn pests and diseases, the unique differential metabolites in the Venn diagram were screened, and a total of 14 differential metabolites were screened out (Table 4). Among them, there are 3 metabolites that can inhibit the growth of mites, and these 3 are intermediate synthetic products of avermectin; there are 11 metabolites that inhibit the growth of fungal pathogens. Figure 5 It can be seen that compared with BC225T1, most of the antifungal and anti-mite differential metabolites of FCW119-1M2 were upregulated, and only two antifungal differential metabolites were downregulated, indicating that compared with BC255T1, the endophytic bacteria FCW119-1M2 has stronger antifungal and anti-tetranychus abilities, which is consistent with the results of the antagonism experiments and field experiments.

[0111] Afterwards, the KEGG pathways of the above 14 unique differential metabolites were analyzed. The results showed that the number of differential metabolites in the pathway Biosynthesis of 12-, 14- and 16-membered macrolides (ko00522) was the largest ( Figure 6 ) are all intermediate synthetic products of avermectin, followed by KEGG pathways such as Biosynthesis of alkaloids derivedfrom terpenoid and polyketide (ko01066) and Sesquiterpenoid and triterpenoidbiosynthesis (ko00909).

[0112] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be practiced over a wide range of parameters, concentrations, and conditions without departing from the spirit and scope of the present invention and without unnecessary experimentation. Although specific embodiments have been given herein, it should be understood that further modifications may be made to the present invention. In summary, this application is intended to encompass any variations, uses, or improvements to the present invention, including those made by conventional techniques known in the art that depart from the scope of the present invention. Applications of the essential features may be made within the scope of the following claims.

Claims

1. Bacillus velezensis 2025-FCW119-1M2, whose deposit number is CGMCC No.33928.

2. The culture of Bacillus Velezii according to claim 1.

3. A composition comprising the Bacillus Velez subtilis according to claim 1 or the culture according to claim 2.

4. A product comprising the Bacillus Velez subtilis of claim 1 or the culture of claim 2 or the composition of claim 3.

5. The product according to claim 4, characterized in that: The product is a biological agent or a biocontrol agent.

6. Use of the Bacillus velezensis of claim 1, the culture of claim 2, the composition of claim 3, or the product of claim 4 in at least one of the following or in the preparation of a product having at least one of the following functions: A1) Prevent plant diseases and insect pests; A2) Treatment of plant diseases and insect pests; A3) Inhibit plant pathogens.

7. A method for preventing plant diseases and insect pests, comprising the steps of: applying the Bacillus Velez subtilis according to claim 1, the culture according to claim 2, the composition according to claim 3, or the product according to claim 4 to plants, plant organs, or plant tissues to cultivate plants and prevent plant diseases and insect pests.

8. A method for treating plant diseases and insect pests, comprising the steps of: applying the Bacillus Velez subtilis according to claim 1, the culture according to claim 2, the composition according to claim 3, or the product according to claim 4 to plants, plant organs, or plant tissues to cultivate plants and achieve the treatment of plant diseases and insect pests.

9. The use according to claim 6 or the method according to claim 7 or 8, characterized in that: The plant pests and diseases are spider mites.

10. A method for inhibiting plant pathogens, comprising the steps of: applying the Bacillus Velezii of claim 1, the culture of claim 2, the composition of claim 3, or the product of claim 4 to plants, plant organs, or plant tissues to cultivate the plants, thereby inhibiting the plant pathogens; Or, a method for inhibiting plant pathogens, comprising the steps of: subjecting the Bacillus velezensis according to claim 1 or the culture according to claim 2 or the composition according to claim 3 or the product according to claim 4 to plant pathogens to achieve inhibition of plant pathogens.

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